3种检测番茄抗根结线虫Mi基因分子标记法的比较  

Comparison of three molecular markers for detecting Mi gene of resistance to root-knot nematode in tomato cultivars

作　　者：戴均涛 张慎璞[2] 王暄 丁修恒 李红梅 DAI Juntao;ZHANG Shenpu;WANG Xuan;DING Xiuheng;LI Hongmei(College of Plant Protection/Key Laboratory of Integrated Management of Crop Diseases and Pests,Ministry of Education, Nanjing Agricultural University,Nanjing 210095,China;Henan Vocational College of Agriculture,Zhengzhou 451450,China)

机构地区：[1]南京农业大学植物保护学院/农作物生物灾害综合治理教育部重点实验室,江苏南京210095 [2]河南农业职业学院,河南郑州451450

出　　处：《南京农业大学学报》2018年第5期848-853,共6页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(31371922)

摘　　要：[目的]Mi基因是目前番茄遗传育种及生产实践中应用最为广泛的根结线虫抗源,通过对3种分子标记法检测番茄中Mi基因的结果进行比较与分析,以期为番茄抗根结线虫遗传育种提供准确、高效的分子鉴定方法。[方法]采用酶切扩增多态性序列(CAPS)、引物对组合以及序列特征扩增区(SCAR)3种分子标记法分别检测16个番茄品种的Mi基因及其基因型,同时采用人工接种法测定番茄品种对南方根结线虫的抗、感性。[结果]针对上述品种,3种分子标记法的检测结果存在着明显的差异,CAPS法检测全部供试番茄均含Mi基因,除‘线虫绝39号’外均为Mi/Mi纯合基因型;而PM3Fb/PM3Rb、PMi F3/PMiR3引物对组合检测和SCAR标记Mi23F/Mi23R检测的结果均显示有9个品种含Mi基因,前者检测表明‘VFN’‘双抗265’和‘双抗228’为Mi/Mi纯合基因型,但后者检测则显示仅‘VFN’和‘线虫绝39号’为Mi/Mi纯合基因型。接种测定结果显示:‘VFN’和‘线虫绝3号’为南方根结线虫免疫品种,‘双抗228’‘仙客1号’‘Sparta’和‘线虫绝39号’为高抗品种,‘双抗265’和‘牟番1号’为抗病品种,其余均为感病品种。[结论]CAPS法容易产生假阳性,不适用于番茄Mi基因的检测,而PM3Fb/PM3Rb、PMi F3/PMiR3的引物对组合和SCAR标记Mi23F/Mi23R这2种方法能够相对准确地检测番茄Mi基因,其中SCAR标记经单次PCR反应即可直接鉴定Mi基因的有无及其基因型,应用更为便捷。[Objectives]The Mi gene is the most widely used genetic resource for resistance to root-knot nematodes in tomato breeding and production practice.In order to provide the molecular techniques for accurate and efficient identificaiton of Mi gene in tomato breeding,three different molecular markers were compared and analysed for the detection of Mi gene in tomato cultivars.[Methods]The Mi gene and its genotype in 16 tomato cultivars were detected using the cleaved amplified polymorphic sequences(CAPS),primer pairs combination and sequence characterized amplified regions(SCAR),respectively.Meanwhile,the resistance of these cultivars to Meloidogyne incognita was evaluated by the artificial inoculation test.[Results]Using three different molecular markers revealed the detection of Mi gene in 16 tomato cultivars showing the appearently differences.The detection by CAPS showed that all cultivars contained Mi gene,and all cultivars,except‘Xianchongjue 39’,were homozygous genotype of Mi/Mi.The detection by the combination of markers PM3Fb/PM3Rb and PMiF3/PMiR3 and by the SCAR marker of Mi23F/Mi23R both showed that 9 cultivars contained Mi gene.The former one detected‘VFN’‘Shuangkang 265’and‘Shuangkang 228’having homozygous genotype of Mi/Mi,however,the latter one detected‘VFN’and‘Xianchongjue 39’having homozygous genotype of Mi/Mi.Inoculation test revealed that the cultivars‘VFN’and‘Xianchongjue 3’were immune to M.incognita,‘Shuangkang 228’‘Xianke 1’‘Sparta’and‘Xianchongjue 39’were highly resistant,‘Shuangkang 265’and‘Moufan 1’were resistant,and the rest cultivars were susceptible.[Conclusions]The detection of CAPS easily producted the false-positive results,which is not suitable for applying in the identification of Mi gene in tomato.The combination of markers PM3Fb/PM3Rb and PMiF3/PMiR3 and the SCAR marker of Mi23F/Mi23R both could be used for detecting the Mi gene in tomato with relatively accuracy.The SCAR marker has the advantage of one step PCR instead of two st

关 键 词：根结线虫 Mi基因 分子标记 基因型 抗性评价 番茄 

分 类 号：S432.4[农业科学—植物病理学]