4.8 Article

Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants

期刊

PLANT JOURNAL
卷 106, 期 3, 页码 817-830

出版社

WILEY
DOI: 10.1111/tpj.15202

关键词

cowpea transformation; embryonic axis; cotyledonary node (cot‐ node); shoot organogenesis; spectinomycin; Agrobacterium; CRISPR; Cas

资金

  1. CSIRO
  2. UGA
  3. LANGEBIO under the Bill and Melinda Gates Foundation (BMGF) [OPP1076280]
  4. Corteva Agriscience
  5. Bill and Melinda Gates Foundation [OPP1076280] Funding Source: Bill and Melinda Gates Foundation

向作者/读者索取更多资源

This study presents a highly efficient transformation system for cowpea using embryonic axis explants, which achieved efficient gene delivery, transgenic selection, and transgenic plant generation. By removing the shoot apical meristem from the explants, direct multiple shoot organogenesis from the cotyledonary node tissue was stimulated.
Cowpea (Vigna unguiculata (L.) Walp.) is one of the most important legume crops planted worldwide, but despite decades of effort, cowpea transformation is still challenging due to inefficient Agrobacterium-mediated transfer DNA delivery, transgenic selection and in vitro shoot regeneration. Here, we report a highly efficient transformation system using embryonic axis explants isolated from imbibed mature seeds. We found that removal of the shoot apical meristem from the explants stimulated direct multiple shoot organogenesis from the cotyledonary node tissue. The application of a previously reported ternary transformation vector system provided efficient Agrobacterium-mediated gene delivery, while the utilization of spcN as selectable marker enabled more robust transgenic selection, plant recovery and transgenic plant generation without escapes and chimera formation. Transgenic cowpea plantlets developed exclusively from the cotyledonary nodes at frequencies of 4% to 37% across a wide range of cowpea genotypes. CRISPR/Cas-mediated gene editing was successfully demonstrated. The transformation principles established here could also be applied to other legumes to increase transformation efficiencies.

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