探索巴黎多肉变种云南花粉操作在改变种子休眠方面的潜力

Meng Wang, Qiuxia Wang, Xiao Wang, Dingkang Wang, Xudong Yin, Yanwen Qiao, Mingkai Ma, Yanli Du, Bin Wang
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引用次数: 0

摘要

云南白药(Paris polyphylla var.yunnanensis)是一种著名的中药材,它有一个独特的生理特征,即花粉成熟后花药会周期性开闭。本研究旨在探讨这一现象对育种的影响。研究人员利用 RNA 测序和甲基化测序技术,仔细观察并比较了花药开闭和寒冷暴露过程中花粉和种子的基因表达谱和甲基化变化。研究人员检查了富集在《京都基因和基因组百科全书》(KEGG)通路中的基因,以确定花粉和种子中易受温度相关甲基化变化影响的基因簇。使用了四种花粉处理模型,即正常对照、"低温保护花粉"、"刚开放花药的花粉 "和 "闭锁花药的花粉",通过人工授粉生产相应的种子。随后,利用 qRT-PCR 验证了在不同处理情况下授粉种子中标记基因表达模式的变化。通过转录组分析,确定了在花药和正常组织之间表现出明显表达差异的基因,以及与低温处理花粉和种子的甲基化变异有关的基因区域。在氧化磷酸化(ko00190)、植物激素信号转导(Ko04075)和玉米素生物合成(ko00908)这三个信号通路中观察到了趋同性。值得注意的是,发现了易受温度诱导甲基化变化影响的基因簇,如 NADH-ubiquinone 氧化还原酶链 5、质膜 ATPase 4、细胞色素 c 氧化酶亚基 2、顺式玉米素 O-葡萄糖基转移酶、ABSCISIC ACID-INSENSITIVE 5-like protein 4 和吲哚-3-乙酸-氨基合成酶(IAAS)。利用各种花粉授粉模型进行的评估显示,五个休眠调节标记基因的表达模式发生了改变:IAAS、蔗糖合成酶(SUS)、赤霉素 2-oxidase (GA2ox)、ABA INSENSITIVE 2 (ABI2)和辅酶抑制蛋白(ARP)。封闭花药处理导致IAAS、SUS、GA2ox和ABI2的表达显著上调,而ARP的表达明显下降,表明种子休眠倾向延长。相反,在低温保护花药模型中,SUS、ARP、GA2ox和IAAS的表达水平降低,而ABI2的表达上调,总体上促进了种子的萌发。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Exploring the potential of Paris polyphylla var. yunnanensis pollen manipulation in modifying seed dormancy
Paris polyphylla var. yunnanensis, a well-known Chinese medicinal herb, shows a unique physiological trait characterized by the cyclic opening and closing of its anthers after pollen maturation. The aim of this study was to explore the implications of this phenomenon on breeding. RNA sequencing coupled with methylation sequencing was used to scrutinize and compare gene expression profiles and methylation alterations in pollen and seeds during anther opening and closing, along with cold exposure. Genes enriched within Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were examined to identify gene clusters susceptible to temperature-related methylation changes in both pollen and seeds. Four pollen treatment models, namely, normal control, “pollen protected from low temperatures,” “pollen from just-opened anther,” and “pollen from close-blocked anther,” were used to produce corresponding seeds via artificial pollination. Subsequently, qRT-PCR was used to validate modifications in the expression patterns of marker genes in pollinated seeds under diverse treatment scenarios. Genes exhibiting significant differences in expression between anthers and normal tissues, along with gene regions linked to methylation variations attributed to low-temperature-treated pollen and seeds, were identified through transcriptomic analysis. Convergence was observed in three signaling pathways: oxidative phosphorylation (ko00190), plant hormone signal transduction (Ko04075), and zeatin biosynthesis (ko00908). Notably, gene clusters prone to temperature-induced methylation changes, such as NADH-ubiquinone oxidoreductase chain 5, plasma membrane ATPase 4, cytochrome c oxidase subunit 2, cis-zeatin O-glucosyltransferase, ABSCISIC ACID-INSENSITIVE 5-like protein 4, and indole-3-acetic acid-amido synthetase (IAAS), were identified. Evaluation using various pollen pollination models revealed altered expression patterns of five dormancy-regulating marker genes: IAAS, sucrose synthase (SUS), gibberellin 2-oxidase (GA2ox), ABA INSENSITIVE 2 (ABI2), and auxin-repressed protein (ARP), in seeds pollinated with pollen from close-blocked anthers, cold-protected pollen, and pollen from freshly opened anthers. The close-blocked anther treatment led to significantly upregulated expression of IAAS, SUS, GA2ox, and ABI2, whereas ARP expression decreased markedly, indicating a propensity toward prolonged seed dormancy. Conversely, in the low-temperature-protected anther model, SUS, ARP, GA2ox, and IAAS exhibited reduced expression levels, whereas the expression of ABI2 was upregulated, overall facilitating seed germination.
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