journal of ornamental plants - webs 4(1)/jopmarch2014 … · journal of ornamental plants, volume...

Effective Use of Synthetic Seed Technology in the Regeneration of Dendrobium White Fairy

Orchid...........................................................................................................................................1-7

Wee Leng Siew, Mei Yun Kwok, Yong Mei Ong, Hui Ping Liew and Boon Keat Yew

In Vitro Flower Bud Formation, Plant Regeneration and Morphogenetic Studies in Local

Scented Cultivar of Rosa indica.................................................................................................9-18

Raageeva Bimal and Niti Kiran

Study of Genetic Diversity among Philodendron Varieties by Randomly Amplified Polymorphic

DNA Markers............................................................................................................................19-26

Achar Devaraja AM, Jakhar Mamta , Jakeer P and Shetty KPV

Growth, Yield and Quality of Carnation (Dianthus caryophyllus L.) Cultivars under Lath House

Conditions.................................................................................................................................27-32

Malik Abid Mehmood, Muhammad Saleem Akhtar Khan and Naveed Ahmad

Utilization of Rice Hull as a New Substrate for Turf Grass Seed Germination in Sod Production

as a Sustainable Approach.......................................................................................................33-37

Mohammad Ali Golestani, Ali Dolatkhahi, Navid Vahdati and Omid Nouri Roudsari

Comparison Tea Extract, 8-Hydroxy Quinoline Sulfate and Rifampicin on the Vase Life of Cut

Chrysanthemum (Denderanthema grandiflorum L. cv. Purple)............................................39-43

Davood Hashemabadi and Hamideh Bagheri

Micropropagation of Rosa canina Through Axillary Shoot Proliferation............................45-51

Mahboubeh Davoudi Pahnekolayi, Ali Tehranifar, Leila Samiei and Mahmoud Shoor

Drought stress impact on some biochemical and physiological traits of 4 groundcovers (Loliumperenne, Potentilla spp, Trifolium repens and Frankinia spp) with potential landscape

usage..........................................................................................................................................53-60

Elham Samieiani and Hossein Ansari

Volume 4, Number 1

March 2014

Journal of Ornamental Plants


It is approved publication of Journal of Ornamental Plants (based on approbation of 61st session

of "Survey and Confirmation Commission for Scientific Journals" at Islamic Azad University dated

on 01/25/2010.

Publisher: Islamic Azad University, Rasht, Iran.

Executive Director: Dr. Ali Mohammadi Torkashvand

Editor-in-Chief: Professor Roohangiz Naderi

Executive Manager: Dr. Shahram Sedaghat Hoor

Editorial Board:

Professor Ramin, A., Isfahan University of Technology, Iran

Professor Abdollah Hatamzadeh, University of Guilan, Iran

Professor Honarnejad, R., Islamic Azad University-Varamin Branch, Iran

Associate Professor Shahram Sedaghathoor, Islamic Azad University, Rasht Branch, Iran

Dr. Davood Hashemabadi, Islamic Azad University, Rasht Branch, Iran

Associate Professor Moazzam Hassanpour Asil, University of Guilan, Iran

Assistant Professor Behzad Kaviani, Islamic Azad University, Rasht Branch, Iran

Professor Nagar, P.K., Institute of Himalayan Bio-Resource Technology, India

Professor Salah El Deen, M.M., Al Azhr University, Egypt

Assistant Editor: Zahra Bagheramiri

Abstracting/Indexing

SID, Index Copernicous, Islamic World Science Citation Center (ISC), Open-J-Gate, Magiran,

EBSCO, Directory of Research Journals Indexing (DRJI), Agricola and Journal Seek, DOAJ.

Journal of Ornamental Plants is an international journal devoted to the publication of original papers

and reviews in the Ornamental plants, Floriculture and Landscape. Articles in the journal deal with

Floriculture and Landscape. The scope JOP includes all Ornamental plants, Floriculture and Landscape.

All articles published in JOP are peer-reviewed. The journal is concerned with Ornamental plants, Flori-

culture, Landscape and covers all aspects of physiology, molecular biology, biotechnology, protected

cultivation and environmental areas of plants.

Publication schedule: The journal publishes: Article on original research in Ornamental plants,

Floriculture, Landscape and related fields that contain new information for solving Ornamental

plants, Floriculture and Landscape problems of world.

Submission of article: Typescripts should be submitted in Journal of Ornamental Plants (IAU-Rasht

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Authors” (published in all issues before submission of their typescripts).

Address: Islamic Azad University, Rasht, Iran.

Telfax: 0131- 4224069, email: [email protected]

Web Site: www. jornamental.com

Effective Use of Synthetic Seed Technology in the Regeneration of Dendrobium White Fairy

Orchid..............................................................................................................................................1-7Wee Leng Siew, Mei Yun Kwok, Yong Mei Ong, Hui Ping Liew and Boon Keat Yew

In Vitro Flower Bud Formation, Plant Regeneration and Morphogenetic Studies in Local Scented

Cultivar of Rosa indica...................................................................................................................9-18Raageeva Bimal and Niti Kiran

Study of Genetic Diversity among Philodendron Varieties by Randomly Amplified Polymorphic DNA

Markers.......................................................................................................................................19-26Achar Devaraja AM, Jakhar Mamta , Jakeer P and Shetty KPV

Growth, Yield and Quality of Carnation (Dianthus caryophyllus L.) Cultivars under Lath House

Conditions....................................................................................................................................27-32Malik Abid Mehmood, Muhammad Saleem Akhtar Khan and Naveed Ahmad

Utilization of Rice Hull as a New Substrate for Turf Grass Seed Germination in Sod Production as a

Sustainable Approach..................................................................................................................33-37Mohammad Ali Golestani, Ali Dolatkhahi, Navid Vahdati and Omid Nouri Roudsari

Comparison Tea Extract, 8-Hydroxy Quinoline Sulfate and Rifampicin on the Vase Life of Cut

Chrysanthemum (Denderanthema grandiflorum L. cv. Purple)....................................................39-43Davood Hashemabadi and Hamideh Bagheri

Micropropagation of Rosa canina Through Axillary Shoot Proliferation.....................................45-51Mahboubeh Davoudi Pahnekolayi, Ali Tehranifar, Leila Samiei and Mahmoud Shoor

Drought stress impact on some biochemical and physiological traits of 4 groundcovers (Loliumperenne, Potentilla spp, Trifolium repens and Frankinia spp) with potential landscape usage.....53-60Elham Samieiani and Hossein Ansari

Content Page

www.jornamental.com

Journal of Ornamental Plants, Volume 4, Number 1: 1-7, March, 2014 1

Effective Use of Synthetic Seed Technology in the

Regeneration of Dendrobium White Fairy Orchid

The synthetic seed technology is becoming popular due to its wide

application in germplasm conservation and for exchanges between countries in

the floriculture trade. In this study, this method was used to study the germination

and conversion capabilities of orchid species Dendrobium White Fairy when

stored at different storage times and with different storage containers. A high

germination percentage of 80% was observed for encapsulated synthetic seeds

up to 150 days of storage in Petri dishes and screw capped polypropylene tubes

and this percentage only began to decline gradually from 150 storage days and

more. Besides that, synthetic seeds stored in polypropylene tubes were shown

to germinate faster and develop into plantlets with longer shoots and roots as

compared to those stored in the Petri dishes. This proved the efficiency and

suitability of polypropylene tubes over Petri dishes as storage containers.

Keywords: Dendrobium, Protocorm-like bodies, Regeneration, Synthetic seeds.

Wee Leng Siew*, Mei Yun Kwok, Yong Mei Ong, Hui Ping Liew and Boon Keat Yew

Department of Life Sciences, School of Applied and Health Sciences, Institute of Technical Education,

Singapore.

*Corresponding author,s email: [email protected]

Abstract

Journal of Ornamental Plants, Volume 4, Number 1: 1-7, March, 20142

INTRODUCTION

In the orchid family, Dendrobium is one of the largest tropical genus that is found in diverse

habitats of Asia, with about 1,200 species. Many of its hybrid species are relatively easy to grow

and due to its wide variety, colourful nature of the flowers and its compact size for some species,

it is very popular in the retail trade, and it is one of the most important orchids in the horticulture

industry (Martin and Madassery, 2006). Dendrobium White Fairy, which is the target of our study,

is a hybrid between Dendrobium Singapore White and Dendrobium Walter Oumae. This tropical

orchid that is locally bred in Singapore has an average height of about 2 to 3 feet and blooms with

about 6 to 12 buds in a star-shaped blossom and a gentle jasmine-like scent. Its elegant appearance

made it very popular as ornamental plants as well as for landscaping purposes and they are widely

exported as cut flowers.

In general, propagation of orchids from seeds is relatively difficult due to its minute seed

size and lack of endosperm, heterozygousity of seeds and the requirement of the seed to associate

with mycorrhizal fungi for germination (Saiprasad, 2001). The method used to germinate orchid

seeds in the orchid growing industry is asymbiotic micropropagation, where seeds are germinated

and cultured aseptically using culture media such as MS media (Murushige and Skoog, 1962) sup-

plemented with a carbon source. Although this method is able to maintain the genetic uniformity

of the plants, it is more suitable for small scale propagation as it is labour intensive and has rela-

tively low multiplication rate (Saiprasad, 2001).

An alternative method, which has been studied and found to be more effective, is the syn-

thetic seed technology. This method is preferred because it combines the advantages of clonal prop-

agation and seed propagation with the possibility of long term storage of the seeds through

encapsulation in a gel-like matrix (Lambardi et al., 2006). In this technique, non-embryogenic veg-

etative propagules such as shoot tips, nodal segments or axillary buds, protocorm-like bodies (PLBs)

or calluses are artificially encapsulated using sodium alginate as the preferred coating agent. These

synthetic hydrogel seeds contain nutrients that will help in the survival and speedy growth of the

embryos into plantlets during their cultivation after storage (Sharma et al., 2013). This cost effective

method can be scaled up and has been shown to be successful in several commercially important

agronomic and horticultural crops, such as bananas and potatoes (Ganapathi et al., 1992; Nyende

et al., 2003), as well as ornamental plants such as lilac and orchids (Lambardi et al., 2006). In or-

chids, PLBs are used for encapsulation and has been shown to successfully germinate and regenerate

into plantlets (Ara et al., 2000; Saiprasad and Polisetty, 2003; Gantait et al., 2012).

In view of this technology, various optimizations have been performed and reported in or-

chid propagation using synthetic seeds, such as the concentration of sodium alginate and calcium

chloride as gelling agents, the stage of PLBs suitable for encapsulation, the storage period and

temperature of the synthetic seeds (Saiprasad and Polisetty, 2003; Mohanraj et al., 2009; Gantait

et al., 2012). The objective of this study is to determine the effects of different storage times and

compare between different storage containers on the germination and regeneration of DendrobiumWhite Fairy orchids using the synthetic seed technology.

MATERIALS AND METHODS

Chemicals, media and solutions

The following chemicals and media were used in this study.

Calcium chloride (MP Biomedicals, Cat. No. 153502); Sodium alginate (MP Biomedicals,

Cat. No. 218295); Murashige & Skoog (MS) Basal media (PhytoTechnology Laboratories, Cat.

No. M519); Phyto Agar (plantMedia, Cat. No. 40100072-2).

For the preparation of encapsulation matrix, 3% (w/v) sodium alginate was prepared in ½

MS liquid media containing 2% sucrose, while 75 mM calcium chloride (CaCl2.2H2O) was pre-

pared in double distilled water.


Plant material, culture media and culture conditions

Green capsule seed pods of Dendrobium White Fairy orchid were obtained via self-polli-

nation. The seeds were germinated aseptically in Magenta GA-7TM vessels with 50 mL of ½ MS

media supplemented with 15% (v/v) coconut water, 2% (w/v) sucrose and 0.6% (w/v) Phyto-agar

(PlantMedia). All media were adjusted to pH 5.2 before autoclaving at 121oC for 20 min. All cul-

tures were incubated and maintained at 25±1oC with a 24-hour photoperiod.

After 4 weeks of cultivation in solid media, the green pin-head-like PLBs formed were then

transferred to ½ MS liquid media supplemented with 15% (v/v) coconut water and 2% (w/v) su-

crose and maintained on a rotary shaker at 200 rpm under the above specified conditions for PLB

induction. PLBs in the liquid media were subcultured every 3 weeks with fresh media until they

were ready to be used for the experiments.

Encapsulation of Dendrobium White Fairy orchid PLBs

Twelve-week old Dendrobium White Fairy orchid PLBs were used in this study. The en-

capsulation procedure was adapted from Lambardi et al., 2006. Encapsulation was carried out by

mixing PLBs (about 3-4 mm diameter) with 3% (v/v) sodium alginate solution. Using a 7 mm di-

ameter cut micropipette tip, aliquots of the alginate solution, each containing one PLB, were asep-

tically pipetted out and gently dropped into 75 mM CaCl2.2H2O solution (Gantait et al., 2012).

The calcium alginate beads that were formed were maintained in the CaCl2.2H2O solution for 30

min in a continuously stirring environment to prevent the beads from adhering to each other. These

synthetic seeds were then decanted from the CaCl2.2H2O solution, rinsed thrice in sterile water

and blotted dry with sterilized filter paper.

Storage of encapsulated PLBs

The experiment aims to observe the effects of different storage time and compare between

different storage containers on the germination and conversion ability of the encapsulated synthetic

seeds. Non-encapsulated, or naked PLBs were used as the experimental control.

Storage time points of 0, 30, 60, 90, 120, 150 and 180 days were selected for this study.

For each time point, three replicates were used and each replicate consists of a set of 40 synthetic

seeds, which were prepared via encapsulation. These synthetic seeds were then divided equally

into 2 groups and each group was stored in either a Petri dish or 15 ml polypropylene (PP) tube.

Both storage containers contain 2 ml of sterile water. The same protocol was conducted for non-

encapsulated PLB controls at 0, 30 and 60-day storage time-points. The control and the encapsu-

lated synthetic seeds were then stored at 25±1oC under dark conditions until the storage time point

is reached.

Assessment of germination and direct conversion of stored synthetic seeds

After each storage time point, the synthetic seeds were taken out of the Petri dish or 15 ml

PP tube and were cultured aseptically in Petri dishes with 30 ml of ½ MS media containing 2%

(w/v) sucrose and 0.6% (w/v) Phyto-agar (Plant Media). The cultures were incubated and main-

tained at 25±1oC with a 16-hour photoperiod. Germination of the seeds was monitored and recorded

based on the number of days taken for the shoot to emerge from the gel matrix (Gantait et al.,2012). The conversion of synthetic seed to plantlet was recorded after 50 days of in vitro culture.

The percentage of germination and conversion into plantlet were then calculated and these results

were compared to the controls.

RESULTS AND DISCUSSION

Encapsulation of Dendrobium White Fairy orchid PLBs

Dendrobium White Fairy orchid PLBs of size ranging from 3 to 4 mm in diameter (12


weeks old) were used because previous studies has shown that this size range is ideal for encap-

sulation and optimum conversion in orchids (Corrie and Tandon, 1993). PLBs smaller than the

size range displayed poor conversion frequencies which could be due to tissue immaturity, thus

the PLBs were unable to withstand encapsulation or require a longer time to emerge from the

gelling matrix (Corrie and Tandon, 1993; Saiprasad and Polisetty, 2003; Poobathy et al., 2009).

Besides that, Saiprasad (2001), Lambardi et al. (2006) and Gantait et al. (2012) documented

that the ideal synthetic seeds were obtained using 3% sodium alginate and 75 mM calcium chloride

as the gelling matrix. This combination creates optimal ion exchange between the sodium and cal-

cium ions, which produces firm, clear and isodiametric beads (Gantait et al., 2012). In addition,

sodium arginate is also ideal because of its low toxicity to the embryo and the gel bead is able to

protect the fragile embryo during handling (Ara et al., 2000). Germination was also best achieved

when half-strength MS media was used in the gelling matrix, providing sufficient nutrients for the

plant tissue (Gantait et al., 2012). Based on the collated findings, the encapsulation protocol for

Dendrobium White Fairy orchid PLBs was adapted (Fig. 1).

Assessment of germination and conversion of encapsulated PLBs

Based on studies by Saiprasad and Polisetty (2003), Lambardi et al. (2006) and Mohanraj

et al. (2009), it was observed that encapsulated PLBs in the Dendrobium and Oncidium orchid

species could generally maintain maximum germination percentage when stored at 4oC for 45 and

60 days, however, there is a decrease in the conversion percentage to plantlets when the storage at

low temperature is prolonged (Lambardi et al., 2006). In a recent paper, it was found that storage

at 25oC showed a higher percentage in germination and in conversion to plantlets as compared to

Fig. 1. Encapsulated and non-encapsulated Dendrobium White Fairy PLBs12-week old Dendrobium White Fairy orchid PLBs (about 3 to 4 mm in diameter) used for the study. Non-encapsulated PLBs (A) and encapsulated PLBs (B) stored in Petri dish with 2mL of sterile water on a filterpaper. Non-encapsulated PLBs (C) and encapsulated PLBs (D) stored in 15ml polypropylene tube (Falcon)containing 2ml of sterile water. Size range of the encapsulated PLBs (about 6mm) and non-encapsulatedPLBs (about 4mm) (E).Calcium arginate beads containing PLBs for the experiments (F).

A

B

C

D

E

F


4oC up to 120 days for a monopodial orchid species (Gantait et al., 2012). Hence, in our study,

25oC was used as our storage temperature.

From table 1, a high germination percentage of more than 80% was observed for encap-

sulated synthetic seeds that were first stored in both Petri dish and screw capped PP tube and

this germination percentage only began to decline after 150 days of storage. There is no major

difference between the encapsulated seeds stored in Petri dish and PP tube at 25oC under dark

conditions and this could be due to optimum storage conditions which allowed the uptake of

nutrients from the half strength MS gel matrix to the seed to sustain its viability. The dark envi-

ronment helped to delay PLB germination by keeping the PLBs at dormant stage. The mainte-

nance of same temperature from storage of synthetic seeds to their cultivation at 25oC also

imposed lesser physical harm to these PLBs as compared to storage at lower temperature due to

the fluctuation in temperature and cold stress during the transfer. This observation was seen in

a research done by Gantait et al. (2012). After 180 days of storage, synthetic seeds that were

stored in the PP tubes were able to achieve 77% germination, while those stored in Petri dish

had dried up and were unable to germinate at all. This could be because screw capped PP tubes

are able to retain moisture content longer, and thus able to maintain their viability and prevent

the synthetic seeds from drying up.

The conversion capability of the stored synthetic seeds into plantlets was also monitored

after their respective storage time points for a period of 50 days. A similar trend was observed for

the conversion percentage of Dendrobium White Fairy synthetic seeds as with the germination re-

sults shown in table 1.

Generally, synthetic seeds stored in PP tubes showed a slightly higher percent of conversion

as compared to those stored in Petri dish and this difference in conversion rate between seeds

stored in the two vessels generally increased with storage time. This could be because in addition

to the PP tube’s ability to retain moisture, the longer storage time also enabled the synthetic seeds

to have ample time to adapt, grow and develop into mature PLBs with the available nutrients in

the gel matrix, so that they are able to develop into complete plantlets faster due to their maturity

stage. Synthetic seeds that were stored in PP tubes were also able to produce more plantlets with

shoots and roots exceeding 5 mm and 10 mm respectively as compared to those that were stored

in Petri dishes (Table1).

Based on the physical observation of the developed plantlets from synthetic seeds (Fig. 2).

Storage

Time (Day)

Percent of Germination Percent of Conversion

Average number of plant-

let with shoot of minimum

5mm length

Average number of

plantlet with root of

minimum 10mm length

Petri Dish 15ml PP

Tube

Petri Dish 15ml PP

Tube

Petri Dish 15ml PP

Tube

Petri Dish 15ml PP

Tube

0

30

60

90

120

150

180

95

100

90

100

98

87

0

95

97

80

95

92

88

77

35

18

8

28

38

33

0

35

24

17

48

60

43

44

4

2

1

5

4

7

0

4

4

1

7

10

9

4

2

1

0

3

2

3

0

2

0

1

4

6

4

3

Table 1. Effect of storage time on the germination and conversion of encapsulated PLBs.

Note: Dendrobium synthetic seeds stored in either Petri dish or 15 ml polypropylene (PP) tube at different storage times

and storage containers were germinated on ½ MS media. Their percentage of germination, percentage of conversion,

average shoot and root lengths were tabulated respectively after the 50-day cultivation period. Encapsulated PLBs at 0-

day storage time point were used as controls.


It was observed that when the encapsulated synthetic seeds were stored for 90 days or

longer, the PLBs appeared yellow and unhealthy when they were first taken out from the dark, but

when the PLBs were cultured onto ½ MS media, they gradually became green in colour and looked

healthier (data not shown). About 15% to 20% of the plantlets which were stored for 90 days or

longer also appeared elongated (Fig. 2C as compared to 2A and 2B). This could be due to gradual

nutrient depletion when the encapsulated seeds were stored at longer time periods.

During the course of our experiments, an additional interesting observation was made as

shown in Fig. 3.

Generally, longer storage period will lead to slower germination rate and lower germination

capability and this was observed for the non-encapsulated seeds that were stored in Petri dishes

for 30 and 60 days as compared to the 0-day control (Fig. 3B). The germination percentage also

declined drastically from the 30-day storage to 60-day storage. The encapsulated PLBs that were

stored in Petri dishes, on the other hand, were able to maintain a high germination percentage as

expected (Fig. 3A).

In contrast, the PLBs stored in screw capped PP tubes maintained a high germination

percentage regardless of its encapsulation status. Both encapsulated and non-encapsulated

PLBs showed no reduction in germination rate as compared to the 0 day control for both the

30-day and 60-day storage time points (Fig. 3). They developed relatively well, just like a

fresh seeds where there is no sign of delay in their germination ability. This could be due to

the presence of water that was added to the tubes at the start of the experiment, which created

a moist environment as well as the greater ability of screw capped PP tubes over Petri dishes

to maintain a humid environment for the developing PLBs. This is crucial so as to prevent

dehydration of the encapsulated seed or PLBs and in turn, preserve its viability (Gantait etal., 2012).

Fig. 2. Phenotypes of synthetic seeds stored in Petri dish at 90-day time point at the end of 50 days of cultivation.

(A) Synthetic seeds which were successfully converted into plantlets. (B) Some seeds grew slower and were

still at the PLB stageat the end of cultivation. (C) About 15% to 20% of the plantlets appeared elongated. For

those stored in screw capped polypropylene tubes, the elongated phenotype was also observed in about

15% to 20% of the plantlets, but only after 120 days of synthetic seed storage.

A B C

A B

Fig. 3. Comparison of germination rate between encapsulated PLBs.

(A) and non-encapsulated PLBs, (B) stored in either Petri dish or 15ml PP tube for 0, 30, and 60

days. Cultivation conditions: 25±1oC, 16-hour photoperiod, 50-day cultivation period.


CONCLUSION

The synthetic seed technology can prove to be an effective method to preserve Orchid seeds

for large scale propagation of these beautiful ornamental species or for import and export purposes

based on other previous studies and our research findings. Our studies showed that DendrobiumWhite Fairy can be stored at 25oC in the dark as encapsulated beads in both Petri dishes and screw

capped PP tubes, in which the later has shown to be more favourable. This study has also proved

that non-encapsulated protocorms can be stored directly in screw capped PP tubes without the need

to encapsulate, with the ability to maintain a high germination percentage of about 80% after 60

days of storage at 25oC in the dark. With these findings, we have shown a more simplified way of

storing and propagating Dendrobium White Fairy PLBs. However, more research would be re-

quired to study and optimize the storage procedure for this orchid species.

ACKNOWLEDGEMENTS

The authors would like to thank Professor Chia Tet Fatt for his valuable technical advice, Mrs

Khoo-Woon Mui Hwang, the Singapore Botanic Gardens, for providing technical training. The study

is financially supported by the Ministry of Education, Singapore (Grant No. MOE2009-IF-1-012).

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www.jornamental.com


In Vitro Flower Bud Formation, Plant Regeneration

and Morphogenetic Studies in Local Scented Cultivar

of Rosa indica

In vitro propagation of rose (Rosa indica) as well as rose breeding is a

valuable economic enterprise and has received highest importance globally

including Indian subcontinent. The development of new cultivars for commerce

is faced with new challenges. In vitro micropropagation protocols of varieties

of roses have been established but very few reports on the clonal propagation

of local cultivars of Indian scented rose has been made. In the present paper

we report the dedifferentiation and redifferentiation of stem nodal, leaf disc,

petals well as ovary explants cultured on MS medium supplemented with

various growth adjuvants such as coconut water (CW 5%-20%v/v), auxins

(IAA, NAA, 2,4-D and IBA) and cytokinins (BAP and Kn) in varying con-

centrations. Vigorous callus formation was observed showing embryogenic

clusters in both the nodal and leaf disc explants on MS medium supplemented

with CW(10% v/v) + 2,4- D (11.3 µM) + BAP (3.55 µM). The proliferation

of 4-7 multiple shoots (mean number of shoots 5 ± 1.24) as well as in vitroflower bud formation on in vitro regenerated rose plants from nodal explants

cultured on MS medium supplemented with CW + 2,4-D + BAP have been

observed. The regenerated shoots treated with ‘Quic Root’ (commercially

available rooting chemical) produced well developed root system.

Keywords: Multiple shoots,‘Quic Root’, Rose, Rosa indica, Tissue culture.

Raageeva Bimal and Niti Kiran*

Biotech Lab, University Department of Botany, B.R.A. Bihar University, Bihar-842001 (India).


Abstract


INTRODUCTION

Plant tissue culture is a novel technique widely used for micropropagation of plants under

aseptic condition. Rose,being the queen of flowers is a symbol of beauty, love and tranquility and

has been a favorite object of research for plant scientists for morphogenetic studies. Well estab-

lished and reliable protocols for micropropagation of rose have been developed for hybrid cultivars

using shoot tip and axillary bud and petal explants (Hasegawa, 1979; Bressan et al., 1982; Burger

et al., 1990; Skirvin et al., 1990; Rout et al., 1991; Chatani et al., 1996; Murali et al., 1996; Pati

et al., 2004, 2005; Zabbarzadeh and Khos-khui, 2005; Senapati and Rout, 2008; Nak-Udom et al.,2009; Kermani et al., 2010; Mamaghani et al., 2010; Ebrahimi and Mohammadi-Nejad, 2011;

Hegde et al., 2011; Mukhambetzhanov et al., 2011; Pawlowska, 2011; Farahani and Shaker, 2012;

Moallem et al., 2012; Shadparvar, 2012; Shirdel et al., 2013; Zeng et al., 2013). Earlier attempts

to induce organogenesis from callus of rose were unsuccessful (Khosh- Khui and Sink, 1982).

Later, adventitious differentiation in callus culture obtained from leaf, stem, internodes and zygotic

embryo segments have been demonstrated by (Tweedle et al., 1984; Lloyd et al., 1988; Burger etal., 1990). Recently, organogenesis and plant regeneration from petal explants have also been re-

ported in hybrid roses (Chatani et al., 1996; Murali et al., 1996; Nanomura et al., 2001). Flower

formation is a very important morphogenetic event depending on the genetic makeup, physiological

stage of explants and environmental factors. In vitro flower initiation is not a very common phe-

nomenon. In vitro induction of flower bud has been reported in different varieties of R.hybrida L.

(Wang et al., 2002; Kanchananpoom et al., 2009), hybrid tea rose cv.‘First Prize’(Vu et al., 2006).

Tissue culture provides a convenient tool to study the biological mechanism of transition from

vegetative to reproductive phase. However, no systematic studies have been made on In vitroflower or bud formation in scented variety of R.indica and also there are very few reports on the

morphogenetic potentials of vegetative and reproductive parts of Indian scented rose (Soomro etal., 2003; Hameed et al., 2006). Rose propagation which is usually practiced by cutting and grafting

is labour intensive (Horn, 1992) and dependent on season. The slow multiplication rate in roses is

another constrain which may be overcome by using the plant tissue culture technique for efficient

and reliable mass production of identical plants of rose throughout the year. In the present paper

multiple shoot, plant and flower bud formation in local scented cultivar of rose (Rosa indica) fol-

lowed by ‘Quic Root’(a commercially available rooting chemical) induced In vitro rooting in shoots

have been established and the protocol has potentials for application in tissue culture nursery both

for raising planting materials as well as rose flower (buds).

MATERIAL AND METHODS

Fresh explants were collected from Botany department of B.R.A. Bihar University. The

explants such as node, inter node, pollinated flower buds, leaf, petals and embryo were washed

thoroughly under running tap water for half an hour followed by washing in liquid detergent for

next half an hour. Later the explants were again washed under running tap water for 10 min. The

explants were surface sterilized with 0.1% HgCl2 solution for 5 min and finally washed two to

three times with double distilled sterile water. These explants were cultured on Murashige and

Skoog’s medium (1962) supplemented with coconut water (CW 10%v/v), IAA (0.57- 28.5 µM),

2,4-D (0.45-22.6 µM), NAA (0.54-26.8 µM), BAP (0.44-22.2 µM), Kn (0.46-23.2 µM), IBA (0.49-

24.60 µM) either singly or in various combinations. The regenerated shoots were rooted in vitroby treating the lower ends of shoots by 'Quic Root' (Ashwin Chemical, Banglore, India) for 10-30

sec and culturing in MS liquid medium (half strength of macro, micro, iron and vitamins) and 3%

sucrose. The cultures were incubated at 25ºC ± 5ºC under cool fluorescent light and 16 h photope-

riod in culture room. The experiments were repeated at least three times to substantiate the repro-

ducibility of the observations and data presented in table 1 show the mean value of three sets of

experiments.


RESULT AND DISCUSSION

In local scented cultivar of R.indica the explant taken from both vegetative and reproductive

organs responded in culture and showed dedifferentiation into callus as well as regeneration of

shoots. The explants cultured on MS medium without any growth adjuvants did not show any re-

sponse.

Table 1 shows relationship between the number of nodal explants responding by prolifer-

ating shoot(s) on different nutrient media supplemented with various growth promoters. The nodal

explants cultured on MS + BA (2.22 µM) showed callus formation from all over the surface of the

explants. However, the nodal explants cultured on MS + BAP (22.20 µM) + Kn (2.32 µM) showed

proliferation of axillary buds, into 2 shoots/node and very poor callus formation was observed.

Rooting in the cultured explants was observed on MS medium supplemented with BAP (22.20

µM) + Kn (2.32 µM), but these roots started dedifferentiating into callus in 4-6 days of culture.

The nodal explants cultured on MS + BAP (11.1 µM) + Kn (4.6 µM) also produced two shoots

per explant but the frequency of response was better. In another experiment, the nodal explants

cultured on MS + BAP (3.55 µM) + 2,4-D (11.31 µM) + CW10%v/v produced 4-7 (mean number

of shoot 5 ± 1.24) multiple shoots (Figs.1-3). However, the nodal explants cultured on MS + 2,4-

D (2.26 µM) + Kn (4.46 µM) and 2,4-D (4.5 µM) + Kn (4.6 µM) promoted the growth of only

one shoot in 7.6% and 16.9% explants, respectively. The regenerated shoots treated with ‘Quic

Root’ and cultured on half strength MS constituents containing 3% sucrose produced well devel-

oped root system and the protocol proved to be highly effective (Figs. 4-6). The ‘Quic Root’ treated

ends turned dark (Fig. 4) and in 2-3 days rupturing of the surface due to bulging at the treated end

was observed (Fig. 5). The root initiation was also observed in 5- 6 days of culture followed by

formation of 3-4 well developed roots in about 12 -15 days of culture (Fig. 6).

The stem pieces (Figs. 7-8), petal (Fig. 9), leaf discs (Fig. 10) and fertilized ovary pieces

(Figs. 11-12) cultured on the same medium (MS + BAP + 2,4-D + CW) showed callus formation.

The regenerated shoots sub-cultured on MS medium containing NAA (5.37 µM) + BAP (22.20

µM) induced formation of flower buds on tiny rose shoots (Plate 2 Figs. 13-14 and 16). The first

flower bud was observed in 35 days of culture, while the other bud appeared in 62 days of culture.

The first one was miniature flower bud with green sepals and enclosing dark violet petals, the buds

measured 2-3 mm in length but did not open. The peduncle of the first bud did not show clearly

formed adnate stipule. The second flower bud was comparatively elongated and showed green

sepals and dark violet petals. The stalk of the bud very clearly showed adnate stipules probably

indicating the transformation of leaf into flower bud (Plate 2, Fig. 16).

The flower bud (Fig. 15) also produced on MS + BAP (0.89 µM) + CW (10%v/v). The IBA

was used for rooting of the regenerated shoots and the shoots were cultured on MS medium sup-

plemented with IBA (4.90 µM) developed roots. Callus formation was observed in 6-8 weeks with

on MS medium supplemented IBA and the callus growth was average. In some of the shoots

Sl.No. Different hormones supplemented in MS basal

medium

Number of responsive ex-

plants in culture in term of

shoot proliferation/130 ex-

plants in each set of medium*

Response in terms of number of

shoot(s)/explants

1.

2.

3.

4.

5.

2,4-D (11.3 µM) + BAP (3.55 µM) + CW (10%)

BAP (22.2 µM) + Kn (2.3 µM)

BAP (11.1 µM) + Kn (4.6 µM) + NAA (5.3 µM)

2,4-D (2.26 µM) + Kn (4.46 µM)

2,4-D (4.5 µM) + Kn(4.6 µM)

60.10±0.9

17.09±0.9

21.18±0.9

10.40±1.6

22.09±1.6

4-7 (mean shoot number 5±1.24)

2

2

1

1

Table 1. Correlation between number of nodal explants responding by proliferating shoot(s) on different nutrient

media containing various growth promoters

*( mean ± SD )


in vitro differentiation of flower bud was observed in presence of IBA but did not blossom. The

nodal explants cultured on MS medium containing 9.80 µM IBA showed profuse callus formation

in 5-6 days as well as also root formation in 20-25 days of culture. When the nodal explants were

cultured on MS medium supplemented with IBA (4.90 µM) and Kn (5.71 µM) both callus forma-

tion and root initiation was observed in 10-12 days of culture. Callus formation was observed in

the nodal explants cultured on MS medium containing IBA (9.80 µM) + CW (10%v/v) in 6-7 days.

The half strength MS medium supplemented with 2.46 µM IBA induced callus formation from

nodal explants in just 2- 3 days of culture. Petal disc cultured on MS medium supplemented with

BAP (0.44 µM) + NAA (1.07 µM) and incubated in dark produced callus in 80% of the explants

but no differentiation or shoot bud was observed.

1 2 3 4

5 6 7 8

9 1210 11

Plate 1. Morphogenetic studies in Rosa indica. Figs.1-3. Multiple shoots developing from nodal explants

on MS + 2,4-D (11.3 µM) + BAP (3.5 µM) + CW (10%v/v). Figs. 4-6. Different stages of ‘Quic Root’ induced

root formation. Figs.7-8. Embryogenic callus showing globular embryoids from nodal explants. Figs.9-12.

Callus proliferation from petals, leaf discs unfertilized and fertilized ovary pieces.

13 14 15 16

Plate 2. Morphogenetic studies in Rosa indica. Fig.13. A regenerated shoot. Figs. 14, 16. Differentiation

of flower buds in vitro. Fig.15. A well developed flower bud was formed on MS + BAP (0.89 µM) + CW

(10%v/v).


R.indica (local cultivar) bearing the most attractive flowers, is strongly scented flowering

plant and is widely used in perfumery, cosmetic products, medicine and food products. In vitromcropropagation protests in any of rose cultivars have been developed in the past using shoot tip

and nodal explants (Rout et al., 1989, 1990; Skirvin et al., 1990; Pati et al., 2005; Zabbarzadeh

and Khosh-Khui, 2005; Sengupta and Rout, 2008; Canali and Kazaz, 2009; Kermani et al., 2010;

Hegde et al., 2011; Pawlowska, 2011; Ebrahimi and Mohmmadi-Nejad, 2011; Farahani and

Shaker, 2012; Moallem et al., 2012; Shadparvar, 2012; Shirdel, 2013; Zeng et al., 2013). However,

the slow rates of multiplication through conventional method of cutting as well as its dependence

on the season are greatest disadvantages for establishment of rose nursery for commerce. In the

present paper we report the establishment of an efficient method for shoot and root regeneration

in tissue culture of local scented cultivar of experimental system R.indica L. 2,4-D, BAP and CW

induced maximum number of 4-7 shoot buds from node each node of the explants.The hormonal

combination BAP + Kn produces only 2 shoots/explants,while MS medium supplemented with

2,4-D + Kn produced only one shoot per nodal explant. Soomro et al. (2003) reported plant re-

generation in R. indica on MS medium supplemented with IBA and NAA. Rout et al. (1991) re-

ported the induction of callus and somatic embryognesis in R.hybrida var.Landora only in the

presence of three adjuvants. Rout et al. (1990) observed that BAP is the most effective growth ad-

juvant for shoot proliferation. They also reported similar observations in R.canina and R.damas-cena. The formation of globular callus along with rooting has been observed in present system

which shows similarity with the observation made by Rout et al. (1990). In R. damascena direct

shoot regeneration from leaf in the presence of TDZ (thidiazuron) and AgNO3 (silver nitrate) was

observed (Pati et al., 2004). In R. hybrida L. cv. ‘Perfume Delight’ 3 shoots were regenerated from

nodal explants on BA+NAA combination (Nak- Udom et al., 2009). However, Senapati and Rout

(2008) reported high frequency of shoot multiplication in IAA + cytokinin supplemented medium

in three cultivars of R. hybrida viz. R. hybrida ‘Cri Cri’, ‘Pariser Charme’ and ‘First Red’. The

rose cultivar’Pareo’ the nodal explants produced multiple shoots on MS medium containing BAP

(Mukhambetzhanov et al., 2011). The proliferation of 4±7 multiple/shoots explants was obtained

on MS + 2,4-D + BAP + CW, while Soomro et al. (2003) reported regeneration of 2-6 shoots per

explants in presence of IBA and NAA. Shirdel et al. (2013) higher concentration of BAP as the

most important factor for multiple shoot induction in R. canina. In present system, BAP alone in-

duced callus formation but when supplemented with 2,4-D and CW proved to be important for

proliferation of multiple shoots. Shadparvar (2012) reported callus mediated shoot formation in

R.miniature ‘Green Ice’ in the presence of IBA and 2iP. In R. foetida, BA + IBA combination was

found essential for highest number of shoots (4.8 mean shoot number per explants) (Ebrahini and

Mohammadi-Nejad, 2011). Farahami and Shaker (2012) recorded maximum no.of 4 shoots per

node on MS + BAP + IBA in miniature. In R. hybrida cv.‘Black Baccara’ Bayanati and Mortazavi

(2013) reported proliferation in the presence of BAP. Kermani (2010) also found BAP inducing

maximum of 4.66 shoots on VS medium in R. persica. In R. damacena, BAP + TDZ containing

MS medium induced maximum number of shoots (Mamaghani et al., 2010). In the present study,

the rose explants (nodal, leaf, ovary and petals) treated individually with different auxins such as

IAA, NAA, 2,4-D, IBA did not show any noticeable morphogenetic effect. Although auxin (2,4-

D) exerts control at transcriptional and translational level (Sengupta and Raghavan,1980 a,b) the

auxin applied individually did not favour redetermination of a cell for morphogenesis in the present

experimental system. The physiological role of IBA in morphogenesis in rose is remarkable. The

half strength MS containing IBA induced callus formation but supported healthy shoot develop-

ment only when IBA was added in the full strength MS medium. Low concentration of IBA has

been found to promote shoot development in Macrotyloma uniflorum (Tejavathi et al., 2010).

Under normal growth conditions differentiation of flower takes place when a plant attains maturity,

since the age of the plant is genetically determined and is species specific. A flower bud or flower


is produced when physico-chemical and environmental milieu are congenial. However, these con-

ditions can be regulated for transformation of vegetative phase of a plant into reproductive phase.

The pioneer work on in vitro flowering was reported in Cassava without hormones in culture

medium (Tang et al.,1983). In vitro inflorescence had been induced from axillary buds of Morus

alba (Naik and Latha, 1996). Pai et al. (1986) induced inflorescence through inflorescence culture.

The precocious flowering in bamboos has been reported through inflorescence culture (Nadgauda

et al., 1990). In vitro flowering from cotyledon cultures of groundnut was reported by Narasimhalu

and Reddy (1984). In vitro flowering was produced on callus derived plantlets of Papaver somnifera

(Yoshikama and Furaya, 1983) and during somatic embryo formation in Brassica nigra (Mehta etal., 1993). Very recently, in vitro flower induction from callus in juvenile explants of sugarcane,

Saccharum officinarum var. CoC 671 has been reported (Virupakshi et al., 2002). A tiny spathe

like formation has been reported in banana from cultured male floral primordia (Bimal and Jha,

2008). In the present experimental system, flower bud formation in the regenerated plant took

place in 6-8 weeks in the presence of hormone IBA or in combination of NAA+BAP. Flowering

from vegetative tissue was observed in vitro on long-term callus culture of sugarcane meristematic

tissue (Virupakshi et al., 2002). It seems probable that osmotic and nutritional stress to the growing

tissue resulted in flower bud formation in 6-8 weeks of culture on the medium without subculturing.

In vitro flower bud or flower formation is not a spontaneous developmental decision of the plant

rather it is the culmination of closely integrated programmed physiological and developmental

changes which evocate flowering (Govil, 2004). The flower buds formed in culture did not open,

it is likely that the appropriate signal to induce opening of the in vitro produced flower buds might

be lacking in the scented variety of R.indica. There are many reports indicating the synergistic ac-

tion of auxin and coconut water (Bapat and Narayan Swami, 1977; Bimal and Jha, 1986). Wang

et al. (2002) studied in vitro flowering in six cultivars of rose and the highest percentage of flower

bud induction (49.1%) was obtained on media supplemented with TDZ and NAA for the cultivar

Orange Parade. TDZ or ZT was the best choice for flower induction in all six cultivars studied.

The culture and subculture duration were also found to be important for flower induction. Age of

plantlets in culture and age of mother plant providing explants influence the induction of flower

formation and generating a consistent system of large scale production. Five Parade rose hybrids

(Binanea Parade, Fiesta Parade, Orange Parade, Scarlet Parade and Viva Parade, from the Nether-

lands) and a bigger red rose cultivar (RF) from Malaysia were tested and a period of 45 days in

induction medium was responsible for highest percentage of plantlets forming flower buds (Wang

et al., 2002). It is likely that prolonged vegetative growth including elongation of stem and forma-

tion of roots in culture might have made transition to the reproductive stage. Vase life of in vitroinduced flower was about one month at room temperature or below 25 ̊C. ‘First Prize’ variety of

rose (hybrid tea) differentiated flower shoots cultured on MS medium + BAP + NAA + 3% sucrose

(Vu et al., 2006). According to them sucrose is a key factor in induction, cytokines increase the

flowering percentage and normal development and reduced inorganic and organic salt concen-

tration in MS medium had a positive effect on in vitro flowering. Kanchanapoom et al. (2009) re-

ported proliferation of 5 shoots per explants on MS + BAP (13.30 µM) + Kn (4.60 µM), followed

by flower induction on MS + BAP (8.90 µM) in 9 weeks in R. hybrida L.cv. ‘Red Master Piece’.

Although in vitro flower bud has been induced, more reliable culture conditions for in vitro flower

induction need to be elucidated. In our experiments flower bud formation in local scented variety

of rose (R. indica) has been observed. Stimulation of cell division by coconut water has been re-

ported earlier (Morel, 1950; Bakely and Steward, 1961). However, in the present study, besides

coconut water the presence of 2,4-D and BAP seems to be a prerequisite for stimulation of cell di-

vision. Although, there are reports on the induction of callus, shoot etc. in the presence of either

auxin or cytokinin individually, auxin or cytokinin did not play a role in any type of morphogenesis

in R. indica var. local scented except IBA. However, IBA in combination with BAP was found to


be the most suitable treatment in damask rose, R. damascena Mill. (Zabbarzadeh and Khosh-Khui,

2005). Sisko (2011) reported genotype dependent rooting response in Rosa spp.The regeneration

of roots is regulated at least in part by auxin and the localization of the differentiation of roots at

the basal end of the cuttings is due to the polar movement of auxin towards the physiologically

lower end. Mukhambetzhanov et al. (2011) reported 100% rooting in rose cultivar’ Pareo’ in the

presence of IBA + IAA. The effect of rooting chemical 'Quic Root' has been studied in R. indicafor the first time. The efficiency of root induction and morphogenesis observed in R. indica was

significant. Altogether 3-7 roots differentiated and attained proper growth. The rooting chemical

has been reported to enhance cambial activity by mobilizing reserve food materials to the site of

root formation (Gurumurithi et al., 1984; Purohit et al., 2008). Although spring or the season when

the fresh flush starts has been found to be a favorable period for rooting experiment, in the present

experiment rooting can be induced round the year.

CONCLUSION

Results obtained in the present study report the establishment of a two step protocol for

rapid micropropagation of the local scented cultivar of R.indica L. and in vitro proliferation of

rose flower buds.The protocol using MS nutrient Medium supplemented with 2,4-D, BAP and CW

has been successfully established for regeneration of multiple shoots from nodal explants followed

by ‘Quic Root’ mediated rooting of rose micro-shoots which has potentials for development of tis-

sue culture nursery supplying tissue culture raised planting material to the market.

ACKNOWLEDGEMENTS

Authors thank Head of the University Department of Botany for facilities. One of us (NK)

thanks UGC for financial assistance (No.F.15-21/12 (SA-II).

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Study of Genetic Diversity among Philodendron Varieties

by Randomly Amplified Polymorphic DNA Markers

In the present study, Randomly Amplified Polymorphic DNA (RAPD)

markers were used to analyze the genetic diversity in 20 varieties of

Philodendron. The polymerase chain reaction was performed with 60 RAPD

primers, out of which 21 primers showed clear amplification as well as more

polymorphism. In total, 354 scorable RAPD loci with 348 polymorphic

bands (98%) were observed. Percentages of polymorphic bands ranged from

80% to 100%. Amplified DNA fragments were used for statistical analysis.

The data was obtained by scoring the bands and analyzed by using STATISTICA

software. A dendrogram was obtained by cluster analysis based on the

presence or absence of band which indicates similarity and diversities. The

cluster analysis grouped the Philodendron varieties based on the growth

habits, morphological characters and also geographical origin. This indicates

that there is an association between the RAPD patterns and the geographic

origin of the varieties used. The genetic diversity among the varieties was

moderate, which might be due to genetically heterozygous and somaclonal

variation.

Keywords: Aroid, Houseplants, Self- heading, Spadix, Tropical forests.

Achar Devaraja AM*, Jakhar Mamta , Jakeer P and Shetty KPV

Biotechnology laboratory, Indo American Hybrid Seeds (I) Pvt Ltd., Bangalore 560 098, India.


Abstract


INTRODUCTION

Philodendron is the second largest genus of flowering plants from the Arum family

(Araceae), mainly found in many diverse habitats in the humid tropical forests of Americas and the

West Indies. None the less, they can also be found in Australia, some Pacific islands, and Asia al-

though they aren't indigenous to these regions. Rather they were introduced or accidentally escaped.

According to TROPICOS (a service of the Missouri Botanical Garden), more than 900 species were

recognized, however Mayo et al. (1997) reported that there are about 400 formally recognized

species, whereas according to Croat (1997) and Boyce and Croat (2012) list there are about 700

and in Brazil it has ca. 165 species (Sakuragui et al., 2012) in several biomes. Compared to other

genera of the family Araceae, Philodendrons have an extremely diverse array of growth habits. The

growth habits were divided into three groups, the first group is veining, and it is grown either in

hanging baskets or container where vines are supported by poles. In the second group growing style

is upright and third group is erect-arborescent type, which appears self-heading when they are young,

but assume more woody and treelike shapes as they mature (McColley and Miller, (1965). However,

majority of Philodendron species leaf growth gradually go through metamorphosis; there is no im-

mediately distinct difference between juvenile and adult leaves (Bell and Bryan, 2008).

Reproduction in Philodendron can occur in two primary ways: through sexual and asexual re-

production. Sexual reproduction is achieved by means of beetles, with many Philodendron species

requiring the presence of a specific beetle species to achieve pollination. However crosses between

Philodendron species may perhaps not possible (McColley and Miller, 1965), due to differences in

the chromosome number (Henny and Chen, 2003). Chromosome counts in the genus are available for

only 10% of the species with a predominance of 2n = 32, 34 and 36, found in P. scandens, P. wend-landii, and P. bipinnatifidum, respectively (Cotias-de-Oliveira1 et al., 1999). However the genetic re-

latedness among the Philodendron varieties has not been studied thoroughly. Therefore in the present

study, RAPD markers were used to investigate genetic divergence among widespread varieties of

Philodendron and strategies for future hybrid development as well as conservation of genetic resource.


Plant material

Twenty Philodendron varieties were collected from Indo American Hybrid Seeds Company

plant tissue culture green house, Bangalore. These varieties were produced through tissue culture

Sl.No. Varieties Growth habit Origin

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

Philodendron xanaduPhilodendron panduriformeMoonshine

Tropical Sunrise (hybrid)

Kerala Hybrid (hybrid)

Philodendron laciniatumPhilodendron ceylonP. selloum var. Gold (hybrid)

Philodendron selloumPhilodendron Emerald DukeRed Emerald (hybrid)

Philodendron wendlandii Royal Queen (hybrid)

Pluto (Hybrid)

Black Cardinal (hybrid)

Philodendron deflexumGreen Emerald (hybrid)

Painted lady (hybrid)

Pink princes (hybrid)

Golden Xanadu (hybrid)

Self- heading

Semi- vining

Self-heading

Self-heading

Semi-vining

Semi-vining

Semi-vining

Self-heading

Self-heading

Semi-vining

Semi-vining

Semi-vining

Semi-vining

Self-heading

Self-heading

Semi-vining

Semi-vining

Semi-vining

Semi-vining

Self-heading

Brazil

North & Western parts of Amazon

Florida

American tropics

west coast of Kerala

South Eastern Brazil

Brazil

Brazil

Brazil

Thailand

Florida

Florida

Florida

unknown

Florida

Brazil (South America)

Florida

Brazil

Florida

Brazil

Table 1. Growth habit and origin of 20 Philodendron varieties used in the present study.


in company laboratory. The growth habit and country of origin is being given in Table 1 and Fig.

1. The genotypic study was conducted during January 2013.

Genomic DNA extraction and primer selection

Total genomic DNA was also extracted from fresh leaves according to the CTAB method

described by Porebski et al. (1997). After isolation of DNA, its quantity was determined spec-

trophotometrically at a wavelength of 260 and 280 nm using Spectrophotometer. The ratio between

absorbance at 260 and 280 nm (260/280) was used to estimate DNA purity. DNA of each cultivar

was diluted to a working concentration of 20 ng/μl for PCR/RAPD analysis.

Randomly five varieties of DNA were used to optimize the RAPD protocols and selected

the suitable primers which exhibit clear amplification. Overall, 60 arbitrary decamer oligonu-

cleotides, from Operon Technologies Inc. (Alameda, California, USA), were tested as single

primers to identify the most promising ones for detecting polymorphism. Eventually 21 primers

were chosen on the basis of their ability to detect the polymorphism and produce the reliable and

easily scorable banding patterns (Table 2).

PCR-amplification and gel electrophoresis

A modified RAPD method based on Williams et al. (1990) was used with a model T-100Tm

thermal cycler (BIO-RAD, USA). The PCR reaction mix included the following: 20 μl reaction

mixture containing 1 × PCR buffer [10 mM Tris HCl (pH 8.3), 50 mM KCl], 1.5 mM MgCl2, 200

μM each deoxynucleotide triphosphate (dNTPs), 0.4 μM of 10-mer primer (Operon Technologies

Inc., Alameda, CA), 1 unit Taq DNA polymerase and 20 ng of template DNA for the amplification

of genomic DNA. The PCR profile started with 94°C for 5 min followed by 45 cycles of denatu-

Fig. 1. Photograph of 20 Philodendron varieties used in the present study.


ration at 94°C for 1 min, annealing at 37°C for 1 min and extension at 72°C for 2 min. A final ex-

tension, 72°C for 8 min was included, followed by soaking at 20°C.

Electrophoresis

The RAPD-PCR products were analyzed directly on 1.5% agarose gels in 0.5X TBE buffer,

visualized by staining with ethidium bromide and transillumination under short-wave UV light.

Data analysis

The clear and unambiguous RAPD bands were scored to the presence (1) and absence (0)

of the corresponding band among the varieties. The marker data subjected to cluster analysis using

the software STATISTICA” (Statsoft, 1996). A dendrogram was constructed for genotypic data

using UPGMA (unweighted pair-group method using arithmetic averages) method. The dissimi-

larity matrix was developed using Squared Euclidean Distance (SED) that estimated all pairwise

differences in the amplification product (Sokal and Sneath, 1973).


RAPD profile and analysis

The 10-mer 60 RAPD primers were tested for genetic analysis of 20 Philodendron varieties.

However, the best 21 primers that rendered polymorphic and reproducible banding pattern were

chosen. The selected few gel profiles of 20 Philodendron varieties are furnished in Fig. 2.

Using twenty one selected primer combinations, a total of 354 scorable RAPD fragments

were generated, out of which 348 (98%) were polymorphic (Table 2). Out of 21 primers, 16 primers

showed 100% polymorphism and four other primers also showed 94 to 98% polymorphism except

one primer OPC08, which showed 80% polymorphism. One of the reasons for this high level of

polymorphism could be the extensive inter species variation in Philodendron. The other explanation

could be that primers with 60 to 70% GC content were used. Fukuoka et al. (1992) observed an

increase in the number of bands with increasing GC content of the primers. They got an average

of 0.8 bands primer-1 with 40%, 6.1 bands with 50% and 8.6 bands with 60% GC content.

Primers Sequence

(5' to 3')

No. of

Bands

No. Polymorphic

bands

% polymorphism

OPA01

OPA03

OPA04

OPA05

OPA09

OPA12

OPA13

OPA15

OPA16

OPA17

OPA18

OPA19

OPC02

OPC05

OPC06

OPC07

OPC08

OPD18

OPF03

OPF13

OPN12

CAGGCCCTTC

AGTCAGCCAC

AATCGGGCTG

AGGGGTCTTG

GGGTAACGCC

TCGGCGATAG

CAGCACCCAC

TTCCGAACCC

AGCCAGCGAA

GACCGCTTGT

AGGTGACCGT

CAAACGTCGG

GTGAGGCGTC

GATGACCGCC

GAACGGACTC

GTCCCGACGA

TGGACCGGTG

GAGAGCCAAC

CCTGATCACC

GGCTGCAGAA

CACAGACACC

Total

16

16

15

19

14

18

13

21

21

18

25

21

15

14

11

15

15

14

12

18

23

354

16

15

15

19

14

18

13

21

21

18

25

21

15

14

11

15

12

14

11

17

23

348

100

94

100

100

100

100

100

100

100

100

100

100

100

100

100

100

80

100

92

94

100

98

Table 2. List of 21 random primers used for generating RAPDs in varieties of Philodendron.


The DNA fragments ranged from 100 to 1500 bp with a majority of polymorphism. The

level of polymorphism from this study is corroborated to that of 43 Philodendron varieties and

five aroid genera, where AFLP technique was used to measure the genetic diversity (63% poly-

morphism). Furthermore Chen et al. (2004a, 2004b) reported 69% polymorphism in Aglaonemaspp. and 71% polymorphism in Dieffenbachia spp., the two other aroid genera.

As expected, genetic distances between the species varieties were comparatively low. Eu-

Fig. 2. RAPD profiles of twenty different varieties of Philodendron using primers

OPA12 (a), OPA13 (b), OPA15 (c), OPD18 (d). M: Molecular weight marker (1000

bp DNA ladder), Lane 1: P. xanadu, 2: P. panduriforme, 3: Moonshine, 4: Tropical

Sunrise (hybrid), 5: Kerala Hybrid (hybrid), 6: P. laciniatum, 7: P. Ceylon, 8: P. sel-loum var. Gold (hybrid), 9: P.selloum, 10: P. Emerald Duke, 11: Red Emerald (hy-

brid), 12: P. wendlandii, 13: Royal Queen (hybrid), 14: Pluto (Hybrid), 15: Black

Cardinal (hybrid), 16: P. deflexum, 17: Green Emerald (hybrid), 18: Painted lady

(hybrid), 19: Pink princes (hybrid), 20: Golden Xanadu (hybrid).

Green xanadu

p.panduriformMoonshine

Tropical Sunrise

Kerala Hybrid

Lacinatum

Ceylon

slm var. Gold

p. selloumEmerald Duke

Red Emerald

wendlandii

Royal Queen

Pluto

Black Cardinal

Deflexum

Green Emerald

Painted lady

Pink princes

Golden Xanadu

0

10

10

11

10

11

11

10

9.9

11

11

10

11

11

11

11

11

11

11

8.9

0

9.7

9.9

9.2

9.4

9.8

9.7

11

9.8

9.6

9.6

9.9

9.8

9.9

11

10

9.5

9.9

10

0

7.3

9.9

9.6

9.7

9.8

9.4

8.8

9.2

8.9

8.9

9.5

8.8

9.6

10

90

9

11

0

9.9

9.8

9.9

11

10

9.3

9.5

9.2

9.3

9.6

9.1

9.3

9.9

9.2

9.2

11

0

9.7

10

9.7

11

10

11

10

10

9.9

10

11

11

10

10

9.7

0

9.7

10

10

10

10

10

10

9.5

10

10

10

11

10

11

0

10

11

9.8

10

9.9

10

10

11

10

10

10

9.8

11

0

9.7

10

10

10

9.7

10

9.8

10

10

9.8

10

10

0

10

11

10

11

11

11

11

11

11

11

11

0

8

8.1

7.9

8.3

8.1

8.9

9.4

8.4

7.9

10

0

7.9

7.9

8.4

8.2

8.4

9.7

6.2

8

10

0

7.8

8.3

8.5

7.8

8.8

8.1

7.7

10

0

8.1

7.1

8.9

9.7

8.1

6.9

10

0

7.9

9.6

9.5

8.4

8.4

10

0

8.8

9.5

8.1

7.6

10

0

8.7

8.1

8.4

11

0

9.8

9

11

0

7.6

9.9

0

10 0

Gre

en x

anad

u

p.pa

ndur

iform

Moonsh

ine

Tro

pic

al S

unri

se

Ker

ala

Hybri

d

Lac

inat

um

Cey

lon

slm

var

. G

old

Em

eral

d D

uke

Red

Em

eral

d

wen

dla

ndii

Royal

Quee

n

Plu

to

Bla

ck C

ardin

al

Def

lexum

Gre

en E

mer

ald

Pai

nte

d l

ady

Pin

k p

rince

s

Gold

en X

anad

u

p. se

lloum

Table 3. Euclidean distance metric based on RAPD marker data showing the relationships between

Philodendron species.


clidean distance metrics estimates of distance for 21 varieties ranged from 7.6 to 11 (Table 3). In

fact most of them are assorted 10 to 11.

Genetic relationship among the varieties

A dendrogram was constructed for 20 Philodendron varieties based on UPGMA using

squared euclidean distance matrix (Fig. 3). The generated three major clusters and revealed max-

imum linkage distance of 108 units. Cluster I, II, and III covered a linkage distance of 108, 102

and 90 units, respectively. The first cluster contains Philodendron xanadu varieties viz. green

xanadu, golden xanadu and P. bipinnatifidum varieties viz. selloum and selloum gold. They are

native to Brazil, multi-lobed and self heading. However P. xanadu was referred as miniature of P.selloum (Devanand et al., 2004), but there is an entirely different plant also called miniature P. sel-loum, which is in fact a unique natural variation of P. bipinnatifidum, however, that plant is rare

(Croat et al., 2002). These two species were differentiated with spathe and spadix. The spathe of P.xanadu is dark violet-purple bordering on red and the spathe of P. bipinnatifidum is basically green

with a white interior. In fact P. bipinnatifidum (syn. P. selloum), is one of the world's most ubiquitous

houseplants which has provided a rich source of variation for breeding different foliage forms.

However in our study grouping of these species in first cluster infers there is close genetic

similarity between species and they may be rather genetically heterozygous.

3 The cluster II contains four varieties and all are scandent (climbing) vine. ‘Kerala hybrid

with pale green color, ovate leaves, P. ceylon with yellow color, ovate leaves, P. laciniatum and

P. panduriforme with light green and dark green lobed leaves respectively. The genetic distance

among them ranges 9.7 to 10 (Table 3). This indicates that they are not genetically distant, even

though they are genetically heterozygous.

Cluster III contains twelve varieties and most of them are climbers except four varieties

which are self headers. As estimated in the distance matrix, cluster analysis placed most of the va-

rieties close to each other showing a high level of genetic relatedness. Among climbers and self

heading varieties, cluster generally corresponds to color and shape of the leaf as well as plants

growth habit. For example tropical sunrise and moonshine are grouped together with a genetic dis-

tance of 0.73 and both are self header, stout or almost no stems with bright red new growth. Black

Cardinal is a hybrid developed from a series of crosses involving P. wendlandii, P. domesticum, P.erubescens, P. imbe, P. fragrantissimum (Devanand et al., 2004) and is grouped with ‘Pink prices

Fig. 3. Unweighted Pair Group Method with Arithmetic Mean (UPGMA) dendro-

gram showing the genetic relationships among the 20 Philodendron varieties using

RAPD markers.


(dark leaves and light pink variegated blotches)’, ‘Emarld Duke (deep green leaves)’ and ‘Royal

Queen’ (deep maroon foliage)’. The grouping of these hybrids could be due to similarity of mor-

phological characteristics as well as narrow genetic base, furthermore it is supported with Euclidean

distance range 7.1 to 8.1(Table 3).

In general agreement, our RAPD-based analysis categorize the three clusters based on va-

rietal growth habit and origin, indicating that, there is an association between the RAPD patterns

and the geographic origin of the varieties used, despite more polymorphism (Fig. 2, Table 2). How-

ever, more genetic polymorphism would infer most of the Philodendrons are open pollinated and

propagated by vegetative means and hence species might be extremely heterozygous. Moreover,

somaclonal variation is another additional factor for the addition of new varieties, since plants

have been cloned heavily through tissue culture. Furthermore, Philodendron species are known to

be highly variable and not every leaf of every species will always appear the same and size is also

not a determining factor when it comes to determining the species. For example the size of any

morphological structure in Anthurium is highly variable because plants often change remarkably

in size over the course of their lives and also in response to the general climatic conditions where

they occur, particularly in geographically widespread species (Croat et al., 2002).

This study reveals the ability of RAPD markers to reliably differentiate between different

Philodendron varieties and their use as a tool for the estimation of genetic diversity. The informa-

tion about genetic similarity will be helpful to avoid the chances of using genetically similar lan-

drace/genotypes and will also be helpful in future breeding programs to select genetically diverse

parents for Philodendrons.

Litrature Cited

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(accessed: 26 March 2013).

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6531-6535.


Growth, Yield and Quality of Carnation (Dianthus caryophyllusL.) Cultivars under Lath House Conditions

Five carnation cultivars (‘Grand Salam’, ‘Nelson’, ‘Kaly’, ‘Cinderella’

and ‘Tempo’) were evaluated with respect to growth, yield and quality char-

acteristics under lath house conditions at Kala Shah Kaku Research Station,

Directorate of Floriculture, Lahore during 2011. Among the cultivars studied,

maximum plant height was recorded in carnation cultivar ‘Grand Salam’

(78.66 cm) closely followed by ‘Kaly’ (78.23 cm) and ‘Cinderella’ (77.96

cm). Number of shoots was highest in cultivars ‘Tempo’ (6.3) and ‘Nelson’

(6.2). Maximum stem thickness was observed in ‘Nelson’ (6.21 mm) and

minimum in ‘Grand Salam’ (3.63 mm). Maximum number of internodes per

stem were found in ‘Nelson’ (12.66) followed by ‘Kely’ (11.33) and ‘Grand

Salam” (11.00). Highest flower yield per plant was recorded in cultivars

‘Tempo’ (6.4) and ‘Nelson’ (6.33). Maximum number of flowers per square

meter were recorded in cultivar ‘Nelson’ (198.3) followed by ‘Tempo’ (189.6).

Hence it can be concluded that cultivars ‘Nelson’ and ‘Tempo’ were found su-

perior with respect to growth and flower yield characteristics under lath

house conditions.

Keywords: Carnation, Cultivars, Lath house, Vase-life, Yield.

Malik Abid Mehmood, Muhammad Saleem Akhtar Khan and Naveed Ahmad*

Directorate of Floriculture (T&R) Punjab, 21-Davis Road, Lahore, Pakistan.


Abstract


INTRODUCTION

Carnation (Dianthus caryophyllus L.) a native to Mediterranean region (Salehi, 2006) is an

important flower crop having great commercial value as a cut flower due to its excellent keeping

quality, wide array of colours and forms (Pralhad, 2009). Carnation, apart from producing cut flow-

ers has also become useful in gardening for bedding, edging, borders, pots, and rock gardens

(Biondo and Noland, 2000; Dole and Wilkins, 2005). From medicinal point of view, the carnation

flowers are considered to be cardiotonic, diaphoretic and alexiteric (Shiragur et al., 2004).

Carnation flowers are sold as cut flowers round the year throughout the world and it is on

the top three cut flowers traded in the international market. The flower quality is maintained in the

long distance transport as they have ability to rehydrate after transportation (Pralhad, 2009). The

demand for carnation as cut flower is gaining momentum with increasing socio-economic standard

of the people.

In Pakistan, the floriculture crops are grown in open fields. Planting after April has to pass

through a great stress due to prevailing high temperature. Though low temperature conditions exist

during winter months but the shortage of light during winter months is the main barrier for its re-

duced and delayed flower production. In addition this, other operations are also known to affect

its flowering but so far, there is no systematic report on its growth and flower behavior to different

climatic conditions.

The performance of carnation varies with region, season, genotypes and growing environ-

ment. In Pakistan, there is a wide fluctuation in temperature, light intensity and humidity which

not only affect the yield and quality of flowers but also limit their availability during particular

period of a year. It is necessary to grow carnation under lath house condition for obtaining good

quality flowers. Testing of the available varieties for suitability and adaptability with respect to

flowering, flower quality and yield parameters are of prime importance. There is need of suitable

varieties to our climatic conditions. Selection of proper variety for producing the desired quantity

and quality of flowers for domestic as well as export market is of greater importance.

At present, sparse information is available about the suitability of potential carnation vari-

eties for cultivation in central Punjab. Hence, the objective of this study was to evaluate suitability

of different carnation cultivars by analyzing their vegetative and reproductive characteristics under

the agro climactic conditions of Lahore, Pakistan.


The present investigation was carried out to study the performance of carnation cultivars

under lath house conditions during 2010-2011. The experiment was carried out at Kala Shah Kaku

Research Station, Directorate of Floriculture, Lahore (31°62'N; 74°9'E). The experiment was laid

out in a complete randomized design (CRD) with three replications. The soil was sterilized with

four 3 % formalin and covered with black polyethylene for 72 hours. The plots were incorporated

with fully decomposed farmyard manure and sand in the ratio of 2:1. Following five cultivars of

standard carnation were used during the study; ‘Grand Salam’, ‘Nelson’, ‘Kaly’, ‘Cinderella’ and

‘Tempo’. The rooted cuttings of five carnation cultivars were procured and were transplanted in

field experimental units of 3.3m x 2.5m, keeping row to row and plant to plant distance 45 and 30

cm, respectively. All the cultural practices such as fertilizers, irrigation, weeding and hoeing were

done uniformly for each treatment. The beds were irrigated regularly to keep the soil moderately

moist. The experiment was carried out under poly-carbonated (6 mm thickness) lath house with

misting and cooling facility.

The physiological characteristics of soil were also determined. The pH value of soil was

7.5 with electrical conductivity 2.0 dS m-1. The organic matter of the soil was 0.9%. Available

phosphorus, calcium and potassium contents of the soil were 55, 220 and 120 mg kg-1of dry soil,

respectively. The soil was mixed with NPK as recommended doses to increase production (Dufault


et al., 1990).

The data on plant height (cm), number of shoots per plant, stem thickness (mm); number

of internodes per stem, yield per plant, number of flowers per square meter and vase life were

recorded. Plant height was measured from the plant crown up to point just below the flower head

and the average plant height was worked out and expressed in centimeter (cm). Carnation flowers

for vase life evaluation were harvested when the outer petals unfold nearly perpendicular to the

stem. The flowers were harvested early in the morning and were immediately placed in fresh water.

Later these flower stalks were cut to have uniform stalk length. After that flowers were kept indi-

vidually in flask containing tap water at 22 oC and 60 % humidity. Each flask had five flower

stems replicated three times. Flowers were observed daily till they were found unfit for containing

in vase. The vase life was expressed in terms of days from the date of harvesting to final observation

(50 % petals wilting).

The climatic data consisting of daily observations of average temperature and relative hu-

midity was recorded during the whole study period (Fig. 1).

The data were subjected to analysis of variance (ANOVA) using Genstat (release 31.1;

Lawes Agricultural Trust, Rothamsted Experimental Station, Rothamsted, UK). The effects of var-

ious treatments were assessed within ANOVA and Fisher’s least significant differences were cal-

culated following a significant (p ≤ 0.05) F test. All the assumptions of analysis were checked to

ensure validity of statistical analysis.


Plant height (cm)

The plant height among the different cultivars of carnation differed significantly. The range

was from 64.96 cm to 78.66 cm. Cultivar ‘Grand Salam’ (78.66 cm) recorded highest plant height

closely followed by ‘Kaly’ (78.23 cm) and ‘Tempo’ was shortest (64.96 cm) (Table 1).

Fig. 1. The average temperature and relative humidity of Kala Shah

Kaku from October 2010 to September 2011.

Varieties Plant height

(cm)

Number of

shoots. plant-1

Stem diameter

(mm)

Number of

internodes. stem-1

Grand Salam

Nelson

Kaly

Cinderella

Tempo

LSD (0.05)

78.66 a

74.40 b

78.23 a

77.96 a

64.96 c

3.23

4.6 c

6.2 a

3.7 d

5.4 b

6.3 a

0.65

3.63 d

6.21 a

5.44 b

4.48 c

5.61 b

0.14

11.00 b

12.66 a

11.33 ab

10.00 b

7.33 c

1.62

Table 1. Growth characteristics of different carnation cultivars under lath house.

Means with different letters in a column differ significantly at p≤0.05.


This variability in plant height among the carnation varieties is mainly due to genetic nature,

growing environmental conditions, production technology and cultural practices. Patil (2001) also

observed same plant height in carnation cultivars ‘Alma’, ‘Sugar Baby’ and ‘Leon’ under low cost

polyhouse. Our results are also in accordance with the results of Sathisha (1997) and Shiragur etal. (2004) who have also recorded vigorous growth in terms of plant height in different carnation

cultivars.

Number of shoots per plant

Cultivars ‘Tempo’ (6.3) and ‘Nelson’ (6.2) recorded more number of shoots as compared

to ‘Cinderella’ (5.4) and ‘Grand Salam’ (4.6). Least number of shoots was counted in cultivars

‘Kaly’ (Table 1). Similar differences with respect to number of shoots per plant were also noticed

by Shiragur (2002). These results are also similar to that of Shahakar et al., (2004).

Stem diameter

Stem thickness may vary from cultivar to cultivar. It gives strength to the plant. Cultivar

‘Nelson’ (6.21 mm) had thicker and stronger stems, while ‘Tempo’ (3.63 mm) had weaker stems.

On the other hand ‘Grand Salam’, ‘Kaly’ and ‘Cinderella’ had stems of moderate thickness (Table

1). Similar variations in stem thickness in different carnation cultivars had been recorded earlier

by Mahesh (1996). Thicker stems indicated higher capacity of strong reserve food material.

Number of internodes per stem

Carnation cultivar ‘Nelson’ possessed the maximum number of internodes per stem (12.66),

followed by ‘Kaly’ (11.33) and ‘Grand Salam’ (11.00) (Table 1). ‘Tempo’ produced minimum

number of internodes per stem (7.33). Variations in number of internodes per stem had been

recorded earlier among different carnation varieties by Hanzel et al., (1954). These differences in

number of internodes per stem may be due to varietals characters.

Yield per plant

There was a significant difference among the cultivars for flower yield per plant. The high-

est number of flowers was recorded in ‘Tempo’ (6.4) followed by ‘Nelson’ (6.3). Cultivar ‘Kaly’

(2.66) produced minimum number of flowers per plant (Fig. 2).

Yield. Meter -2

Maximum number of flowers per square meter was recorded in carnation cultivar ‘Nelson’

(198.3). The next superior variety was ‘Tempo’ (189.6), whereas cultivar ‘Kaly’ recorded minimum

number of flowers per square meter (135.6) (Fig. 2). The increased flower yield might be attributed

Fig. 2. Yield and yield component data of five carnation cultivars

under lath house conditions.


to more number of leaves resulted in production and accumulation of maximum photosynthetic

material which ultimately resulted in production of more number of flowers with bigger sized

flowers. Similar variation in carnation with respect to flower yield was also observed by Ryagi

(2007) and Shahakar et al. (2004).

Vase life

Carnation cultivars differ in the length of the vase life of cut flowers which is one of the

characteristics determining the commercial value of the ornamental flowers. Thus, it is of economic

importance to know the vase life of cut carnation flowers of different cultivars. There was signif-

icant difference among the carnation cultivars with respect to shelf life. Shelf life was highest in

cultivar ‘Tempo’ (8.2 days). The next superior cultivar was ‘Nelson’ (6.3 days) and ‘Grand Salam’

(5.2 days). The cultivar ‘Kaly’ and ‘Cinderella’ (4.0 days) were on par to each other (Fig. 3).

The vase life is one of the important traits which decide its economic value. This variation

in vase life among the varieties might be attributed to the variations in accumulation of carbohy-

drates since these varieties could produce more number of leaves and indicated positive and sig-

nificant correlation between these characters. Variation in vase life could also be attributed to fact

that, the variation in ability to produce ethylene among the different cultivars. Similar variation

for vase life in different cultivars was also reported previously in carnation by Pathania (2000),

Singh et al. (2007), Shahakar et al. (2004) and Patil (2001). The increased ethylene production

promotes the in-rolling of petals resulting in wilting of the flower. The time of onset of ethylene

production and the amount of ethylene produced in the flowers vary with the carnation cultivar,

and thus influence their vase life (Nukui et al., 2004).

CONCLUSIONS

The above mentioned findings indicated that considering the important characteristics, ‘Nel-

son’ and ‘Tempo’ were found superior with respect to growth, yield and vase life characteristics.

These cultivars are suitable for commercial cultivation under lath house conditions.

Literature Cited

Biondo, R.J. and Noland, D.A. 2000. Floriculture: from greenhouse production to floral design.

Inter-State Publ., Danville, Illinois. 662 pp.

Dole, J.M. and Wilkins, H.F. 2005. Floriculture: principles and species, 2nd edition. Pearson Prentice

Hall, Upper Saddle River, New Jersey. 347 pp.

Dufault, R.J., Phillips, T.L. and Kelly, J.W. 1990. Nitrogen and potassium fertility and plant populations

influence field production of gerbera. Hort Science, 25(12): 1599 1602.

Hanzel, R.J., Nelson, K.S. and Kiplinger, D.C. 1954. Floral initiation and development in the carnation

Fig. 3. Vase life of five carnation cultivars grown under lath house

conditions.


variety in Northland. Journal of American Society of Horticultural Science, 65: 455-462.

Mahesh, K. 1996. Variability studies in carnation (Dianthus caryophyllus L.). MSc. (Agri.) Thesis,

Univ. Agri. Sci., Bangalore (India).

Nukui, H., Kudo, S., Amashita, A.Y. and Satoh, S. 2004. Repressed ethylene production in the gynoecium

of long-lasting flowers of the carnation ‘White Candle’: Role of gynoecium in carnation

flower senescence. Journal of Experimental Botany, 55: 641-650.

Pathania, N.S., Sehgal, O.P. and Gupta, Y.C. 2000. Pinching for flower regulation in Sim Carnation.

Journal of Ornamental Horticulture, 3(2): 14-17.

Patil, R.T. 2001. Evaluation of standard carnation (Dianthus caryophyllus L.) cultivars under protected

cultivaton. M. Sc. (Agri.) Thesis, Univ. Agric. Sci., Dharwad (India).

Pralhad, G.C. 2009. Evaluation of carnation (Dianthus caryophyllus L.) varieties under greenhouse

condition. M.Sc. Thesis, Dept. Horti., Univ. Agri. SCI., Dharwad.

Ryagi, V.Y., Mantur, S.M. and Reddy, B.S. 2007. Effect of pinching on growth, yield and quality

of flower of carnation varieties grown under polyhouse. Karnataka Journal of Agricultural

Science, 20(4): 816-818.

Salehi, H. 2006. Can a general shoot proliferation and rooting medium be used for a number of

carnation cultivars? African Journal of Biotechnology, 5(1): 25-30.

Sathisha, S. 1997. Evaluation of carnation (Dianthus caryophyllus L.) cultivars under low cost

green house. M.Sc. (Agri.) Thesis, Univ. Agric. Sci., Bangalore (India).

Shahakar, A.W., Golliwar, V.J., Bhuyar, A.R., Dharmik, Y.B., Kadu, R.B. and Gondane, S.U. 2004.

Growth, flowering quality and yield of carnation cultivars under polyhouse condition. Journal

of Soils and Crops, 14(2): 305-307.

Shiragur, M. 2002. Performance of standard carnation (Dianthus caryophyllus L.) cultivars under

protected conditions for second flush. M.Sc. (Agri.) Thesis,Univ. Agric. Sci., Dharwad (India).

Shiragur, M., Shirol, A.M., Reddy, B.S. and Kulkarni, B.S. 2004. Performance of standard carnation

(Dianthus caryophyllus L.) cultivars under protected cultivation for vegetative characters.

Journal of Ornamental Horticulture, 7(3-4): 212-216.

Singh, K., Singh, P. and Kaoor, M. 2007. Effect of vase pulsing solutions on keeping quality of

standard carnation cut flower. Journal of Ornamental Horticulture, 10(1): 20-24.


Utilization of Rice Hull as a New Substrate for Turf Grass

Seed Germination in Sod Production as a Sustainable Approach

Sod culture is one of the important methods in establishing and

repairing of turf grass, especially in sport fields. Nowadays, a mixture of sand

and peat are commonly used in sod production in Iran. Because peat media is

expensive, it seems necessary to find an alternative medium. Rice hull, tea

waste and leaf compost as economical organic material that are available in

huge loads in north of Iran. The objective of this study was to evaluate the

utilization of low cost organic matter on seed germination and uniformity of

turf grass in sod production. Therefore a completely randomized design with

3 replications and 6 treatments: 1- mixture of leaf compost and sand (1:1)

(v.v), 2- tea compost and sand (1:1) (v.v), 3- sand, 4- mixture of rice hull and

sand (3:1) (v.v), 5- mixture of treatments 1, 2, 3, 4 and 6- petri dish as control

was carried out in the experimental field of agricultural school of Islamic

Azad University, Chaloos branch during summer 2008. Turf grass species

used was Lolium prenne. According to the results, it was revealed that the

effect of different substrates was significant on seed germination percentage

(p≤0.05). Results showed that mixture of rice hull and sand (3:1) (v.v)

increased germination percentage over other treatments. This was probably

related to high water retaining capacity and well aeration of rice hull.

Furthermore, the lowest and highest uniformity rate was related to mixture

treatment and sand media, respectively. It generally seems that these waste

products can be used for this purpose economically and sustainably.

Keywords: Agricultural waste, Rice hull, Seed germination, Sod turf grass production, Uniformity.

Mohammad Ali Golestani1*, Ali Dolatkhahi2, Navid Vahdati2 and Omid Nouri Roudsari3

1Department of Landscape Architecture, Faculty of Agriculture, Branch, Islamic Azad University, Chalous,

Iran.2Ph.D. Student, Department of Horticultural Sciences, Ferdowsi University of Mashhad, Mashhad, Iran.3Department of Agroecology, Environmental Sciences Research Institute, Shahid Beheshti University,

Evin Street, Tehran, Iran.


Abstract


INTRODUCTION

Sustainable agriculture is a system of farming which empowers the farmer to work with

natural processes to conserve resources such as soil and water, whilst minimizing waste and envi-

ronmental impact (Mason, 2003). Nowadays, most efforts have been directed towards using re-

newable resources. Sod is the established turf that is harvested with roots and soil attached to it

and transplanted from its place of origin to grow in another place (Pessarakli, 2008). Sodding may

be a quick and simple solution to repair of sports-field damage and thick-cut sod is used if there

is little time for rooting (Pessarakli, 2008). Sod production could use large quantities of solid-

waste compost as an end product, thus benefiting recycling efforts (Cisar and Snyder, 1992). Or-

ganic amendments, including municipal and animal sources of bio-solids, can be applied to im-

prove soil physical and chemical properties and turf grass establishment, growth, and quality

(McCoy, 1998).

Producing sod on soilless media spreads over plastic has important advantages over con-

ventional sod production (Roberts et al., 2001). Conventional methods of grass sod production on

soil require considerable time and expense, producing sod in a soilless growing medium spread

over plastic can be accomplished more efficiently in much less time (Decker and Decker, 1988).

Aeration, moisture and nutrition of medium composition have a major role in growth and optimum

quality (Bruce et al., 2000; Brien and Barker, 1995). Organic materials are among the kinds of

compositions. The main medium for sod and roll production in Iran is a mixture of sand and peat.

Peat is an expensive medium in Iran, so finding a suitable alternative for this purpose seems to be

necessary. Rice hull, tea factory waste and leaf compost which are found in large volumes in the

north of Iran and are very cheap and also easily available. Using these types of material will addi-

tionally help natural recycling. McCoy (1992), showed on a sport lawn, increasing organic material

to 3.5% (v/v %) can increase water holding capacity up to 2 folds. Cisar and Synder (1992) re-

ported, lawns that grew on urban compost showed a better quality and were ready for transport

earlier. Hensler et al. (1998) also reported successful sod production on a debris Knaf web. Results

of Mirtchall et al. (1994) research on compost and saw dust, showed the positive effect of these

constituents on lawn density and growth. The considerable effects of leaf compost and animal ma-

nure on shoot dry weight increase of some cultivars was hence observed (Hensler et al., 1998;

Mirtchall et al., 1994). Sod production duration on plastic with use of organic material in compar-

ison to its conventional soil, decreased from 2 years to 7-10 weeks for Tall fescue (Festuca auran-dinaceae) (Decker, 2001; Decker, 1989). Disease infections were also declined to use of organic

material (Bruce et al., 2000; Munster et al., 2004; Bertran et al., 2004, Garcia-Gomez et al., 2002).

The main aim of this research was to evaluate the impact of rice hull, tea factory waste and leaf

compost as a cheap, biodegradable and good alternative media for sod lawn production and seed

germination, germination rate and establishment.

MATERIAL AND METHODS

This experiment was planned as a completely randomized design with 6 treatments ( ½ leaf

compost + ½ sand, ½ tea compost + ½ sand, sand, ¾ rice hull + ¼ sand, mixture of leaf compost

+ tea compost + sand + rice hull and petri dish as control) (Table 1) and 3 replicates and it was

Water holding

capacity (%)

Total porosity

(%)

Bulk density

(g/cm-3)

Particle density

(g/cm-3)

Media

Leaf compost

Tea compost

Sand

Rice hull

190

324.1

210

410

88

95

56

79

0.062

0.07

0.12

0.081

0.78

1.53

1.75

0.36

Table 1. Physical characteristics of the media used for sod production in this experiment.


conducted in experimental field of Islamic Azad University, Chaloos branch in 2008. Loliumperenne was used for sod production. Germination percentage was calculated according to equa-

tions below:

Germination in the various treatments was described by the final germination percentage

(FGP = Σn, where n is the number of seeds that germinated at each counting) and germination rate

[GR = Σ (n/D), where D is the number of days to germinate].

Data were analyzed as factorial ANOVAs using SPSS. Where significant (p≤0.05) treatment

effects were determined by ANOVA, data means comparison were separated by the LSD test.

For visual quality of sods, the uniformity was evaluated. This is a difficult procedure and

for this purpose visual observations are usually used. Visual observations therefore are usually

done by 10 lawn experts. Different aspects of determination number range were 1-9 which 1

showed the least and 9 the best quality, and numbers more than 6 are considered acceptable for

lawn and turf grass (Patton et al., 2010). After determining of each parameter, scores are added

and then the mean is reported as the qualification rate of the turf grass.


The present study revealed that different substrates for sod can impact uniformity trait and

seed germination percentage. Results showed that seed germination percentage in media with ¾

rice hull and ¼ sand (v/v) were significantly higher compared to other media (p<0.05) (Table 2

and Fig. 1).

It is interesting to note that germination percentage in medium with rice hull and sand was

higher than other treatments of pure sand medium and petri dish. The principal reason for this

result may be a high water retention capacity of rice hull and also it’s higher aeration from others.

The results showed that mixtures of sand and leaf compost have the potential to inhibit germination

of cool-season turf grass (Lolium prenne). Turf grass quality is determined by uniformity, smooth-

ness, texture, and color. Among these, the uniformity of stand is very important for the sod grower

(Pessarakli, 2008). Our results showed that different substrates had significantly affected the uni-

formity (p≤0.01) (Table 2 and Fig. 2).

Mean squares

Source of variation df Germination (%)

Treatments

Error

5

12

508.444*

4.830

Mean squares

Source of variation df Germination (%)

Treatments

Error

4

45

17.98**

0.466

Table 2. Analysis of variance for turf grass to evaluate effects of different growing media on studied traits.

*: Significant at p≤0.05, **: Significant at p≤0.01.

Fig. 2. Effect of different growing media on

uniformity of turf grass.Fig.1. Effect of different growing media on seed

germination percentage of turf grass.


Sod produced on full mixed treatment (sand + rice husk + tea compost + compost leaf)

showed acceptable quality than other treatment and appeared to handle well when harvested. In

this experiment, the lowest uniformity was related to growing sod on sand beds. One reason for

this may be that sand requires special care and regular watering, and usually does not provide

under field conditions. It is important that increasing the amount of organic material greatly in-

creased uniformity of turf grass. There are many reports that increasing of organic matter increased

water holding capacity and therefore the quality of the grass (McCoy, 1992; Hensler et al., 1998).

McCoy (1992) reported that using of various organic materials in sand-based root zone mixes can

increase nutrient retention and maintain a stable supply of nutrients which plants can use between

regular fertilizer applications. Producing sod by rice hull could reduce the costs associated with

expensive substrate materials such as peat. Recycling waste for sod production play a role impor-

tant in reducing the potential source of waste and preserving agricultural soils (Cisar and Snyder,

1992). According to the results of this experiment, it is concluded that mixture of sand and rice

hull is the best growing media for sod production. Because of rice hull as a low cost organic matter

is very abundant in the north of Iran, it can be recommended for utilization for sod production as

an alternative substrate.

CONCLUSION

Sod culture is one of the important methods in establishing and repairing of turf grass, es-

pecially in sport fields. Nowadays, a mixture of sand and peat are commonly used in sod production

in Iran. Rice hull, tea factory waste and leaf compost are economical organic material and there is

great resource of them in the north of Iran, and considered suitable alternatives. Germination per-

centage in medium with rice hull and sand was highest among treatments. The principal reason

for this may be a high water retention capacity of rice hull and also its higher aeration.

Literature Cited

Bertran, E., Sort, X., Soliva, M. and Trillas, I. 2004. Composting winery waste: sludges and grape

stalks. Bioresource Technology. 95: 203–208.

Brien, O. and Barker, A.V. 1995. Evaluation of fresh and year – old solid waste composts for production

of wild flower and grass sods on plastic compost. Science and Utilization. 3(4) 69-77.

Bruce, R., Decker, H., Ganahl, L. and Yarmark, E. 2000. Biosolid residues as soilless media for

growing creeping bent grass. American Society for Horticultural Science. 1:1.

Cisar, J.L. and Synder, G.H. 1992. Sod production on a solid-waste compost over plastic. HortScience.

27: (3) 219-222.

Decker, H. F. 1989. Growing sod over plastic: turf in five weeks. Landscape Management, 28:68–70.

Decker, H.F. 2001. Producing sods over plastic in soilless media. Horticultural Review, Vol. 27.

Decker, H.F. and Decker, J.M. 1988. Lawn care: A handbook for professionals. Prentice-Hall, Inc.,

Englewood Cliffs, N.J. 304 pp.

Garcia-Gomez, A., Bernal, M.P. and Roig, A. 2002. Growth of ornamental plants in two composts

prepared fromagroindustrial wastes. Bioresource Technology. 83: 81–87.

Hensler, K.L., Baldwin, B.S. and Gootley, J.M. 1998. Kenaf – based fiber mat as a substrate for

establishing soilless sod. Hort. Technology. 8: (2) 171 – 175.

Mason, J. 2003. Sustainable Agriculture. Landlinks Press, Australia. 205 PP.

McCoy, E.L. 1992. Quantitative physical assessment of organic material used in sport turf rootzone,

mixes. Agronomy Journal, 84: (3) 375-381.

McCoy, E.L. 1998. Sand and organic amendments influences on soil physical properties related

to turf establishment. Agronomy Journal, 90: 411–419.

Mirtchall, W.H., Molnar, C.J. and Barton, S.S. 1994. Using Compost to grow wildflower sod. Bio

Cycle. 35: (2) 62-63.


Munster, C.L., Hanzlik, J.E., Vietor, D.M., White, R.H. and McFarland. A. 2004. Assessment of

manure phosphorus export through turfgrass sod production in Erath County, Texas. Journal

of Environmental Management. 73: 111–116.

Patton, A., Richardson, M., Karcher, D. and Trappe, J. 2009. 2007 NTEP Bermudagrass Trial- Year

3 results. Arkansas Turfgrass Report, Arkansas Agriculture Experiment Station Research

Service, 579:20-24.

Pessarakli, M. 2008. Hand book of turf grass management and physiology. CRC Press, Tucson,

Arizona, U.S.A. 690 pp.

Roberts, B.R., Decker, H.F., Ganahl, L.M. and Yarmark, E. 2001. Biosolid residues as soilless

media for growing creeping bent grass sod. Hort. Technology. 11: (3) 451– 455.

www.jornamental.com


Comparison Tea Extract, 8-Hydroxy Quinoline Sulfate

and Rifampicin on the Vase Life of Cut Chrysanthemum

(Denderanthema grandiflorum L. cv. Purple)

Maintaining quality and longevity of cut flowers is one of the critical

issues in the floriculture industry, especially in cut flowers; and one of the

most important problems in chrysanthemum, as one of the most popular cut

flowers. Therefore, an experiment was conducted based on completely ran-

domized design as pulse treatment with the three factors: tea extracts with 4

levels (5, 10, 20, and 40 %), 8-hydroxy quinoline sulfate with 4 levels (100,

200, and 400 mg l-1), and rifampicin with 3 levels (100, 200, and 400 mg l-1)

on spray chrysanthemum (cv. Purple) with three replications. According to

the results, the 20% tea extract treatment and 100 mg l-1 8-hydroxy quinoline

sulfate treatment showed the highest vase life, petal protein, total chlorophyll,

water absorption, and petal carotenoid.

Keywords: Petal protein, Petals carotenoid, Solution uptake, Total chlorophyll, Vase life.

Davood Hashemabadi1* and Hamideh Bagheri2

1 Department of Horticulture, Rasht Branch, Islamic Azad University, Rasht, Iran.2 Young Researchers Club, Rasht Branch, Islamic Azad University, Rasht, Iran.


Abstract


INTRODUCTION

Chrysanthemum (Denderanthema grandiflorum) from Asteraceae family is one of the most

important cut flowers in the world (Edmond et al., 1957). Cut chrysanthemum is a non – climac-

teric flower with relatively long life and low ethylene production (Bartoli et al., 1996). Senescence

of chrysanthemum is its reaction to the changes that occur in the level of carbohydrates (Adachi

et al., 1999) and ethylene has not significant impact in this process. Decrease in quality of cut

chrysanthemum is mostly due to its leaves' yellowing (Halevy and Mayak, 1981) which is accom-

panied by the degradation of chlorophyll that causes earlier senescence of petals compared to petals

(van Leperen et al., 2001). Thus, delay in petals' wilting and, consequently delay in senescence,

will elongate the vase life of cut chrysanthemum (Petridou et al., 2001). Jiao (2009) in cut gladiolus

said that antibiotics enhanced vase life and water relations and increase wet and dry weights. Cang

et al. (2010) studied on the influence of tetracycline, sucrose and 8-hydroxy quinoline sulfate on

the vase life of Gladiolus grandiflorus and found that 500 mg l-1 of 8-hydroxy quinoline sulfate

and tetracycline enhanced the quality of cut flowers through improving the fresh weight and water

relations. In this research, effects of different levels of tea extracts, 8-hydroxy quinoline sulfate

and rifampicin will compared for maintaining quality of cut chrysanthemum cv. ‘Purple’.


Cut chrysanthemum were harvested at commercial stage from a greenhouse in Mahallat

and were transfered to the postharvest laboratory. This experiment was conducted in completely

randomized design as pulse treatment with three factors of tea extract (5, 10, 20 and 40%), 8-hy-

droxy quinoline sulfate (100, 200 and 400 mg l-1) and rifampin (100, 200 and 400 mg l-1) in three

replications. Vase life room conditions was 12 hours day length, 20±2 °C, 60 to 70% RH and 12

μmol s-1 m-2 light intensity and traits such as: vase life, petals carotenoid, petals protein, total chloro-

phyll content and water uptake were measured. Vase life measured as flower wilting (Ferrant etal., 2002). For measurement of petal̕s protein, in 5th day sampling was done and protein content

was evaluated by Bradford (1976) method. In 5th of experiment, sampling for total chlorophyll

and petal carotenoids was done and these traits measured by Mazumdar and Majumdar (2003)

method. Data were analyzed using SPSS statistical software and mean comparison was performed

by LSD test.


Vase Life

Analysis of variance showed that effect of different compounds on vase life were significant

(p ≤ 0.05). Mean comparisons showed that the 20% tea extract with 18.86 days increased the vase

Treatment Vase life

(days)

Water uptake

(ml g-1 FW)

Total chlorophyll content

(mg g-1 FW)

Petals protein

(%)

Petals Carotenoids

(μg g-1FW)

(Tea extract 5%) E1

(Tea extract 10%) E2



(100 mg l-18- HQS) H1

(200 mg l-18- HQS) H2

(400 mg l-18- HQS) H3

(Rifampin100 mg l-1) R1

(Rifampin 200 mg l-l) R2

(Rifampin 400 mg l-1) R3

Control

14.7200 ab

17.5633 a

18.8600 a

17.7000 a

17.8733 a

15.5733 ab

17.4267 a

18.2133 a

16.1067 ab

16.5967 ab

9.6433 b

14.7200 ab

17.5633 a

18.8600 a

17.7000 a

17.8733 a

15.5733 ab

17.4267 a

18.2133 a

16.1067 ab

16.5967 ab

9.6433 b

14.7200 ab

17.5633 a

18.8600 a

17.7000 a

17.8733 a

15.5733 ab

17.4267 a

18.2133 a

16.1067 ab

16.5967 ab

9.6433 b

14.7200 ab

17.5633 a

18.8600 a

17.7000 a

17.8733 a

15.5733 ab

17.4267 a

18.2133 a

16.1067 ab

16.5967 ab

9.6433 b

14.7200 ab

17.5633 a

18.8600 a

17.7000 a

17.8733 a

15.5733 ab

17.4267 a

18.2133 a

16.1067 ab

16.5967 ab

9.6433 b

Table 1. Mean comparison of different concentrations of tea extract, 8-hydroxyquinoline sulfate and rifampin on the

measured traits.

*Similar letters in each column indicate not significant difference at 1% and 5% (LSD test).


life for 9.2 days as compared to the control (9.64 days) (Table 1). All treatments were better than con-

trol, but difference between them was not significant.

One of the most important problems in postharvest physiology of cut flowers, is the vascular

blockage. This problem might be due to air embolism or growth of bacteria. Another reason for vas-

cular blockage is reaction of the plant to cutting. Certain enzymes are stimulated in reaction to cutting

and sent to the cut area to block it (Loubaud and Van Doorn, 2004). Therefore, antibacterial compounds

enhance water relations and prevent wilting and delay senescence through preventing microorganisms'

accumulation in the vessels. Also, blockage of the vessels in cut chrysanthemum can be stimulated by

the plant itself. According to the obtained results, biocides have not only prevented the vascular ob-

struction caused by the microorganisms, but also prevented the blockage stimulated by the plant itself

and these results are consistent with reports by Kim and Lee (2002).

Petal’s Protein

The influence of various levels of treatments on the petal protein content was significant in

1% probability. Mean comparisons showed that the 20% tea extract with 8.74% and 200 mg l-1 8-hy-

droxy quinoline sulfate with 10.32% were the best treatments compared to the control (1.62%) (Table

1). The senescence of cut flowers has the hormonal regulatory mechanism and this process is involved

in changing the physical and biochemical features in cellular membrane (Buchanan Wollaston, 1997).

One of the reasons for beginning senescence in plant tissues, is increasing the activity of ROS like

H2O2 that causes senescence by degrading proteins, lipids, and nucleic acids. Maintaining of the petals'

protein can be due to decreasing in activities of enzymes that decompose protein and increasing the

water absorption which maintened the stability of membrane and prevent protein degradation (Sood

and Nagar, 2003; Lerslerwonga et al., 2009).

Solution uptake

The effect of various levels of treatments on water uptake was significant in the statistical level

of 5%. Mean comparisons showed that the 20% tea extract with 0.77 ml g-1 FW and the 100 mg l-1

8-hydroxy quinoline sulfate with 0.71 ml g-1 FW, and the 100 mg l-1 rifampicin with 0.64 ml g-1 FW

had highest water uptake and control (0.34 ml g-1 FW) had lowest water uptake. Cut flowers' wilting

during vase life might be caused by insufficient water absorption due to vascular blockage that can be

the result of growth of bacteria and accumulation of microorganisms in vessels.

Stem end blockage and inhibition of water relations, decreased vase life of cut flowers (Silva,

2003). Therefore, providing energy and preserving the water absorption power of the plant are two

main factors for extending vase life of cut flowers, so these compounds maintain constant flow of water

in the cut stems by controlling the activity of microorganisms (such as bacteria and fungi) and preventing

the vessel blockage (Monshizadeh et al., 2011; Figueroa et al., 2005). These results are consistent with

Jalili Marandi et al. (2011) and Burt (2004). Similar results were reported by Zadeh Bagheri et al.(2011) about the positive effect of herbal essences and antibiotic compounds that are consistent with

results of the present study. Kim and Lee (2002) reported that 8-hydroxy quinoline sulfate, as a common

antibiotic, has germicidal properties and improves longevity by enhancing water absorption. Nabigol

et al. (2006) found that antiseptic, anti-ethylene and antibiotics compounds significantly improve the

water absorption in plants; and their results are consistent with results of this study.

Petal’s Carotenoids

The effect of different treatments on amount of the petal’s carotenoid was significant (p≤0.01).

Results showed that the 20% tea extract with 5.95 μg g-1 FW and 100 mg l-1 rifampicin with 3.28 μg

g-1 FW and 100 mg l-1 8-hydroxy quinoline sulfate with 5.39 μg g-1 FW, were the best treatments

(Table1). Petal’s discoloring is one of the postharvest problems for cut flowers that reduces quality of

flowers and this problem has significant effect on senescence. Carotenoids and anthocyanins are two

great important pigments in cut flowers (Amarjit, 2000; Hasanpour Asil and Karimi, 2010). Antibac-


terial compounds prevent from degredation of flavonoids and enhance flowers' freshness by improving

the water uptake.

Total Chlorophyll

The effect of treatments on the total chlorophyll was significant in 5% probability. The 20%

tea extract with 3.19 mg g-1 FW and the 100 mg l-1 8-hydroxy quinoline sulfate with 4.57 mg g-1 FW

were the best treatments compared to control (2.32 mg g-1 FW) (Table1). These compounds prevent

senescence and wilting by their antibacterial property and reducing the pH of the environment; they

extend the vase life by controlling vascular obstructions and enhancing the water absorption (Elgimabi

and Ahmad, 2009; Edrisi, 2009). Controlling activity of chlorophyll decomposing enzymes is one of

the other reasons for improvement of total chlorophyll by the above mentioned compounds. These re-

sults are consistent with the results of the studies by Ferrante et al. (2002).

CONCLUSION

The 20% tea extract with 18.86 days increased the vase life of cut chrysanthemum compared

to controls (9.64 days) by 9.2 days.

ACKNOWLEDGMENT

The authors would like to thanks Islamic Azad University Rasht Branch, Specific Dr. Ali Mo-

hammadi Torkashvand (Research Office Manager) for financial supports.

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www.jornamental.com


Micropropagation of Rosa canina Through Axillary Shoot

Proliferation

In vitro propagation of rose has played a very important role in rapidmultiplication of species with desirable traits and production of healthy anddisease-free plants. Micropropagation using nodal segments of Rosa caninaunder different combinations of BAP, GA3 and NAA on MS and VS, mediumwas investigated. The results showed that the highest shoot proliferation wasobtained on VS medium containing 2 mg/L BAP without any GA3 and NAA.Furthermore the highest root regeneration obtained in half strength VSmedium. The present investigation recommended a practiciable in vitro plantprotocol for R. canina as an important step for successful implementation ofbiotechnological techniques for rose improvement in Iran.

Keywords: Micropropagation, Native species, Plant growth regulators, Rosa canina.

Abbreviations: MS (Murashige and Skoog), VS (Vander Salm), BAP (Benzyl Amino Purine), NAA(Naphtalene Acetic Acid), GA3 (Gibberellic Acid).

Mahboubeh Davoudi Pahnekolayi1*, Ali Tehranifar2, Leila Samiei3 and Mahmoud Shoor4

1Msc student, Department of Horticultural Science, Ferdowsi University of Mashhad.2Department of Horticultural Science, Ferdowsi University of Mashhad.3Ornamental Plant Department, Reasearch Center for Plant Sciences, Ferdowsi University of Mashhad.4Department of Horticultural Science, Ferdowsi University of Mashhad.


Abstract


INTRODUCTION

Rosa canina is a medicinal plant that its fruits have many important medicinal propertiesbut unknown for many people especially in Iran. However, this crop can be produced commerciallyand its orchads can be established such as other fruit trees (Sharafi, 2010؛ Luncna, 2005). Rosacanina belongs to Rosaceae family. Dog rose (Rosa canina) has been used as a rootstock for orna-mental roses (Khosh-Khui and Sink, 1982) while considered as a medicinal plant as well. Rosacanina L. (dog rose, rose hip, briar rose) is one of wild roses appreciated for their vitamin C en-riched fruits being beneficial to human health (Kazaz et al., 2009). More than 200 species are pres-ent in the genus Rosa (Wissemann, 2003) from which 14 wild species are present in Iran.Traditionally, most roses are heterozygous and do not breed true to type. Therefore, they are prop-agated by vegetative methods. Since most rose species are difficult to root, conventional propa-gating methods are very slow, time consuming, and tiring. Tissue culture on the other hand isbecoming increasingly popular as an alternative to the conventional plant propagation methods(Roberts and Schum, 2003). Micropropagation has five major advantages compared to the con-ventional methods of plant propagation: (i) it is an valuable aid in the multiplication of elite clonesof intractable/recalcitrant species؛ (ii) it is important in terms of multiplying plants throughout theyear, with control over most facts of production؛ (iii) it is possible to generate pathogen-free plantseven from explants of infected mother plants؛ (iv) plant materials such as male sterile, fertilitymaintainer and restorer lines can be cloned؛ and (v) it enables the production of a large number ofplants in a short time from a selected number of genotypes (Jafarkhani Kermani et al., 2011). Theobjective of the study was to investigate the best hormonal compound and media for micropropa-gation of Rosa canina in vitro condition.


Sterilization of explants

After removing chilling requirement of buds in February 2012, axillary buds of dog rose (grownat the botanical garden of Ferdowsi University of Mashhad, Iran) were cut and placed under runningtap water (for 1 h) and were decontaminated with 70% ethanol (for 30 s), and sodium hypochlorite(2.5%) (for 15 min.). Then all explants were washed three times with sterile distilled water.

Shoot proliferation stage

In this experiment, shoot explants with 3 adventitious buds and 2 cm in size were transferredto 2 different media: MS and VS medium. 20 treatments with 8 replications were considered foreach medium in this stage. Different hormonal composition of BAP (0, 0.5, 1, 1.5, 2 mg/L), GA3(0, 0.5 mg/L) and NAA (0, 0.1 mg/L) were considered for both media. The shoot number, shootlength, percentage of shoot regeneration, leaf number and percentage of green leaves were recordedafter 60 days. The pH of all media was adjusted to 5.8 using 1.0 N potassium hydroxide (KOH) or1.0 N hydrochloric acid (HCl), before adding 8 g/L plant agar. Media were autoclaved for 15 min.at 121 ̊̊C and 1.2 kgf/cm pressure. All the in vitro cultures were placed under 16/8 h light/darkcycle in a culture room maintained at ±24 ̊̊C.

Rooting stage

For rooting, new shoots (1-2 cm height) were transferred to 2 media of VS and ½ VS withdifferent hormonal composition of IBA (0, 0.3, 0.6, 0.9 mg/L) and NAA (0, 0.3, 0.6, 0.9 mg/L).Root length, root number and percentage of root induction were recorded after 30 days.

Acclimatization of plantlets

The plantlets were cultivated for 5 weeks in plastic glasses containing steril mixture of peat:perlite (3:1). The plantlets were put in culture room under a 16-hour photoperiod, at a tempreture


of 23/25 ̊C (night/day) and 80% relative humidity. When a growth cycle was over, the shoot sur-vival percentage was evaluated.

Experimental design and statistical analysis

The shoot proliferation experiment was performed in a factorial based completely randomdesign with 40 treatments and 8 replications. Rooting experiment was carried out in a factorialbased completely random design with 16 treatments and 15 replications. Analysis of variance wasperformed and comparisons of means were conducted using LSD test. All analysis were regardedas significant if p value was less than 0.05. The percentage data was transformed using angulartransformation (Arc Sin√%) before carrying out ANOVA.

RESULTS

Shoot proliferation

The results showed that there was significant difference of effect of type and concentrationof PGRs on vegetative traits of R. canina in proliferation stage (p≤0.05). Maximum number ofnew leaves (19.96) was produced on the medium containing 2 mg/L BAP (Fig. 1A), whereas themaximum percentage of green leaves were obtained on the media containing 1.5 mg/L BAP

Fig. 1. Effect of type and concentration of PGRs on vege-

tative traits of R.canina in proliferation stage, showing؛ A)

Average number of new leaves, B) Average percentage

of green Leaves, C) Average number of axillary shoots,

D) Average shoot percentage, E) Average shoot

length(cm).

A

CD

E

B


(84.84%) (Fig. 1B). The maximum number of axillary shoots (4.21) and shoot percentage (41.10%)were observed in 2 mg/L BAP (Fig. 1C, D) and the highest increase in plant height (1.27 cm) wasobserved in the medium containing 1.5 mg/L BAP (Fig. 1E). Also, there was significant differenceof type of medium on vegetative traits. The lowest shoot multiplication was observed on mediumMS while the highest shoots were formed on medium VS (Table 1).

Kim et al. (2003) indicated that in vitro shoot proliferation and multiplication are largelybased on media formulations containing cytokinins as major plant growth regulators, although lowconcentrations of auxins or GA3 are also essential. The results of the present study demonstratedthat inclustion of 0.5 mg/L GA3 to the culture media didn’t increase the number of axillary shootsand stem height in all of the BAP concentrations. The results showed that in some of the traits thedifferences between the treatments were not significant, however, the maximum number of axillaryshoots were significantly higher in the VS medium containing 2 mg/L BAP. AS the concentrationof BAP was lower than 2 mg/L, a reduced growth rate was noted. Also, in this experiment 2 typesof MS and VS media were used and the results showed that the best plantlets (in proliferation androoting stage) were in VS medium because of the alleviating Fe deficiency in leaves.

Root initiation and acclimatization

Results of ANOVA on effect of different media on root number, root length and rootingpercentage revealed significant difference (p≤0.01) (Table 2). The results showed that there was

Medium Culture Root number Root length (cm) Rooting percentage (%)

VS½ VS

1.01 b2.28 a

0.922 b1.85 a

14.03 b24.75 a

Table 3. Effect of different media on some traits of R. canina in rooting stage.

Concentration (mg/L) Root number Root length (cm) Rooting percentage (%)

00.30.60.9

0.77 b1.4 ab2.14 a1.83 a

1.42 a1.27 a1.38 a1.48 a

12.29 b17.78 ab23.08 a20.86 a

Table 4. Effect of type and concentrations of PGRs on rooting traits of R.canina.

Table 2. Analysis of variance (ANOVA): Effect of type and concentrations of different plantgrowth regulators (PGRs) and medium on some traits of R. canina in rooting stage (p≤0.05).

Source of Variation Degree of

Freedom

Root

number

Root

length

Rooting

percentage (%)

PGRs typePGRs concentrationMedium CulturePGRs type * ConcentrationPGRs type * Medium cultureConcentration * Medium culturePGRs type * concentration * Medium CultureError

1313133

224

13.43 *14.46 *85.12 **11.69 *

36.85 **19.03 **5.23 ns

4.54

0.74 ns

0.48 ns

45.88 **0.85 ns

18.07 **9.15 *3.33 ns

3.44

882.25 *871.28 *

6036.00 **704.061 *2523.17 **1312.36 **324.209 ns

295.96

Different letters show significant differenced according LSD Test (p≤0.05).

Medium Culture Shoot percentage Grean leaves percentage Shoot length (cm) Leaf number Soot number

Murashige & Skoog (MS)Van der Salm (VS)

26.25a28.21a

46.29b56.49a

0.768 b0.993 a

8.37b10.37a

2.15a2.4a

Table 1. Effect of different media on some vegetative traits of R. canina in proliferation stage.


significant difference between PGRs concentrations and types in root number, rooting percentagebut there was no difference in root length (p≤0.05). The highest root length, root number and root-ing percentage was recorded in ½ VS medium (Table 3). Also, maximom average of root number(2.14) and maximum average of rooting percentage (23.08%) were observed in 0.6 and 0.9 mg/Lconcentration (Table 4). The results showed that there was no difference between IBA and NAAin root length with control, but they had significance difference in root number and rooting per-centage. Therefore, the PGRs concentrations had more importance than the type in rooting stage.

The resulting shoots cultured on VS medium containing ½ and full strength of VS macro,micro elements and vitamins showed variable response to rooting afters 4 weeks of culture. Thebest results were obtained on VS medium containing ½ strength of VS macro, micro salts and vi-tamins. The rooted plants were not difficult to acclimatize at ±24 ̊C and relative humidity of 80%during initial stages of development gradually reduced to 40% after 4 weeks of culture and wastransferred to the greenhouse for flowering.

DISSCUSION

Tissue culture techniques are used extensively for growing plants commercially. Thisprocess involves growth of new plants from small pieces of plant tissue in a nutrient medium insterile conditions. Under sterile conditions, plants can be induced to rapidly produce new shootsand these can be subdivided to produce more plants. (Dixon and Gonzales, 1993). In this study,according to the responses of explants cultured on different medium, VS and VS/2 medium wasrelatively the best and the most appropriate treatment for shoot and root formation. A successfulmicropropagation protocol proceeds through a series of stages, each with a specific set of require-ments. These are (i) initiation of aseptic cultures, (ii) shoot multiplication, (iii) rooting of macroshoots and (iv) hardening and field transfer of tissue culture raised plants (Kumar Pati et al., 2006).For initiation of aseptic cultures, a thorough knowledge of the physiological status and the sus-ceptibility of the plant species to different pathological contaminants are required. A review of lit-erature showed that surface sterilization of rose is problematic and different approaches have beenadopted to achieve sterilization. Other researchers have reported a high percentage of bud breakon hormone-free medium within 10-12 days, but the growth rate was very low in roses (Rout andJain, 2004). In our study the effects of different concentrations of BAP (0 to 2 mg/L) and GA3 (0,0.5 mg/L) and NAA (0, 0.1 mg/L) were obvious on bud break and growth but on MS and VSmedium without growth regulators the growth rate was lower.

In vitro shoot multiplication relies largely on medium formulations containing BAP as themajor PGRs in combination with a low concentration of NAA (Xing et al., 2010). In the presentstudy, 2 mg/L BAP was the optimum treatment for in vitro multiplication of Rosa canina. Thisresult is similar to the findings on optimal BAP concentration (4.4 – 13.2 µM) (Carelli and Echev-errigaray, 2002). BAP is needed for proliferation of canina rose plants, but a high concentration isundesirable: at a concentration higher than 2.2 µM it will lead to multiplication of shoots, whichis not beneficial to shoot elongation which is not in line with our results. When the culture mediumcontains NAA at higher concentration (0.1 mg/L), the bud can form more callus from the base se-lection, which will greatly affect the young seedlings absorption of water and nutrition, and thusinhibit its growth. The inhibition is especially obvious when NAA concentration is increased to0.1 mg/L. Auxin is a rooting hormone and application of synthetic auxin i.e. IBA might have in-creased the biosynthesis of indole acetic acid (IAA) or could act as synergistic to IAA. The anotherpossible reason for higher rooting and early root initiation by IBA might be involvement of IBAin ethylene biosynthesis (Arteca, 1990) and it has been suggested that auxin induced ethylene mayinduce adventitious root formation instead of action of auxin itself (Mudge, 1989). A high con-centration of GA3 (0.5 mg/L) always finally caused the nigrescence and death of young shoots,which appeared waterlogged that is in line with Xing et al. (2010). The best results in rooting stage


achieved with 0.6 or 0.9 mg/L IBA/NAA in ½ VS medium. Hyndman et al. (1982) recommendeddecreasing medium salt concentration from MS medium generally increase rooting for rose prop-agation. The same results were reported in R. hybrida cv. Peace (Kirichenko et al., 1991) and R.rugosa (Xing et al., 2010). Our study yielded a practical protocol for efficient axillary bud multi-plication from Rosa canina explants. Here we demonstrate high-efficiency micropropagation ofR. canina for the first time. Micropropagation of R. canina provides an opportunity to harvestvirus-free material and conserve important germplasm resources in vitro. It furnishes material forgenetic transformation and foe employing molecular techniques in breeding.

CONCLUSION

In conclusion, the present investigation recommended a practicable, in vitro propagationprotocol for R.canina. At the proliferation stage, maximum number of axillary shoots was achievedin the medium containing 2 mg/L BAP. At the rooting stage, 0.6 and 0.9 mg/L IBA/NAA were ef-fective. Also the VS medium with additive Fe was better than MS medium in all stages of micro-propagation of this plant.

Fig. 2. Different stages of micropropagation of Rosa canina: A) Proliferation stage B) Root-ing stage C) Acclimatization stage.

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www.jornamental.com


Drought Stress Impact on Some Biochemical and

Physiological Traits of 4 Groundcovers (Lolium perenne,

Potentilla spp, Trifolium repens and Frankinia spp) with

Potential Landscape Usage

Stress management is considered as an important factor in nowadays

landscape. Therefore this research was conducted using a factorial experiment

based on a randomized completely design with regulated deficit irrigation at

four levels (100, 75, 50, and 25% of lawn irrigation requirement) and three

replicates to evaluate some biochemical and physiological traits on four

groundcover plants (Lolium perenne, Potentilla spp., Trifolium repens and

Frankinia spp.) with the potential use in landscape. Results showed that

Frankinia spp. and 75% lawn irrigation requirement bears the highest proline

content with same amount (0.84 mg/g fresh weight) and on the other hand

highest (32.61 mg/g fresh weight) and lowest (15.95 mg/g fresh weight)

chlorophyll content was shown in control (irrigation treatments) and Frankiniaspp., respectively. For total soluble carbohydrates content, highest amount

(1.54 mg/g fresh weight) belongs to Lolium perenne whereas the lowest (0.79

mg/g fresh weight) belongs to Trifolium repens. Trifolium repens also had the

most DPPH free radical-scavenging assay (75.05%) among species. In general,

regulated deficit irrigation affects different biochemical characteristics of ex-

amined groundcover plants significantly, but as a managing method applying

it to below the damaging point for landscape plants can be a good method for

water consumption management in this manner.

Keywords: Antioxidant activity, Chlorophyll, Proline content, Total soluble carbohydrate content.

Elham Samieiani1and Hossein Ansari2

1MSc student, Department of Horticultural Science, Ferdowsi University of Mashhad.2Associate Professor of Agricultural enjeeniring, Ferdowsi University of Mashhad.


Abstract


INTRODUCTION

For maintenance of urban landscaping, frequent watering increases the costs and does not

collaborate with sustainable use of water resources. Low water-requiring or ‘‘water-wise’’urban

landscaping is important policy tool in arid/semiarid areas in Iran. Water-wise landscaping can

maintain acceptable appearance during drought when designed with more drought-tolerant plants

than turf grasses and when irrigation is determined by plant water needs (Mee et al., 2003). Ap-

propriate species selection for water-wise landscaping can be improved with knowledge of drought

tolerance and water stress response mechanisms among candidate species (Kjelgren et al., 2000).

Plants in nature are continuously exposed to several biotic and abiotic stresses. Among the

environmental stresses, drought is one of the most severe stresses for plant growth and productivity.

Water stress affects virtually every aspect of plant physiology and metabolism. Drought stress re-

duces both nutrient uptake by the roots and transport from roots to the shoots, due to restricted

transpiration rates and impaired active transport and membrane permeability (Yuncai and Schmid-

halter, 2005). Drought stress reduced dry matters of plants by reduction in the area of the leaf,

height of plant and lateral stem number (Aliabadi Farahani et al., 2009).

Osmotic adjustment is a mechanism to maintain water relations under osmotic stress. It in-

volves the accumulation of a range of osmotically active molecules/ions including soluble sugars,

sugar alcohols, proline, organic acids, calcium, potassium, etc. (Farooq et al., 2009). The production

of reactive oxygen species (ROS) is linear with the severity of drought stress, which leads to en-

hanced peroxidation of membrane lipids and degradation of nucleic acids, and both structural and

functional proteins. The severity of this damage largely depends on the status of antioxidant systems,

since plants develop antioxidants to remove toxic reactive oxygen species and protect the plant cells

from lipid peroxidation and inactivation of enzymes that occur under stress (Smirnoff, 1993).

In recent years turf grass has been extensively used in urban greenspaces. Turf grass is a

high water demanding plant and needs many operations for maintaining, it is essential to optimize

its surface in dry regions. For partial replacement of lawns, the suitable option is ground cover

plants that along with their easiness in maintaining conditions require less water.

In a comparative study between sport turfgarass and Frankenia thymifolia L., the annual

maintenance costs of F. thymifolia in 100 m2 was almost twice less than sport lawns and it required

approximately 80% less water compared to sport lawns (Shooshtarian and Tehranifar, 2010). Acar

and Var (2001) studied compatibility and ornamental potential of 19 ground cover plant species,

endemic in Trabazon Province (Turkey). They recommended two species of Sedum spurium and

Thymus praecox for application in urban green space because of their acclimation and high levels

of coverage. Dou et al. (2004) recommended three species of ground cover plants native to Yunnan

Province (China) among 205 species. Those three species were recommended regarding with

growth habit and ornamental features to be used in urban landscape of tropical regions.

Water is becoming more and more rare resource in arid and semi arid regions of Iran that char-

acterized by little rainfall, high solar radiation and high temperatures in the summer. In recent years,

the normal seasonal droughts that have occurred in Iran have caused local and state government to enact

water conservation ordinances. Urbanization and increases in population, however, are seriously threat-

ening sustainable natural resources. At present, non-renewable groundwater resources are being depleted

to an alarming extent. As high-quality water supply becomes limited, the use of saline water with high

salt levels for landscape irrigation is being encouraged. Despite the voluminous research on the sensi-

tivity of plants to water stress, relatively little attention has been paid to landscape plants. Several orna-

mental plants have long been considered to be very drought resistant but their actual water needs and

the reasons behind their adaptations are not well known. Understanding the responses of plants to water

deficits is led to establish methods for improving water use efficiencies, and to select appropriate plants

and sites in planning landscape (Mohammadian et al., 2005). The aim of this study was to evaluate the

physiological and biochemical responses of 4 ornamental ground cover species under drought stress.


MATERIAL AND METHOD

Plant material and experimental condition

This research was conducted using a factorial experiment based on a completely randomized

design with regulated deficit irrigation at four levels (100, 75, 50, and 25% of lawn irrigation require-

ment) and three replicates to evaluate some biochemical and physiological indexes on four groundcover

plants namely Lolium perenne, Potentilla spp., Trifolium repens and Frankinia spp. with the potential

use in landscape. For this purposes transplants of three species were planted and Lolium perenne seeds

were sowed in 48 plastic pots filled with simple horticultural soil and marketed peat. Water stress treat-

ments started after two months of well water irrigation at the stage of establishment.

Leaf relative water content

Leaf Relative Water Content (LRWC) was calculated based on the method of Schonfeld et al.,(1988). Leaves were first removed from the stem and then weighed to obtain fresh mass (FM) at the

harvest stage. In order to determine the turgid mass (TM), leaves were floated in distilled water inside

a closed Petri dish for 6 hours. The leaf samples were weighed after gently wiping the water from the

leaf surface with tissue paper, then the leaf samples were placed in an oven at 80 ̊C for 48 h, in order

to obtain dry mass (DM). All mass measurements were made using an analytical scale, with a precision

of 0.0001 g. Values of FM, TM and DM were used to calculate LRWC using the following equation:

LRWC (%) = [(FM–DM)/(TM–DM)] ×100.

Proline content

The proline content was estimated by the method of Bates et al. (1973).The plant material

was homogenized in 3% aqueous sulfosalicylic acid and the homogenate was centrifuged at 1000

rpm. Supernatant was used for the estimation of proline content. The reaction mixture consisted

of 2 ml supernatant, 2 ml ninhydrin acid and 2 ml of glacial acetic acid, which was boiled at 100º

C for 1 h. After termination of the reaction in ice bath, the reaction mixture was extracted with 4

ml of toluene and the absorbance was read at 520 nm. The proline concentration was determined

using calibration curve. The results were calculated based on DW.

Chlorophyll content

Leaf samples were selected randomly from the plants and homogenized in a mortar in ace-

tone. The extract was centrifuged at 5000 rpm for 5 min. Absorbance of the supernatant was

recorded at 663, 645 and 450 nm spectrophotometrically (Techcomp 8500 II, South Korea). Chloro-

phyll (Chl) content was determined following the method of Arnon (1949).

Electrolyte leakage

Electrolyte leakage which is used to assess membrane permeability was determined ac-

cording to Lutts et al. (1996). Leaf discs (1 cm in diameter) from two randomly chosen plants per

plot were taken from the middle portion of youngest fully developed leaf and then were placed in

individual stoppered vials containing 20 ml of distilled water after three washes with distilled water

to remove surface contamination. After incubating the samples at room temperature on a shaker

(150 rpm) for 24 h, the electrical conductivity (EC) of the bathing solution (EC1) was determined.

The same samples were then placed in an autoclave at 121 ̊C for 20 min. and a second reading

(EC2) was determined after cooling solution to room temperature. The electrolyte leakage was

calculated as EC1/EC2 and expressed as percent.

Total soluble carbohydrate content

Total soluble carbohydrates were extracted according to a modified procedure described by

Wardlaw and Willenbrink (1994). 0.5 g of dry leaves was homogenized with 5 ml of 95% ethanol. One-


tenth ml of alcoholic extract preserved in refrigerator mixed with 3 ml anthrone (150 mg anthrone, 100

ml of 72% sulphuric acid,W/W). The samples placed in boiling water bath for 10 minutes. The light

absorption of the samples was estimated at 625 nm using a PD-303 model spectrophotometer. Contents

of soluble sugar were determined using glucose standard and expressed as mg g-1DW of leaves.

DPPH free radical-scavenging activies

The free radical scavenging ability was measured using the protocols described by Shi-

mada et al. (1992) and Yoshiki et al. (2001). Briefly, an aliquot of 4 ml of the methanolic ex-

tract (4, 6 and 8 mg/ml) was added to 1 ml of 10 mM DPPH (2, 2-diphenyl-1- picrylhydrazyl)

solution freshly prepared in methanol. The mixture was left in the dark for 30 min, and decol-

orization of DPPH donated H+ was followed by measuring the absorbance at 517 nm. DPPH

radical-scavenging activity was calculated from the absorption according to the following

equation:

DPPH radical-scavenging activity % = [(Acontrol - Asample) / Acontrol)] × 100.

Statistical Analysis

Analysis of variance (ANOVA) for all the variables was carried out using the JMP8 soft-

ware. Treatment means were compared using the protected least significant difference (LSD) test

at p≤0.05 level.


Results of this experiment showed significant effects of water stress on different studied

traits and also among different species. Mean comparison are shown in table 1.

Leaf relative water content

The results showed that water stress effect was significant (p≤0.01), but interaction effects

were not significant. Trifolium repens (41.6%) had higher relative water content as compared to

other species (Table 1). On the other hand, no signifiacant difference is observed in water stress

treatments up to 50% of lawn requirement, and a sharp decline occurs from then. The rate of RWC

in plant with high resistance against drought is higher than others. In other words, plant having

higher yields under drought stress should have higher RWC. Decrease in RWC in plants under

drought stress may depend on plant vigor reduction and have been observed in many plants (Liu

et al., 2002). Under water deficit, cell membrane subjects to changes such as penetrability and de-

crease in sustainability (Blokina et al., 2003).

Proline content

Water stress, species and their interaction effects were highly significant in proline content

Proline

(mg/g DW)

Carbohydrate

(mg/g FW)

Chlorophyll con-

tent (mg/g FW)

DPPH scav-

enging (%) RWC EL

Plant species

Water stress

Lolium perennePotentilla spp.Trifolium repensFrankinia spp.0

25%

50%

75%

0.54b

0.51b

0.54b

0.89a

0.41d

0.54c

0.65b

0.84a

1.54a

1.38a

0.79b

0.99b

1.2a

1.4a

1.2a

0.89b

31.3a

31.2a

27.99a

15.9b

32.6a

30.9a

21.4b

21.5b

38.9c

57a

46.8b

51.1ab

7.8c

53.8b

63.89a

68.5a

32.7bc

29c

41.6a

36.2ab

35.7a

38.5a

39.3a

26b

29.5b

12.4d

17.9c

49.69a

20.2c

22.7bc

26.6b

39.9a

Table 1. Mean comparison of studied traits in species and water stress levels.

Digits with different letters in each column are significantly different at P≤5%.


(p≤0.01). Frankinia spp showed the highest proline content (with 0.84 mg/g FW) among species.

Under water stress, the highest amount of proline content were observed in 25% lawn requirement

treatment and potentila spp. species (Fig. 1.).

Among amino acids, the accumulation of proline is frequently reported in many plants or

tissues in response to a variety of abiotic stresses (Beltrano et al., 1997). The higher proline content

could be due to enhance activity of ornithine aminotransferase (OAT) and pyrroline 5-carboxylate

reductase (P5CR), the enzyme involved in proline biosynthesis as well as the inhibition of proline

oxidase, proline catabolising enzymes (Debnath, 2008).

Chlorophyll content

Results from leaf chlorophyll content measurements showed a significant difference be-

tween water stress treatments and species (p≤0.01) and interaction effects was significant at 5%

level. Frankinia spp showed the lowest chlorophyll content (15.9 mg/g FW) and 50% and 75%

lawn requirement also showed the lowest chlorophyll amounts (Fig. 2). Our results are in agree-

ment with Nyachiro et al. (2001), who described a significant decrease of chlorophyll a and b

caused by water deficit in six Triticum aestivum cultivars. Under water deficit conditions, photo-

synthetic rate of all wheat cultivars was significantly reduced. It is well evident that reduction in

photosynthetic rate occurs due to stomatal closure under water deficit conditions which may limit

CO2 diffusion into the leaves (Nyachiro et al., 2001).

Electrolyte leakage

Effects of drought stress, species and their interaction effects were significant on electrolyte

leakage (p≤0.01). Frankinia spp. and Potentila sp scored the highest (49.6 %) and lowest (12.4%)

respectively. 75% lawn requirement (39.9 %) also bears the highest electrolyte leakage water stress

treatments. Electrolyte leakage increases linearly with the severity of drought stress in all 4 species.

Water stress can increase reactive oxygen species synthesis (ROS) that produce proteins,

membrane lipids and photosynthetic pigments degradation with a loss of cell membrane stability

Fig. 1. Interaction effects of water stress and species on proline content.

Fig. 2. Interaction effects of water stress and species on chlorophyll content.


(Navari-Izzo et al., 1997; Beltrano et al., 1997). Tolerance drought strategy could be associated to

integrity cell membrane preservation and its rapid reparation (Oliver, 1991).

Total soluble carbohydrate content

According to the results of this study, applying water stress significantly affected total sol-

uble carbohydrate content of plants significantly (p≤0.01). Interactions were not significant. Plant

species are significantly different for this trait.The highest and lowest total soluble carbohydrate

content (1.54 and 0.79 mg/g FW) was observed in Lolium perenne and Trifolium repens, respec-

tively. The most water stress showed the lowest amount of this trait (0.89 mg/g FW). The tolerance

mechanism in water-deficit may be associated with accumulation of osmoprotectants such as pro-

line and soluble sugars. Accumulations of total soluble carbohydrates increase the resistance to

drought stress in plants (Keyvan, 2010). Earlier reports mentioned that sugars protect the cells dur-

ing drought by the following mechanism; the hydroxyl groups of sugars may substitute for water

to maintain hydrophilic interactions in membranes and proteins during dehydration. Thus, sugars

interact with proteins and membranes through hydrogen-bonding, thereby preventing protein de-

naturation (Al- Rumaih and Al- Rumaih, 2007).

DPPH free radical-scavenging activities

Water stress, species and their interaction effects were highly significant in DPPH free radical-

scavenging activities (p≤0.01). The highest (68.5 %) and lowest (7.8 %) amounts of free radical scav-

enging activities were obtained from sever water stress and control treatments, respectively (Table 1).

Fig 4. Interaction effects of water stress and species on DPPH free radical-scavenging assay.

Fig. 3. Interaction effects of water stress and species on electrolyte

leakage.

Fig. 4. Interaction effects of water stress and species on DPPH free

radical-scavenging assay.


Water deficit stress induced the generation of reactive oxygen species (ROS) (Shao, 2008).

It is now widely accepted that these cytotoxic ROS are responsible for various stress-induced dam-

ages to macromolecules mainly lipid membrane peroxidation which could be determined by meas-

uring malon-dialdehyde (MDA). Enhancement in the production, ability and capacity of

antioxidants may play an important role in metabolic stress tolerance and appeared to play a role

in the protection of cellular machinery against damage by reactive oxygen species (Sairam and

Srivastava, 2001; Guo et al., 2006).

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Yuncai, H. and Schmidhalter, U. 2005. Drought and salinity: A comparison of the effects of drought

and salinity. Journal of Plant Nutrition and Soil Science, 168: 541-549.

ـــت ـــبز از اهمی ـــای س ـــم در فض ـــور مه ـــک فاکت ـــوان ی ـــروزه به عن ـــش ام ـــت تن مدیریویـــژه ای برخـــوردار اســـت. به همیـــن منظـــور مطالعـــه ای بصـــورت آزمایـــش فاکتوریـــل در قالـــب ـــا 4 تیـــار آبیـــاری تنظیـــم شـــده )25، 50، 75 و 100 درصـــد نیـــاز آبـــی طـــرح کامـــال تصادفـــی بچمـــن( و 3 تکـــرار بـــرای ارزیابـــی تعـــدادی از ویژگی هـــای بیوشـــیمیایی و فیزیولوژیکـــی 4 ـــال ).Potentilla spp(، شـــبدر ســـفید ـــه )Lolium perenne(، پوتنتی ـــوم پرن ـــاه پوششـــی لولی گیــای ــتفاده در فضـ ــیل اسـ ــا پتانسـ ــا ).Frankinia spp( بـ )Trifolium repens( و فرانکینیـســـبز انجـــام گرفـــت. نتایـــج نشـــان داد کـــه فرانکینیـــا و 75 درصـــد نیـــاز آبـــی چمـــن، باالتریـــن میـــزان پرولیـــن )0/84 میلی گـــرم در هـــر گـــرم وزن تـــر( را داشـــت و از ســـوی ـــرم ـــر گ ـــرم در ه ـــل )32/61 میلی گ ـــزان کلروفی ـــرین می ـــی بیش ـــرات تک ـــن اث ـــر در بی دیگوزن تـــر( در گیاهـــان بـــدون تنـــش آبـــی و کمریـــن میـــزان کلروفیـــل )15/95 میلی گـــرم ـــدارت ـــوای کربوهی ـــرای محت ـــد. ب ـــاهده ش ـــا مش ـــاه فرانکینی ـــر( در گی ـــرم وزن ت ـــر گ در هکل، بیشـــرین مقـــدار )1/54 میلی گـــرم در هـــر گـــرم وزن تـــر( بـــه لولیـــوم پرنـــه تعلـــق ـــق ـــر( متعل ـــرم وزن ت ـــر گ ـــرم در ه ـــدار )0/79 میلی گ ـــن مق ـــه کمری ـــی ک ـــت در حال داشـــای آزاد ـــردن رادیکال ه ـــی ک ـــزان خنث ـــن می ـــفید باالتری ـــبدر س ـــود. ش ـــفید ب ـــبدر س ـــه ش بـــده روی ـــم ش ـــاری تنظی ـــت. در کل، آبی ـــا داش ـــن گونه ه ـــد( را در بی DPPH )75/05 درصـــا ـــود. ام ـــی دار ب ـــش شـــده معن ـــان پوششـــی آزمای ـــیمیایی گیاه ـــف بیوش ـــات مختل خصوصیـــرای ـــی ب ـــد روش خوب ـــاه، می توان ـــه گی ـــتانه خســـارت ب ـــا آس ـــی ت ـــن روش مدیریت ـــرد ای کارب

ـــد. ـــبز باش ـــای س ـــان فض ـــرف آب در گیاه ـــت م مدیری

دهــیـکـچ

تاثیـر تنـش خشـکی روی برخـی ویژگی هـای بیوشـیمیایی و فیزیولوژیکی 4 ،)Potentilla spp.( پوتنتیـا ،)Lolium perenne( گیاه پوششـی لولیـوم پرنـهشـبدر سفید )Trifolium repens( و فرانکینیا ).Frankinia spp( با پتانسیل

اسـتفاده در فضای سبزالهام سمیعیانی1* و حسین انصاری2

1 دانشجوی کارشناسی ارشد، گروه علوم باغبانی، دانشگاه فردوسی مشهد

2 دانشیار گروه کشاورزی، دانشگاه فردوسی مشهد

تاریخ تایید: 9 بهمن 1392 تاریخ دریافت: 17 شهریور 1392 [email protected] :ایمیل نویسنده مسئول *

کلیــد واژگــان: فعالیت آنتی اکسیدانی، کلروفیل، محتوای پرولین، محتوای قندهای کل محلول.

مجله گیاهان زینتیwww.jornamental.com قابل دسترس در سایت

شماره استاندارد بین المللی چاپ: 6433-2251 شماره استاندارد بین المللی آنالین: 2251-6441

مجله گیاهان زینتی، سال چهارم، شماره 1، )1392(8

ـــی دارد. ـــای گیاه ـــع گونه ه ـــر رسی ـــی در تکثی ـــش مهم ـــه ای رز نق ـــاد درون شیش ازدی

ایـــن روش منجـــر بـــه تولیـــد صفـــات مطلـــوب، تولیـــد گیاهـــان ســـامل و عـــاری از

ـــای ـــا غلظت ه ـــره و ب ـــای گ ـــتفاده از منونه ه ـــا اس ـــادی ب ـــز ازدی ـــود. ری ـــاری می ش بی

ـــج ـــط کشـــت MS و VS انجـــام شـــد. نتای ـــی از BAP، GA3 و NAA روی محی مختلف

ــاوی دو ــط کشـــت VS حـ ــاره روی محیـ ــرآوری شاخسـ ــرین پـ ــه بیشـ ــان داد کـ نشـ

ــرین ــن بیشـ ــد. همچنیـ ــل شـ ــدون GA3 و NAA حاصـ ــر BAP بـ ــرم در لیـ میلی گـ

ــتورالعمل ــه یـــک دسـ ــن مطالعـ ــد. ایـ ــه در محیـــط VS/2 بدســـت آمـ ــد ریشـ تولیـ

ـــم ـــه مه ـــک مرحل ـــوان ی ـــه ای Rosa canina به عن ـــت درون شیش ـــرای کش ـــردی ب کارب

ـــر رز ـــربد تکثی ـــت پیش ـــوژی جه ـــک بیوتکنول ـــک تکنی ـــز و ی ـــر موفقیت آمی ـــرای تکثی ب

ـــرد. ـــه ک ـــران ارائ در ای

دهــیـکـچ

مجله گیاهان زینتیwww.jornamental.com قابل دسترس در سایتشماره استاندارد بین المللی چاپ: 6433-2251 شماره استاندارد بین المللی آنالین: 2251-6441

ریزازدیادی گیاه Rosa canina از طریق پرآوری شاخه های جانبی

محبوبه داودی پهنه کالیی 1*، علی تهرانی فر2، لیال سامعی3 و محمود شور41 دانشجوی کارشناسی ارشد، گروه علوم باغبانی دانشکده کشاورزی دانشگاه فردوسی مشهد

2 گروه علوم باغبانی دانشکده کشاورزی دانشگاه فردوسی مشهد

3 گروه گیاهان زینتی، مرکز تحقیقات علوم گیاهی، دانشگاه فردوسی مشهد

4 گروه علوم باغبانی دانشکده کشاورزی دانشگاه فردوسی مشهد

تاریخ تایید: 1 بهمن 1392 تاریخ دریافت: 14 مهر 1392 [email protected] :ایمیل نویسنده مسئول *

کلیــد واژگــان: Rosa canina، گونه های بومی، ریز ازدیادی، تنظیم کننده های رشد گیاهی

کوتاهه ها: MS )موراشیگی و اسکوگ(، VS ) وان در سامل(، BAP )بنزیل آمینو پورین(، NAA )نفتالین استیک اسید(، GA3 )اسید جیربلیک(.

7 مجله گیاهان زینتی، سال چهارم، شماره 1، )1392(

حفظ کیفیت و دوام گل هاي بریده یکی از مسائل مهم صنعت تولید گل و گیاه

به ویژه در زمینه گل هاي شاخه بریده می باشد و مهمرين مشكل داودی به عنوان

یکی از با ارزش ترين گل هاي شاخه بريده، نگهداری پس از برداشت آن مي باشد. لذا

بدین منظور آزمایشی بر پایه طرح کامال تصادفی به صورت تیار پالس با سه فاکتور

سولفات کینولین 8-هیدروکسی درصد(، و40 20 ،10 ،5( غلظت های با چای عصاره

با ریفامپیسین بیوتیک آنتی و ) لیر در میلی گرم )100، 200 و 400 با غلظت های

غلظت های )100، 200 و 400 میلی گرم در لیر( و شاهد بر روی داودی خوشه ای )رقم

بنفش( در سه تکرار مورد آزمون قرار گرفت. نتایج نشان داد که تیار 20 درصد عصاره

چای و تیار 100 میلی گرم در لیر 8- هیدروکسی کینولین سولفات باالترین عمرگلجایی،

پروتئین گلربگ، کلروفیل کل، جذب آب و کاروتنوئید گلربگ را نشان دادند.

دهــیـکـچ

تاثیـر عصـاره چـای، 8- هیدروکسـی کینولیـن سـولفات و ریفامپیسـین Denderanthema بـر عمـر پس از برداشـت گل شـاخه بریـده داودی

grandiflorum L.cv. Purple

داود هاشم آبادی1* و حمیده باقری21 استادیار گروه باغبانی، دانشگاه آزاد اسالمی واحد رشت

2 فارغ التحصیل کارشناسی ارشد علوم باغبانی، دانشگاه آزاد اسالمی واحد رشت و عضو باشگاه پژوهشگران جوان واحد رشت، رشت، ایران

تاریخ تایید: 10 اسفند 1392 تاریخ دریافت: 26 آبان 1392 [email protected] :ایمیل نویسنده مسئول *

کلیــد واژگــان: پروتئین گلبرگ، جذب محلول، عمرگلجایي، کاروتنوئید گلبرگ، کلروفیل کل




پــرورش چمــن رول یکــی از روش هــای مهــم در اســتقرار و تعمیــر چمــن به خصوص

در زمین هــای ورزشــی اســت. امــروزه، از مخلــوط ماســه و پیــت در تولیــد چمــن رول در

ــران اســتفاده می شــود. چــون پیــت گــران اســت، به نظــر می رســد یــک بســر کاشــت ای

جایگزیــن بایــد معرفــی شــود. شــلتوک برنــج، پســاند چــای و کمپوســت بــرگ ترکیبــات

آلــی اقتصــادی و مقــرون بــه رصفــه هســتند کــه در زمیــن شــال ایــران قابــل دســرس

هســتند. هــدف از ایــن مطالعــه ارزیابــی اســتفاده از ترکیبــات آلــی ارزان قیمــت بــرای

جوانه زنــی بــذر چمــن و تولیــد رول یکنواخــت اســت. بنابرایــن یــک طــرح کامــال تصادفــی

بــا 3 تکــرار و 6 تیــار: 1- مخلــوط کمپوســت بــرگ و ماســه )1:1 حجمــی(، 2- کمپوســت

ضایعــات چــای و ماســه )1:1 حجمــی(، 3- ماســه، 4- مخلــوط شــلتوک برنــج و ماســه )3:1

ــه آزمایشــی دانشــگاه ــاهد در مزرع ــای 1، 2، 3، 4 و 6- ش ــوط تیاره ــی(، 5- مخل حجم

آزاد اســالمی واحــد چالــوس در تابســتان 1387 انجــام شــد. گونــه چمــن مــورد اســتفاده

ــرهای ــر بس ــه اث ــد ک ــخص ش ــده مش ــت آم ــج به دس ــق نتای ــود. طب Lolium prenne ب

کاشــت روی درصــد جوانه زنــی معنــی دار بــود )p≥0/05(. نتایــج نشــان داد کــه مخلــوط

شــلتوک برنــج و ماســه )3:1 حجمــی( درصــد جوانه زنــی را بیــش از بقیــه تیارهــا افزایــش

داد. ایــن امــر احتــاال به خاطــر توانایــی نگهــداری بــاالی آب و تهویــه مطلــوب در شــلتوک

ــزان یکنواختــی به ترتیــب در تیــار ــوده اســت. به عــالوه کمریــن و بیشــرین می ــج ب برن

ــوق را ــات ف ــه ضایع ــی به نظــر می رســد ک ــد. به طــور کل ــوط و ماســه مشــاهده ش مخل

می تــوان به صــورت اقتصــادی بــرای تولیــد چمــن رول اســتفاده کــرد.

دهــیـکـچ

کاربــرد شــلتوک برنــج بــه عنــوان بســتر جدیــد جوانه زنــی بــذر چمــن در تولیــد چمــن رولــی، رویکــرد نوینــی از کشــاورزی پایــدار

محمد علی گلستانی1*، علی دولتخواهی2، نوید وحدتی2 و امید نوری رودسری31 گروه فضای سبز، دانشکده کشاورزی، دانشگاه آزاد اسالمی، واحد چالوس

2 دانشجوی دکتری، گروه علوم باغبانی، دانشگاه فردوسی مشهد، مشهد، ایران3 گروه اگرواکولوژی، موسسه تحقیقات علوم محیطی، دانشگاه شهید بهشتی، تهران، ایران

تاریخ تایید: 18 دی 1392 تاریخ دریافت: 10 آبان 1392 [email protected] :ایمیل نویسنده مسئول *

کلیــد واژگــان: ضایعات کشاورزی، شلتوک برنج، جوانه زنی برنج، تولید چمن رول، یکنواختی



ــو’( از نظــر رشــد، ــی’، ‘ســیندرال’ و ‘متپ ــد ســالم’، ‘نلســون’، ‘کال ــج رقــم میخــک )‘گران پن

عملکــرد و کیفیــت زیــر ســایه بان در ایســتگاه تحقیقاتــی گیاهــان زینتــی »کاال شــاه کاکــو«

در الهــور در ســال 2011 مــورد ارزیابــی قــرار گرفتنــد. بیــن ایــن ارقــام، حداکــر ارتفــاع

ــیندرال’ ــانتی مر( و ‘س ــی’ )78/23 س ــانتی مر(، ‘کال ــالم’ )78/66 س ــد س ــم ‘گران در رق

ــو’ )6/3( و ــام ‘متپ ــاخه در ارق ــداد ش ــرین تع ــد. بیش ــاهده ش ــانتی مر( مش )77/96 س

‘نلســون’ )6/2( بــود. حداکــر قطرســاقه در ‘نلســون’ )6/21 میلی مــر( و حداقــل آن

در ‘گرانــد ســالم’ )3/63 میلی مــر( بــود. حداکــر تعــداد میانگــره در هــر ســاقه در رقــم

‘نلســون’ )12/66( و ســپس رقــم ‘کالــی’ )11/33( و ‘گرانــد ســالم’ )11( مشــاهده شــد.

ــد ــو’ )189/6( تولی ــم ‘نلســون’ )198/3( و ‘متپ ــع در رق حداکــر تعــداد گل در مــر مرب

ــو’ از نظــر رشــد و ــه رقم هــای ‘نلســون’ و ‘متپ ــت ک ــوان نتیجــه گرف ــن می ت شــد. بنابرای

ــد. ــه بهــر بودن ــر ســایه بان از بقی گلدهــی و عملکــرد زی

دهــیـکـچ

رشد، عملکرد و کیفیت ارقام میخک ).Dianthus caryophyllus L( های تولید شده زیر سایه بان

مالک عابد محمود، محمد سالم اخترخان و نوید احمد*مدیریت گلکاری پنجاب، الهور، پاکستان

تاریخ تایید: 17 بهمن 1392 تاریخ دریافت: 6 دی 1392 [email protected] :ایمیل نویسنده مسئول *

کلیــد واژگــان: میخک، ارقام، سایه بان، عمر گلجایی، عملکرد




در ایـن تحقیـق از مارکرهـای RAPD بـرای آنالیـز تنـوع ژنتیکـی بیـن 20 واریتـه ی

فیلودنـدرون اسـتفاده شـد. PCR بـا 60 پرایمـر RAPD، به جز 21 پرایمری که چند شـکلی

بیشـری نشـان داده بودنـد، انجـام شـد. در کل، 354 جایـگاه RAPD قابـل دسـته بندی بـا

348 بانـد چنـد شـکل )98 درصـد( مشـاهده شـد. درصـد باندهـای چنـد شـکلی بیـن 80 تا

100 درصـد بـود. قطعـات DNA تکثیر شـده بـرای آنالیز آماری اسـتفاده شـدند. داده ها به

کمـک عالمت گـذاری باندهـا بدسـت آمدنـد و بـا نـرم افـزار STATISTICA آنالیز شـدند.

یـک دندروگـرام بوسـیله آنالیـز خوشـه ای بـر پایـه ی وجـود یـا عدم وجـود باند بدسـت آمد

کـه نشـان دهنده ی تشـابه و تنـوع اسـت. آنالیـز خوشـه ای واریته هـای فیلودنـدرون را بـر

اسـاس عـادات رشـد، خصوصیـات مورفولوژیکـی و منشـاء جغرافیایـی گروه بنـدی کـرد. این

امـر نشـان می دهـد کـه نوعی ارتبـاط بین الگوهـای RAPD و منشـا جغرافیایـی آن واریته

وجـود دارد. تنـوع زنتیکـی بیـن واریته هـا در حـد متوسـط بـود کـه ایـن امر شـاید به خاطر

هروزیگوتـی و تنوع سـوماکلونال باشـد.

دهــیـکـچ

مطالعـه روی تنـوع ژنتیکـی موجـود بیـن واریته هـای فیلودنـدرون بـه کمک RAPDمارکرهـای

آچار دواراجا*، جاخار مامتا، جاکر پی و شیتی کی پی ویآزمایشگاه بیوتکنولوژی، بذرهای هیبرید ایندو آمریکن، بنگلور، هندوستان

تاریخ تایید: 10 اسفند 1392 تاریخ دریافت: 28 آذر 1392 [email protected] :ایمیل نویسنده مسئول *

کلیــد واژگــان: خانواده شیپوریان، گیاهان آپارتمانی، بی نیاز به قیم، اسپادیکس، جنگل های گرمسیری



بــا ارزش Rosa indica و اصــالح رز یــک راه حــل تکثیــر درون شیشــه ای

ــت رســیده اســت. ــن اهمی ــه باالتری ــد ب ــان و کشــور هن ــه در جه اقتصــادی اســت ک

ــادی ــد اســت. پروتوکل هــای ریزازدی ــک مبحــث جدی ــد ی ــام تجــاری جدی گســرش ارق

ــر ــی و معط ــام محل ــال ارق ــر کلون ــا روی تکثی ــت ام ــده اس ــه ش ــام رز تهی ــرای ارق ب

رز در هنــد گزارش هــای ناچیــزی وجــود دارد. در ایــن مطالعــه متایز یابــی مجــدد

منونه هــای گیاهــی از گره هــای ســاقه، دیســک بــرگ، گلربگ هــا و تخمــدان را روی

محیــط کشــت MS غنــی شــده )تکمیــل شــده( بــا ترکیبــات گوناگــون مثــل آب نارگیــل

)NAA, IAA,توفــوردی, IBA( اکســین ها ،)CW بــه میــزان 5 تــا 20 درصــد حجمــی(

ــت. ــرار گرف ــه ق ــورد مطالع ــف م ــای مختل ــیتوکنین ها )Kn, BAP( در غلظت ه و س

ــا مشــاهده خوشــه های امربیوژنــی در منونه هــای گــره ســاقه ــوس قــوی ب تشــکیل کال

ــل )10% حجمــی( + ــا آب نارگی ــی شــده ب ــرگ روی بســر کشــت MSغن و دیســک ب

ــا توفــوردی )11/3 میکرومــول( + BAP )3/55 میکرومــول( بدســت آمــد. پــرآوری 4 ت

ــه گل در ــکیل جوان ــز تش ــاره 1/24±5( و نی ــن شاخس ــف )میانگی ــاره مضاع 7 شاخس

ــد شــده از ریزمنونه هــای گــره ســاقه ای کــه رشایــط درون شیشــه ای روی رزهــای تولی

روی MS غنــی شــده بــا BAP + توفــوردی + CW کشــت شــده بودنــد، مشــاهده شــد.

شاخســاره های تولیــد شــده در تیــار بــا ‘Quic Root’ )مــواد شــیمیایی قابــل دســرس

ــد. ــد کردن ــرای ریشــه زایی(، سیســتم ریشــه ای بســیار مناســبی تولی و تجــاری ب

دهــیـکـچ

تشـکیل جوانـه گل، باززایی گیـاه و مطالعات مورفولوژیـک در یکی از ارقام معطر محلی رز )Rosa indica( در شـرایط کشـت درون شیشه ای

راگیوا بیمال و نیتی کیران* آزمایشگاه بیوتکولوژی، گروه گیاهشناسی، دانشگاه بیهار، بیهار، هندوستان

تاریخ تایید: 24 دی 1392 تاریخ دریافت: 1 مهر 1392 [email protected] :ایمیل نویسنده مسئول *

کلیــد واژگــان: شاخساره چندگانه، ‘Quic Root’، رز، Rosa indica، کشت بافت




در گلـکاری صنعـت در گسـرده کاربـرد به علـت مصنوعـی بـذر تکنولـوژی

نگهـداری از ژرم پالسـم و تبـادل آن بیـن کشـورهای مختلـف از اهمیـت و محبوبیـت

خاصـی برخـوردار شـده اسـت. در این آزمایش کاربـرد ایـن روش روی توانایی جوانه زنی

و نگهـداری از ارکیـده ی دندربیـوم رقـم ‘وایـت فیـری’ در مـدت زمان هـا و ظـروف

مختلـف نگهـداری، مـورد مطالعـه قـرار گرفته اسـت. درصـد جوانه زنی بـاالی 80 درصد

در بـذوری مشـاهده شـد کـه به صـورت مصنوعـی کپسـوله شـده بودند و بیـش از 150

روز در پری دیش هـا و لولـه پروپیلنـی در بسـته قـرار داشـتند و ایـن درصـد به تدریج از

روز یک صـد و پنجاهـم بـه بعـد رشوع به کاهش کـرد. بعالوه، بـذور مصنوعی نگهداری

شـده در لوله هـای پروپیلنـی رسیعـر جوانـه زدنـد و تبدیـل بـه گیاهچـه شـدند و در

مقایسـه با بذور درون پری دیش، شاخسـاره و ریشـه ی طویل تری داشـتند. این آزمایش

کارایـی و سـازگاری لوله هـای پروپیلنـی را نسـبت بـه پری دیش هـا به عنـوان ظـروف

نگهـداری ثابـت کرد.

دهــیـکـچ

Dendrobium کاربــرد موثــر تکنولوژی بــذر مصنوعــی در تکثیر ارکیــدهرقــم ‘وایــت فیری’

وی لنگ سیو*، می یون کوک، یونگ می اونگ، هیو پینگ لیو و بون کیت ییوگروه علوم زیستی، مدرسه علوم کاربردی و سالمتی، موسسه آموزش فنی، سنگاپور

تاریخ تایید: 24 بهمن 1392 تاریخ دریافت: 25 مهر 1392 [email protected] :ایمیل نویسنده مسئول *

کلیــد واژگــان: Dendrobium، اندام های شبه پروتوکرن، بذور مصنوعی، تکثیر و باززایی



www.jornamental.comThe Journal of Ornamental Plants, is an open access journal that provides rapid publication of manuscripts

on Ornamental plants, Floriculture and Landscape. Journal of Ornamental Plants is published in English,

as a printed journal and in electronic form.

All articles published in Journal of Ornamental Plants are peer-reviewed. All manuscripts should convey im-

portant results that have not been published, nor under consideration anywhere else. Journal of Orna-

mental Plants will be available online around the world free of charge at http://www.jornamental.com.

In addition, no page charge are required from the author(s). The Journal of Ornamental Plants is pub-

lished quarterly by Islamic Azad University, Rasht Branch, Rasht, Iran.

Manuscript Submission

Please read the “Instructions to Authors” before submitting your manuscript. Submit manuscripts as e-

mail attachment to Dr. Ali Mohammadi Torkashvand, Executive Director of Journal of Ornamental Plants,

at [email protected]. Electronic submission of manuscripts is strongly encouraged, provided that

the text, tables, and figures are included in a single Microsoft Word 2003 file. A manuscript acknowledg-

ment including manuscript number will be emailed to the corresponding author within 72 hours.

Please do not hesitate to contact meif you have any questions about the journal. We look forward to

your participation in the Journal of Ornamental Plants.

Address: Islamic azad University, Rasht Branch

Horticultural Department,

Agriculture Faculty,

Rasht,

Iran.

P.O.Box 41335-3516

Email: [email protected]

URL: http:// www.jornamental.com

Topics and Types of PaperJournal of Ornamental Plants is an international journal to the publication of original papers and reviews

in the Ornamental plants, Floriculture and Landscape fields. Articles in the journal deal with Ornamental

plants, Floriculture and Landscape. The scope of JOP includes all Ornamental plants, Floriculture and

Landscape. The journal is concerned with Ornamental plants, Floriculture and Landscape and covers

all aspects of physiology, molecular biology, biotechnology, protected cultivation, and environmental areas

of plants. The journal welcomes the submission of manuscripts that meet the general criteria of signif-

icance and scientific excellence, and will publish:

● Research articles

● Short Communications

● Review

Papers are welcome reporting studies in all aspects of Ornamental plants, Floriculture and Landscape

including:

Any Novel Approaches in Plant Science

Biotechnology

Environmental Stress Physiology

Genetices and Breeding

Photosynthesis, Sources-Sink Physiology

Postharvest Biology

Seed Physiology

Soil-Plant-Water Relationships

Modelling

Published by:Islamic Azad University, Rasht Branch, Iran


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