Construction of a cotton VIGS library for functional genomics study.
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To facilitate cotton functional genomics studies and identify cotton genes mediating biotic and abiotic stress responses, we report here a detailed protocol of construction of a VIGS library from diploid cotton Gossypium raimondii. Sequencing of the representative colonies revealed that the library covers a significant percentage of unique cotton genes. Based on the number of colonies in the primary library, it is estimated that this VIGS library is about 50 coverage of predicted cotton protein-coding genes. As a proof-of-concept experiment, we further describe a protocol to identify the genetic determinants involved in cotton drought tolerance using this VIGS library. The cotton VIGS library established here provides a unique opportunity to identify essential cotton genes in various stress responses and developmental growth regulation.