摘要
应用本室合成、克隆的马铃薯卷叶病毒（Ｐｏｔａｔｏｌｅａｆｒｏｌｌｖｉｒｕｓ，ＰＬＲＶ）中国分离株的外壳蛋白（ＣＰ）基因，通过致瘤农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍｔｕｍｅｆａｃｉｅｎｓ）介导，以马铃薯叶园片为转化材料，转化了马铃薯Ｄｅｓｉｒｅｅ、Ｆａｖｏｒｉｔａ、虎头和乌盟６０１，并获得了上述四个栽培种的转基因植株，成功地建立了一种简单、快速和效率高的转化体系，卡那霉素抗性、ＰＣＲ扩增目的基因、Ｓｏｕｔｈｅｍ和Ｎｏｒｔｈｅｍｂｌｏｔ分析证明，ＰＬＲＶ外壳蛋白基因已经稳定整合进入转基因马铃薯的染色体组中，并在转录水平上得到表达。窄缝转移分析（Ｓｌｏｔｂｌｏｔａｎａｌｙｓｉｓ）表明，每个转化体四倍体基因组中含有１－５个ＣＰ基因拷贝。转基因植物的接种试验证明，转基因工程植株中的ＰＬＲＶ滴度比未转基因的对照植株显著低。蚜虫传播试验证明，转基因植株可减少蚜虫在田间传播ＰＬＲＶ的效率，限制了ＰＬＲＶ的传播，减少田间植物发病的机会。
he coat protein gene of potato leafroll virus(PLRV)chinese isolate was intioduced intothe commercial potato cultivars Desiree,Favorita, Wuineng-601 and 'Tiger head' viaAgrobacterium tumefaciens mediated gene transfer. Kanamycin resistance, PCR amplification,Southern and Northern blot analysis of transgenic potato plants showed that the coat proteingene of PLRV was integrated into the genome of potato plants and expressed at the level oftranscription. The transgenic lines of 4 potato cultivars were obtained. Slot blot analysisshowed that in transgenic plants of Desiree there were 1-5 copies of PLRV CP gene inpotato tetraploid genome. Inoculation tests of transgenic lines with PLRV by aphids showedthat the PLRV titer in transgenic potato lines was decreased by 67-86% in comparison withnontransgenic potato plants, indicating that in transgenic potato plants, PLRV multiplicationwas considerably reduced。Tuber yield analysis showed that the average tuber yield per plantof most transgenic clones was increased when compared with control potato plants. Aphidtransmission tests revealed that the aphid transmission effciency from transgenic plants to otherplants was decreased,thus the capacity of field transmission of PLRV by aphids is reduced.
出处
《病毒学报》
CAS
CSCD
北大核心
1995年第4期342-350,共9页
Chinese Journal of Virology
基金
国家自然科学基金
内蒙自然科学基金
关键词
基因工程
马铃薯
马铃薯卷叶病毒
抗病性
Engineered resistance, Potato leafroll virus, Coat protein gene, Transgenic
