A Genotype-Independent, Simple, Effective and Efficient in Planta Agrobacterium-Mediated Genetic Transformation Protocol

Abstract

Crop improvement under changing climatic conditions is required to feed the growing global population. The development of transgenic crops is an attractive and conceivably the most effective approach for crop improvement with desired traits in varying climatic situations. Here, we describe a simple, efficient and robust in planta Agrobacterium-mediated genetic transformation method that can be used in most crops, including rice, wheat and cotton, and particularly in tissue culture recalcitrant crops, such as chickpea and pigeon pea. The protocol was successfully used for the development of transgenic chickpea and pigeon pea lines for resistance against pod borer. Transgenic lines in chickpea, pigeon pea and wheat were also developed for salt stress tolerance. These lines exhibited improved salt tolerance in terms of various physio-biochemical parameters studied. Since the protocol is rapid, as no tissue culture step is involved, it will significantly contribute to the improvement of most crops and will be of interest for plant biologists working with genetic engineering or genome editing.