陆地棉GhSHA1基因的特征分析及其功能验证

doi:10.6048/j.issn.1001-4330.2025.04.003

摘要/Abstract

摘要：

【目的】分析陆地棉GhSHA1基因的特征并验证其功能,为棉花理想株型育种提供基因资源和一定的理论参考。【方法】以陆地棉品种新陆早33号为材料,以拟南芥SHA1基因为探针通过BLAST对比,得到陆地棉GhSHA1基因,进一步利用VIGS、qRT-PCR和过表达技术,探究GhSHA1基因对棉花株高发育的影响。【结果】GhSHA1基因在棉花茎杆中特异性高表达,GhSHA1是一种偏好以A/T密码子结尾的疏水性跨膜蛋白编码基因。利用VIGS技术在陆地棉中沉默GhSHA1基因,沉默植株与注射空载植株相比,株高显著降低(P<0.05)。构建GhSHA1过表达载体转化拟南芥后发现,过表达株系与野生型相比,株高极显著增高(P<0.01)。同时,拟南芥GhSHA1过表达植株中,株高及顶端分生组织发育相关基因WUS表达极显著升高(P<0.01),CLV3基因表达显著降低(P<0.05),但CLV1和CLV2基因表达无明显变化。【结论】GhSHA1基因通过增强SAM相关基因的表达,从而促进棉花株高发育。

关键词: 棉花; 株高; VIGS; 过表达

Abstract:

【Objective】 The plant height of cotton can affect crop yield by affecting plant density and community photosynthesis efficiency. Therefore, cultivating ideal plant types of cotton is of great significance for improving cotton yield. 【Methods】 In current study, the GhSHA1 genes of upland cotton were obtained by BLAST comparison with the SHA1 genes of Arabidopsis thaliana. The effects of GhSHA1 genes on plant height development of cotton were investigated by VIGS, qRT-PCR and overexpression techniques. 【Results】 The results showed that GhSHA1 gene were highlyexpressed in the stem. GhSHA1 was a hydrophobic transmembrane protein with a preference for codons ending in A/T. GhSHA1 gene was silenced by VIGS technology in upland cotton, and the plant height of silenced plants was significantly lower than that of empty pTRV2:00 plants (P<0.01). GhSHA1 overexpression vector was constructed and transformed into Arabidopsis thaliana. Compared with the control wild type, the plant height of the overexpressed lines was significantly increased (P<0.01). Meanwhile, GhSHA1 overexpression in Arabidopsis thaliana promoted the expression of WUS gene (P<0.01), decreased the expression of CLV3 gene (P<0.01),but had no effect on the expression of CLV1 and CLV2. 【Conclusion】 In summary, the results indicates that the GhSHA1 gene promotes cotton plant height development by enhancing the expression of SAM related genes. The study of this provides theoretical basis and technical support for improving the growth and development status of cotton.

Key words: cotton; plant height; VIGS; overexpression

中图分类号:

S562

李虎情, 邵冬南, 张翼, 刘峰, 张新宇, 李艳军, 孙杰, 杨永林, 薛飞. 陆地棉GhSHA1基因的特征分析及其功能验证[J]. 新疆农业科学, 2025, 62(4): 800-806.

LI Huqing, SHAO Dongnan, ZHANG Yi, LIU Feng, ZHANG Xinyu, LI Yanjun, SUN Jie, YANG Yonglin, XUE Fei. Bioinformatics analysis and functional verification of GHSHA1 gene in Gossypium hirsutum L.[J]. Xinjiang Agricultural Sciences, 2025, 62(4): 800-806.

图/表 4

表1 引物序列

Tab.1 Primer sequence

引物名称 Primer name	引物序列(5’ -3’) Primer sequence(5’ -3’)	用途 Application
GhSHA1	F: GAATTCTTTTGGGCATCACTCCTCCC	VIGS
GhSHA1	R: GGTACCTCACCACTGGCAGTCCTTTC	VIGS
GhSHA1	F:TCTGTGGAAATCCTGCAAAAAC	qRT-PCR
GhSHA1	R:TCACTTAGTCGTTGCCTCCAAA	qRT-PCR
AtActin	F:GGTAACATTGTGCTCAGTGGTGG	qRT-PCR(内参)
AtActin	R:AACGACCTTAATCTTCATGCTGC	qRT-PCR(内参)
GhUBQ7	F:GAAGGCATTCCACCTGACCAAC	qRT-PCR(内参)
GhUBQ7	R:CTTGACCTTCTTCTTCTTGTGCTTG	qRT-PCR(内参)
AtCLV1	F:CATCGCCCCAGAGTATGCAT	qRT-PCR
AtCLV1	R:CAGTCAACCTCGGGTCAACA	qRT-PCR
AtCLV2	F: TCCAACGCATCAAGCTCGGG	qRT-PCR
AtCLV2	R:ACGCCTCTCGTCCGTCTCTT	qRT-PCR
AtCLV3	F:TGTCCGGTCCAGTTCAACAA	qRT-PCR
AtCLV3	R:CTCCCGAAATGGTAAAACCG	qRT-PCR
AtWUS	F:ACAAGCCATATCCCAGCTTCA	qRT-PCR
AtWUS	R:CCACCGTTGATGTGATCTTCA	qRT-PCR

图1 GhSHA1 基因的组织特异性表达分析与多重序列比对

Fig.1 Tissue-specific expression analysis and multiple sequence alignment of gene GhSHA1

图2 GhSHA1 基因沉默下棉花株高的变化注:;A、GhSHA1 基因VIGS植株株高;B、pTRV2::GhSHA1基因沉默后株高变化;C、pTRV2::GhSHA1基因沉默表达效率

Fig.2 Changes in cotton plant height under bene GhSHA1 silencing Note:A、GhSHA1 gene VIGS Plant height;BpTRV2::GhSHA1 Chantes in Plant height after gene silencing;C、pTRV2::GhSHA1 gene silencing expression efficiency

图3 GhSHA1 基因过表达下棉花株高的变化

Fig.3 Changes in plant height of cotton plants with overexpression of gene GhSHA1

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