Transformation and Overexpression of Primary Cell Wall Synthesis-Related Zinc Finger Gene Gh_A07G1537 to Improve Fiber Length in Cotton

Abdul Razzaq; Muhammad Mubashar Zafar; Pengtao Li; Ge Qun; Xiaoying Deng; Arfan Ali; Abdul Hafeez; Muhammad Irfan; Aiying Liu; Maozhi Ren; Haihong Shang; Yuzhen Shi; Wankui Gong; Youlu Yuan

doi:10.3389/fpls.2021.777794

Transformation and Overexpression of Primary Cell Wall Synthesis-Related Zinc Finger Gene Gh_A07G1537 to Improve Fiber Length in Cotton

Front Plant Sci. 2021 Nov 5:12:777794. doi: 10.3389/fpls.2021.777794. eCollection 2021.

Authors

Abdul Razzaq^{1

2}, Muhammad Mubashar Zafar¹, Pengtao Li³, Ge Qun¹, Xiaoying Deng¹, Arfan Ali⁴, Abdul Hafeez¹, Muhammad Irfan⁵, Aiying Liu¹, Maozhi Ren¹, Haihong Shang^{1

6}, Yuzhen Shi¹, Wankui Gong¹, Youlu Yuan^{1

6}

Affiliations

¹ State Key Laboratory of Cotton Biology, Key Laboratory of Biological and Genetic Breeding of Cotton, The Ministry of Agriculture, Institute of Cotton Research, Chinese Academy of Agricultural Science, Anyang, China.
² Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore, Pakistan.
³ School of Biotechnology and Food Engineering, Anyang Institute of Technology, Anyang, China.
⁴ FB Genetics, Four Brothers Group, Lahore, Pakistan.
⁵ Department of Biological Sciences, Forman Christian College, A Chartered University, Lahore, Pakistan.
⁶ School of Agricultural Sciences, Zhengzhou University, Zhengzhou, China.

Abstract

Molecular interventions have helped to explore the genes involved in fiber length, fiber strength, and other quality parameters with improved characteristics, particularly in cotton. The current study is an extension and functional validation of previous findings that Gh_A07G1537 influences fiber length in cotton using a chromosomal segment substitution line MBI7747 through RNA-seq data. The recombinant Gh_A07G1537 derived from the MBI7747 line was over-expressed in CCRI24, a genotype with a low profile of fiber quality parameters. Putative transformants were selected on MS medium containing hygromycin (25mg/ml), acclimatized, and shifted to a greenhouse for further growth and proliferation. Transgene integration was validated through PCR and Southern Blot analysis. Stable integration of the transgene (ΔGh_A07G1537) was validated by tracking its expression in different generations (T₀, T₁, and T₂) of transformed cotton plants. It was found to be 2.97-, 2.86-, and 2.92-folds higher expression in T₀, T₁, and T₂ plants, respectively, of transgenic compared with non-transgenic cotton plants. Fiber quality parameters were also observed to be improved in the engineered cotton line. Genetic modifications of Gh_A07G1537 support the improvement in fiber quality parameters and should be appreciated for the textile industry.

Keywords: CSSLs; cotton biology; fiber length; genetic transformation; zinc finger.