番茄抗病基因Ty-1的CAPS标记及检测

CAPS Marker and Detection of Resistant Gene Ty-1 in Tomato

为了获得番茄抗病基因Ty-1的分子标记,利用特异引物对3份抗病纯合材料(基因型Ty-1/Ty-1)和3份感病纯合材料(基因型ty-1/ty-1)进行PCR扩增,抗病、感病材料均产生400bp的PCR扩增片段,感病和杂合抗病材料的酶切产物存在RsaⅠ酶切位点,酶切后分别产生了350、50bp以及400、350、50bp的片段,而抗病纯合材料的扩增产物无此酶切位点,酶切后仍为400bp的片段。该标记能够区分抗病材料、感病材料及杂合抗病材料,是与番茄黄化曲叶病毒病抗病基因Ty-1紧密连锁的共显性标记。利用该标记对33份番茄F1进行检测,有20份材料含有抗病基因Ty-1,13份材料不含抗病基因Ty-1。利用该标记对国内的24份番茄自交系进行检测,24份材料均不含抗病基因Ty-1。经反复验证,结果准确可靠,该标记可以用于对番茄抗病基因Ty-1的筛选鉴定。

In order to obtain molecular marker of resistant gene Ty-1 in tomato(Lycopersicon esculentum Mill.),3 resistant homozygous lines(Ty-1/Ty-1)and 3 homozygous susceptible lines(ty-1/ty-1)were amplified by specific primer.Both resistant and susceptible lines produced about 400 bp PCR fragment.After digestion with enzyme Rsa I,susceptible genotypes and heterozygous resistant genotypes could produce fragment about 350,50 bp and 400,350,50 bp,respectively.While the resistant genotypes still presented 400 bp fragment.This marker could distinguish resistant and susceptible lines,was a co-dominant marker tightly linked to Ty-1 gene.33 F1 hybrids were detected using this marker,and the results showed that 20 hybrids contained Ty-1 resistant gene and 13 hybrids did not.This marker was also used to detect 24 important inbred lines and all of them did not contain Ty-1 gene.The repeated verification proved that Ty-1 resistant gene could be identified by this marker.