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Cloning and Expression Analysis of Eight Upland Cotton Pentatricopeptide Repeat Family Genes

机译:八个陆地棉五肽重复序列家族基因的克隆与表达分析

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The pentatricopeptide repeat (PPR) gene family is one of the largest gene families in plants. Most PPR genes are localized in mitochondria and chloroplasts functioning in regulation of plant growth and development, fertility restoration for cytoplasmic male sterility (CMS), and stress defense. In this study, using in silico cloning and PCR amplification with degenerate primers based on Arabidopsis PPR genes, we cloned eight new full-length PPR genes encoding protein sequences ranging from 458 to 875 amino acids, with 8 to 16 repetitive PPR elements in upland cotton and all of them lack introns. Expression analysis revealed that eight PPR genes were differently expressed in roots, stems, leaves, and floral buds. As for GhI12, its expression in floral buds at days 3-5 was significantly higher in line 777R (restorer line) than in line 777A (CMS line). Further tests with real-time PCR showed that GhI12 expression peaked at day 3 in 777R, followed by a gradual decline, while its expression fluctuated in 777A, peaking at day 5 and day 13. In addition, Gh155c17 and GhI12 were upregulated under salt stress. This is the first report of upland cotton PPR genes involved in salt stress response.
机译:五肽重复序列(PPR)基因家族是植物中最大的基因家族之一。大多数PPR基因位于线粒体和叶绿体中,可调节植物的生长和发育,恢复胞质雄性不育(CMS)的繁殖力和防御胁迫。在这项研究中,我们使用基于拟南芥PPR基因的简并引物进行计算机克隆和PCR扩增,我们在陆地棉上克隆了8个新的全长PPR基因,它们编码的蛋白质序列范围为458至875个氨基酸,具有8至16个重复的PPR元素他们都缺少内含子。表达分析表明,八个PPR基因在根,茎,叶和花蕾中的表达不同。至于GhI12,其在第3-5天在花蕾中的表达在777R系(恢复系)中明显高于在777A系(CMS系)中。实时PCR的进一步测试显示,GhI12表达在777R的第3天达到峰值,然后逐渐下降,而其表达在777A中波动,在第5天和第13天达到峰值。此外,在盐胁迫下Gh155c17和GhI12上调。 。这是陆地棉PPR基因参与盐胁迫反应的首次报道。

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