Mgl-1 BA Research Articles

In vitro culture of nodal cuttings in kiwifruit

This study aimed to determine the effects of plant growth regulators and different medium on in vitro propagation of ˈHaywardˈ and ˈZespri Goldˈ kiwifruits. Micro-cuttings were used as explant sources and cultured in MS and WPM medium containing different concentrations of BA and GA3. Subcultures were carried out at intervals of 4-6 weeks. Contamination rate (%), blackening rate (%), callus development rate (%), shoot rate (%), and propagation coefficient (plantlet/explant) were determined. The micropropagation experiment was carried out according to the randomized plot design. The arc-sin transformation was applied to the percentage values before analysis. Significance levels of the means were compared with the LSD test. It was determined that 0.1% mercury chloride application increased the sterilization efficiency. As the BA concentration in the medium increased, the rate of callus formation of the explants increased. While there was no difference in shoot rates of explants cultured on MS and WPM medium, the propagation rate was higher in MS medium. The propagation coefficient of the ˈHaywardˈ kiwi cultivar was 3.08 in MS medium containing 1.0 mgL-1 BA + 0.5 mgL-1 GA3. MS medium can be recommended as a basic medium for in vitro kiwif propagation.

Enhanced production of lupeol through elicitation in in vitro shoot cultures of snake grass (Clinacanthus nutans)

Clinacanthus nutans (Acanthaceae), generally known as ‘snake grass’, has diverse uses in customary system of herbal medicine. The species is endowed with various bioactive compounds exhibiting extensive pharmacological properties. The present investigation focused on elicitor-intervened in vitro shoot biomass cultivation and scale-up production of the anti-cancerous compound ‘lupeol’, one of the foremost constituents in this species. For the augmented production of lupeol, the shoot cultures were elicited with various concentrations of yeast extract (YE), chitosan and methyl jasmonate (MeJA). Maximum shoot biomass yield and production of lupeol was detected in MS medium supplemented with 1.0 mgl-1 BA and 400 mgl-1 YE. The petroleum ether extracts of selected samples upon TLC analysis proved Rf values corresponding to lupeol. HPTLC analysis revealed that the sample treated with YE displayed relatively higher amount (975.50 ng) of lupeol than the in vivo plant (713.69 ng). Hence the in vitro shoot culture system with elicitor (YE) treatment propose an appropriate method for the elevated synthesis of lupeol which can be scaled up via bio-reactor technology in doing so profiting the pharmaceutical appliances.

Establishment of an efficient in vitro propagation system for Holostemma annulare (Roxb.) K. Schum. using SH medium

Holostemma annulare (Roxb.) K. Schum. (Asclepiadaceae) is an important medicinal herb and its root tubers are an important ingredient of the drug ‘jivanti’. Here, a more reliable procedure of in vitro propagation of the plant is discussed using SH nutrient medium formulation that facilitates the in vitro multiplication and establishment without callusing from the basal region of shoots that was prominent in the earlier protocols. Surface sterilization of the nodal explants with 10% (v/v) NaOCl for 12 minutes followed by 0.1% (w/v) HgCl2 for 8 minutes exhibited 90% shooting response. The in vitro initiated shoots subcultured to SH medium fortified with 2.0 mgl-1 BA in combination with 0.5 mgl-1 Kinetin and 1.0 mgl-1 IBA produced maximum 7.16±0.15 shoots. Better root induction of the in vitro raised shoots was noticed in SH medium augmented with 0.01 mgl-1 NAA that induced a mean number of 5.65±0.23 thin, long, white coloured roots. The plantlets were transferred to polythene bags containing river sand:garden soil (1:1) for acclimatization, further transplanted in garden soil and river sand mixture (3:1) in earthern pots in the greenhouse and subsequently to the field got established at 80% efficiency. They produced tuberous roots after 4 months of field establishment. Thus, the study standardized an improved system of in vitro propagation of H. annulare using SH medium and this protocol can be extended for long-term conservation methods such as cryoconservation and also for the in vitro production of bioactive metabolites.

Effects of shoot tip size on in vitro regeneration and virus elimination of grapevine cv. superior seedless

In this study, we investigate the relation between the explant size and the success of virus elimination in ‘Superior Seedless’ grape variety. Plants were tested by prior to thermotherapy and in vitro culture by using Double Antibody Sandwich-Enzyme-Linked Immunosorbent Assay (DAS-ELISA) and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and verified as infected by different viruses (Grapevine leafroll associated virus -1+3, Grapevine leafroll associated virus -4 Strains, Grapevine rupestris stem pitting associated virus). Different size of shoot tip explants (0.1-0.5-1-5-10 mm) were excised and cultured on MS medium supplemented with 1 mgl-1 BA. All of the explants bigger than 0.1 mm 100% regenerated while 70% of the 0.1 mm explants showed complete regeneration and turned into a whole plant. Plants regenerated from 10 mm shoot tip explants have all three viruses (GLRaV-1+3, GLRaV-4 strains and GRSPaV) as founded in mother plant. Plants regenerated from 5 mm explants have two of them (GLRaV-1+3, GLRaV-4 strains) and plants regenerated from 1 mm explants have one (GLRaV-1+3). Plants regenerated from 0.1 mm explants were found completely virus-free.

EVALUATION OF INHIBITION OF PROLIFERATION OF SK-MEL-28 CELL LINES BY ACMELLA CILIATA (KUNTH) CASS. CELL BIOMASS

Acmella ciliata (Kunth) Cass., a medicinally important plant in the family Asteraceae, contains many phytochemicals like alkaloids, tannins, saponins, flavonoids, including a highly mportant alkylamide ‘Spilanthol’ which accounts for most of its pharmacological applications. The plant has rich commercial value because of its traditional phytomedicinal uses. The present study focused on the anticancer potentiality of cell biomass of A. ciliata obtained through cell suspension culture. The purplish friable callus raised in MS medium containing 0.5 mgl-1 BA and 1.0 mgl-1 NAA were cultured in liquid MS medium with same hormone combination. The cells after 28 days of culture were collected and quantification of spilanthol using HPLC revealed that the harvested cells from cell suspension culture of A. ciliata recorded 173.702 µgg-1 spilanthol content in them. The analysis of antiproliferative effect of the methanolic extract of the cell biomass revealed that it is capable of inhibiting or reducing the growth of SK-MEL-28 cell lines and a lower concentration i.e., 24.144 µgml-1 of extract was needed to inhibit the growth by 50%. The study suggested the effectiveness of the cell biomass extract for the inhibition of uncontrolled growth of this melanoma cells and thus throw light on the possibility of formulating an anticancerous drug from this resource thereby benefiting the pharmaceutical industry.

Hypericum adenotrichum SPACH‘UN KALLUS İNDÜKSİYONU VE ADVENTİF SÜRGÜN REJENERASYONU

In this study, A standart protocol to callus formation and adventitious shoot regeneration of Hypericum adenotrichum Spach via direct or indirect organogenesis has been described.Callus induction was carried out by using leaves which were collected from their native environment. The maximum callus induction frequency has been observed on MS media containing 4 mgL-1 BA and 0.2 mgL-1 NAA. These calli were not induced to shoot regeneration. Shoot formation was obtained by transferring the calluses from BA-containing media to MS medium containing 0.5 mgL-1 KIN. Maximum shoot number was observedin transferred calluses from callus induction medium containing 3 mgL-1 BA to media containing 0.5 mgL-1 KIN.Direct shoot formation developed on leaf explants of H. adenotrichum on MS media containing KIN alone. The highest direct shoot development was observed on MS medium with 1 mgL-1 KIN.

Organogenesis of Tomato (Lycopersicon esculentum Mill.) cv. Sandra and Rocky by Hypocotyl, Cotyledon and Leaf Explants

In vitro regeneration protocols were established for two Tomato (Lycopersicon esculentum Mill.) cultivars (Sandra and Rocky) grown in Kurdistan region using hypocotyl, cotyledonous leaves and leaf explants. The explants excised from 12-15 days old of in vitro plantlets cultured on MS medium supplemented with different concentrations and combinations of plant growth regulators. High organogenesis percentage (100%) was achieved from hypocotyl grown in MS medium augmented with 1.0mgl-1 BA+ 0.5mgl-1 kinetin, which record approximately 3.75 in Sandra and 4.09 in Rocky cultivar as average number of shoots produced from each explant. While the highest organogenesis percentage (100%) was formed from cotyledon leaves using 1.5 mgl-1 BA+0.5mgl-1 kinetin that gave around 3.5 in Sandra cultivar as average number of shoots / explant. In contrast, the highest organogenesis was found on MS medium supplemented with 2.0mgl-1 BA+1.0 mgl-1 kinetin and the average number of shoots produced from each explant were approximately 3.17 in Rocky cultivar. The results of combination between 5.0mgl-1 BA with (0.0, 0.2, 0.5 and 1.0)mgl-1 NAA, indicated that the highest organogenesis percentage was recorded from hypocotyl (91.67 % with 6.5 shoots/explant and 66.67% with 4.0 shoots/ explant) in Sandra and Rocky cultivars respectively grown on MS medium supplemented with 5.0 mgl-1 BA alone. While the highest organogenesis percentage (100%) was induced from cotyledonous leaves in both cultivars (3.67 shoots for Sandra and 3.08 for Rocky) on MS medium supplemented with 5.0mgl-1 BA +0.2mgl-1 NAA. Meanwhile, the highest organogenesis percentage was obtained from leaf explants for both cultivars on the combination of 1.0mgl-1 BA +0.5mgl-1 kinetin, (77.78% and 66.67%) in Sandra and Rocky cultivars respectively. The regenerated plants were successfully transplanted to the field for acclimatization

Acquirement of CRY8DB Transgenic Tall Fescue (Festuca arundinacea Schreb.) by Agrobacterium tumefaciens to Develop Resistance Against Pentodon idiota Herbest.

Tall fescue plants are widely exposed to white grubs. Regarding the rate of damage caused by the white grubs to tall fescue and difficulty of its ecological and economical control, production of resistant cultivars is a priority. In this experiment, for the first time, we report production of transgenic lines resistant to white grub using CR8DB gene. For this, mature seeds were placed on MS basal medium with 0-15mgL-1 2,4-D for callogenesis and 0-1.75mgL-1 BA for regeneration. 'Asterix' (54.11%) in 7.5 and 'Talladega' (52.53%) in 10mgL-1 2,4-D showed maximum callogenesis. Regeneration percentage was higher in 0.5mgL-1 BA. Agrobacterium tumefaciens strain LBA4404 harbouring binary vector pCAMBIA 1301 with CRY8DB gene, which contains HPTII gene and uidA and various types MS media were used for transformation of calli. The highest percentage of gus enzyme activity and hygromycin resistance in calli was related to the modified medium type 11. The PCR and RT-PCR analysis was done to confirm the presence and expression of the target gene in transgenic 5 lines in 'Asterix' and 3 lines in 'Talladega'. According to bioassay, larvae mortality of 91.66% was observed in transgenic plants, whereas it was 15.52% in control plants.

Establishment of tissue culture and acclimatization method for in vitro mass propagation of Echeveria laui and Echeveria elegans

The objective of this study was to investigate the suitable parts for callus induction and optimal concentrations of growth regulators contained in the medium affecting shooting and rooting Echeveria laui and Echeveria elegans for in vitro mass production. To determine the suitable plant parts for callus induction, the leaves were divided into upper, medium and bottom parts and cultured on MS medium at different concentrations with 0 ~ 2 mgL-1 NAA and 0 ~ 4 mgL-1 BA. The upper and middle parts of leaves both showed 100% callus formation rate with NAA 1 mgL-1 and BA 1 mgL-1 treatment in E. laui. The middle parts of leaves showed 83.3% callus formation rate at NAA 2 mgL-1 and BA 4 mgL-1 treatment in E. elegans. The shoot induction rate from callus was highest at NAA 0.1 mgL-1 and BA 3 mgL-1 treatment in E. laui and NAA 0.3 mgL-1 in E. elegans. In addition, the number of shoots formation was 10.4 shoots high in NAA 1 mg L-1 and BA 1 mgL-1 treatment in E. laui and 12.0 shoots in most effective NAA 1 mgL-1 and BA 0.1 mg L-1 treatment in E. elegans. In the case of acclimatization of regenerated plant, growth characteristics did not show any significant difference (35 ~ 55%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate considered plant height and appearance preference of E. laui and E. elegans. It was established that the optimization of culture condition was responsible for the mass propagation in vitro cultures of E. laui and E. elegans.

Micropropagation response of three grape vines (Vitis vinifera L.) cultivars to ferrous, nitrate and growth regulators

Three Grapevine (Vitis vinifera L.) cultivars, Kamaly, Taify and Rash Mew were propagated by tissue culture technique and a reproducible and reliable protocol was developed through this investigation. Single nodes of grapevine were initiated on MS medium after getting healthy aseptic cultures by their surface sterilization with 0.15% HgCl2 for 6 minutes. The shoots were multiplied by the addition of BA, Fe and N. The best multiplication parameters were obtained from the Kamaly cultivar grown on MS medium supplemented with 3.0 mgl-1 BA and 10 mgl−1 Fe. Whereas, the culture of Taify grape cultivar on MS medium supplemented with 10 mgl−1 N produced the longest shoots (3.00 cm). The best rooting results were obtained for Taify and Rash Mew cultivars especially when grown on full MS salts strength media supplemented with 1.0 mgl−1 NAA. A high survival rate of gradually acclimatized Rash Mew plantlets was obtained (100%). The best culture mixture for acclimatization stage was composed of peatmoss and perlite.

An Efficient Protocol Of Stevia rebaudiana Regeneration For Large-Scale Production

Stevia rebaudiana is considered a valuable natural sweetener and medicinal plant containing sweetener compounds in its leaves. Nowadays, plant drugs are frequently considered to be free from side effects and less toxic than the synthetic one, for that people and some pharmacological companies are heading to the herbal medicine. Thus, to meet the increased demand for raw material of this medicinal herb, I described in this study an efficient regeneration protocol using shoot tip and nodal explants and different PGRs on MS medium for healthy biomass production. The optimum shoot growth with a high number without callus formation was obtained using MS medium in the presence of 1.0 mgL-1 BA after 6 weeks which shows (4.06-4.46) shoot buds/explant after 6 weeks. The combination of 1.0 mg L-1 BA with 0.5 mg L-1 IBA was found to be best combination for shoot multiplication that recorded 6.66 shoots/explant with 4.46 cm shoot length. Healthy elongated shoots were harvested and then cultured on half strength MS basal salts medium without vitamins medium in the presence of different concentrations of IBA and NAA individual or in combination with IAA or 2,4-D. Moreover, the optimum root growth with high roots number (4.46) with (76.6 %) root formation was observed using 1.0 mg L-1 IBA on half strength MS medium. Therefore, well-developed healthy in vitro rooted plantlets after 6 weeks were successfully acclimatized and they grew as normal plants in the greenhouse.

In Vitro Regeneration of Iranian Melon (Cucumis Melo L. ‘Samsoori’) Using Antibiotic and Benzyl adenine Micropropagation of Cucumis Melo L. ‘Samsoori’

Cucumis melo L. is one of the most economically important horticultural crops worldwide. However, low plant regeneration frequency of this plant during genetic transformation is the major hurdle for applying biotechnological approaches. Hence, this study aimed to evaluate the effect of 6-benzyladenine (BA), cefotaxime (CTX), kanamycin (KAN), and indole-3-acetic acid (IAA) on the regeneration of cotyledonary petioles generated from 6-day-old in vitro grown seedlings. Results showed that application of 1.5 mgl-1 BA plus 250 mgl-1 CTX and 1 mgl-1 BA with 1000 mgl-1 CTX formed the most efficient media for plant regeneration. The highest callus production was recorded on medium containing 1 mgl-1 BA with 250 mgl-1 CTX and 1.5 mgl-1 BA with 750 mgl-1 CTX. Medium containing 500 mgl-1 CTX plus 0.1 mgl-1 IAA efficiently induced both root and leaf formation. All regenerated plants were died by adding 100 mgl-1 kanamycin therefore this level considered as threshold level for kanamycin application.. Overall, the results indicated that presence of BA plays an essential role for melon regeneration and cefotaxime can be considered as an auxiliary agent.

Induksi Tetraploid Pada Tanaman Jambu Biji Merah (Psidium guajava L.) secara In Vitro

ABSTRACT Some commercial varieties of Guava seedless are triploid and rarely found naturally. A triploid variety with less seeds and better yield potential can be achieved by crossing tetraploid to diploid variety. Guava tetraploid plants can be synthetically induced by using oryzalin or colchicine to double the chromosome from its diploid. This research was aimed to obtain tetraploid lines by studying the effects of oryzalin in germination and in vitro growth of Guava. Seed from Red Guava were cultured in liquid medium MS + 2 mgL-1 BA with adding oryzalin according to the treatment, then seed planted in MS solid medium. Seed explants were exposed to Oryzalin 0 (controls), 15, 30 dan 60 ?M with exposure time are 23, 36 and 49 days or 3, 5, and 7 weeks. Results from polyploid induction were 8 tetraploid (5.48%) and 9 (6.16%) mixoploid shoots in vitro. High est tetraploid shoots were obtained from treatments by exposing the seed explant to 15 – 30 ?M oryzalin for 23 – 36 days. Oryzalin treatments inhibit germination and in vitro growth of Guava until 1st subculture. After second subculture, tetraploid or mixoploid shoots quantitatively showed no difference respons on in vitro growth with its diploid. Keywords: guava, tetraploid, seed explant, in vitro germination, polyploid

In vitro conservation protocol of Ceropegia bulbosa: An important medicinal and threatened plant species of Western Rajasthan

In vitro regeneration protocol has been standardized for highly medicinal and threatened succulent Ceropegia bulbosa Roxb. The paper focuses on morphogenic response of nodal explant when cultured on MS media. Murashige and Skoog medium supplemented with 6-benzyladenine (BA) (2.0 mgl-1) was found optimum for axillary shoot bud induction with 83.4 % response. Further shoots were multiplied through repetitive (3-4 times) transfer of the original explant and by subculture of the in vitro generated shoots. Maximum number of shoots 5.7±0.78 with shoot length of 3.6±0.82 cm was achieved on MS medium augmented with combination of 0.25 mgl-1 BA + 0.25 mgl-1 KN + 0.1 mgl-1 IAA and additives (50.0 mgl-1 ascorbic acid, 25 mgl-1 each of citric acid, arginine and adenine sulphate). For ex vitro rooting, pulse treatment of IBA 250 mgl-1 for 3 min was found optimum. The rooted shoots were successfully hardened in the green house condition (RH 75-80% at 26-28˚C) and about 80 % shoots were transferred to the garden.

In vitro micropropagation of 4 Hungarian bryophytes as potential sources for pharmacologically active substances

The plant kingdom is a rich source of pharmacologically active biomolecules, and bryophytes represent a relatively unexploited taxon within plants. From this point of view, Hungarian mosses are very promising, since only a small percent of the 659 species has been studied in detail. The aim of this project is to analyse Hungarian bryophyte species as potential sources of molecules for anticancer drug and antibiotic research. Important steps during the research are to identify bryophyte species with bioactivity, to isolate and to identify their active components and to characterize their effects in detail. Based on an in vitro screening of 58 species, the most promising species are further studied. In order to ensure sustainability of further research, in vitro micropropagation studies - of species regarded as the most promising - are being carried out. Optimization of surface sterilization (using 10% CaCl2O2 or 10% commercial Domestos) of Brachythecium rutabulum, Oxyrrhynchium hians, Tortula muralis and Campylopus introflexus is completed. In the selected bryophytes selection of appropriate explants (shoot tips or lateral shoots), optimization of substrates (1/2 MS without sucrose) for in vitro cultivation were carried out. Various explants of bryophyte species, different surface sterilization methods, various culture media with different plant hormone combinations and without hormones (1 mg l-1 NAA/ 1 mg l -1 BA/ 1 mg l-1 2, 4 D/ 1 mg l-1 BA+ 0.1 mg l-1 NAA/ 2 mg l-1 BA/ 1 mg l-1 NAA) were tested for optimal growth and morphogenesis. New moss shoots or protonema developments, depending on the hormonal supply, were observed. Growth response was recorded in every selected species during in vitro cultivation. The research was supported by the grant NKFI OTKA 115796 (supervisor Dezső Csupor, “Hungarian bryophytes as potential sources for pharmacologically active substances”). Work of Boglarka Toth was supported by the UNKP-17-2 New National Excellence Program of the Ministry of Human Capacities.

Initiation phase of meristem culture of Prunus avium cvs. "Hajyusefi" and "Zard"

This trial was carried out to determine the best medium and plant growth regulator concentrations on the initiation of meristem tips from sweet cherry trees cvs. Hajyusefi and Zard. Meristems were isolated from actively growing plants in the spring and cultured on three kinds of basal media (MS, WPM and QL) supplement with three different concentrations of BA (0, 0.5 and 1 mgL-1), 0.1 mgL-1 GA3 and 0.1 mgL-1 IBA. Explants were soaked in antioxidant solutions (100 mgL-1 ascorbic acid and 150 mgL-1 citric acid for 1 hours). The meristem explants (0.5-0.7 mm) cultured in medium and maintained in a growth room at a 16h - light/8 hr-dark with a light intensity of 2000-3000 lux from white fluorescent light. After six weeks, survival and necrosis ratios and leaf number were studied. Result showed that the highest survival rate and the low necrosis rate (83.3%) was in WPM medium supplemented with 0.5 mgL -1 BA for Hajyusefi cultivar. Necrosis occurred more often in vitro condition. Necrosis rate was high in MS (100-83.3%) for Zard and Hajyusefi cultivars, respectively. The most Leaf number (11) observed in WPM medium supplemented with 0.5 mgL-1 BA in Hajyusefi cultivar and the

The effect of plant growth regulators and different types of explants on in vitro regeneration of sweet pepper (Capsicum annuum L.)

Present study was conducted to standardize a protocol for in vitro plant regeneration in Capsicum annuum L. cv California Wonder. Three different types of explants (Hypocotyl, Cotyledon and Cotyledonary petiole) from 8day-old seedling were used. Various concentrations of BA (2, 3 mgL-1), IBA (0.1, 0.2 mgL-1) and NAA (0.1, 0.2 mgL -1 ) were used for shoot regeneration. The highest number of shoot per explant was obtained from cotyledonary petiole on a MS medium supplemented with 3 mgL-1 BA and 0.2 mgL-1 IBA. Effective in vitro shoot elongation was achieved on half strength MS medium containing 0.1 mgL-1 BA in combination with 0.5 mgL-1 GA3. As well as adding activated charcoal (3 gL -1 ) and citric acid along with ascorbic acid caused eliminate phenolic material and prevent shoot necrosis in sweet pepper. Elongated shoots were rooted on half strength MS medium supplemented with 1 mgL-1 IBA.

English

The objective of the present was to enhance the frequency of plant regeneration in date palm (Phoenix dactylifera L.) cv. Hillawi. Explants were incubated on Murashige and Skoog (MS) medium supplemented with 1 mgl-1 6-benzyladenine (BA) and different concentrations (0.1 to 2.0 mgl-1) of thidiazuran (TDZ), or free of BA and TDZ (control treatment). The results indicate that the Maximum response (66.67%) was observed on medium supplemented with 1.0 mgl-1 BA and 0.5 mgl-1 TDZ, producing an average of 4.2 and 18.2 buds per culture after 16 and 24 week from culture, respectively. TDZ at concentrations higher than 0.5 mgl-1 resulted in suppressed buds formation, where a decrease in the number of buds was noticed when the concentration of TDZ was increased from 0.5 to 2 mgl-1. Regarding the activity of antioxidant enzyme peroxidase during budding of date palm cv. Hillawi, (The chemical analyses of peroxidase compounds were spectrophotometrically performed) the applied concentration of 0.5 mgl-1 TDZ with 1 mgl-1 BA enhanced peroxidase activity, where peroxidase activity was associated with increased number of buds formation. Histological studies revealed that adventitious buds were formed directly from epidermal cells without callus formation, and adventitious buds were developed from meristematic cells in shoot tip tissues. Shoots were elongated on 0.5 mgl-1 GA3+ 0.1 mgl-1 NAA MS media and rooted on MS media supplemented with 0.2 mgl-1 of α-naphthaleneacetic acid (NAA). Rooted shoots were successfully acclimatized and established in a mixture of peat moss and perlite (2:1) with 80% success. Key words: Date palm, thidiazuran (TDZ), adventitious buds, peroxidase (POD), histology.

In vitro plant regeneration of two cucumber (Cucumis sativum L.) genotypes: Effects of explant types and culture medium

The effect of different phytohormone concentrations on callusogenesis and organogenesis in two cucumber genotypes were studied. It was established that the rate of plant regeneration depends on genotype, explant type and culture medium. Hypocotyls were found to be more responsive than cotyledons in morphogenesis. In vitro planlet-regenerants have been obtained in hypocotyls explants on culture medium with 1.0 and 2.0 mgL-1 BA for cultivar Gergana and in 1.0 and 3.0 mgL-1K-line 15B. Induction of regeneration in cotyledons were established only in cultivar Gergana on culture medium supplemented with 3.0 mgL-1 BA and in combination of 0.5 mgL-1IAA.

English

The present investigation was carried out with a view to standardize an in vitro culture technique for mass propagation of an endangered medicinal plant Rauwolfia serpentinaBenth. Shoot tip was used as explant for initial culture. The explants were cultured on MS (Murashige and Skoog, 1962) medium supplemented with different concentration and combination of NAA and BA for primary shoot proliferation. The best shoot proliferation was observed in MS medium containing 0.1 mg L-1 NAA and 2.5 mgL-1 BA, where 92% of plants showed proliferation. For rooting, half strength MS medium supplemented with 0.4 mgL-1NAA and 0.1 mgL-1 IBA showed maximum root formation (91%). After acclimatization and transplantation, 90% of the in vitro derived plants survived in ex vivo condition.   Key words: Rauwolfia serpentina, in vitro culture, naphthalene acetic acid, benzyl adenine, indole buteric acid, MS medium.