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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....

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Detalles Bibliográficos
Autores principales: Das Bhowmik, Sudipta Shekhar, Cheng, Alam Yen, Long, Hao, Tan, Grace Zi Hao, Hoang, Thi My Linh, Karbaschi, Mohammad Reza, Williams, Brett, Higgins, Thomas Joseph V., Mundree, Sagadevan G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6498970/
https://www.ncbi.nlm.nih.gov/pubmed/31105725
http://dx.doi.org/10.3389/fpls.2019.00524
Descripción
Sumario: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.