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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 a third generation molecular marker based on genome sequencing and has been applied in the genetic research of some crops. In this study, Brassica rapa L.ssp.pekinensis hybrid variety Yuxin 4 and its parents and 8 one-half and one-half parents were used as experimental materials. The InDel marker and primer combinations The purity of Chinese cabbage hybrids was rapidly identified. The results showed that three pairs of 104 pairs of InDel primers (BrID10667, BrID90107 and BrID90147) were polymorphic in Yuxin 4 and their parents and their bands were clear and stable. The three pairs of InDel primers were used for two batches of hybrids Testing, the first batch of materials measured purity of all 98.00%. The purity of the second batch of materials tested were 100.00%, 98.30% and 99.10%, respectively, which were highly consistent with the purity of 100.00% in the field. The concordance reached 99.13%. Three pairs of specific InDel primers were selected for InDel analysis of Yuhsin-4 and its four heresy and four of the half-parent crosses, respectively, and no pair of primers were able to amplify Yuxin-4 and its Four daughters and four half-breeds are also distinguished at the same time. However, the primer pairs BrID90107 and BrID10667 annealed at the same temperature of 55 ℃ for all three pairs of primers, but their positions were different. Using primer combination BrID90107 and BrID10667, we could compare Yuxin 4 with its parents and four parents Four half-breeds are completely separated. The experimental results showed that the InDel marker could quickly, accurately and economically identify the purity of Chinese cabbage hybrids. At the same time, it was proved that the combination of primers could be more effective in identifying the biological hybridization in Chinese cabbage hybrid seeds, indicating that InDel labeling technology is effective in Chinese cabbage Hybrid in-house rapid detection of purity has a wide range of applications.