Abstract
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.
Highlights
Chickpeas are an important grain legume with an annual global production of 14.2 million tons in 2014 (FAOSTAT, 2016)
Micro-injured Transformed half embryo explants (THEE) grown under light emitting diode (LED) light generated 2.5fold more shoots than those grown under fluorescent light (39.2 ± 1.4% and 15.7 ± 4.8% respectively) (Table 1)
These results indicate that micro-injury negatively affected shoot regeneration of THEE grown under fluorescent light but not under LED light
Summary
Chickpeas are an important grain legume with an annual global production of 14.2 million tons in 2014 (FAOSTAT, 2016). Global chickpea productivity does not meet current demand because of several abiotic and biotic constraints. Conventional breeding and enhanced management practices have addressed several chickpea production constraints as well as improved grain quality, the lack of diverse germplasms for certain traits, sexual incompatibility and a high degree of autogamy continue to limit chickpea yields (Van Rheenen et al, 1993; Somers et al, 2003; Atif et al, 2013). Recent advances in gene editing allow the precise manipulation of a crop’s genome without the integration of foreign DNA. To apply these powerful tools for crop improvement, an efficient regeneration and Robust Genetic Transformation of Chickpea transformation system is a priority. For most crops, transformation and regeneration remain difficult even after more than 30 years of technological advances (Altpeter et al, 2016)
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