Shah Jasmine M, Raghupathy Vengoji, Veluthambi Karuppannan
Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, 625 021, Tamil Nadu, India.
Biotechnol Lett. 2009 Feb;31(2):239-44. doi: 10.1007/s10529-008-9856-5. Epub 2008 Oct 16.
The 42-kDa endochitinase (cht42) gene from the mycoparasitic fungus, Trichoderma virens, driven by CaMV 35S promoter, was introduced into rice by Agrobacterium-mediated transformation. Eight transgenic plants harboring single copies of complete T-DNA were identified by Southern blot analysis. Homozygous transgenic plants were identified for five lines in the T(1) generation by Southern blot analysis. Homozygous T(2) plants constitutively accumulated high levels of the cht42 transcript, showed 2.4- to 4.6-fold higher chitinase activity in total leaf extract and 1.6- to 1.8-fold higher chitinase activity in the extracellular fluid. Infection assays performed on the homozygous T(2) plants with Rhizoctonia solani showed up to 62% reduction in the sheath blight disease index.
由花椰菜花叶病毒35S启动子驱动的来自寄生真菌绿色木霉的42 kDa内切几丁质酶（cht42）基因，通过农杆菌介导的转化方法导入水稻。通过Southern杂交分析鉴定出8株携带单拷贝完整T-DNA的转基因植株。通过Southern杂交分析在T(1)代中鉴定出5个纯合转基因株系。纯合的T(2)植株组成型积累高水平的cht42转录本，总叶提取物中的几丁质酶活性高2.4至4.6倍，细胞外液中的几丁质酶活性高1.6至1.8倍。用立枯丝核菌对纯合T(2)植株进行的感染试验表明，纹枯病病害指数降低了62%。
