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从小麦中克隆一个新基因并转化以提高其耐盐性的特性分析

Characterization of a Novel Gene From and Its Transformation in Wheat to Enhance Salt Tolerance.

作者信息

Yang Zhifen, Mu Yuanhang, Wang Yiqin, He Fang, Shi Luxi, Fang Zhongming, Zhang Jun, Zhang Qingqin, Geng Guangdong, Zhang Suqin

机构信息

College of Agriculture, Guizhou University, Guiyang, China.

Guizhou Subcenter of National Wheat Improvement Center, Guiyang, China.

出版信息

Front Plant Sci. 2022 Apr 4;13:830848. doi: 10.3389/fpls.2022.830848. eCollection 2022.

Abstract

Late embryogenesis-abundant (LEA) proteins are critical in helping plants cope with salt stress. "Y1805" is a salt-tolerant We identified a "Y1805"-specific LEA gene that was expressed highly and sensitively under salt stress using transcriptome analysis. The novel group 2 LEA gene () was cloned from "Y1805." contained a 453 bp open reading frame encoding an 151-amino-acid protein that showed maximum sequence identity (77.00%) with by phylogenetic analysis. It was mainly found to be expressed highly in the roots by qRT-PCR analysis and was located in the whole cell. Forty-eight candidate proteins believed to interact with TtLEA2-1 were confirmed by yeast two-hybrid analysis. These interacting proteins were mainly enriched in "environmental information processing," "glycan biosynthesis and metabolism," and "carbohydrate metabolism." Protein-protein interaction analysis indicated that the translation-related 40S ribosomal protein SA was the central node. An efficient wheat transformation system has been established. A coleoptile length of 2 cm, an cell density of 0.55-0.60 OD, and 15 KPa vacuum pressure were ideal for common wheat transformation, with an efficiency of up to 43.15%. Overexpression of in wheat "1718" led to greater height, stronger roots, and higher catalase activity than in wild type seedlings. conferred enhanced salt tolerance in transgenic wheat and may be a valuable gene for genetic modification in crops.

摘要

胚胎后期丰富(LEA)蛋白在帮助植物应对盐胁迫方面至关重要。“Y1805”是一种耐盐品种。我们通过转录组分析鉴定了一个在盐胁迫下高表达且敏感表达的“Y1805”特异性LEA基因。从“Y1805”中克隆了新的第2组LEA基因()。该基因包含一个453 bp的开放阅读框,编码一个151个氨基酸的蛋白质,通过系统发育分析显示与的最大序列同一性为77.00%。通过qRT-PCR分析发现它主要在根中高表达,并且定位在整个细胞中。通过酵母双杂交分析确认了48个被认为与TtLEA2-1相互作用的候选蛋白。这些相互作用的蛋白主要富集在“环境信息处理”、“聚糖生物合成与代谢”和“碳水化合物代谢”中。蛋白质-蛋白质相互作用分析表明,与翻译相关的40S核糖体蛋白SA是中心节点。已经建立了高效的小麦转化系统。对于普通小麦转化,2 cm的胚芽鞘长度、0.55 - 0.60 OD的细胞密度和15 KPa的真空压力是理想的,转化效率高达43.15%。在小麦“1718”中过表达导致其株高更高、根系更强且过氧化氢酶活性高于野生型幼苗。在转基因小麦中赋予了增强的耐盐性,可能是作物遗传改良的一个有价值的基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4023/9014267/3c0ef538c6b1/fpls-13-830848-g001.jpg

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