A method and application of genome ssr molecular marker development of sugarcane cultivars
A cultivar and genome technology, applied in biochemical equipment and methods, recombinant DNA technology, DNA/RNA fragments, etc., can solve the problems of backward construction of sugarcane SSR molecular marker genetic linkage map, limited number of SSR molecular markers, and difficult development, etc. To achieve the effect of promoting the improvement of sugarcane genetics and breeding level, enriching polymorphism information, and protecting legitimate rights and interests
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Embodiment 1
[0022]A total of 4,460 sequences were downloaded from the Sugarcane Genome Center of the French Agricultural Research Institute (http: / / sugarcane-genome.cirad.fr) using the above method, 27,241 SSR sites were found, and 22,932 pairs of SSR primers were successfully designed. The specific operations are as follows :
[0023] From 22,932 pairs of designed primers, search for SSR primers with TG and CA motif types, randomly select 50 pairs, and use 4 Saccharum materials (cultivar R570, cultivar ROC1, tropical species LA purple and cut hands) species SES208) for amplification efficiency verification (see Table 1). The amplification results showed that: a total of 45 pairs of primers could amplify clear amplification bands, the remaining 5 pairs of primers had no amplification bands or the amount of amplified products was weak, and another 35 pairs of primers showed multiple The polymorphic rate was 70% (35 / 50), among which there were 28 pairs of primers of TG repeat type and 7 pa...
Embodiment 2
[0027] In order to further verify the polymorphism of the SSR primer pairs identified in this study, 20 pairs of SSR primers with high polymorphism were selected from the 35 pairs of primers screened above, and the 18 backbone parents in my country (whose blood relationship comes from Tropical species, 2-4 species of sarcasm, large-stem wild and Indian species) (see Table 2), 2 sugarcane ancestors (sarcasm SES 208 and tropical species LA purple) and 4 world Genetic diversity analysis and SSR primer polymorphism evaluation of important sugarcane cultivars (LCP85-384, R570, ROC 16 and ROC 22). The results showed that: 20 pairs of primers showed obvious polymorphisms on 24 sugarcane experimental materials, 95 alleles were amplified in total, 1-7 alleles were amplified by each pair, and each pair of primers amplified on average 4.75 alleles were identified. figure 2 The PCR amplified electrophoresis patterns of 2 SSR primers on 24 sugarcane materials tested were displayed, and the...
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