Robust Genetic Transformation System to Obtain Non-chimeric Transgenic Chickpea

Chickpea transformation is an important component for the genetic improvement of this crop, achieved through modern biotechnological approaches. However, recalcitrant tissue cultures and occasional chimerism, encountered during transformation, hinder the efficient generation of transgenic chickpeas. Two key parameters, namely micro-injury and light emitting diode (LED)-based lighting were used to increase transformation efficiency. Early PCR confirmation of positive in vitro transgenic shoots, together with efficient grafting and an extended acclimatization procedure contributed to the rapid generation of transgenic plants. High intensity LED light facilitate chickpea plants to complete their life cycle within 9 weeks thus enabling up to two generations of stable transgenic chickpea lines within 8 months. The method was validated with several genes from different sources, either as single or multi-gene cassettes. Stable transgenic chickpea lines containing GUS (uidA), stress tolerance (AtBAG4 and TlBAG), as well as Fe-biofortification (OsNAS2 and CaNAS2) genes have successfully been produced

<i>In vitro </i>regeneration of <i>Begonia rubrovenia</i><span style="mso-bidi-font-style:italic"> var. <i>meisneri</i> C.B. Clarke— A rare and endemic ornamental plant of Meghalaya, India</p>

300-303In vitro plant regeneration from leaf and petiole explants of Begonia rubrovenia meisneri C.B. Clarke, an endemic and rare plant of Meghalaya, has been established. Multiple shoots were induced on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-benzyl aminopurine (BAP) and thidiazuron (TDZ). In vitro produced leaf and petiole sections (secondary explants) exhibited enhanced shoot proliferation in MS medium supplemented with either BAP or TDZ in different concentrations (0.05-5.0 mg/L). A maximum, 65 shoots/petiole were formed on MS+0.1 mg/L TDZ. The in vitro-raised shoots formed roots in MS medium containing 0.1 mg/L indole 3-acetic acid (IAA), with a maximum of 13.8 roots/shoot. About 73.33% of the 500 plantlets transferred acclimatized successfully within 4 wk in a glasshouse. On being transferred to the field, all the acclimatized plantlets survived after 8 wk. </p