A direct method for genetically transforming rice seeds modelled with FHVB2, a suppressor of RNAi

作者：Das Sudhanshu Sekhar; Sanan Mishra Neeti^*

来源：Plant Cell, Tissue and Organ Culture, 2015, 120(1): 277-289.

DOI：10.1007/s11240-014-0604-3

• Rice seeds
• Transformation
• In planta
• RNAi suppressors
• FHVB2

RNA interference (RNAi) is a novel method of gene regulation and one of the potent host defense mechanisms against viruses. It however acts as a deterrent in transgene-technology by constraining the expression of the introduced gene. The virus-encoded suppressors have the ability to restrict host RNAi to promote pathogenicity. They thus have tremendous potential to ameliorate low transgene expression and have important applications in the biofarming sector. Unfortunately the suppressors severally reduce plant regeneration potentials in the standard procedures. In this study, we report a simple, fast and efficient method for in planta transformation of rice seeds that can be used for over-expressing FHVB2, a well-characterized suppressor of RNAi. The protocol involves agro-inoculation of embryos without vacuum infiltration or injury followed by their growth ex vitro. Following transformation the transgene integration, expression and stable inheritance was confirmed. We observed that the FHVB2 transgenic survival in this methodology was 15-fold higher compared to that in available callus-based methods. The protocol has the potential to be extended for transforming rice with any gene as exemplified by the use of control constructs.

• 出版日期2015-1