一年生辣椒(Cpsicum annuum L.)与中华辣椒(Cpsicum chinense Jacquin)DNA甲基化多样性分析  

Surveying DNA Methylation Diversity in the Capsicum annuum L.and Cpsicum chinense Jacquin

作　　者：李涛[1] 徐小万[1] 李颖[1] 李明珠[1] 王恒明[1] 徐晓美[1] 罗少波[1] 

机构地区：[1]广东省农科院蔬菜研究所,广州510640

出　　处：《分子植物育种》2014年第2期306-315,共10页Molecular Plant Breeding

基　　金：农业部948项目(2012-Z55);广东省科技计划项目(2010B020304001);广东省农科院院长基金项目(201108);国家大宗蔬菜产业技术体系广州综合试验站(2012B061800053)共同资助

摘　　要：本研究调查了一年生辣椒和中华辣椒种的DNA甲基化多样性。选取了24个一年生辣椒栽培种和6个中华辣椒栽培种作为研究对象，采用MSAP（methylation—sensitiveamplificationpolymorphism）技术对其基因组CCGG位点的甲基化多样性进行了分析。结果表明，从63对MSAP引物中筛选出5对重复性好、条带清晰的引物对，对30份辣椒种质基因组DNA进行MSAP扩增，得到939条带谱，其中多态性条带937个，多态性比例高达99．79％。DNA甲基化模式分析表明，类型Ⅰ为非甲基化带型（3721），类型Ⅱ为半甲基化带型（3152），类型Ⅲ为全甲基化带型（3839），辣椒甲基化模式主要以全甲基化为主，一年生辣椒（capsicumannltumL．）和中华辣椒（CapsicumchinenseJacquin）甲基化条带平均分别为319和217，其扩增位点的甲基化率分别77．24％和63．64％，差异达极显著水平。基于相似性系数的UPGMA法聚类分析，在0．68相似水平可以将30个种质分为3个类，第1类：No．10-No．13和No．15共5个种质，其中No．10为一年生辣椒，其它4个为中华辣椒；第2类：No．2-No．9、No．14、No．16-No．30，其中No．14和No．17为中华辣椒；第3类：只有No．1，为一年生辣椒。应用MSAP未能将一年生辣椒和中华辣椒区分开来，表明辣椒表观遗传十分丰富。Nei’s基因多态性指数（h）和Shannon信息指数（I）在2个栽培种中分别为0．2040和O．3298、0．1864和0．2975，一年生辣椒的表观遗传多样性稍高于中华辣椒。DNA甲基化多样性作为标志遗传多样性的一种信号源，本研究结果为深入探讨辣椒种群分化及物种进化奠定了基础。Intra- and inter-population methylation diversity at CCGG sites were surveyed in the genomes of C. annuum L. and C. chinense Jacquin using the MSAP technique. The results showed that 5 EcoR Ⅰ ＋Hpa Ⅱ and EcoR Ⅰ ＋MspⅠ primer combinations from 63 primer combinations for amplified the gene DNA of 30 peppers. Totally 939 bands including 937 polymorphic bands were produced, with a polymorphism rate of 99.79%. In NDA pattern analysis, type I （3 721 bands） was no methylation （EcoR Ⅰ ＋Hpa Ⅱ/EcoR Ⅰ ＋MspⅠ ）. Type 11 （3 152 bands） was hemi-methylation bands digested with EcoR Ⅰ ＋Hpa Ⅱ. Type Ⅲ（3839 bands） was fully methylation bands digested with EcoR Ⅰ ＋MspⅠ . Fully methylation was major methylation type in the two pepper species. The average methylation bands of C. annuum L. （319 bands ） was higher than C. chinense Jacquin （217 bands）, and thepercentage ofmethylation was 77.24%, 63.64% respectively. Difference reached an extremely significant leve. All of the materials could be separated into three groups at about 0.683 similarity value with UPGMA, including the first group （No. 11, No. 12, No. 13, No. 15-Capsicum c hinense Jacquin and No. 1 O-Capsicum annuum L.）, the second group （No.2-No.9, No.14, No.16-No.30）, No.14 and No.17 are Capsicum chinense Jacquin, the third group （No. 1-Capsicum annuum L.）. The MSAP cannot distinguish C. annuum L. and C.chinense Jacquin, and it was showed that the Cpsicum spp. had high epigenetic diversity. Nei＇s gene diversity （h） and Shannon information index （I） of Capsicum annuum L. （0.204%, 0.329 8） were higher than those of Capsicum ehinense Jacquin （0.186 4, 0.297 5）. We deduced that there was higher epigenetic diversity in Capsicum annuum L. than Capsicum ehinense Jacquin. DNA methylation diversity was an informative indicator of genetic diversity in Cpsieum spp.. And we provided a theoretical basis for evaluating the Cpsicum spp. germplasms and breeding the fine varieties.

关 键 词：辣椒 表观遗传多样性 DNA甲基化 MSAP 

分 类 号：S641.3[农业科学—蔬菜学]