Liau Chia-Hui, Lu Jian-Cheng, Prasad Venkatesh, Hsiao Hsin-hao, You Su-Juan, Lee Jent-turn, Yang Ning-Sun, Huang Hsiang-En, Feng Teng-Yung, Chen Wen-Huei, Chan Ming-Tsair
Institute of BioAgricultural Sciences, Academia Sinica, Taiwan 11529, Republic of China.
Transgenic Res. 2003 Jun;12(3):329-36. doi: 10.1023/a:1023343620729.
Genetic engineering to date has not been used to introduce disease resistance genes into the orchid gene pool. The ferredoxin-like protein gene originally isolated from sweet pepper is thought to function as a natural defense against infection due to its antimicrobial properties. Hence it was reasoned that introduction of this gene might produce Oncidium plants resistant to Erwinia carotovora, the causal agent for the soft rot disease. An expression vector containing sweet pepper ferredoxin-like protein (pflp) cDNA, hph and gusA coding sequence was successfully transformed into protocorm-like bodies (PLBs) of Oncidium orchid, using Agrobacterium tumefaciens strain EHA105. A total of 17 independent transgenic orchid lines was obtained, out of which six transgenic lines (beta-glucuronidase (GUS) positive) were randomly selected and confirmed by Southern, northern and western blot analyses. A bioassay was conducted on the transgenic lines. Transgenic plants showed enhanced resistance to E. carotovora, even when the entire plant was challenged with the pathogen. Our results suggest that pflp may be an extremely useful gene for genetic engineering strategies in orchids to confer resistance against soft rot disease.
迄今为止，基因工程尚未被用于将抗病基因导入兰花基因库。最初从甜椒中分离出的铁氧还蛋白样蛋白基因，因其抗菌特性，被认为具有天然的抗感染防御功能。因此，人们推断导入该基因可能会培育出对胡萝卜软腐欧文氏菌（软腐病的致病因子）具有抗性的文心兰植株。利用根癌农杆菌EHA105菌株，将含有甜椒铁氧还蛋白样蛋白（pflp）cDNA、hph和gusA编码序列的表达载体成功转化到文心兰类原球茎（PLB）中。共获得了17个独立的转基因兰花株系，从中随机选取6个转基因株系（β-葡萄糖醛酸酶（GUS）呈阳性），并通过Southern、Northern和Western印迹分析进行了确认。对转基因株系进行了生物测定。即使整株植物受到病原菌的挑战，转基因植株对胡萝卜软腐欧文氏菌的抗性也有所增强。我们的结果表明，pflp可能是兰花基因工程策略中用于赋予软腐病抗性的一个极其有用的基因。
