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In this study, 32 were Brassica genus vegetables germplasm for materials, including 11 copies of Brassica vegetables, 11 Cabbage, 10 brassicas, using three methods to extract Brassica vegetable seed DNA, to explore a simple fast, efficient, and applies to the SSR molecular marker analysis of DNA extraction methods; further Brassica vegetable germplasm identification and analysis using 31 SSR primer pairs, trying to find the specificity of the three categories of Brassica vegetables interspecific fragment and clear the Brassica vegetable germplasm kinship; the last cloning and sequencing Brassica genus vegetables interspecific specific fragment and sequence analysis, the main results are as follows: 1. comparison of three different methods (SDS method, CTAB method, NaOH method) extracted 22 Brassica genus DNA quality vegetable seeds, found that: SDS extraction of the seeds of the best DNA quality, high integrity of OD260/OD280 average of 1.873, with considerable effect CTAB method leaves the DNA (OD260/OD280 ratio 1.802-1.906); CTAB method followed, in its OD260/OD280 average 1.600; worst quality of DNA of NaOH extracted seed of OD260/OD280 values ??are low 1.3 2. SSR markers, three categories of vegetables in 32 Brassica vegetables (cabbage (Brassica rapa) cabbage (Brassica oleracea), mustard (Brassica juncea)) species specificity intraspecific polymorphic sexual and kinship analysis. From 31 pairs of primers screened 21 polymorphic primers amplified a total of 113 clear polymorphic bands. The primer MF4 SSRRAS46, P004 and NA3 Brassica vegetables, cabbage, vegetables, brassicas specific bands, 10 confirmatory material verification, the result is exactly the same. 21 pairs of polymorphic primers average PIC value of 0.604, the primer B01 can distinguish five cabbage varieties, the 8 cabbage the variety, JU5 can zone mustard varieties separately. Hierarchical cluster analysis of the 22 copies of Brassica case the vegetable germplasm into two groups: cabbage vegetables for a class, Chinese cabbage and mustard vegetables a class, the bootstrap values ??up to 100%. Similar coefficient of 0.29, cabbage vegetables and brassicas were clustered into two subcategories. Principal coordinates analysis is consistent with the system cluster analysis results. Brassica vegetable species specific fragment was cloned and sequenced, and further for sequence analysis, primer MF4 in Brassica specific fragment size 140bp, cabbage-specific primers SSRRAS46 194bp fragment size, NA3 and P004 specificity in the mustard fragment size is 207bp and 213bp. BLAST, we can see the sequence in the sequence with the original sequence homology of 98%, 100%, and also high homology with other sequences, up to 85% -96%.
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