Jute is a crop of commercial importance that is widely cultivated for its bast fiber production but susceptible to many diseases that results in major economic loss. New genes can be introduced into this plant through Agrobacterium mediated genetic transformation for its genetic improvement, which is dependent on the availability of suitable in vitro techniques. An efficient regeneration system has been developed for in vitro culture of jute (Corchorus capsularis) from the distal cut ends of cotyledonary petioles. High frequency shoot regeneration was obtained on Murashige and Skoog (MS) nutrient agar medium supplemented with 0.5 mg/l NAA, 0.5 mg/l BAP and 36 g/l sucrose. On transfer to soil, the regenerated plantlets survived and appeared to be morphologically similar to the normal seed-grown plants. They developed pods and set fertile seeds. Histological analysis revealed de novo origin of shoot buds in the in vitro cultured cotyledonary petioles. Parameters affecting transformation were optimized by assaying GUS activity in these regenerable tissues after cocultivation with Agrobacteria. These tissues appear to be susceptible for infection and transformation by Agrobacterium carrying uid (GUS INT) and nptII genes, as well as shoot multiplication. The cells at the cut end of the petioles were found competent to take up the DNA, which was monitored by transient GUS gene expression. EHA105 at 0.3 O.D and LBA4404 at 0.5 O.D were found to be compatible in giving optimal levels of transient GUS expression.
SummaryEfficient in vitro propagation of Ceropegia candelabrum L. (Asclepidaceae) through somatic embryogenesis was established. Somatic embryogenesis depended on the type of plant growth regulators in the callus-inducing medium. Friable callus, developed from leaf and internode explants grown on Murashige and Skoog (MS) medium supplemented with 4.52 mM 2,4-dichlorophenoxyacetic acid (2,4-D), underwent somatic embryogenesis. Compared to solid media, suspension culture was superior and gave rise to a higher number of somatic embryos. Transfer of the friable callus developed on MS medium containing 4.52 mM 2,4-D to suspension cultures of half-or quarter-strength MS medium with lower levels of 2,4-D (0.23 or 0.45 mM) induced the highest number of somatic embryos, which developed up to the torpedo stage. Somatic embryogenesis was asynchronous with the dominance of globular embryos. About 100 mg of callus induced more than 500 embryos. Upon transfer to quarter-strength MS agar medium without growth regulators, 50% of the somatic embryos underwent maturation and developed into plantlets. Plantlets acclimatized under field conditions with 90% survival.
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