番茄抗性基因Ty―1的PCR快速检测.docVIP

番茄抗性基因Ty―1的PCR快速检测 　　摘要：利用国内外已发表的10对标记引物对抗番茄黄化曲叶病毒病纯合系“JZ-108”（Ty-1/Ty-1）、感病纯合系“1712”（ty-1/ty-1）及杂交F1代的Ty-1抗性基因进行PCR扩增筛选，筛选出一对特异性引物SSR47，在抗病纯合材料中产生750 bp的扩增片段，感病材料中产生640 bp的片段，抗病杂合材料中同时产生750 bp和640 bp的扩增片段，标记结果与田间鉴定完全一致，证明该标记能够区分抗病材料、感病材料及杂合抗病材料，是与抗番茄黄化曲叶病毒病基因Ty-1紧密连锁的共显性标记。利用该标记对“JZ-108×1712”F2代的48个单株进行检测，有8株为抗病纯合基因型，19株为感病纯合基因型，21株为抗病杂合基因型，其中抗病纯合株与抗病杂合株田间表现均为抗病。经反复验证，结果准确可靠，该标记可用于对番茄抗病基因Ty-1的快速筛选鉴定。 　　关键词：番茄黄化曲叶病；Ty-1；PCR 　　中图分类号：Q785 文献标识号：A 文章编号：1001-4942（2014）03-0005-05 　　AbstractThe resistant gene Ty-1 of homozygous resistant lines “JZ-108”（Ty-1/Ty-1）， homozygous susceptible lines “1712” （ty-1/ty-1）and their F1 generation was amplified by PCR method using 10 pairs of marker primers published at home and abroad. The results showed that SSR47 was the specific primer which had a 750 bp amplification fragment in homozygous resistant lines， a 640 bp fragment in susceptible lines and both 750 bp and 640 bp fragments in heterozygous resistant lines. The detection results were completely consistent with those of field identification. So this co-dominant marker， tightly linked to Ty-1 gene， could distinguish homozygous and heterozygous resistant lines and susceptible lines. Forty-eight individuals from F2 generation of “JZ-108×1712” were detected. There were 8 homozygous resistant lines， 19 homozygous susceptible lines and 21 heterozygous resistant lines. Both homozygous and heterozygous resistant lines displayed resistance to tomato yellow leaf curl disease in the field. The replicated stable results proved that SSR47 could be used for rapid identification of Ty-1 resistant gene in tomato. 　　Key wordsTomato yellow leaf curl disease；Ty-1；PCR 　　番茄（Solanum lycopersicum）是中国种植面积最大的蔬菜作物之一，它品种多、产量高、营养丰富、用途广泛，深受消费者的喜爱。目前病毒病是番茄生产中最常见的病害之一，其分布极为广泛，危害严重，容易造成番茄品质下降、产量锐减。其中番茄黄化曲叶病毒病（Tomato Yellow Leaf Curl Virus Disease， TYLCVD）影响最为严重，已成为世界番茄生产的限制性因素[1]，一旦发病很难被控制。 　　番茄黄化曲叶病毒病于20世纪30年代末在以色列首次被发现[2]，是一类由烟粉虱（Bemisia tabaci）传播的双生病毒（Geminiviruses），为菜豆金色花叶病毒属（Begonurvints）[3]