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利用 BSA-seq 和转录组测序精细定位棉花株高的主 QTL。

Fine-mapping of a major QTL controlling plant height by BSA-seq and transcriptome sequencing in cotton.

机构信息

State Key Laboratory of Cotton Bio-breeding and Integrated Utilization, Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang, 455000, China.

State Key Laboratory of North China Crop Improvement and Regulation, Key Laboratory for Crop Germplasm Resources of Hebei, Hebei Agricultural University, Baoding, 071000, China.

出版信息

Theor Appl Genet. 2024 Sep 9;137(10):217. doi: 10.1007/s00122-024-04714-w.

Abstract

GhSOT (GH_D05G3950) plays a negative role in regulating plant height development by modulating the GA signaling. Plant height is an important indicator affecting mechanical harvesting for cotton. Therefore, understanding the genes associated with the plant height is crucial for cotton breeding and production. In this study, we used bulk segregant analysis sequencing to identify a new quantitative trait locu (QTL) called qPH5.1, which is linked to plant height. Local QTL mapping using seven kompetitive allele-specific PCR (KASP) markers and linkage analysis successfully narrowed down qPH5.1 to ~ 0.34 Mb region harbored five candidate genes. Subsequently, RNA sequencing (RNA-seq) analysis and examination of expression patterns revealed that GhSOT exhibited the highest likelihood of being the candidate gene responsible for the plant height at this locus. Seven SNP site variations were identified in the GhSOT promoter between the two parents, and Luciferase experiments confirmed that the promoter of GhSOT from cz3 enhances downstream gene expression more effectively. Additionally, suppression of GhSOT in cz3 resulted in the restoration of plant height, further emphasizing the functional significance of this gene. Application of exogenous gibberellin acid (GA) significantly restored plant height in cz3, as demonstrated by RNA-seq analysis and exogenous hormone treatment, which revealed alterations in genes associated with GA signaling pathways. These results reveal GhSOT is a key gene controlling plant height, which may affect plant height by regulating GA signaling.

摘要

GhSOT(GH_D05G3950)通过调节 GA 信号通路在调控植物株高发育中起负调控作用。株高是影响棉花机械收获的一个重要指标。因此,了解与株高相关的基因对棉花的育种和生产至关重要。在本研究中,我们使用 bulk segregant analysis sequencing 鉴定了一个新的与株高相关的数量性状位点(QTL),称为 qPH5.1。利用七个竞争性等位基因特异性 PCR(KASP)标记进行局部 QTL 作图和连锁分析,成功地将 qPH5.1 缩小到大约 0.34 Mb 区域,该区域包含五个候选基因。随后,RNA 测序(RNA-seq)分析和表达模式研究表明,GhSOT 最有可能是该基因座控制株高的候选基因。在两个亲本之间,在 GhSOT 启动子中发现了七个 SNP 位点变异,荧光素酶实验证实来自 cz3 的 GhSOT 启动子能更有效地增强下游基因的表达。此外,在 cz3 中抑制 GhSOT 导致株高的恢复,进一步强调了该基因的功能意义。外源赤霉素(GA)的应用显著恢复了 cz3 的株高,这可以通过 RNA-seq 分析和外源激素处理得到证实,这些处理改变了与 GA 信号通路相关的基因。这些结果表明 GhSOT 是控制株高的关键基因,它可能通过调节 GA 信号通路来影响株高。

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