利用InDel标记鉴定大白菜杂交种豫新四号种子纯度  

Purity Identification of Chinese Cabbage(Brassica rapa L.ssp. pekinensis) Hybrid Yuxin No.4 by InDel Markers

作　　者：薛银鸽[1,2] 原玉香[2] 张晓伟[2] 董森森 姚秋菊[2] 蒋武生[2] 田保明[1] 张强[2] 赵艳艳[2] 魏小春[2] 

机构地区：[1]郑州大学生命科学学院,郑州450001 [2]河南省农业科学院园艺研究所,郑州450008

出　　处：《农业生物技术学报》2014年第4期449-456,共8页Journal of Agricultural Biotechnology

基　　金：国家大宗蔬菜产业技术体系(No.CARS-25-G-27);农业部黄淮地区蔬菜科学观测实验站(河南)(No.10205020);"十二五"科技支撑计划(No.2012BAD02B01-3);"十二五"863计划(No.2012AA100202-7);河南省科技创新杰出青年(No.124100510018)

摘　　要：InDel(insertion deletion length polymorphism)是基于基因组测序的第三代分子标记,已在一些作物的遗传研究中得到运用。本研究以大白菜(Brassica rapa L.ssp.pekinensis)杂交种豫新四号及其亲本和8个同父异母、同母异父的杂交组合为实验材料,利用InDel标记和引物组合方法对大白菜杂交种纯度进行了快速鉴定。结果表明,从104对InDel引物中筛选到3对(BrID10667、BrID90107和BrID90147)在豫新四号及其亲本间表现多态性且带型清晰稳定,这3对InDel引物对两批杂交种进行检测,第一批材料所测纯度全部为98.00%。第二批材料所测纯度分别为:100.00%、98.30%和99.10%,与田间鉴定结果 100.00%的纯度高度一致,吻合度达99.13%。利用筛选出来的3对特异性InDel引物分别对豫新四号及其4个同母异父和4个同父异母的杂交组合进行InDel分析,没有一对引物能够将豫新四号以及其4个同母异父和4个同父异母杂交组合同时区分开来。然而在这3对引物中引物BrID90107和BrID10667退火温度相同均为55℃,但是各自扩增产物位置不同,应用引物组合BrID90107和BrID10667就可以将豫新四号与其亲本及4个同父异母和4个同母异父的杂交组合完全区分开。实验结果显示,InDel标记可以快速、准确、经济地鉴定大白菜杂交种纯度,同时证明,利用引物组合的方法可以更有效地鉴别大白菜杂交种子中的生物学混杂,表明InDel标记技术在大白菜杂交种纯度室内快速检测中具有广泛的应用前景。InDel（insertion deletion length polymorphism） is the third generation of molecular markers which is based on genome sequencing. It has already applied to the genetic studies. In this research, we use F1 of Chinese cabbage （Brassica rapa L. ssp. pekinensis） （Yuxin No.4）, its parents and eight half-blooded hybrid combinations as to be experimental material.Using the third generation of molecular markers - InDel and primer combination method to identify Chinese cabbage hybrid＇s purity rapidly. The results showed that there were three pairs primers （BrID10667, BrID90107 and BrID90147） presented stable codominance among 104 pairs of screened InDel primers in Yuxin No.4. The purity of first batch of materials for three primers was 98.00%. The purity of second batch of materials for three primers was 100.00%, 98.30%, 99.10%, respectively. The results tested in field is 100%,which was highly consistent with the results tested in indoor. The match index of two results were up to 99.13%. The results showed that the InDel markers, as an accurate and cost- efficient means, could be suitable for rapid purity identification of Chinese cabbage hybrid seeds. Using three pairs of specificity InDel primers which were screened out from 104 pairs of InDel primers to analyse Yuxin No.4 and eight other half-blood cross combinations, respectively. None of this three pairs of primers could distinguish Yuxin No.4 and eight other half-blood cross combinations at the same time. However, in this three primers, the annealing temperature of primer BrID90107 and primer BrID10667 was the same, but their position of amplification production was different. So we used multiplex primers BrID90107 and BrID10667 to identify Yuxin No.4 from their parental lines and eight other half-blood cross combinations. The results showed that multiplex primers methods could be used for rapid indoor identification of Chinese cabbage hybrid purity. Using the method of multiplex primers to test different hybrid lines could identify the contaminatio

关 键 词：大白菜 INDEL标记 纯度鉴定 豫新四号杂交种 

分 类 号：S565.903.2[农业科学—作物学]