Secondary embryogenesis from rapeseed microspore-derived embryos (MDEs) was studied in three Brassica napus L. cultivars Global, PF₇₀₄ and Option. The best results in terms of secondary embryogenesis percentage obtained in cultures of Global and PF₇₀₄ MDEs (75.88 and 65.97 %, respectively) and PF₇₀₄ produced the highest number of secondary embryos per each primary embryo (14.91 ± 2.18). After optimization of physical parameters, rapeseed hypocotyls of MDEs were bombarded with microcarriers coated with a plasmid containing GUS reporter gene. The highest levels of transient GUS expression were obtained using bombardment with gold particles of 1.6 µm, at helium pressure of 9.3 MPa, a bombardment distance of 9 cm, chamber vacuum pressure of 7.1 × 10^-6 kPa and single bombardment in bombardment medium containing 0.4 M mannitol.

The effects of exogenous salicylic acid (SA) on photosystem 2 (PS 2) in barley (Hordeum vulgare L.) seedlings were investigated. SA pretreatment provided protection against subsequent osmotic stress. The highest protective effect of 0.25 mM SA was confirmed by determination of chlorophyll fluorescence, electrolyte leakage, malonyldialdehyde contents, PS 2 mRNAs and proteins. SA pretreatment increased reactive oxygen species (ROS), decreased net photosynthetic rate and stomatal conductance immediately, but prevented ROS accumulation during subsequent osmotic stress by activating antioxidant enzymes. Elimination of H₂O₂ during SA pretreatment inhibited almost all above mentioned SA effects. Therefore, SA pretreatment enhanced osmotic stress tolerance in barley seedlings mainly through ROS signals, rather than SA itself. The only SA-dependent and ROS-independent effect of exogenous SA on PS 2 was reduction of non-photochemical quenching.

Long-term culture establishment and efficient in vitro regeneration protocol for Sansevieria cylindrica Bojer ex Hook was developed using leaf derived callus and nodule culture. Profuse callus induction on leaf discs was achieved on Murashige and Skoog (MS) medium supplemented with 10 µM indole-3-butyric acid (IBA), while a high frequency of nodulation was induced on 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) containing media. Shoot regeneration ability from cultured tissues occurred at varying degrees on all media. Through callus culture a maximum of 17.6 ± 0.14 shoots per culture was formed on medium containing 5µM 6-benzyladenine (BA) and 2 µM α-naphthaleneacetic acid (NAA). Among nodule cultures, the 2,4-D generated nodules were more proliferative and regenerative as compared to 2,4,5-T induced nodules and a maximum of 25 ± 0.16 shoots per culture was produced on a medium containing 5 µM BA plus 1 µM NAA. The regenerated shoots were successfully rooted on a semi-solid half strength MS medium containing 5 µM IBA with an average root number 3.5 ± 0.18 and root length 6.5 ± 0.14 cm. The regenerative ability of callus tissues was steady upto one year, while the nodules retained the totipotency to regenerate on optimal medium even after 3 years of subculturing. The histological sections of nodules confirm the typical anatomy exhibiting the vascular elements in bundles with well demarcated cortex and epidermal covering.

The effect of various hormonal combinations on callus formation and regeneration of shoot and root from leaf derived callus of Acanthophyllum sordidum Bunge ex Boiss. has been studied. Proteins and activity of antioxidant enzymes were also evaluated during shoot and root organogenesis from callus. Calli were induced from leaf explants excised from 30-d-old seedlings grown on Murashige and Skoog medium containing 4.52 µM 2,4-dichlorophenoxyacetic acid + 4.65 µM kinetin. Maximum growth of calli and the most efficient regeneration of shoots and roots occurred with 2.69 µM 1-naphthalene acetic acid (NAA), 2.69 µM NAA + 4.54 µM thidiazuron and 2.46 µM indole-3-butyric acid. Protein content decreased in calli and increased significantly during regeneration of shoots from callus. Superoxide dismutase activity decreased in calli comparing to that of seedlings, then increased in regenerated shoots and roots. High catalase activity was detected in seedlings and regenerated shoots, whereas high peroxidase activity was observed in calli and regenerated roots.

In pot experiments performed on maize seedlings chilled at 5 °C, leaf injury was diminished by the application of elevated temperature (1 or 5 h at 15 or 20°C, "warm breaks" treatment) in a dose-dependent manner. The lower the injury count, the higher the catalase (CAT) activity. In a separate experiment, the application of 100 % relative humidity also protected the plants from chilling injury and water loss, increased their gas exchange and variable to maximum chlorophyll fluorescence ratio (F_v/F_m), but did not influence CAT activity. Another protective environmental factor, elevated atmospheric CO₂ concentration [700 µmol(CO₂) mol^-1(air)] diminished CAT activity inhibition, but only in plants of chilling-resistant cultivar. The positive impact of specific environmental factors accompanying chilling is not obviously related to the suppression of the inhibition of CAT activity, although the enzyme is considered as chilling-sensitive.

A novel method of organogenesis in neem (Azadirachta indica A. Juss.) from unfertilized ovaries is described. The Murashige and Skoog's (MS) medium with 9 % sucrose, 1 µM 2,4-dichlorophenoxyacetic acid (2,4-D) and 5 µM 6-benzylaminopurine (BAP) was the best for callus induction from unfertilized ovaries. However, further proliferation of callus occurred better on MS medium supplemented with 0.5 µM 2,4-D either alone or in combination with 4.5 µM kinetin. Maximum shoot regeneration (78 %) was observed when calli, induced from ovaries of 4 mm size flower buds and proliferating on MS + 0.5 µM 2,4-D, were subcultured to MS medium containing 5 µM BAP. Histological analysis revealed that 4 mm sized flower bud corresponds to a 2-nucleate stage of embryo sac. The shoots were then multiplied by forced axillary branching on MS medium supplemented with 1.0 µM BAP and 250 mg dm^-3 casein hydrolysate. The shoots could be rooted on 1/4 strength MS medium supplemented with 0.5 µM indole-3-butyric acid (IBA) at a frequency of 79 %. Cytological analysis by root tip squash preparations revealed that all the plantlets were diploids. These plants were subsequently hardened and established in soil with transplantation rate of 81.8 %.

Expansins, found in the cell wall, have the unique ability to induce immediate cell wall extension. In this study, a β-expansin gene (TaEXPB23) isolated from wheat (Triticum aestivum L.) coleoptiles was transformed to tobacco (Nicotiana tabacum) to investigate its role in plant growth and development. TaEXPB23 was preferentially expressed in wheat coleoptile and a close correlation between TaEXPB23 expression and coleoptile growth was observed. The over-expression of TaEXPB23 in tobacco also resulted in accelerating growth of leaves and internodes at earlier developmental stages, and it was involved in regulating plant development.

Two rice (Oryza sativa L.) cultivars differing in chilling sensitivity, Changbaijiu (chilling-tolerant) and Zhongjian (chilling-sensitive) were pre-treated with 0.5, 1.0 and 2.0 mM salicylic acid (SA) for 24 h before chilling at 5°C for 1 d. Chilling induced SA accumulation, particularly conjugated SA in both leaves and roots of the two rice cultivars. After SA administration, SA accumulated in the roots of both cultivars at a concentration-dependent manner, whereas only a slight increase was observed in their leaves. Conjugated SA accounted for most of the increase. The beneficial effect of SA treatment on protecting rice seedlings from chilling injury was not observed at any concentration in either cultivar. Pre-treatment with SA even decreased their chilling tolerance confirmed by increased electrolyte leakage and lipid peroxidation. Further, most of the activities of antioxidant enzymes decreased or remained unchanged in leaves and roots of SA pre-treated seedlings after chilling. These results implied that down-regulation of antioxidant defence might be involved in the reduction of chilling tolerance in SA-pre-treated plants.

A cDNA-encoding γ-tocopherol methyltransferase (γ-TMT) from Arabidopsis thaliana was overexpressed in deoduck (Codonopsis lanceolata L.) to improve the tocopherol composition. Deoduck (T₂) containing the γ-TMT transgene was produced by Agrobacterium-mediated transformation. Transgene expression was confirmed by polymerase chain reaction and RNA gel blot analysis. The transgenic plants produced more leaves than control plants. In addition, the transgenic plants showed higher levels of the CSOD, CTRX, CAPX, CNADP ⁺-IDCH, and CSO transcripts and higher SOD-like activity compared with the control plants.

Black gram [Vigna mungo (L.) Hepper] cv. IPU 94 plants grown in sand culture with deficient zinc (0.1 µM Zn) nutrition and those deprived of normal (1 µM) Zn supply at the initiation of flowering, showed decrease in dry matter production and especially seed yield. These plants showed a decrease in the size of anthers and stigmatic heads, pollen producing capacity of the anthers and stigmatic exudations. Zn deficiency caused structural alterations in exine and retarded germination of pollen grains and tube growth. The pollen extracts and stigmatic exudates of the Zn-deficient plants showed increase in activity of acid phosphatase isoforms and inhibition of esterase isoforms. Zn deficiency led to decrease in number of pods, seeds per pod and seed mass, altered seed coat topography and reduced seeds germinability. Low seed yield under Zn deficiency is attributed to a role of Zn in pollen function, as also in pollen-pistil interaction conducive to fertilization and development of seeds.

The influence of cytokinin thidiazuron (TDZ) and auxin indole-3-acetic acid (IAA) on in vitro shoot organogenesis of fifteen Rhododendron genotypes was investigated and a protocol for high frequency adventitious shoot regeneration from leaf explants was developed. High genotypic variation was observed and regeneration frequencies ranged from 0 to 100 %. Genotype Ovation had the highest number of shoots (26.4 per explant) after 12 weeks on medium with 0.57 µM IAA and 1.20 µM TDZ, but only 65 % of explants regenerated. Catawbiense Grandiflorum had 17.7 shoots per explant and 75 % regeneration on medium with 5.70 µM IAA and 0.45 µM TDZ and Van Werden Poelman had 14.3 shoots per explant and 100 % regeneration on medium with 0 57 µM IAA and 0.45 µM TDZ.

Effects of high night temperature on the lipid and protein compositions in the tonoplasts isolated from the leaves of two Crassulacean acid metabolism (CAM) plants, Ananas comosus (pineapple) and Kalanchoë pinnata were studied. The results showed that the phospholipids/protein ratios in the tonoplasts isolated from pineapple and K. pinnata leaves decreased from 1.82 to 1.21 and 2.63 to 1.50, respectively, as the night temperature increased from 20 to 37 °C. Under high night temperature, relative amount of total unsaturated fatty acids in K. pinnata was increased by 6 %, which was mainly caused by increased C18:2 and C18:3, whereas unsaturated fatty acids, C18:2 and C18:3 in pineapple did not show significant change. The distribution patterns of tonoplast proteins in the two CAM species were different between normal and high night temperature and in K. pinnata, especially those with molecular mass ranging from 66.2 to 97.4 KDa. Compared with normal night temperature, more proteins were found in pineapple, but no difference was found in K. pinnata. Thus, above result indicated that the pineapple tonoplasts could keep higher rigidity under high night temperatures compared to the K. pinnata.

The effects of amino acids (arginine, aspargine, cysteine, glutamine, glycine and proline) and polyamines (putrescine and spermidine) on embryogenesis and plant regeneration from cultured anthers of Guizotia abyssinica (L. f.) Cass. cv. Ootacamund was studied. Supplementation of amino acids (0.5-5.0 mM) to the induction medium individually and in combination, improved embryo yield. B5 medium supplemented with 2 mM proline, 10 µM 2,4-dichlorophenoxyacetic acid, 2 µM kinetin and 0.2 M sucrose induced highest number of embryos (63 per 60 anthers cultured). Addition of polyamines (5-200 µM) to the same medium also enhanced the rate of embryogenesis.

An experiment was taken up to find out possibilities of manipulating the in vitro flowering in moth bean. Abscisic acid (ABA) and proline both alone and in combination influenced days to flower induction, number of flowers per plant, number of pods per plant and seeds per pod. Frequency of flowering plants approached 100 % at 1 and 3 µM ABA and 800 µM proline. The range of flowering period (3 to 23.6 d) has also been influenced by various treatments.

A quantitative trait loci (QTL) approach was applied to dissect the genetic control of the common wheat seedling response to osmotic stress. A set of 114 recombinant inbred lines was subjected to osmotic stress from the onset of germination to the 8^th day of seedling development, induced by the presence of 12 % polyethylene glycol. Root, coleoptile and shoot length, and root/shoot length ratio were compared under stress and control conditions. In all, 35 QTL mapping to ten chromosomes, were identified. Sixteen QTL were detected in controls, 17 under stressed conditions, and two tolerance index QTL were determined. The majority of the QTL were not stress-specific. In regions on five chromosome arms (1AS, 1BL, 2DS, 5BL and 6BL) the QTL identified under stress co-mapped with QTL affecting the same trait in controls, and these were classified as seedling vigour QTL, in addition to those expressed in controls. Tolerance-related QTL were detected on four chromosome arms. A broad region on chromosome 1AL, including five QTL, with a major impact of the gene Glu-A1 (LOD 3.93) and marker locus Xksuh9d (LOD 2.91), positively affected root length under stress and tolerance index for root length, respectively. A major QTL (LOD 3.60), associated with marker locus Xcdo456a (distal part of chromosome arm 2BS) determined a tolerance index for shoot length. Three minor QTL (LOD < 3.0) for root length and root/shoot length ratio under osmotic stress were identified in the distal parts of chromosome arms 6DL (marker locus Xksud27a) and 7DL (marker locus Xksue3b). Selecting for the favourable alleles at marker loci associated with the detected QTL for growth traits may represent an efficient approach to enhance the plants' ability to maintain the growth of roots, coleoptile and shoots in drought-prone soils at the critical early developmental stages.

6-Methoxy-2-benzoxazolinone (MBOA) inhibited germination of rice (Oryza sativa L.), wheat (Triticum aethiopicum Jakubz), rye (Secale cereale L.), onion (Allium cepa L.), wild oat (Avena fatua L.), barnyard grass [Echinochloa crus-galli (L.) Beauv.], ryegrass (Lolium rigidum Gaudin), cress (Lepidium sativum L.), lettuce (Lactuca sativa L.), tomato (Lycopersicum esculentum Mill.), carrot (Daucus carota L.) and amaranth (Amaranthus retroflexus L) and the inhibition increased with increasing MBOA concentrations. MBOA also inhibited the induction of α-amylase in these plant seeds and the inhibition increased with increasing MBOA concentrations. There were variations in sensitivity of these plant species to MBOA, and species of family Poaceae (barnyard grass, wild oat, rice, rye, ryegrass, and wheat) were less sensitive to MBOA than the other plant species.

A bacterial artificial chromosome (BAC) library of banana (Musa acuminata) was used to select BAC clones that carry low amounts of repetitive DNA sequences and could be suitable as probes for fluorescence in situ hybridization (FISH) on mitotic metaphase chromosomes. Out of eighty randomly selected BAC clones, only one clone gave a single-locus signal on chromosomes of M. acuminata cv. Calcutta 4. The clone localized on a chromosome pair that carries a cluster of 5S rRNA genes. The remaining BAC clones gave dispersed FISH signals throughout the genome and/or failed to produce any signal. In order to avoid the excessive hybridization of repetitive DNA sequences, we subcloned nineteen BAC clones and selected their 'low-copy' subclones. Out of them, one subclone gave specific signal in secondary constriction on one chromosome pair; three subclones were localized into centromeric and peri-centromeric regions of all chromosomes. Other subclones were either localized throughout the banana genome or their use did not result in visible FISH signals. The nucleotide sequence analysis revealed that subclones, which localized on different regions of all chromosomes, contained short fragments of various repetitive DNA sequences. The chromosome-specific BAC clone identified in this work increases the number of useful cytogenetic markers for Musa.

An efficient micropropagation protocol was developed for elite male and female genotypes of Simmondsia chinensis using nodal segments. Bud initiation was found to be best on Murashige and Skoog's (MS) medium supplemented with 4.44 µM 6-benzylaminopurine (BAP) and 88.8 µM adenine. Upon sub-culture, 10-15 shoots per explant were obtained when 4.44 µM BAP and 74.0 µM adenine were incorporated in the medium. Increase in KNO₃ concentration in the medium improved shoot multiplication rate and in vitro flowering in 20 % of male cultures. Elongated shoots were harvested, pulse treated for 48 h on liquid medium supplemented with 49.0 µM indole-3-butyric acid, 5.40 µM α-naphthaleneacetic acid and 5.71 µM indole-3-acetic acid for root induction and rooting (92 %) was achieved on hormonal free half-strength MS medium supplemented with 1.37 µM chlorogenic acid, 1 % activated charcoal and 2 % sucrose. After successful hardening, plantlets were transferred to greenhouse with 99 % establishment.

A highly reproducible system for efficient plant regeneration from protoplast via somatic embryogenesis was developed in cotton (Gossypium hirsutum L.) cultivar ZDM-3. Embryogenic callus, somatic embryos and suspension culture cells were used as explants. Callus-forming frequency (82.86 %) was obtained in protoplast cultures from suspension culture cells in KM₈P medium with 0.45 µM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.93 µM kinetin (KIN), 1.5 % glucose and 1.5 % maltose. Protocolonies formed in two months with plating efficiency of 14 %. However, the callus-forming efficiencies from other two explants were low. The calli from protoplast culture were transferred to somatic embryo induction medium and 12.7 % of normal plantlets were obtained on medium contained 3 % maltose or 1 % of each sucrose + maltose + glucose, 2.46 µM indole-3-butyric acid (IBA) and 0.93 µM KIN. Over 100 plantlets were obtained from protoplasts derived from three explants. The regenerated plants were transferred to the soil and the highest survival rate (95 %) was observed in transplanting via a new method.

A rapid and efficient method for the regeneration of Eucommia ulmoides Oliver has been developed. The ability of hypocotyl segments to produce adventitious buds varied depending upon their position, seedling age and culture medium. The most adventitious buds were induced from the hypocotyl segments near to the root of 2-week-old seedlings on the Murashige and Skoog (MS) basal medium supplemented with 10 µM benzylaminopurine (BAP). However, the MS medium with 6 µM BAP and 6 µM gibberellic acid (GA₃) was most suitable for shoot multiplication and further growth. Shoots longer than 2 cm cultured on the half-strength MS medium supplemented with 0.5 µM naphthaleneacetic acid (NAA) produced a maximum number of roots per shoot. Regenerated plantlets could be successfully acclimatized.

It was observed that the unpollinated flowers of Cymbidium pendulum (Roxb.) Sw. and C. aloifolium (L.) Sw. stayed fresh for 20 and 18 d, respectively, but attained senescence in 8 and 7 d, respectively, after pollination. The higher content of total soluble sugars, reducing sugars and free amino acids was observed in all the floral organs of pollinated flowers than in unpollinated ones. Pollination also up-regulated the activity of hydrolytic (α-amylase, β-amylase, invertase) and proteolytic enzymes (proteases) in floral organs. Amongst floral organs, the lip and perianth possessed highest contents of metabolites. Application of auxin inhibitor (0.25 µM triiodobenzoic acid) and ethylene inhibitor (0.25 µM AgNO₃) to the pollinated flowers partially prevented the process of senescence.

Carnation plantlets (Dianthus caryophyllus L.) cultured in vitro often develop morphological and physiological anomalies, a phenomenon called hyperhydricity, which impairs their survival ex vitro. When the agar concentration of the growth medium was increased (from 0 to 12 g dm^-3), thereby reducing water availability, the hyperhydricity of those adventitious shoots regenerated from carnation petals decreased. This was accompanied by a progressive fall in the water content of shoots (94.9 to 91.4 %), fresh mass (from 57.2 to 1.8 mg), number of leaf parenchyma cell layers (from 9.3 to 7.7), and the size of these cells (from 968 to 254 µm²). However, the number of regenerated shoots also decreased (17.7 in 2 g dm^-3 agar to 4.3 in 12 g dm^-3). Similarly, in ventilated tubes, which exhibit a lower relative humidity than tightly closed tubes, shoot organogenesis diminished up to 28 %, in tandem with shoot water content. Thus, relative humidity and water availability in culture vessels do not only influence shoot hyperhydricity in carnations, but also greatly affect adventitious shoot organogenesis.

Inheritance of 10 morphological and quantitative traits related to plant and fruit development and resistance to the pathogen Phytophthora capsici was studied in an intraspecific cross between a non-pungent, susceptible Capsicum annuum parent (cv. Americano) and a wild, pungent and resistant line (Serrano Criollo de Morelos-334). Data were obtained from the segregation of 166 F₂ plants and 50 F₃ plants in four years. Three of the traits analyzed (necrosis length, leaf width and leaf length) exhibited a transgressive segregation. A multiple linear regression analysis was applied in order to establish a relationship between necrosis length and some of the morphological traits measured such as length and width of leaf, length, diameter and mass of fruit, capsaicin content in fruits, and presence of hair on leaves and stems. The results identified a linear dependence between necrosis length (as an inverse measurement of resistance) and leaf width, fruit diameter and hair presence in the stem. Pungency was not related with resistance.

To dissect differences in both lipid accumulation and composition and the role of these modifications during drought stress, four grapevine cultivars exhibiting differential tolerance to drought were subjected to water shortage. Tolerant cultivars, Kahli Kerkennah and Cardinal, exhibited higher leaf water potential (Ψ_w), and lower lipid peroxidation compared to the sensitive cultivars Guelb Sardouk and Superior Seedless during stress. Total lipid amounts increased during stress only in the leaves of the tolerant cultivars. Drought induced increases in the ratios digalactosyldiacylglycerol/monogalactosyldiacylglycerol and phosphatidylcholine/phoshatidylethanolamine of almost all the drought stressed cultivars. Moreover, the overall analysis of the composition of fatty acids revealed that a linolenic acid was prevalent in grapevine and the unsaturation level of lipids increased under water stress in all the cultivars. Specific adjustments in the lipid composition during stress could compromise stress tolerance.

Phylogenetic analysis was conducted based on sequences of the internal transcribed spacer region (ITS) of nuclear ribosomal DNA in 17 species of Kengyilia, together with those of 18 species from Pseudoroegneria, Agropyron, Roegneria and Douglasdeweya by the maximum parsimony, maximum likelihood and neighbor-joining distance methods. The results indicate that species of Kengyilia had close affinities to species of Douglasdeweya and Agropyron. The species in Kengyilia was identified as two subgroups with regard to geographic distribution, indicating that species from the same distribution had a closer phylogenetic relationship. The genus Kengyilia was found as a ligament-group between Roegneria and Agropyron. The ITS sequence is a useful tool for studying the phylogeny of closely related species.

Different lines of cell suspension cultures of Taxus × media Rehd. and Taxus floridana Nutt. were cryopreserved with a two-step freezing method using a simple and inexpensive freezing container instead of a programmable freezer. Four to seven days old suspension cell cultures were precultured in growth medium supplemented with 0.5 M mannitol for 2 d. The medium was then replaced with cryoprotectant solution (1 M sucrose, 0.5 M glycerol and 0.5 M dimethylsulfoxide) and the cells incubated on ice for 1 h. Before being plunged into liquid nitrogen, cells were frozen with a cooling rate of approximately -1 °C per min to -80 °C. The highest post-thaw cell viability was 90 %. The recovery was line dependent. The cryopreservation procedure did not alter the nuclear DNA content of the cell lines. The results indicate that cryopreservation of Taxus cell suspension cultures using inexpensive freezing container is possible.

Waterlogging is a serious problem, which affects crop growth and yield in low lying rainfed areas. The main cause of damage under waterlogging is oxygen deprivation, which affect nutrient and water uptake, so the plants show wilting even when surrounded by excess of water. Lack of oxygen shift the energy metabolism from aerobic mode to anaerobic mode. Plants adapted to waterlogged conditions, have mechanisms to cope with this stress such as aerenchyma formation, increased availability of soluble sugars, greater activity of glycolytic pathway and fermentation enzymes and involvement of antioxidant defence mechanism to cope with the post hypoxia/anoxia oxidative stress. Gaseous plant hormone ethylene plays an important role in modifying plant response to oxygen deficiency. It has been reported to induce genes of enzymes associated with aerenchyma formation, glycolysis and fermentation pathway. Besides, nonsymbiotic-haemoglobins and nitric oxide have also been suggested as an alternative to fermentation for maintaining lower redox potential (low NADH/NAD ratio), and thereby playing an important role in anaerobic stress tolerance and signaling.

An efficient micropropagation protocol was established for Capsicum chinense Jacq. cv. Umorok, a pungent chilli cultivar. Shoot-tip explants were cultured on Murashige and Skoog (MS) medium containing cytokinins (22.2-88.8 µM 6-benzylaminopurine, BAP, 23.2-93.0 µM kinetin, Kin, or 22.8-91.2 µM zeatin, Z) alone or in combination with 5.7 µM indole-3-acetic acid (IAA). Maximum number of shoots were induced on medium containing 91.2 µM Z or 31.1 µM BAP with 4.7 µM Kin. The separated shoots rooted and elongated on medium containing 2.5 or 4.9 µM indole-3-butyric acid (IBA). Axillary shoots were induced from in vitro raised plantlets by decapitating them. The axillary shoot-tip explants were used for further multiple shoot buds induction. A maximum of about 150 plantlets were obtained from a single seedling. Hardened and acclimatized plantlets were successfully established in the soil.