• Journal of Plant Biotechnology
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• v.7 no.4
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• pp.233-240
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• 2005

Slices of embryonic axis of mature pea (Pisum sativum L. cv. Green Arrow) seeds were used as explant. Transformation of explants was done via Agrobacterium tumefaciens bearing vector pBI-P5CS construct. The best results for inoculation of explants were obtained when they were immersed for 90 s at a concentration of $6{\times}10^8$ cell $ml^(-1)$ of bacterial suspension. Transformed pea plants were selected on $50\;mg\;l^(-1)$ kanamycin and successful transformants were confirmed by PCR and blotting. Transgenic plants were further analyzed with RT-PCR to confirm the expression of P5CS. Transgenic plants and non-transgenic plants were treated with different concentrations of NaCl 0 (control), 100, 150 and 200 mM in culture medium. Measurement of proline content indicated that transgenic plants produced more amino acid proline in response to salt in comparison with non-transgenic plants. Photosynthetic efficiency in transgenic plants under salt-stress was more than that of non-transgenic plants.

• Journal of Plant Biotechnology
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• v.7 no.4
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• pp.211-218
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• 2005

Genetic transformation was carried out by using biolistic gun method. The hypocotyl derived embryogenic calli (explants) of cotton (Gossypium hirsutum L.) cv. Cocker-312 were transformed with a recombinant pGreen II plasmid, in which both, bar (selection marker) and GUS (${\beta}$-glucuronidase) reporter genes were incorporated. Explants were arranged on osmoticum-containing medium (0.5M mannitol) 4 hours prior to and 16 hours after bombardment that was resulted into an increase about >80% for GUS stable expression. 3 days after bombardment, GUS assay was performed, which exhibited, $18.36{\pm}1.00$ calli showed blue spots. The transformed embryogenic calli were cultured on selection medium (@ 6 mg/L basta) for 3 months. The putative transgenic plants were developed via selective somatic embryogenesis (@1.50 mg/L basta); maximum $27.58{\pm}1.25$ somatic embryos were obtained while $17.47{\pm}1.00$ embryos developed into plantlets (@ 0.75mg/L basta). In five independent experiments, up to 7.24% transformation efficiency was recorded. The presence of the transgenes was analyzed by using PCR and southern hybridization analysis. The transgenic plants were developed with in 6-7 months, but mostly transformants were abnormal in morphology.

• Journal of Korean Society of Forest Science
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• v.86 no.2
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• pp.128-134
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• 1997

An effective in vitro multiplication method was developed for clones of Populus davidiana Dode. Ten different media were tested for their effect on shoot multiplication. Both MS and LP medium with 0.2mg/l BAP appeared to be the best for the shoot multiplication with the rate of 9 shoots per explant. There were significant differences among the clones in both multiplication rate and shoot growth. While some clones did not require BAP to promote shoot formation, others did. More than 60% of in vitro shoots rooted on the half-strength GD medium containing 0.2mg/l IBA.

• Journal of Forest and Environmental Science
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• v.25 no.2
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• pp.119-126
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• 2009

An efficient in vitro plant regeneration system was developed through shoot proliferation from axillary buds of Tectona grandis (L.f), APNBV-1 (Andhra Pradesh North Badrachalam Venkatapuram-1) clone. Multiple shoots of high quality were produced in vitro from axillary bud explants. An average of 4.39 shoots/explant were obtained on Murashige and Skoog's (MS) medium supplemented with plant growth regulators (PGRs) benzyl amino purine (BA), kinetin (KN), indole acetic acid (IAA), gibberillic acid ($GA_3$), growth adjuvants casein hydrolysate (CH), adenine sulphate (Ads) and antioxidants ascorbic acid, polyvinyl pyrrollidine (PVP). Eighty five percent of rooting was observed in ex vitro rooting media containing IBA and vermiculite. In ex vitro rooting, single shoots with 2 to 3 nodes were subjected to IBA of different concentrations at different periods of time intervals. Direct rooting in vermiculite at 500 ppm concentration of IBA resulted in 4.3 number of roots with 2 cm length. Minimum response of rooting and length of roots were recorded at 100 ppm concentration of IBA. Planlets were transferred to plastic bags for short acclimatization stage in green house where they survived at 95%.

• Journal of Forest and Environmental Science
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• v.24 no.2
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• pp.79-88
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• 2008

Twenty-eight of 44 Indian Ceropegia species are endemic and their survival is threatened. As a step towards conservation, we implied in vitro methods for the sustainable propagule production in C. hirsuta, C. lawii, C. maccannii, C. oculata and C. sahyadrica. Effects of explant, growth regulators, sucrose and photoperiod were studied. High frequency microtuber production was achieved with the seedling-apical buds, grown on MS medium containing 4-6 mg $1^{-1}$ BAP, 3-8% (w/v) sucrose, under continuous illumination. Each microtuber, when subcultured proliferated to form a cluster of secondary microtubers. Every primary and secondary microtuber bore at least one shoot-bud and a root primordium. Each tuber (formed with any of the significantly effective treatments) weighed more than 500 mg, enough to plant directly in non-sterilized soils. Microtubers could be produced and proliferated round the year. Proliferation could be solely attributed to in vitro procedures as these plants bear solitary tubers in vivo. Microtubers could be sprouted in vitro to prepare ready to pot plantlets. As, this novel method succeeded for all five species, though they belong to different eco-physiological backgrounds, we recommend its implementation in the conservation programs for a broader range of Ceropegia species, supported by other integrated strategies.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.198-204
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• 2005

Seeds of neem were collected from different parts of India and analyzed for their azadirachtin content by High Performance Liquid Chromatography (HPLC). In order to assess the effects of genotypic and geographical variation on azadirachtin content in cell cultures, callus development was attempted in the seeds containing high and low concentration of azadirachtin. The concentration of azadirachtin in callus cultures was significantly affected by the explant source. Seed kernels with higher azadirachtin content produced higher azadirachtin content in callus cultures and lower azadirachtin content was seen in callus cultures produced from seed kernels with low azadirachtin content. The protocol for development of elite stock culture of Azadirachta indica was established with the objective of selecting a high azadirachtin-producing cell line. The highest azadirachtin-producing cell line was selected and the effects of different media and illumination conditions on growth and azadirachtin production were studied in shake flask suspension culture. Detailed batch growth kinetics was also established. These studies provided elite starter culture and associated protocols for cultivation of A. indica plant cell culture in the bioreactor.

• Journal of Crop Science and Biotechnology
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• v.11 no.2
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• pp.107-110
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• 2008

A high-efficiency, reproducible somatic embryogenesis system for strawberry cultivar Chandler was developed. Thirty-one somatic embryos per explant(max no.) were recorded in leaf discs which were cultured on medium containing MS salts+$B_5$ vitamins+2% glucose+4.0 mg $1^{-1}$TDZ(Thidiazuron) and incubated at $10{\pm}1^{\circ}C$ under darkness for one week followed by three weeks under 16-h photoperiod. The scanning electron microscopic(SEM) ontogeny revealed the normal development of somatic embryos from globular to heart-shaped and dissection microscopy from torpedo-shaped to cotyledonary-stage embryos. The maximum germination percentage of 48% could be obtained on MS medium containing kinetin(1.0 mg $1^{-1}$) and the maximum survival percentage(79%) of plantlets after four weeks was found to be in the mixture of vermiculite, peatmoss, and soilrite(1:1:1).

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.5
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• pp.356-360
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• 2005

Mature embryos were aseptically excised with a scalpel and sliced in fragments measuring 0.5 mm in diameter (sliced mature embryo fragment; 4 ${\~}$ 5 fragments/one embryo). Sliced mature embryo fragments of six wheat cultivars were cultured to develop an efficient method of callus induction and plant regeneration. Callus derived from sliced mature embryo fragments showed a good capacity to embryogenesis and regeneration. Furthermore sliced mature embryo fragments decreased contamination from fungi and bacteria. The high efficiency of callus induction were obtained Keumkangmil and Bob­white. For plant regeneration, selected embryogenic calli were transferred to two types regeneration media. An average number of green spots per callus was 4 to 5 in regeneration media after about one week. Percentage of plant regeneration showed high in regeneration medium containing 0.1 mg/l 2,4-D and 5 mg/l zeatin. Especially, Keumkangmil ($27.5\%$) and Bobwhite ($33.3\%$) showed high regeneration efficiency. This regeneration system from sliced mature embryo fragments may provide an effective and convenient explant for plant transformation studies.

• Korean Journal of Plant Tissue Culture
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• v.21 no.4
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• pp.227-231
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• 1994

The experiments were conducted to obtain culture conditions on optimal cultural conditions for propagation of a topical orchid, Haemaria discolor, through shoot tip cultrue in vitro. Survival percentage of shoot tip explant, in initial culture media was higher in H$_3$P$_4$ medium than in Murashige and Skoog's medium. Explants, cultured in H$_3$P$_4$ medium containing 1.0 mg/L kinetin, showed more shoot elongation when compared with those on other media. When the distal part of stem (pseudobulb) was longitudinally sectioned into half and cultured in H3P4 medium containing 0.1 mg/L 2ip, 0.1, or 1.0 mg/L kinetin, shoots from axillary bud were much more elongated under light condition.

• Korean Journal of Plant Tissue Culture
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• v.21 no.4
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• pp.221-225
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• 1994

This experiment was conducted to identify the optimal in vitro propagation condition of Cnidii rhizoma (Cnidium officinale Makino). It was effective to reduce contamination and improve regeneration of shoot when shoot tips taken in July were cultured in 1/2 strength Murashige and Skoog medium supplemented with 500 mg/L carbenicillin disodium 1.0 mg/L BA and 1.0mg/L $GA_3$followed by surface sterilization of explant source in solution of 1% sodium hypochlorite for 20 minutes. When shoot tips were 쳐cultured in 1/2 strength MS medium with 0.5 mg/L BA and 60 g/L sucrose, shoot elogation and subsequent multiplication of the formed shoot were favorable than in other media. Regenerants were well rooted in 1/2 strength MS medium containing 3.0 mg/L NAA.