小麦AL型细胞质雄性不育系与同型保持系穗花发育时期代谢物变化比较研究

doi:10.3724/SP.J.1006.2025.51016

摘要/Abstract

摘要：

研究小麦AL型细胞质雄性不育系与同型保持系穗花发育不同时期代谢组物质变化的关系，为外源激素诱导细胞质雄性不育育性转换提供应用理论依据。选用不育系AL-22A和同型保持系AL-22B为试验材料，用国际法测定不育系套袋自交结实率。在穗发育二棱期、小花原基分化期、雌雄蕊原基分化期、药隔期、四分体期、单核中位期、单核靠边期、二核期、三核期等9个时期取穗样检测。采用液相色谱-质谱联用仪测定5种植物内源激素含量、过氧化物酶(POD)活性、活性氧(ROS)含量、ATP含量及非靶向代谢组学分析代谢物变化。不育系套袋自交结实率为0，达到全不育水平；在穗分化多个发育时期不育系与保持系比较，ABA含量高于保持系，IAA、ZR、GA3含量低于保持系，MeJA含量在穗分化二棱期至四分体期低于保持系，单核期以后高于保持系；在激素平衡关系中，IAA/ABA比值在不育系穗发育各期表现低于保持系，不育系激素比值异常增高或降低均可能引发细胞质雄性不育，其中ABA和MeJA含量增高对激素比值变化的影响最大；生理代谢物ATP和ROS在不育系和保持系穗发育各期表现差异不显著，在穗分化二棱期至四分体期保持系POD活性显著高于不育系，二核期和三核期不育系POD活性高于保持系；非靶向代谢组研究发现，雌雄蕊原基分化期和单核中位期不育系代谢物下调数目多于保持系，其他时期不育系上调代谢物数目多于下调数目，并且达到差异显著。对不育系和保持系穗发育各时期非靶向代谢物表达数目进行聚类分析，可将其分为早期(I~Ⅲ)、中期(Ⅳ~Ⅵ)、晚期(Ⅶ~Ⅸ)3大类群，穗发育不同时期代谢物数目的差异导致了小麦的育性变化；代谢通路富集分析发现，代谢物差异主要集中在小花分化期、药隔期、四分体期、单核靠边期、三核期，代谢物主要以羧酸代谢、次级代谢为主。对小麦育性发育影响较大的时期为早中期(Ⅱ~Ⅳ)和中后期(Ⅵ~Ⅷ)，不同内源激素之间的平衡关系和代谢差异物数量异常可能是导致小麦细胞质雄性不育的主要原因。本研究为进一步揭示小麦细胞质雄性不育穗花发育的生理代谢机制以及探索AL型细胞质雄性不育系从不育转变为生理性可育提供了理论和应用依据。

关键词: 小麦, AL型, 细胞质雄性不育, 内源激素, 代谢组

Abstract:

To investigate the relationship between the AL-type cytoplasmic male sterile (CMS) wheat line and the metabolome of its maintainer line during spikelet development, and to provide a theoretical foundation for the use of exogenous hormones in regulating fertility restoration, the AL-22A sterile line and the AL-22B homotypic maintainer line were used as experimental materials. Self-seed setting rate was determined using international standard methods. Spike samples were collected at nine developmental stages: young panicle differentiation, double ridge, floret primordium differentiation, pistil and stamen differentiation, chamber formation, tetrad, mononuclear median, uninucleate, binucleate, and trinucleate stages. The levels of endogenous hormones, peroxidase (POD), reactive oxygen species (ROS), and ATP were measured using liquid chromatography-mass spectrometry (LC-MS), and metabolites were analyzed through non-targeted metabolomics. The self-seed setting rate of the sterile line was zero, indicating complete sterility. Compared to the maintainer line, the sterile line showed higher levels of abscisic acid (ABA) and lower levels of indole-3-acetic acid (IAA), zeatin riboside (ZR), and gibberellic acid (GA3). Methyl jasmonate (MeJA) content was lower in the sterile line from the double ridge to tetrad stages but higher after the mononuclear median stage. The IAA/ABA ratio was consistently lower in the sterile line, suggesting that hormone imbalance may contribute to sterility, particularly the elevated levels of ABA and MeJA. No significant differences in ATP and ROS levels were observed between the two lines across developmental stages. However, POD activity was significantly higher in the maintainer line from the double ridge to tetrad stages, while the sterile line showed distinct POD levels at the binucleate and trinucleate stages. Non-targeted metabolomic analysis revealed that the sterile line had more downregulated metabolites during the pistil and stamen differentiation and mononuclear median stages, while other stages showed more upregulated than downregulated metabolites, with significant differences. Clustering analysis of metabolite expression patterns across developmental stages grouped samples into early (I–III), middle (IV–VI), and late (VII–IX) phases. Stage-specific differences in metabolite expression were associated with fertility changes. Pathway enrichment analysis indicated that differential metabolites were primarily concentrated in the floret primordium differentiation, chamber formation, tetrad, uninucleate, and trinucleate stages, mainly involving carboxylic acid metabolism and secondary metabolism. The stages most critical to fertility development occurred in early to mid (II–IV) and mid to late (VI–VIII) phases. These results suggest that the interplay between endogenous hormone balance and abnormal metabolite accumulation may be key factors in CMS. This study provides both theoretical insights and practical references for understanding the physiological and metabolic mechanisms of spikelet development in CMS wheat and for developing strategies to promote fertility restoration in AL-type sterile lines.

Key words: wheat, AL-type, CMS, endogenous hormones, metabolome

孔德真, 桑伟, 聂迎彬, 李伟, 徐红军, 李江博, 刘鹏鹏, 田笑明. 小麦AL型细胞质雄性不育系与同型保持系穗花发育时期代谢物变化比较研究[J]. 作物学报, 0, (): 0-.

KONG De-Zhen, SANG Wei, NIE Ying-Bin, LI Wei, XU Hong-Jun, LI Jiang-Bo, LIU Peng-Peng, TIAN Xiao-Ming. Comparative analysis of metabolite changes during young panicle development in wheat AL type cytoplasmic male serile line and homologous maintainers[J]. Acta Agronomica Sinica, 0, (): 0-.

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