Clonal Propagation of Strawberry (Fragaria x ananassa Duch.) through In vitro Runner Tip Culture through Incorporation of Growth Hormones

Runner tips explants of strawberry give rise to multiple shoots when cultured on MS medium supplemented with different concentrations and combinations of BAP with KIN or NAA or GA3.The highest response of shoot multiplication was obtained on MS containing 2.5 mgl-1 BAP + 0.5 mgl-1 Kin + 0.5 mgl-1 GA3. The maximum frequency of rooting (83%) and highest number of roots (3.49) was produced in medium containing 1.0 mgl-1 IBA. The well grown rooted plantlets were acclimatized and successfully established in autoclaved vermiculate soil and as well as natural condition. Using our established protocol, it is also possible to provide large numbers of micropropagated plantlets of this cultivars to produce high quality strawberry fruit for commercial cultivation practices.

Direct multiple shoots proliferation of black night shade (Solanum nigrum l.) from shoot tip explants induced by thidiazuron .

The present investigation was undertaken in Black Night Shade (Solanum nigrum L.) which is an important medicinal plant. Direct multiple shoots proliferation was achieved from shoot tip. The shoot tips were cultured on MS medium fortified with Thidiazuron (TDZ) (1.0-7.0 mg/L) for multiple shoot induction. Multiple shoots proliferation was best observed at 3.0 mg/L TDZ from the shoot tip explants within three weeks of culture. Shoot number per explant ranged between 2 and 10. Individual shoots were aseptically excised and sub cultured in the same media for shoot elongation. The elongated shoots were transferred to Indole Acetic Acid/Indole Butyric Acid (IAA/IBA) (0.5mg/L–2.0mg/L) for root induction. Rooting was observed within two weeks of culture. Rooted plantlets were successfully hardened under culture conditions and subsequently established in the field conditions. The recorded survival rate of the plants was 86%. Plants looked healthy with no visually detectable phenotypic variations.

In vitro regeneration and ploidy level analysis of Eulophia ochreata Lindl.

Various parameters including explant-type, medium compositions, use of phytohormones and additives were optimized for direct and indirect regeneration of E. ochreata, a medicinal orchid under threat. Protocorm-like-bodies (PLBs) proved to be the best explants for shoot initiation, proliferation and callus induction. Murashige and Skoog’s (MS) medium containing 2.5 mg L-1 6-benzylaminopurine (BAP), 1.0 mg L-1 kinetin (Kin) and additives (adenine sulfate, arginine, citric acid, 30 mg L-1 each and 50 mg L-1 ascorbic acid) was optimal for shoot multiplication (12.1 shoots and 7.1 PLBs per explant with synchronized growth), which also produced callus. Shoot number was further increased with three successive subcultures on same media and ~40 shoots per explant were achieved after 3 cycles of 30 days each. Additives and casein hydrolysate (CH) showed advantageous effects on indirect shoot regeneration via protocorm-derived callus. Optimum indirect regeneration was achieved on MS containing additives, 500 mg L-1 CH, 2.5 mg L-1 BAP and 1.0 mg L-1 Kin with 30 PLBs and 6 shoots per callus mass (~5 mm size). The shoots were rooted (70% frequency) on one by fourth-MS medium containing 2.0 mg L-1 indole-3-butyric acid, 200 mg L-1 activated charcoal and additives. The rooted plantlets were hardened and transferred to greenhouse with 63% survival rate. Flow-cytometry based DNA content analysis revealed that the ploidy levels were maintained in in vitro regenerated plants. This is the first report for in vitro plant regeneration in E. ochreata.

Regeneration of Shoots from Callus of Ceropegia pusilla Wight and ARN.

This study was undertaken to evaluate the most suitable concentration of plant growth regulators and suitable explants for callus induction and subsequent organogenesis of an endangered medicinal plant Ceropegia pusilla. The best performance of callus induction and morphogenesis was found on MS medium supplemented with 6- benzylaminopurine (BA) and naphthalene acetic acid (NAA), from node and internode the maximum account of callus initiation was recorded on MS medium supplemented with BA+NAA (2.22μM + 5.37 μM) and the maximum % of callus induced on subculture is 5.47  0.68. Rooting was best achieved on MS medium augmented with IBA (2.46μM). The maximum number of roots (4.72  0.66 cm) and root length (4.8  0.49 cm) were recorded). Plantlets regenerated in vitro with well-developed shoot and roots were successfully acclimatized in pots containing a mixture of decomposed coir waste, perlite and compost 1: 1:1 ratio and grown in a shade house with 81  3.16 percent survival rates.

An efficient in vitro plant regeneration of Dipteracanthus prostratus (Poir.) Nees.- a medicinal herb

Objective: This is the first attempt for an efficient plant regeneration protocol through in vitro direct organogenesis for a valuable medicinal plant, Dipteracanthus prostratus using nodal segment. Methods: Multiple shoots were induced from nodal explants cultured on Murashige and Skoog medium supplemented with kinetin (KIN), 6-benzylaminopurine (BAP) and 1-Naphthaleneacetic acid (NAA). Results: Maximum shoot responses (80%) were obtained with kinetin at 1.0 mg-1. The rate of shoot multiplication was maintained in subsequent subculture on similar fresh culture medium. The highest shoot length (3.96cm) was obtained with seventy three percentages of shoots at 0.2 mg-1 NAA along with 1.0 mg-1 kinetin. Maximum length of root (3.63cm) was formed at 0.5 mg-1 IBA with significant responses (80%). Rooted plantlets were then transferred to perforated plastic cups and grown in the green house at 80% survival rate. Conclusions: The highest survival rate was noticed and this plant developmental protocol could be used for large- scale regeneration of D. prostratus.