Functional Verification of GhbHLH149-like Gene in Salt Tolerance in Upland Cotton

doi:10.7668/hbnxb.20196467

Abstract

Abstract:

To further investigate the function of cotton basic helix-loop-helix(bHLH)transcription factors in salt stress response,we introduced the cotton bHLH transcription factor GhbHLH149-like(previously identified by our team)into wild-type Arabidopsis thaliana via Agrobacterium-mediated transformation,and verified the function of GhbHLH149-like transcription factor in Arabidopsis.Protein structure prediction indicated that the secondary structure of GhbHLH149-like protein is mainly composed of α-helice and random coil,and it was a non-transmembrane hydrophilic protein.Quantitative Real-time PCR(qRT-PCR)results showed that the expression level of the GhbHLH149-like gene in transgenic Arabidopsis was significantly higher than wild-type.The results of phenotypic and physiological index assays indicated that the seed germination rate and root length of transgenic seedlings were significantly higher under salt stress,and the growth performance of transgenic seedlings was markedly better.Meanwhile,the contents of hydrogen peroxide(H₂O₂)and superoxide anion(O₂^-)were significantly lower,whereas the superoxide dismutase(SOD)activity was significantly higher in the transgenic lines.Taken together,GhbHLH149-like gene can significantly enhance the salt tolerance of Arabidopsis,presumably by participating in reactive oxygen species(ROS)scavenging.This study lays a theoretical foundation for the in-depth dissection of the molecular mechanism of cotton GhbHLH149-like gene in salt stress response.

Key words: Upland cotton, GhbHLH149-like, Protein sequence analysis, Salt stress, Function verification

CLC Number:

LIU Di, ZHEN Junbo, JIANG Mengmeng, LIU Linlin, FENG Xiaoqing, CHI Jina. Functional Verification of GhbHLH149-like Gene in Salt Tolerance in Upland Cotton[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(3): 68-74. doi: 10.7668/hbnxb.20196467.

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Figures/Tables 7

Tab.1 Primer of qRT-PCR

引物名称 Primer name	引物序列(5'-3') Primer sequence(5'-3')
AtUBQ5-F	GACGCTTCATCTCGTCC
AtUBQ5-R	CCACAGGTTGCGTTAG
qGhbHLH149-like-F	ACAGGGTTCTCGCCGTTTT
qGhbHLH149-like-R	TATCGGGCTTCCTCAACCTG

Fig.1 The secondary structure(A)and tertiary structure(B)prediction of GhbHLH149-like protein

Fig.2 Amino acid sequence analysis of GhbHLH149-like A.Phosphorylation site prediction;B.Protein transmembrane domain prediction;C.Protein hydrophilicity/hydrophobicity prediction.

Fig.3 Expression analysis of GhbHLH149-like in Arabidopsis thaliana transgenic lines A.Detection of transgenic positive plants:M.D2000 plus;CK-.Negative control;CK+.Positive control;B.Detection of gene expression levels in transgenic positive plants.Different lowercase letters indicated significant difference(P<0.05).The same as Fig.4-6.

Fig.4 Analysis of germination rate of Arabidopsis thaliana under salt stress A.Seed germination assay between transgenic lines and wild type;B.Analysis of germination rate.

Fig.5 Analysis of root length of Arabidopsis thaliana under salt stress A.Root elongation assay between transgenic lines and wild type;B.Quantitative analysis of root length.

Fig.6 Phenotypes and physiological indices analysis of Arabidopsis thaliana under salt stress A.Phenotypes of Arabidopsis thaliana under salt stress;B-D.Physiological indices analysis.

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