Construction of a cotton VIGS library for functional genomics study.
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.
Department of Biochemistry and Biophysics, Institute for Plant Genomics and Biotechnology, Texas A&M University, 401 Joe Routt Blvd, College Station, TX, 77843, USA.
Maoying
M
Li
Fangjun
F
Li
Ping
P
He
D055864:Droughts; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D020868:Gene Silencing; D018745:Genome, Plant; D003368:Gossypium / Q000235:genetics / Q000254:growth & development*; D017422:Sequence Analysis, DNA; D013312:Stress, Physiological