Liu Xiao-Hong, Zhang Hong-Wei, Liu Xin, Liu Xin-Jie, Tan Zhen-Bo, Rong Ting-Zhao
Beijing Agro-biotechnology Research Center, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100089, China.
Sheng Wu Gong Cheng Xue Bao. 2005 Jan;21(1):144-8.
The MDMV (Maize Dwarf Mosaic Virus, MDMV) CP (Coat Protein, CP) gene was cloned by RT-PCR method and introduced into the embryonic calli derived from immature embryos of elite inbred 18-599hong and 18-599bai via particle bombardment. Bombarded calli were selected on selection medium containing 5-10 mg/L (PPT) Bialaphos. From resistant calli, 79 plantlets were regenerated. 18 of 79 were grown and harvested. The results of Southern blotting and PCR analysis demonstrated that MDMV CP have been integrated into the genome of the transgenic plants. PCR-positive progeny plants were artificially inoculated with MDMV strain B, and the average chlorosis of the functional leaves of each plant was investigated. The typical symptoms were observed from the leaves of the control inbreds. while, the presence of the MDMV CP gene provided resistance to inoculation with MDMV strain B.
采用RT-PCR方法克隆了玉米矮花叶病毒(MDMV)的外壳蛋白(CP)基因,并通过粒子轰击法将其导入优良自交系18-599红和18-599白未成熟胚来源的胚性愈伤组织中。在含有5-10mg/L双丙氨膦(PPT)的选择培养基上筛选轰击后的愈伤组织。从抗性愈伤组织中再生出79株植株。79株中有18株生长并收获。Southern杂交和PCR分析结果表明,MDMV CP已整合到转基因植物的基因组中。对PCR阳性后代植株人工接种MDMV B株,调查每株植株功能叶的平均黄化情况。对照自交系的叶片出现典型症状。而MDMV CP基因的存在使植株对接种MDMV B株具有抗性。