Rohini V K., Sankara Rao K
Department of Biochemistry, Indian Institute of Science, 560012, Karnataka, Bangalore, India
Plant Sci. 2001 Apr;160(5):889-898. doi: 10.1016/s0168-9452(00)00462-3.
Fertile transgenic plants of peanut (Arachis hypogaea L.) cv. TMV-2 expressing tobacco chitinase and neomycin phosphotransferase (npt II) genes were generated using an Agrobacterium tumefaciens (LBA4404/pBI121-pBTex)-mediated transformation system. A tissue culture-independent method wherein embryo in the mature seed is inoculated and reared into single plant transformant was used for transformation. Southern blot analysis of genomic DNA isolated from T(0) transformants and progeny plants (T(1)) demonstrated that the transgenes are stably integrated in the genome of transgenic peanut plants and inherited by the offspring. The expression of the heterologous chitinase gene driven by CaMV 35S promoter led to a high level of activity in some of the transgenic plants. Small-scale field tests indicated increased ability of these plants to resist the fungal pathogen Cercospora arachidicola (the causal organism of the leaf spot or Tikka disease of peanut), which is an important peanut pathogen. These results suggest that a heterologous chitinase gene was functional in peanut and expressed in healthy plants. The study also shows that peanut plants containing transgenically increased activity of chitinase were resistant to attack by the fungal pathogen C. arachidicola to different degrees. The strategy employed may be useful for the control of other fungal diseases of the crop.
利用根癌农杆菌(LBA4404/pBI121-pBTex)介导的转化系统,培育出了表达烟草几丁质酶和新霉素磷酸转移酶(npt II)基因的花生(Arachis hypogaea L.)品种TMV-2的可育转基因植株。采用一种不依赖组织培养的方法进行转化,即对成熟种子中的胚进行接种并培育成单株转化体。对从T(0)代转化体和子代植株(T(1))中分离的基因组DNA进行Southern杂交分析表明,转基因稳定整合到转基因花生植株的基因组中并可由后代遗传。由CaMV 35S启动子驱动的异源几丁质酶基因在一些转基因植株中表现出高水平的活性。小规模田间试验表明,这些植株抵抗花生重要病原菌花生尾孢菌(花生叶斑病或蒂卡病的致病生物)的能力增强。这些结果表明,异源几丁质酶基因在花生中具有功能并在健康植株中表达。该研究还表明,几丁质酶活性转基因增强的花生植株对花生尾孢菌的侵染具有不同程度的抗性。所采用的策略可能有助于防治该作物的其他真菌病害。