ABSTRACT
A novel combination of plant growth regulators comprising indole-3-butyric acid (IBA), 6-benzylaminopurine (BA) and gibberellic acid (GA3) in Murashige and Skoog basal medium has been formulated for in vitro induction of both shoot and root in one culture using cotyledonary node explants of guar, (Cyamopsis tetragonoloba). Highest percentages of shoot (92%) and root (80%) induction were obtained in the medium containing (mg/L) 2 IBA, 3 BA and 1 GA3. Shoot regeneration from the cotyledonary node explants was observed after 10-15 days. Regeneration of roots from these shoots occurred after 20 to 25 days. The regenerated plantlets showed successful acclimatization on transfer to soil. This protocol is expected to be helpful in carrying out various in vitro manipulations in this economically and industrially important legume.
Subject(s)
Cyamopsis/drug effects , Cyamopsis/growth & development , Gibberellins/pharmacology , Indoles/pharmacology , Kinetin/pharmacology , Plant Development/drug effects , Plant Growth Regulators/chemistry , Plant Growth Regulators/pharmacology , Plant Leaves/drug effects , Plant Leaves/growth & development , Plant Roots/drug effects , Plant Roots/growth & development , Plant Shoots/drug effects , Plant Shoots/growth & developmentABSTRACT
Development of a reproducible, versatile and efficient in vitro plant regeneration system is highly warranted for Indian soybean varieties for their mass multiplication in view of their commercial significance. Accordingly a protocol for direct shoot organogenesis in soybean variety JS 335 has been developed. Using cotyledonary node explants significant organogenic responses, mean shoot number and shoot length were observed when these were incubated on MS medium supplemented with 0.89 µM Benzyladenine (BA) and 5 μg/L triacontanol (TRIA) where in 9.3 ± 0.5 shoots were obtained. TRIA at 5 μg /L able to produce 6.8 ± 0.5 shoot buds in presence of 0.98 µM IBA and 0.89 µM BA. Highest mean shoot buds (14.0 ± 0.5 and 9.0 ± 0.5) and mean shoot length (4.6 ± 0.3 and 10.0 ± 0.7) were obtained when cotyledonary node and shoot tip explants were cultured on MS medium containing 0.14 µM gibberellic acid (GA3), 0.89 µM BA and 5 μg/L TRIA. Moreover, TRIA supported highest mean root number (6.3±0.5) and root length (21.5 ± 0.57 cm). Field survival of in vitro derived plants of TRIA treatment was 70% and the overall growth and seed yield was also significantly better than control plants. This protocol may be used for improving the in vitro regeneration of soybean variety JS 335 for transformation studies.
ABSTRACT
An efficient, highly reproducible protocol for multiple shoot induction and plant regeneration of Pongamia pinnata has been successfully developed using cotyledonary node explants. This study also demonstrates that preconditioning of explant stimulates production of multiple shoots from cotyledonary nodes of P. pinnata. The highest direct shoot regeneration (90 percent) with an average of 18.4 +/- 3.1 shoots/explant were obtained when cotyledonary node explants were excised from seedlings germinated on Murashige and Skoog (MS) media supplemented with benzyladenine (BA) 1 mg l-1, and subsequently cultured on MS media with 1 mgl-1 thidiazuron (TDZ). Scanning electron microscope observations of cotyledonary node (CN) explants excised from pre-conditioned and normal seedlings, revealed larger buds with rapid development in BA-preconditioned CN explants. The addition of adenine sulphate significantly increased the average number of shoots per explant. The highest direct shoot regeneration (93 percent) with an average of 32.2 +/- 0.93 shoots/explant was obtained from BA-preconditioned CN when cultured on MS media supplemented with 1 mg l-1 TDZ and 200 mg l-1 adenine sulphate (ADS). Repeated shoot proliferation was observed from BA preconditioned CN explants up to 3 cycles with an average of 15 shoots/explant/cycle when cultured on MS media supplemented with 1 mg l-1 TDZ and 150 mg l-1 L-glutamine, thus producing 45 shoots/CN explant. Shoots were elongated on hormone free MS media and rooted on 1/2 MS media supplemented with 1 mg l-1 of IBA. Rooted shoots were successfully acclimatized and established in soil with 80 percent success. The highly regenerative system developed in this investigation for this important tree could be a useful tool for genetic transformation.