Richa Kamboj, Tiwari Ila M, Devanna B N, Botella Jose R, Sharma Vinay, Sharma Tilak R
National Research Centre on Plant BiotechnologyNew Delhi, India.
Department of Bioscience and Biotechnology, Banasthali UniversityBanasthali, India.
Front Plant Sci. 2017 Apr 25;8:596. doi: 10.3389/fpls.2017.00596. eCollection 2017.
Sheath blight disease (ShB), caused by the fungus Kühn, is one of the most destructive diseases of rice ( L.), causing substantial yield loss in rice. In the present study, a novel rice chitinase gene, was cloned from QTL region of 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 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 . 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.
纹枯病（ShB）由真菌库恩氏菌引起，是水稻（Oryza sativa L.）最具毁灭性的病害之一，会导致水稻大幅减产。在本研究中，从耐纹枯病水稻品系特特普的QTL区域克隆了一个新的水稻几丁质酶基因，并通过对纹枯病敏感的粳稻品系台北309（TP309）进行遗传转化来进行功能验证。利用分子和功能方法对转化体进行了表征。使用一组针对花椰菜花叶病毒35S启动子、几丁质酶和基因的特异性引物进行PCR分子分析，证实了转基因植物中存在转基因，这通过Southern杂交进一步得到验证。此外，对转基因植物的qRT-PCR分析表明，转基因表达与转化体中纹枯病抗性水平之间存在良好的相关性。功能互补试验证实了几丁质酶介导的抗性在所有对纹枯病具有不同程度增强抗性的转基因TP309植物中的有效性。因此，本研究从水稻QTL区域克隆并表征的新几丁质酶基因在水稻抗纹枯病分子植物育种计划中将具有重要用途。
