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OsPGIP1 在水稻纹枯病抗性中的功能分析。

Functional analysis of OsPGIP1 in rice sheath blight resistance.

机构信息

National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan, People's Republic of China.

出版信息

Plant Mol Biol. 2015 Jan;87(1-2):181-91. doi: 10.1007/s11103-014-0269-7. Epub 2014 Dec 9.

Abstract

As one of the most devastating diseases of rice, sheath blight causes severe rice yield loss. However, little progress has been made in rice breeding for sheath blight resistance. It has been reported that polygalacturonase inhibiting proteins can inhibit the degradation of the plant cell wall by polygalacturonases from pathogens. Here, we prokaryotically expressed and purified OsPGIP1 protein, which was verified by Western blot analysis. Activity assay confirmed the inhibitory activity of OsPGIP1 against the PGase from Rhizoctonia solani. In addition, the location of OsPGIP1 was determined by subcellular localization. Subsequently, we overexpressed OsPGIP1 in Zhonghua 11 (Oryza sativa L. ssp. japonica), and applied PCR and Southern blot analysis to identify the positive T0 transgenic plants with single-copy insertions. Germination assay of the seeds from T1 transgenic plants was carried out to select homozygous OsPGIP1 transgenic lines, and the expression levels of OsPGIP1 in these lines were analyzed by quantitative real-time PCR. Field testing of R. solani inoculation showed that the sheath blight resistance of the transgenic rice was significantly improved. Furthermore, the levels of sheath blight resistance were in accordance with the expression levels of OsPGIP1 in the transgenic lines. Our results reveal the functions of OsPGIP1 and its resistance mechanism to rice sheath blight, which will facilitate rice breeding for sheath blight resistance.

摘要

作为水稻最具破坏性的疾病之一,纹枯病会导致严重的水稻减产。然而,在培育水稻抗纹枯病方面进展甚微。据报道,多聚半乳糖醛酸酶抑制蛋白可以抑制病原菌来源的多聚半乳糖醛酸酶对植物细胞壁的降解。在这里,我们通过原核表达和纯化了 OsPGIP1 蛋白,并通过 Western blot 分析进行了验证。活性测定证实了 OsPGIP1 对 Rhizoctonia solani 的 PGase 的抑制活性。此外,通过亚细胞定位确定了 OsPGIP1 的位置。随后,我们在 Zhonghua 11(Oryza sativa L. ssp. japonica)中过表达了 OsPGIP1,并应用 PCR 和 Southern blot 分析鉴定了具有单拷贝插入的 T0 转基因植株。对 T1 转基因植株的种子进行萌发试验,以筛选出纯合的 OsPGIP1 转基因系,并通过定量实时 PCR 分析这些系中 OsPGIP1 的表达水平。对 R. solani 接种的田间试验表明,转基因水稻的纹枯病抗性得到了显著提高。此外,转基因系中 OsPGIP1 的表达水平与纹枯病抗性水平一致。我们的结果揭示了 OsPGIP1 的功能及其对水稻纹枯病的抗性机制,这将有助于培育抗纹枯病的水稻。

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