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Construction of a Cotton VIGS Library for Functional Genomics Study

  • Maoying Li
  • Fangjun Li
  • Ping He
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1287)

Abstract

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.

Key words

Cotton (Gossypium spp.) Virus-induced gene silencing Functional genomics Agrobacterium cDNA library 

Notes

Acknowledgments

The cotton work in He’s lab is supported by USDA NIFA (2012-67013-19433). F. L. was supported by the China Scholarship Council.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Maoying Li
    • 1
    • 2
  • Fangjun Li
    • 1
    • 2
  • Ping He
    • 1
  1. 1.Department of Biochemistry and Biophysics, Institute for Plant Genomics and BiotechnologyTexas A&M UniversityCollege StationUSA
  2. 2.State Key Laboratory of Plant Physiology and Biochemistry, Centre of Crop Chemical Control, College of Agronomy & BiotechnologyChina Agricultural UniversityBeijingChina

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