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Abstract：
Sulfotransferases (SOTs) (EC 2.8.2.-) are sulfate regulatory proteins in a variety of organisms that have been previously shown to be involved in regulating a variety of physiological and biological processes, such as growth, development, adaptation to land, stomatal closure, drought tolerance, and response to pathogen infection. However, there is a lack of comprehensive identification and systematic analysis of SOT in cotton, especially in G. barbadense. In this study, we used bioinformatics methods to analyze the structural characteristics, phylogenetic relationships, gene structure, expression patterns, evolutionary relationships, selection pressure and stress response of SOT gene family members in G. barbadense. In this study, a total of 241 SOT genes were identified in four cotton species, among which 74 SOT gene members were found in G. barbadense. According to the phylogenetic tree, 241 SOT protein sequences were divided into five distinct subfamilies. We also mapped the physical locations of these genes on chromosomes and visualized the structural information of SOT genes in G. barbadense. We also predicted the cis-acting elements of the SOT gene in G. barbadense, and we identified the repetitive types and collinearity analysis of SOT genes in four cotton species. We calculated the Ka/Ks ratio between homologous gene pairs to elucidate the selective pressure between SOT genes. Transcriptome data were used to explore the expression patterns of SOT genes, and then qRT-PCR was used to detect the expression patterns of GBSOT4, GBSOT17 and GBSOT33 under FOV stress. WGCNA (weighted gene co-expression network analysis) showed that GB_A01G0479 (GBSOT4) belonged to the MEblue module, which may regulate the resistance mechanism of G. barbadense to FOV through plant hormones, signal transduction and glutathione metabolism. In addition, we conducted a VIGS (virus-induced gene silencing) experiment on GBSOT4, and the results showed that after FOV inoculation, the plants with a silenced target gene had more serious leaf wilting, drying and cracking than the control group, and the disease index of the plants with the silenced target gene was significantly higher than that of the control group. This suggests that GBSOT4 may be involved in protecting the production of G. barbadense from FOV infection. Subsequent metabolomics analysis showed that some flavonoid metabolites, such as Eupatorin-5-methylether (3\'-hydroxy-5,6,7,4\'-tetramethoxyflavone, were accumulated in cotton plants in response to FOV infection.
摘要：
磺基转移酶(SOT)(酶代码EC2.8.2。-）是先前已被证明参与调节多种生理和生物过程的多种生物体中的硫酸盐调节蛋白，比如增长，发展,适应土地,气孔关闭，耐旱性,以及对病原体感染的反应。然而,缺乏对棉花中SOT的全面识别和系统分析，尤其是在G.Barbadense.在这项研究中,我们使用生物信息学方法来分析结构特征，系统发育关系，基因结构，表达模式，进化关系，G.barbadenseSOT基因家族成员的选择压力和应激反应。在这项研究中,在四个棉花品种中总共鉴定出241个SOT基因，其中在G.barbadense中发现了74个SOT基因成员。根据系统发育树,241个SOT蛋白序列分为五个不同的亚家族。我们还绘制了这些基因在染色体上的物理位置，并可视化了G.barbadense中SOT基因的结构信息。我们还预测了G.barbadense中SOT基因的顺式作用元件，并对4种棉花的SOT基因进行了重复类型和共线性分析。我们计算了同源基因对之间的Ka/Ks比率，以阐明SOT基因之间的选择压力。转录组数据用于探索SOT基因的表达模式，然后使用qRT-PCR检测FOV胁迫下GBSOT4，GBSOT17和GBSOT33的表达模式。WGCNA(加权基因共表达网络分析)显示GB_A01G0479(GBSOT4)属于MEblue模块,这可能是通过植物激素调节G.barbadense对FOV的抗性机制，信号转导和谷胱甘肽代谢。此外,我们在GBSOT4上进行了VIGS(病毒诱导基因沉默)实验,结果表明,目标基因沉默的植物叶片枯萎更严重，干燥和开裂比对照组，目的基因沉默植物的病情指数明显高于对照组。这表明GBSOT4可能参与保护G.barbadense的生产免受FOV感染。随后的代谢组学分析表明,一些类黄酮代谢产物,如Eupatorin-5-甲基醚（3'-羟基-5,6,7,4'-四甲氧基黄酮，在响应FOV感染的棉花植物中积累。
