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| DNA Fingerprint Construction for 72 Chinese Cabbage (Brassica rapa ssp. pekinensis) Hybrids |
| LIU Shuan-Tao, ZHANG Zhi-Gang, LI Qiao-Yun, WANG Rong-Hua, WANG Li-Hua, WANG Shu-Bin, ZHAO Zhi-Zhong* |
| Shandong Branch of National Center for Vegetables Improvement/Vegetable and Flower Institute, Shandong Academy of Agricultural Sciences/Shandong Key Laboratory for Biology of Greenhouse Vegetables/Huang-Huai-Hai Region Scientific Observation and Experimental Station of Vegetables (Shandong), Ministry of Agriculture and Rural Affairs, Ji'nan 250100, China |
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Abstract In production practice, the identification of Chinese cabbage (Brassica rapa ssp. pekinensis) hybrids mostly rely on field planting and phenotypic identification, which is inefficient and inaccurate. Under the condition of more and more mature molecular marker research technology, it is necessary to establish a rapid, efficient and accurate identification technology of Chinese cabbage hybrids based on molecular marker technology. Considering this, a total of 86 pairs of insertion/deletion (InDel) primers were used to identify 40 Chinese cabbage hybrids by PCR amplification and polymorphism detection. Powermarker V3.0 software were used to analyze the polymorphic results. MEGA 7.0 software were performed for cluster analysis of hybrids. In order to screen core primers suitable for DNA fingerprint construction of different kind of Chinese cabbage hybrids, the percentage of heterozygous state for each pair of primers were compared in every group. The results showed that, 189 polymorphic loci were obtained, with an average of 2.2 alleles for each primer combination. The heterozygosity index ranged from 0 to 0.648 2 and averaged at 0.304 2. The major allele frequency ranged from 0.391 9 to 1.000 0 and averaged at 0.701 1. The number of alleles detected by each pair of primers ranged from 1 to 4, of which 74 pairs of primers could only detect 2 alleles. Then, cluster analysis revealed that, based on producing area, the 40 hybrids were clearly divided into imported and domestic varieties, among which the domestic varieties could be further divided into folded-, cylindric-, and fluffy-type varieties.The number of loci with heterozygous state ≥50% and their distribution in genome was significantly different in the 4 groups. Finally, 5 pairs of primers with heterozygous state ≥50% and allele number equal to 2 were selected from each group for construction of core primers. Other 32 Chinese cabbage hybrid varieties which were sold in market in recent years were detected with core primers. The detection results of pure loci were assigned a binary numeral 0 or 1 according to the mobility ratio of electrophoresis from small to large, while heterozygous loci were assigned letter H. All primers were ordered according to chromosome number they located from A01 to A10 and primers on the same chromosome were ordered according to their physical position from small to large. DNA fingerprint of 72 hybrids were constructed. The selected core primers will be universal and compatible with DNA fingerprint construction of new hybrid cultivars created in future. The results could also provide reference for gemplasm screening and hybrids identification of Chinese cabbage.
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Received: 11 June 2020
Published: 01 February 2021
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Corresponding Authors:
* zhaozhizhong454@163.com
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