Key Laboratory of Oasis Eco-agriculture, College of Agriculture, Shihezi University, Shihezi 832000, Xinjiang, China.
CSIRO Agriculture and Food, GPO Box 1700, Canberra 2601, Australia.
Int J Biol Macromol. 2024 Oct;278(Pt 2):134856. doi: 10.1016/j.ijbiomac.2024.134856. Epub 2024 Aug 19.
Male sterility is used in the production of hybrid seeds and can improve the breeding efficiency of cotton hybrids. Reactive oxygen species is closely associated with the tapetum and pollen development, but their relationship in cotton male fertility remains unclear. In this study, we comprehensively compared the cytology and proteome of the anthers from an Upland cotton (Gossypium hirsutum) material, Shida 98 (WT), and its nearly-isogenic male sterile line Shida 98A (MS). Cytology indicated delayed PCD in the tapetum and defects in microspores in MS anthers. And further studies revealed disruption of ROS homeostasis. Proteomic analysis identified proteins with differential abundance mainly being related to redox homeostasis, protein folding, and apoptotic signaling pathways. GhAPX1 interacted with GhHSP70 and played a crucial role in the development of cotton anthers. Exogenous application of HSP70 inhibitor increased HO content and decreased the activity of APX1 and pollen viability. The GhAPX1 mutants generated by CRISPR/Cas9-mediated gene editing exhibited premature degradation of the tapetum, significant decrease in pollen viability, and significant increase in HO content. Altogether, our results imply HSP70 and APX1 being the key players jointly regulating male fertility by mediating ROS homeostasis. These results provide insights into the proteins associated with male fertility.
雄性不育被用于杂交种子的生产，可以提高棉花杂种的育种效率。活性氧与绒毡层和花粉发育密切相关，但它们在棉花雄性育性中的关系尚不清楚。在这项研究中，我们综合比较了陆地棉（Gossypium hirsutum）材料石达 98（WT）及其近等基因雄性不育系石达 98A（MS）花药的细胞学和蛋白质组学。细胞学表明，MS 花药绒毡层的程序性细胞死亡（PCD）延迟，小孢子缺陷。进一步的研究揭示了 ROS 平衡的破坏。蛋白质组学分析鉴定出的差异表达蛋白主要与氧化还原平衡、蛋白质折叠和细胞凋亡信号通路有关。GhAPX1 与 GhHSP70 相互作用，在棉花花药发育中起关键作用。HSP70 抑制剂的外源应用增加了 HO 含量，降低了 APX1 的活性和花粉活力。通过 CRISPR/Cas9 介导的基因编辑产生的 GhAPX1 突变体表现出绒毡层的过早降解、花粉活力显著降低和 HO 含量显著增加。总之，我们的结果表明 HSP70 和 APX1 是通过调节 ROS 平衡共同调节雄性育性的关键因子。这些结果为与雄性育性相关的蛋白质提供了新的见解。
