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Novel Chitinase Gene LOC_Os11g47510 from Indica Rice Tetep Provides Enhanced Resistance against Sheath Blight Pathogen Rhizoctonia solani in Rice

机译:来自In稻Tetep的新型几丁质酶基因LOC_Os11g47510提供了对水稻纹枯病病原体根瘤菌的增强抗性

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摘要

Sheath blight disease (ShB), caused by the fungus Rhizoctonia solani Kühn, is one of the most destructive diseases of rice (Oryza sativa L.), causing substantial yield loss in rice. In the present study, a novel rice chitinase gene, LOC_Os11g47510 was cloned from QTL region of R. solani tolerant rice line Tetep and used for functional validation by genetic transformation of ShB susceptible japonica rice line Taipei 309 (TP309). The transformants were characterized using molecular and functional approaches. Molecular analysis by PCR using a set of primers specific to CaMv 35S promoter, chitinase and HptII genes confirmed the presence of transgene in transgenic plants which was further validated by Southern hybridization. Further, qRT-PCR analysis of transgenic plants showed good correlation between transgene expression and the level of sheath blight resistance among transformants. Functional complementation assays confirmed the effectiveness of the chitinase mediated resistance in all the transgenic TP309 plants with varying levels of enhanced resistance against R. solani. Therefore, the novel chitinase gene cloned and characterized in the present study from the QTL region of rice will be of significant use in molecular plant breeding program for developing sheath blight resistance in rice.
机译:由真菌Rhizoctonia solaniKühn引起的鞘枯病(ShB)是水稻(Oryza sativa L.)最具破坏性的疾病之一,导致水稻的大量减产。在本研究中,从茄红耐性水稻品系Tetep的QTL区域克隆了水稻几丁质酶新基因LOC_Os11g47510,并通过对ShB易感粳稻台北309(TP309)的遗传转化用于功能验证。使用分子和功能方法表征转化体。使用一组对CaMv 35S启动子,几丁质酶和HptII基因具有特异性的引物,通过PCR进行的分子分析证实了转基因植物中转基因的存在,这一点已通过Southern杂交进一步验证。此外,对转基因植物的qRT-PCR分析显示,转基因中转基因表达与鞘枯病抗性水平之间具有良好的相关性。功能互补测定法证实了几丁质酶介导的抗性在所有转基因TP309植物中具有不同水平的对茄茄根瘤菌的增强抗性。因此,本研究从水稻的QTL区域克隆并鉴定的新型几丁质酶基因将在分子植物育种程序中发展水稻的抗白叶枯病的能力。

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