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大豆疫霉感染期间大豆中一个诱导表达的病程相关蛋白10基因(GmPR10)的分离与鉴定

Isolation and characterization of a pathogenesis-related protein 10 gene (GmPR10) with induced expression in soybean (Glycine max) during infection with Phytophthora sojae.

作者信息

Xu Pengfei, Jiang Liangyu, Wu Junjiang, Li Wenbin, Fan Sujie, Zhang Shuzhen

机构信息

Soybean Research Institute, Key Laboratory of Soybean Biology of Chinese Education Ministry, Northeast Agricultural University, Harbin, 150030, Heilongjiang, People's Republic of China.

出版信息

Mol Biol Rep. 2014 Aug;41(8):4899-909. doi: 10.1007/s11033-014-3356-6. Epub 2014 Apr 16.

Abstract

In previous study, a cDNA library enriched for mRNAs encoding ESTs that increased in abundance during infection with Phytophthora sojae was constructed by suppression subtractive hybridization from leaf tissues of a high resistant soybean, and an EST homologous to the class 10 of pathogenesis-related (PR) proteins was identified to be up-regulated by microarray and real-time PCR. Here, the full-length cDNA (termed GmPR10, GenBank accession number FJ960440; ADC31789.1) of the EST was isolated by rapid amplification of cDNA ends, and contains an open reading frame of 474 bp. The GmPR10 protein included a "P-loop'' motif. The constitutive transcript abundance of GmPR10 in soybean was the highest in leaves, followed by roots and stems. Further analysis showed that GmPR10 mRNA abundance was increased during infection with P. sojae following leaf treatments with gibberellin (GA3), hydrogen peroxide (H2O2), salicylic acid (SA), and abscisic acid (ABA). The dialytically renatured GmPR10 protein significantly inhibited P. sojae hyphal growth and exhibited RNase activity. Transgenic tobacco and soybean plants overexpressing GmPR10 showed increased resistance to P. nicotianae Breda and P. sojae, respectively. These results suggest that the GmPR10 protein plays an important role in host defense against P. sojae infection. To the best of our knowledge, this is the first report on the functional characterization of a PR10 protein from soybean in defense against P. sojae.

摘要

在先前的研究中,通过抑制性消减杂交技术,从高抗大豆的叶片组织构建了一个富含编码在大豆疫霉感染期间丰度增加的ESTs的cDNA文库,并通过微阵列和实时PCR鉴定出一个与病程相关(PR)蛋白第10类同源的EST上调。在此,通过cDNA末端快速扩增分离出该EST的全长cDNA(命名为GmPR10,GenBank登录号FJ960440;ADC31789.1),其包含一个474bp的开放阅读框。GmPR10蛋白包含一个“P环”基序。大豆中GmPR10的组成型转录本丰度在叶片中最高,其次是根和茎。进一步分析表明,在用赤霉素(GA3)、过氧化氢(H2O2)、水杨酸(SA)和脱落酸(ABA)处理叶片后,大豆疫霉感染期间GmPR10 mRNA丰度增加。透析复性的GmPR10蛋白显著抑制大豆疫霉菌丝生长并表现出RNase活性。过表达GmPR10的转基因烟草和大豆植株分别对烟草疫霉和大豆疫霉表现出增强的抗性。这些结果表明,GmPR10蛋白在宿主抵御大豆疫霉感染中起重要作用。据我们所知,这是关于大豆中PR10蛋白在抵御大豆疫霉方面功能特征的首次报道。

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