Wang Hui-Zhong, Zhao Pei-Jie, Xu Ji-Chen, Zhao Huai, Zhang Hong-Sheng
National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China.
Yi Chuan Xue Bao. 2003 Jan;30(1):70-5.
Virus disease is a major cause that affects the quality and output of watermelon which is an important fruit in summer. So it is really urgent to develop disease resistance plants. But it takes a long time to breed such plants in conventional ways, and it is very difficult to get ideal result. With the development of plant genetic engineering, new ways have been found to breed plants with disease resistance. By using plant transgenic technique, much progress was been made in plant improvement. There are many successful cases of transgenic plants against corresponding virus disease through transferring coat protein gene. This paper reports the results of inheritance, segregation, expression of WMV-2 coat protein gene in inbred transgenic watermelon and its resistance to virus. Through PCR analysis of inbred plants, we found WMV-2 coat protein gene in the genome of progeny R1 separated with 3:1. After successive selection and identification of 4 generations, 8 transgenic pure lines with almost the same agronomic traits were obtained from 3 independent transformants of T7, T11 and T32. The result of Western blotting shows all 3 different transgenic lines of R4T7-1, R4T11-3 and R4T32-7 can produce coat protein. Disease resistance experiment on transgenic plants with WMV-2 shows that, compared with the control groups, transgenic plants can delay the disease infection and reduce the incidence and the symptoms of virus disease. And the transgenic line R4T32-7 expressed high resistance to infection by WMV-2, which lays a foundation for breeding of disease resistant varieties through plant transgenic technique.
病毒病是影响西瓜品质和产量的主要因素,西瓜是夏季的重要水果。因此,培育抗病植株迫在眉睫。但用传统方法培育抗病植株耗时较长,且很难取得理想效果。随着植物基因工程的发展,已找到培育抗病植株的新方法。通过植物转基因技术,植物改良取得了很大进展。通过转入外壳蛋白基因培育出抗相应病毒病的转基因植物已有很多成功案例。本文报道了WMV-2外壳蛋白基因在转基因西瓜自交系中的遗传、分离、表达及其抗病毒特性。通过对自交植株的PCR分析,我们发现子代R1基因组中的WMV-2外壳蛋白基因以3:1的比例分离。经过连续4代的筛选和鉴定,从T7、T11和T32这3个独立转化体中获得了8个农艺性状几乎相同的转基因纯系。蛋白质免疫印迹结果表明,R4T7-1、R4T11-3和R4T32-7这3个不同的转基因株系均能产生外壳蛋白。对转WMV-2基因植株的抗病性实验表明,与对照组相比,转基因植株能延缓病害感染,降低病毒病的发病率和症状。转基因株系R4T32-7对WMV-2感染表现出高抗性,这为通过植物转基因技术培育抗病品种奠定了基础。