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1.
A highly efficient, stable, and cost-effective micropropagation protocol for the conservation of a medicinal plant Turnera ulmifolia L. was established from nodal tissues via multiple axillary shoot proliferation on using Murashige and Skoog’s (MS) liquid nutrient medium. To begin with, nodal explants were placed on agar gelled medium amended with 2.0 mg L?1 6-benzylaminopurine (BAP) and 0.1 mg L?1 indole-3 acetic acid (IAA) for shoot induction. Subsequently, elongation of regenerated shoots could be possible on liquid MS medium supplemented with 0.5 mg L?1 BAP and Kin (kinetin) each along with 0.1 mg L?1 IAA where high frequency of regeneration in terms of number of shoots (47.2 shoots/explant) was achieved. Furthermore, long and healthy shoots (4?5 cm in length) were rooted on agar gelled half-strength of MS medium supplemented with 2.0 mg L?1 indole-3 butyric acid (IBA). Finally, in vitro regenerated plantlets were gradually acclimatized in the greenhouse and transferred to the field successfully.  相似文献   

2.
Abstract

A micropropagation process was developed for Anogeissus latifolia-a tree of fragile ecosystems. Multiple shoots were regenerated from cotyledonary node and epicotyl explants on Murashige and Skoog's (MS) medium containing 0.1 mgl-1 indole-3-acetic acid (IAA) + 1.5 mgl -l 6-benzylaminopurine (BAP) + additives (25 mgl -l each of adenine sulphate, L-arginine, ascorbic-, citric acids and 1.0 mM L-asparagine) + 200 μM Fe-EDTA (ferric-ethylenediaminetetraacetic acid) salt. The shoots differentiated in vitro were subcultured and repeatedly transferred onto fresh medium but with 1.0 mgl-1 of BAP to achieve 4-5 fold rate of shoot multiplication. After every 4th week of culture, from each culture bottle 8-10 shoots could be harvested for rooting. The shoots produced in culture were rooted in vitro on half strength MS medium with 1.0 mgl -l either of IBA (indolebutyric acid) or NAA (naphthaleneacetic acid). The shoots were pulse treated with combination (100 mgl -l each) of IBA and NOA (2-naphthoxy acetic acid) rooted ex vitro. The ex vitro root induction method is highly efficient and plantlets so generated could be acclimatized and pot transferred. The process developed can be used for large scale production of plants of A.  相似文献   

3.
An efficient in vitro process for rapid clonal propagation of a 40-year-old tree of Azadirachta indica employing nodal stem segments was developed. Season of collection and maturity of explants showed direct influence on bud-break. Nodal stem segments collected during the month of April gave best response. Maximum bud-break (78.6–81%) was obtained when middle order nodes (3rd or 4th node from apex) were taken. Amongst different cytokinins used, 6-benzylaminopurine (BAP) at the concentration of 1.11 μM was found most effective in inducing multiple shoots, whereas inorganic and organic constituents of the medium influenced growth and general condition of proliferating shoots. On an average 3.1 shoots per explant were regenerated in modified Murashige and Skoog medium supplemented with 1.11 μM BAP, 1.43 μM indole-3-acetic acid (IAA) and 81.43 μM adenine hemisulphate. Isolated shoots were rooted in presence of 2.46 μM indole-3-butryic acid (IBA). Root induction took place in 8–10 days with 100% rooting. The in vitro-raised plantlets were successfully transplanted in potted soil and finally grown under field conditions with 100% survival. The genetic fidelity of such in vitro-raised field-grown plants was ascertained by random amplified polymorphic DNA (RAPD) markers. Furthermore, the azadirachtin content of in vitro-cloned plants was found comparable to the mother tree proving their chemical stability also. The protocol developed holds good for in vitro cloning of mature elite neem trees.  相似文献   

4.
This report describes a protocol for the in vitro shoot induction and plant regeneration from epicotyl explants of Cassia angustifolia on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA), Kinetin and 2-iP (0.5–10.0 μM). MS medium supplemented with BA (5.0 μM) was the most effective in inducing adventitious shoots and growth. The highest rate of shoot multiplication was achieved on MS medium supplemented with BA (5.0 μM) and Indole-3-acetic acid (IAA, 1.0 μM). The nodal segments excised from the shoots regenerated from BA (5.0 μM) and IAA (1.0 μM) were used as explants for next three round of micropropagation. The number of shoots significantly increased at successive round of micropropagation. For rooting, MS medium supplemented with 2.0 μM indole-3-butyric acid proved to be better than that supplemented with IAA or α-naphthalene acetic acid. The in vitro raised plantlets with well developed shoot and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse. About 52 plants (85 %) survived out of 60 plants transferred in garden soil.  相似文献   

5.
This report describes in vitro shoot induction and plant regeneration from nodal segments of Balanites aegyptiaca on Murashige and Skoog (MS) medium fortified with 6-benzyladenine (BA), thidiazuron (TDZ) and kinetin (Kin) (0.5–20.0 μM). MS medium supplemented with BA (12.5 μM) was the most effective in inducing bud break and growth and also in initiating multiple shoot proliferation. However, the optimal level of TDZ supplementation to the culture medium was 5.0 μM. Shoot cultures were established by repeatedly subculturing the original nodal explants on the same medium. Highest number of shoots (11.5 ± 0.7) and shoot length (5.0 ± 0.2 cm) were achieved when cultures were subcultured on MS medium supplemented with 12.5 μM BA and 1.0 μM α-naphthalene acetic acid (NAA). The shoots regenerated from TDZ supplemented medium when subcultured to hormone free MS basal medium considerably increased the rate of shoot multiplication and shoot length by the end of fifth subculture. Rooting of the shoots was achieved on MS medium augmented with 1.0 μM indole-3-butyric acid (IBA) plus 0.5% activated charcoal followed by their transfer to half strength MS basal medium. The in vitro raised plantlets with well developed shoots and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse with 70% survival rate. The results of this study provide the first successful report on in vitro direct plant regeneration of B. aegyptiaca.  相似文献   

6.
Vegetative propagation techniques are recognized as indispensable tools for mass multiplication of important multipurpose trees adopted in different agroforestry systems. Albizia procera, one among important species, is difficult to propagate commercially either by stem / root cuttings or layering. A study was undertaken to develop procedure for its in vitro regeneration through organogenesis. Explants collected from 15±2 yr-old mature plus trees and from 15 days old juvenile seedlings were regenerated with exogenous application of different hormones. Epicotyl and hypocotyl explants excised from juvenile seedlings showed higher callusing than axillary bud and shoot tip explants derived from mature trees. Benzylaminopurine (BA) at 3 μg/l was most effective, which induced hundred percent callusing in epicotyl and hypocotyl explants in 1/2 Murashige and Skoog (MS) medium. Callus originated from axillary buds and apical shoot tips of mature trees failed to form organs, however callus derived from epicotyl and hypocotyl explants proliferated and formed de novo shoots and leaflets. A concentration of 3 μg/l of BA was found effective for shoot proliferation. Shoots grew vigorously in 2 μg/l gibberellic acid (GA3) treatment and rooted in 1/2 MS medium supplemented with indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA). Rooting was most successful on medium supplemented with 6 μg/l IBA alone on which 93.3% of the shoots formed roots. Sand or vermiculite supplemented with 4 ml of yoshida solution proved as best hardening media, which recorded 70-80% survival of plantlets. One year old tissue culture raised plants had comparatively more height, collar diameter, biomass, and root shoot ratio than plants raised from cuttings and seeds of the same age. The procedures enumerated provide a basis for the development of in vitro techniques for rapid multiplication of A. procera. This revised version was published online in June 2006 with corrections to the Cover Date.  相似文献   

7.
A highly reproducible and efficient in vitro shoot regeneration system was developed in a potential medicinal plant, Albizia lebbeck using root explants. Root explants from 15 day-old-aseptic seedlings were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations (0.5, 2.5, 5.0, 7.5 and 10.0 μM) of 6-Benzyladenine (BA), Kinetin (Kn), 2-Isopentenyl adenine (2-iP) singly as well as in combination with α-Naphthalene acetic acid (NAA) (0.1, 0.5, 1.0, 1.5 and 2.0 μM). The highest rate of shoot multiplication (16.0 ± 1.87 for the average shoot number and 5.16 ± 0.38 cm for shoot length) was achieved on MS medium supplemented with 7.5 μM BA and 0.5 μM NAA. The effects of medium type, medium strength, pH and subculture on shoot induction and proliferation were also tested. An average of 21.6±2.87 shoots per explants could be obtained following this protocol. Rooting was achieved on microshoots using half strength MS medium with 2.0 μM Indole-3-butyric acid (IBA) after four weeks of culture. The in vitro raised healthy plantlets were successfully established in earthen pots containing garden soil and grown in greenhouse with >80% survival rate.  相似文献   

8.
A micropropagation method for Jaal (Salvadora persica)—a tree of arid horticulture and forestry has been developed. Nodal segments of fresh shoot sprouts originated from axillary buds obtained from a plant around 35–40 years old lopped plant were used as explants for establishment of in vitro cultures. Surface-sterilized explants produced optimum number of shoots through activation of axillary buds on Murashige and Skoog’s (MS) medium containing 8.88 μM BA (6-benzyladenine) + additives (25 mgl−1 each of adenine sulphate, arginine, citric acid, 50 mgl−1 ascorbic acid). The shoot multiplication was influenced by the successive transfer of the mother explants for 4–5 passages. The maximum number (23.1 ± 0.73 shoots per explant) of shoots were regenerated on MS supplemented with 1.11 μM BA + 1.16 μM Kn (Kinetin) + 0.54 μM NAA (α-naphthalene acetic acid). About 90% shoots pulse-treated with a combination of 2460.27 μM Indole-3-butyric acid (IBA) + 494.56 μM NOA (2-naphthoxy acetic acid) were rooted ex vitro on soilrite within 15–18 days. Over 80% cloned plantlets were hardened successfully in a green house and transferred to polybag/pots.  相似文献   

9.
Mroginski  Erika  Rey  Hebe Y.  Mroginski  Luis A. 《New Forests》2003,25(3):177-184
In vitro regeneration of complete plants from nodal single-bud segments of 2-year-old Australian Cedar (Toona ciliata) trees were obtained under defined nutritional and environmental conditions. Explants were dissected from plants obtained by germination of seeds and growth in pots in a greenhouse. The best medium for shoot regeneration was that of Murashige and Skoog at 1/4 strength with 3% sucrose (1/4 MS), supplemented with 0.1 mg/l IBA and 0.5 mg/l BAP. Rooting of regenerated shoots was observed in MS medium with 0.1 mg/l IBA. Using mature tree material was more difficult. Forced flushing was used to induce shoot development on branches of a 10-year-old tree. Nodal segments of these epicormic shoots formed shoots in vitro on 1/4 MS + 0.01 mg/l IBA + 5 mg/l BAP, but rooting was never observed.  相似文献   

10.
In vitro propagation technique ofGmelina arborea multipurpose and a fast growing tree species was studied. Nodal segment including axillary bud was used as a explant. They were cultured on MS media containing various concentrations (0–10 mg/l) of BAP alone or in combination with 0.002 mg/l of IBA. Nodal segments showed axillary bud proliferation in almost all media tested. MS media containing 0.22 mg/l of BAP alone and 2 mg/l of BAP in combination with 0.002 mg/l of IBA were effective for inducing multiple shoots and shoot elongation. MS medium supplemented with 0.02 mg/l of NAA and 1 mg/l of IBA gave the best result for rooting. The regenerated plantlets were potted and acclimatized successfully in a growth chamber and then moved to the green house. Adventitious shoots production from stem explants that were taken from regenerated plantletin vitro was also discussed. Stem segments were tested for their morphogenetic potential on MS media with various combinations and concentrations of BAP, zeatin and TDZ. Successfull result was obtained on MS media supplemented with 2 mg/l of BAP and 1 mg/l of zeatin or supplemented with 0.5 mg/l of BAP and 0.5 mg/l of TDZ. The shoots obtained on MS media containing 2 mg/l of BAP and 1 mg/l of zeatin rooted on MS media containing 0.02 mg/l of NAA and 1 mg/l of IBA, and plantlets were successfully obtained. A part of this paper was presented at the 109th Annual Meeting of the Japanese Forest Society (1998).  相似文献   

11.
In vitro clonal multiplication of Pterocarpus santalinus L. was achieved using mature nodal explants of a 10-year-old elite quality tree. Combinations of serial transfer technique and incorporation of antioxidants (250 mg/l L-ascorbic acid and 50 mg/l citric acid) into the culture medium helped to minimize medium browning and improve explant survival during shoot sprouting. About 70% of explants were sprouted on Murashige and Skoog (MS) liquid medium containing 4.4 μM 6-benzyladenine (BA). The explant harvest period also influenced the bud break and shoot sprouting in nodal explants. The combination of 4.4 μM BA and 2.2 μM thidiazuron (TDZ) was found to be the most suitable growth regulator for obtaining the highest percentage of nodal segment sprouting (74%–75%), the number of secondary shoots per primary shoot (two or three), the shoot length (5–6 cm), the number of new nodal segments generated per active explant (four or five), and the multiplication coefficient (3.5) within 6 weeks. Repeated subculturing of nodal explants obtained from shoot cultures enabled continuous production of healthy axillary shoots. At the end of the sixth passage, about 90% of nodal explants produced five or six healthy green shoots, each being about 6.6 cm long with six or seven nodes. Multiplication coefficient was also increased from the first subculture (5.4) to the sixth subculture (8.3). The best rooting response was achieved on solidified half-strength MS medium supplemented with 4.9 μM indole-3-butyric acid (IBA). About 70% of the micropropagated plantlets were established successfully in 20-cm pots containing a mixture of soil and farmyard manure (4 : 1 ratio) and formed new leaflets.  相似文献   

12.
Santalum album is known as East Indian sandalwood. It is the most economically important tree harvested for heartwood oil, and India is among the chief exporters of sandalwood and its products. Multiple shoots were induced from nodal shoot segments derived from a 50- to 60-year-old candidate plus tree (CPT) on Murashige and Skoog (MS) medium supplemented with 0.53 μM α-naphthaleneacetic acid (NAA) and 11.09 μM 6-benzylaminopurine (BA). In vitro differentiated shoots were multiplied on MS medium with 0.53 μM NAA, 4.44 μM BA, and additives: 283.93 μM ascorbic acid, 118.10 μM citric acid, 104.04 μM cystine, 342.24 μM glutamine, and 10% (v/v) coconut milk. New shoots were harvested repeatedly for up to three subculture passages on fresh medium at 4-week intervals. Microshoots treated with 98.4 μM indole-3-butyric acid (IBA) for 48 h produced roots on growth-regulator-free, quarter-strength MS basal salts medium with vitamin B5 and 2% sucrose. In vitro root induction was achieved from microshoots pulsed with 1230 μM IBA for 30 min in soilrite rooting medium. The percentage of rooting in soilrite was higher than that for agar medium, and in vitro raised plants were established in the field and showed normal growth.  相似文献   

13.
An in vitro regeneration system was developed using organogenic callus derived from in vitro grown cotyledonary explants of Gleditsia caspica Desf., an important leguminous tree. Murashige and Skoog (MS) basal medium augmented with 0.2 g L?1 myo-inositol and various concentrations of either 2,4-dichlorophenoxyacetic acid (2,4-D), naphthaleneacetic acid, or indole-3-butyric acid (IBA) alone as well as combined with cytokinins was used for callus induction. The highest frequency of organogenic yellowish-white and nodular callus (93 %) was obtained from explants grown on medium supplemented with 13.5 μM 2,4-D and 4.4 μM benzyladenine (BA). The yellowish-white and nodular callus when transferred to MS medium supplemented with BA (2.2–17.7 μM) or kinetin (KT; 2.3–18.8 μM) solely or in combination with 2.3 μM 2,4-D produced several microshoots after 5 weeks culture. The calli cultured on MS medium with 4.4 μM BA singly showed superior growth response and produced both maximum shoot regeneration (94 %) and the highest mean number (4.3) of microshoots per callus. Transfer of regenerated microshoots onto modified MS basal medium fortified with 5.8 μM gibberellic acid and 4.4 μM BA resulted in the maximum number of internodes per shoot and the highest shoot elongation after a period of 6 weeks. Optimum rooting of 90 %, an average 6.1 roots per shoot, and a mean root length of 3.6 cm was observed when half-strength MS medium was supplemented with 9.8 μM IBA and 0.92 μM KT. The regenerated healthy plants with well-developed shoots and roots showed a survival rate of 77 % after acclimatization and transplanting to garden soil for a 10-week hardening period under ex vitro conditions.  相似文献   

14.
An efficient system was developed for direct plant regeneration from in vitro-derived leaf explants of Embelia ribes Burm. f., a vulnerable medicinal woody climber of the Western Ghats of India. The in vitro procedure involved three steps that included induction of shoot initials from leaf tissue, regeneration and elongation of shoots from the shoot initials, and rooting of shoots. The induction of shoot initials was achieved on Murashige and Skoog (MS) solid medium supplemented with different concentrations of thidiazuron (TDZ). The best medium for shoot induction was MS with 0.272 μM TDZ. Numerous shoot primordia developed within 2–3 weeks on the leaf margin as well as on the midrib region, without any callus phase. In the second step, the shoot clumps separated from the leaf explant on transfer to MS basal medium, resulting in the differentiation of 90% of the shoot initials into well-developed shoots. The 2- to 3-cm-long shoots rooted on half-strength MS basal medium supplemented with 4.90 μM indole-3-butyric acid (IBA) and 3% (w/v) sucrose in the third stage. The rooted plants could be established in soil with 70% success. This protocol could be utilized for in vitro propagation and conservation of this important threatened medicinal plant.  相似文献   

15.
The in vitro adventitious shoot differentiation in leaflet explants of an adult tree differed from that of leaflet explants of seedlings of Albizia procera(Roxb.)Benth. reported previously elsewhere. The leaflet explants from an adult tree passed through an initial callus phase for30 days on MS medium supplemented with 3 % sucrose,2.5 l M 2,4-D followed by a subsequent adventitious shoot differentiation phase for another 30 days on half MS medium supplemented with 0.25 l M each of BA and IBA.The regeneration rate of in vitro adventitious shoots in explants from the adult tree, i.e.1.66 shoots/callus, was lower than that from seedlings, i.e. [10 shoots/callus,which was reported elsewhere. Correspondingly, the activities of nitrate reductase and peroxidase, and endogenous phenol content remained very low during in vitro adventitious shoot differentiation in leaflet explants of an adult tree possibly due to lower availability of competent stem(juvenile) cells for the process.  相似文献   

16.
A method for rapid in vitro propagation of Cassia siamea Lam. using cotyledonary node explants, excised from 14-day old aseptic seedlings, has been established. Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kn) and thidiazuron (TDZ) singly or in combination with auxins was used for regeneration studies. Among the single treatment of three cytokinins BA at 1.0 μM was found to be optimum for direct shoot regeneration as it induced an average of 8.20 ± 0.66 shoots per explant. The regeneration frequency further enhanced with the application of auxin along with optimal BA concentration. The highest frequency for shoot regeneration (90%), the maximum number of shoots per explant (12.20 ± 0.73) and the maximum shoot length (6.40 ± 0.07) cm were obtained on the medium consisted of MS + 1.0 μM BA + 0.5 μM NAA. Successful in vitro rooting was induced from cut end of the microshoots when placed on half-strength MS + IBA (2.5 μM). The regenerated shoots with well developed root system were successfully acclimatized and established in pots containing sterilized garden soil and garden manure (1:1) and grown under greenhouse conditions with 85% survival rate.  相似文献   

17.
An efficient method for cultivation of Vitex negundo L. through axillary shoot (collected from micropropagated plants) proliferation has been successfully developed, which can be employed at a commercial scale. Axillary shoot induction was most successful using nodal explants for propagation on Murashige and Skoog (MS) medium supplemented with various concentrations of single cytokinin or in various combination with auxins. We obtained the maximum percentage (97.6 ± 1.45) response with highest number (16.4 ± 0.60) of shoots per explant on MS medium supplemented with 6-benzyladenine (BA) and ??-naphthalene acetic acid (NAA) at concentrations of 5.0 and 0.5 ??M, respectively. Shoot regeneration frequency was optimized by manipulating pH and using various media. MS medium and pH 5.8 was found to be the optimum for maximum regeneration. Nodal explants from in vitro regenerated microshoots too developed shoots, thus making the process recurrent. Shootlets with 4?C5 nodes were utilized for in vitro rooting, and best response was evaluated on MS medium supplemented with 1.0 ??M indole-3-butyric acid (IBA). The well-developed micropropagated plants were acclimatized (97%) successfully within 2 weeks in soilrite and planted ex vitro in normal garden soil, where they grew well without any morphological and genetic variations. The present regeneration process not only favoured the rapid multiplication but also expressed the regeneration capability of micropropagated plants.  相似文献   

18.
A method for efficient in vitro regeneration of Leucaena leucocephala cv. K636 was developed using immature zygotic embryos as the explant. Multiple shoot induction was achieved by culturing the explants on half-strength Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ) (0.03 to 1.5 μM). The maximum number of shoots per explants was achieved in 0.26 μM TDZ-supplemented media. TDZ concentration significantly influenced the induction of multiple shoots as well as the shoot-length. TDZ at 0.35 μM or higher concentrations resulted in abnormal stunted shoots. Full-strength MS media devoid of any plant growth regulator were used for shoot elongation. In vitro root induction was achieved in half-strength MS media supplemented either with 0.54 μM 1-naphthaleneacetic acid (NAA) or with 14.76 μM indole-3-butyric acid (IBA) in combination with 0.23 μM kinetin (Kn). Media supplemented with 14.76 μM IBA in combination with 0.23 μM Kn induced twofold–threefold more roots than media supplemented with 0.54 μM NAA. However, the average root length was lower in 14.76 μM IBA in combination with 0.23 μM Kn than in 0.54 μM NAA. Regenerated plants were maintained under normal condition after hardening. Plants, which were rooted on media supplemented with 14.76 μM IBA in combination with 0.23 μM Kn showed higher survival rate during hardening than those rooted on 0.54 μM NAA supplemented media.  相似文献   

19.
Abstract

Multiple shoots and plantlets were developed in vitro from cotyledonary nodal segments of in vitro raised seedlings of Anogeissus rotundifolia (syn. A. sericea var. nummularia)-a rare and endemic tree species of the Thar Desert. About 15-20 shoots differentiated from a single cotyledonary node within four weeks on Mu-rashige and Skoog's (MS) basal medium containing 0.1 mg l?1 indole-3-acetic acid (IAA) + 2.0 mg l?1 6-benzylaminopurine (BAP) + additives (25 mg l?1 each of adenine sulphate, L-arginine, and citric acid, and 50 mg l?1 of ascorbic acid) at 26 ± 2°C temperature and 36 μmol m?2 s?1 photon flux density with a 12 h/day photoperiod. The shoots produced in vitro were further multiplied by subculturing on fresh medium. The original cotyledonary nodal segment was repeatedly transferred (5 to 6 times) onto fresh medium containing 1.0 mg l ?1 BAP + 0.1 mg I ?1 IAA + additives to yield fresh crops of multiple shoots. These shoots were rooted on half-strength MS medium supplemented with 0.1 mg l?1 indole-3-butyric acid (IBA). Plantlets were transferred to pots containing sand-dune soil and ver-miculite it the ratio of 4:1 (v/v) and hardened in a growth chamber for two weeks and finally transferred to a greenhouse. From a single cotyledonary node about 1500 plantlets could be developed within four months. The method developed is useful for mass multiplication and for the conservation of germplasm of Anogeissus rotundifolia.  相似文献   

20.
Abstract

Micropropagation protocols for Dendrocalamus asper using nodal shoots and seeds culture are described. Multiple shoots were induced through forced axillary branching. Ninety-five percent of the nodal shoot explants taken from juvenile primary and lateral branches, produced multiple shoots through axillary buds activation within 2 to 8 weeks on Murashige & Skoog's (MS) medium supplemented with 0.1-15 mg/l benzyladenine (BA). The cultured seeds also produced multiple shoots (1-20) within 6 weeks on this medium. The multiple shoot differentiation was influenced by the concentration of BA in the medium. The in vitro generated shoots were excised and subculture on MS + 3.0 mg/l BAP for further shoot multiplication. Fifteen to 20 fold rate of shoot multiplication was achieved by regular subculturing. These shoots were multiplied for more than 3 years without loss of vigor. Ninety-five percent of the shoots were rooted, when propagules (each consisting of cluster of 3 shoots) were transferred on to MS medium with 3.0 mg/l NAA or 10 mg/l indole-3-butyric acid (IBA).

To date, 18,000 plants (through axillary bud initiated from nodal ex-plants) and 6,000 plants from seed culture have been hardened and acclimatized. 12,000 plants have been field transferred.  相似文献   

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