Not AvailableThe present study describes an efficient in vitro culture protocol for
direct plantlet regeneration and Agrobacterium- mediated genetic
transformation of Corchorus capsularis L. cultivar JRC 517. In vitro
morphogenetic capacity of different explants was evaluated. Nodal
explants with immature axillary buds showed maximum in vitro
culture response (95%) with plantlets induction when cultured on MS
with 0.1 mg l-1
IAA and 0.1 mg l-1 Kin. A. tumefaciens strain
LBA4404 harbouring a binary vector pBI121, containing gusA
reporter gene under the transcriptional control of Cauliflower Mosaic
Virus (CaMV) 35S promoter and NOS terminator was used in
addition with neomycin phosphotransferase (npt-II) as plant selection
marker gene. Different parameters viz. O.D600.- of Agrobacterium cell
suspension: 0.3; one day preculture; one min explants dipping, vacuum
infiltration for 10 min at 600 mm Hg pressure; 0.001 ml l-1
concentration of non-ionic surfactant (Tween 20) and two days cocultivation
with 100 µM acetosyringone (AS) were found to be
optimum treatment to achieve maximum number of stable genetic
transformants (~3.6%). The putative transformants were screened on
MS medium supplemented with 50 mg l-1
kanamycin (Kan50) and
their transient expression was confirmed through GUS histochemical
assay of the reporter gene and PCR analysis. The survivor plants were
grown under Kan50 selection pressure, and rooted successfully.
Regenerated plantlets were acclimatized, hardened and transplanted to
green house. Stable integration of the transgene into the recipient
genome was confirmed by PCR using compatible primers of gusA and
nptII, and through Southern hybridization. The transgenic plants
showed normal morphology and most of them followed 3:1 ratio of
Mendelian inheritance for a single dominant locus. In vitro direct
shoot regeneration protocol from nodal explants with concurrent
transgenic development deemed to be successfully involving
economically important gene/s and trait enrichment in jute.Not Availabl
