通过棉花多组学和功能验证的方法深入解析GhCYP和GhTPS基因在棉酚生物合成途径中的作用

doi:10.1016/j.jia.2024.09.017

Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (5): 1671-1687.DOI: 10.1016/j.jia.2024.09.017

通过棉花多组学和功能验证的方法深入解析GhCYP和GhTPS基因在棉酚生物合成途径中的作用

收稿日期:2024-05-28 修回日期:2024-09-24 接受日期:2024-07-02 出版日期:2025-05-20 发布日期:2024-09-24

Insights into the role of GhCYP and GhTPS in the gossypol biosynthesis pathway via a multiomics and functional-based approach in cotton

Teame Gereziher Mehari^1*, Marijana Skorić^2*, Hui Fang¹, Kai Wang¹, Fang Liu³, Tesfay Araya⁴, Branislav Šiler², Dengbing Yao^1#, Baohua Wang^1#

¹School of Life Sciences, Nantong University, Nantong 226019, China
²Department of Plant Physiology, Institute for Biological Research “Sinisa Stankovic” – National Institute of the Republic of Serbia, University of Belgrade, Belgrade 11108, Serbia
³State Key Laboratory of Cotton Bio-breeding and Integrated Utilization/Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang 455000, China
⁴Department of Soil, Crop and Climate Sciences, University of the Free State, P.O. Box 339, Bloemfontein 9300, South Africa

Received:2024-05-28 Revised:2024-09-24 Accepted:2024-07-02 Online:2025-05-20 Published:2024-09-24
About author:Teame Gereziher Mehari, E-mail: fiamieta21@gmail.com; Marijana Skorić, E-mail: mdevic@ibiss.bg.ac.rs; #Correspondence Baohua Wang, E-mail: bhwang@ntu.edu.cn; Dengbing Yao, E-mail: yaodb@ntu.edu.cn * These authors contributed equally to this study.
Supported by:
We appreciate financial help from the National Key R&D Program of China (2021YFE0101200), the Key Research and Development Project of Jiangsu Province, China (Modern Agriculture, BE2022364), the State Key Laboratory of Cotton Bio-breeding and Integrated Utilization Open Fund, China (CB2024A06). We appreciate the support of the Ministry of Science, Technological Development and Innovation of the Republic of Serbia (451-03-66/2024-03/200007).

摘要/Abstract

摘要：

本研究以一对棉花近等基因系中棉所12有腺体和中棉所12无腺体为材料，通过转录组测序确定与棉酚生物合成相关的潜在基因和代谢途径。我们发现了超过2.35亿个clean reads和1184个差异表达基因（differentially expressed genes，DEGs）。随后，我们进行了加权基因共表达网络分析，发现含有GhTPS（GH_D09G0090）和GhCYP（GH_D05G2016）枢纽基因的白色和黄色模块与棉酚含量有很强的相关性。使用RT-qPCR、病毒诱导的基因沉默（virus-induced gene silencing，VIGS）和靶代谢产物分析证明了GhTPS和GhCYP基因的重要性。与野生型相比，这些基因的沉默导致感染两周后叶片和茎上的腺体减少。此外，通过靶向代谢物分析共鉴定出152种代谢物。差异代谢产物筛选显示，与对照组相比，TRV:GhTPS和TRV:GhCYP植株中分别有12种和18种显著不同的代谢产物；与对照组比较，代谢产物的积累减少。靶代谢产物分析表明，棉酚生物合成的最终产物半棉酚的含量也降低了，这表明这些基因在棉酚生物合成途径中发挥了作用。此外，记录到有腺体和无腺体品系之间棉酚含量存在显著差异。本研究的结果揭示了棉酚含量与GhTPS和GhCYP枢纽基因之间的密切联系，表明它们在棉酚生物合成途径中的作用是减少半棉酚的积累，这可能为棉花棉酚生物合成通路的调控检查点提供新的理解。

Abstract:

Two cotton research institute (CRI) near-isogenic lines, CRI-12 glanded and CRI-12 glandless, were used to pinpoint potential genes and metabolic pathways linked to gossypol biosynthesis through transcriptome sequencing. We discovered more than 235 million clean reads and 1,184 differentially expressed genes (DEGs). Consecutively, we conducted a weighted gene co-expression network analysis and found a strong correlation between white and yellow modules containing GhTPS (GH_D09G0090) and GhCYP (GH_D05G2016) hub genes with the gossypol content. Importance of the GhTPS and GhCYP genes was demonstrated using RT-qPCR, virus-induced gene silencing (VIGS), and target metabolite analysis. Silencing these genes resulted in fewer glands on both leaves and stems two weeks after the infection compared to the wild type. In addition, 152 metabolites were identified through targeted metabolite profiling. Differential metabolite screening revealed 12 and 18 significantly different metabolites in TRV:GhTPS and TRV:GhCYP plants vs. the control group, respectively, showing a reduction in the accumulation of metabolites compared to the control. Content of hemigossypol, the final product of gossypol biosynthesis, was also reduced, as revealed by target metabolite analysis, suggesting the role of these genes in the gossypol biosynthetic pathway. Furthermore, a highly significant difference in gossypol content between the glanded and glandless lines was recorded. Findings of this study reveal a strong link between the gossypol content and GhTPS and GhCYP hub genes, suggesting their role in the gossypol biosynthetic pathway to reduce the accumulation of hemigossypol, which may offer new comprehension into the regulatory checkpoints of the gossypol biosynthesis pathway in cotton.

Key words: cotton , gossypol , near-isogenic lines , transcriptome , target metabolomics , correlation

Teame Gereziher Mehari, Marijana Skorić, Hui Fang, Kai Wang, Fang Liu, Tesfay Araya, Branislav Šiler, Dengbing Yao, Baohua Wang. 通过棉花多组学和功能验证的方法深入解析GhCYP和GhTPS基因在棉酚生物合成途径中的作用[J]. Journal of Integrative Agriculture, 2025, 24(5): 1671-1687.

Teame Gereziher Mehari, Marijana Skorić, Hui Fang, Kai Wang, Fang Liu, Tesfay Araya, Branislav Šiler, Dengbing Yao, Baohua Wang. Insights into the role of GhCYP and GhTPS in the gossypol biosynthesis pathway via a multiomics and functional-based approach in cotton[J]. Journal of Integrative Agriculture, 2025, 24(5): 1671-1687.

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通过棉花多组学和功能验证的方法深入解析GhCYP和GhTPS基因在棉酚生物合成途径中的作用

Insights into the role of GhCYP and GhTPS in the gossypol biosynthesis pathway via a multiomics and functional-based approach in cotton

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