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A novel VIGS method by agroinoculation of cotton seeds and application for elucidating functions of GhBI-1 in salt-stress response

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Abstract

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A VIGS method by agroinoculation of cotton seeds was developed for gene silencing in young seedlings and roots, and applied in functional analysis of GhBI-1 in response to salt stress.

Abstract

Virus-induced gene silencing (VIGS) has been widely used to investigate the functions of genes expressed in mature leaves, but not yet in young seedlings or roots of cotton (Gossypium hirsutum L.). Here, we developed a simple and effective VIGS method for silencing genes in young cotton seedlings and roots by soaking naked seeds in Agrobacterium cultures carrying tobacco rattle virus (TRV)-VIGS vectors. When the naked seeds were soaked in Agrobacterium cultures with an OD600 of 1.5 for 90 min, it was optimal for silencing genes effectively in young seedlings as clear photo-bleaching phenotype in the newly emerging leaves of pTRV:GhCLA1 seedlings were observed at 12–14 days post inoculation. Silencing of GhPGF (cotton pigment gland formation) by this method resulted in a 90% decrease in transcript abundances of the gene in roots at the early development stage. We further used the tool to investigate function of GhBI-1 (cotton Bax inhibitor-1) gene in response to salt stress and demonstrated that GhBI-1 might play a protective role under salt stress by suppressing stress-induced cell death in cotton. Our results showed that the newly established VIGS method is a powerful tool for elucidating functions of genes in cotton, especially the genes expressed in young seedlings and roots.

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Acknowledgements

The authors thank professor Libo Shan, Texas A&M University, and professor Zhaohu Li, China Agricultural University, for providing the pTRV-VIGS vectors. This work is supported by the National Science Foundation in China [31501351]; Youth Scientific Research Foundation of Shandong Academy of Agricultural Science [2015YQN04]; the National Project of Modern Agricultural Industry Technology System in China [CARS-15-05]; the Taishan Scholars Program of Shandong Province [ts201511070] and the State Key Laboratory of Cotton Biology Open Fund.

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  1. Jingxia Zhang and Furong Wang contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Cotton Breeding and Cultivation in Huang-Huai-Hai Plain, Ministry of Agriculture, Cotton Research Center of Shandong Academy of Agricultural Sciences, Jinan, 250100, China

    Jingxia Zhang, Furong Wang, Chuanyun Zhang, Yu Chen, Guodong Liu & Jun Zhang

  2. State Key Laboratory of Cotton Biology, Henan Key Laboratory of Plant Stress Biology, College of Life Science, Henan University, Kaifeng, 475004, China

    Fushun Hao

  3. Nanjing Agricultural University, Nanjing, 210095, China

    Junhao Zhang

  4. College of Life Science, Shandong Normal University, Jinan, 250014, China

    Yanxiu Zhao & Jun Zhang

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  1. Jingxia Zhang
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  2. Furong Wang
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  3. Chuanyun Zhang
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  4. Junhao Zhang
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  5. Yu Chen
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  9. Jun Zhang
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Correspondence to Fushun Hao or Jun Zhang.

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Communicated by Chun-Hai Dong.

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Zhang, J., Wang, F., Zhang, C. et al. A novel VIGS method by agroinoculation of cotton seeds and application for elucidating functions of GhBI-1 in salt-stress response. Plant Cell Rep 37, 1091–1100 (2018). https://doi.org/10.1007/s00299-018-2294-5

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  • DOI: https://doi.org/10.1007/s00299-018-2294-5

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