单 位：
甘肃农业大学动物科学技术学院;甘肃农业大学草业学院草业生态系统教育部重点实验室中-美草地畜牧业可持续发展研究中心
关键词：
单倍型组合;matK基因;rbcL基因;DNA条形码
摘 要：
DNA条形码技术是基于物种种内特异性和种间多样性利用一个或几个标准的DNA片段(DNA barcode)构建生物鉴别数据库。本研究根据Gen Bank中豆科牧草matK和rbcL基因的核苷酸序列,设计4对通用引物对豆科五个主要牧草属(苜蓿属Medicago,车轴草属Trifolium,红豆属Onobrychis,小冠花属Coronilla,野豌豆属Vicia)的六种牧草进行扩增。扩增产物进行测序和分析,筛选出六种牧草的四个标记位点5'端和3'端保守序列,并对各标记位点保守区内的核苷酸进行单核苷酸多态性(SNPs)的单倍型分析,数据表明:matK1、matK2、matK3均有5个单倍型,五个牧草属有其特有单倍型;rbcL基因有6个单倍型,车轴草属白三叶和红三叶有其特有单倍型。根据matK(matK1,matK2,matK3)和rbcL基因筛选的四个标记位点为六种牧草建立了相对应的特异DNA识别码。说明matK和rbcL组合后可作为豆科六种牧草的潜在条形码。
译 名：
Establishment of DNA Barcode of Leguminosae Forages' Six Species
作 者：
Li Yongqing;Zhao Shengguo;Jiao Ting;Faculty of Animal Science and Technology, Gansu Agriculture University;College of Pratacultural Science, Gansu Agricultural University,Key Laboratory of Grassland Ecosystem;
关键词：
Haplotype combination;;matK gene;;rbcL gene;;DNA barcoding
摘 要：
DNA barcoding is an emerging technology based on specificity within a species and diversity between species to build biological identification database by using one or more standard DNA barcode. In the study, we designed four universal primers to amplify six species in five major genera(Medicago, Trifolium, Onobrychis,Coronilla, Vicia) of leguminosae forages, according to nucleotide sequence of leguminosae forages' matK and rbcL genes in Gen Bank. 5' end and 3' end conservative fragments in four marked fragments were selected by amplification products' sequencing and analyzing. Single nucleotide polymorphism(SNP) haplotype analysis was conducted on each marker loci's conservative fragments' nucleotide sequences and the results showed that there were 5 haplotypes among matK1, matK2, and matK3 respectively. By the same token, rbcL gene has 6 haplotypes, Trifolium repens Linn and Trifolium pretense Linn have unique haplotypes. According to the haplotypes about matK(matK1,matK2 and matK3) and rbcL, we had constructed the DNA barcoding for six species leguminosae forages. It proved that the combination of matK and rbcL could be used as candidate DNA barcode for six species leguminosae forages.
