A SNP site closely associated with pepper fruit orientation and its general molecular marker, acquisition method and application
A technology of molecular markers and application methods, applied in the fields of pepper genetics, breeding and molecular biology, can solve the problems of few molecular genetic mechanisms and the like
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Embodiment 1
[0049] This embodiment provides a SNP site closely associated with the orientation of pepper fruit, and the SNP site corresponds to base 37458043 of chromosome 12 of version 2.0 of the "Zunla 1" reference genome. Can be converted to CAPS markers.
[0050] This embodiment also provides a universal molecular marker CaUP12 closely related to the orientation of capsicum fruit, said molecular marker is transformed from genotyping primers designed according to the above SNP loci.
[0051] The primers mentioned above include forward primer CaUP12-F and reverse primer CaUP12-R, and the forward and reverse primers are as follows:
[0052] Forward primer CaUP12-F: 5'-CCAAGTCCCTAGATGGTGGTG-3', (SEQ ID NO.1);
[0053] Reverse primer CaUP12-R: 5'-TGCACAAGGACGTAGGTGTC-3', (SEQ ID NO. 2).
[0054] The above-mentioned universal molecular marker CaUP12 closely associated with the orientation of capsicum fruit is obtained by the following method, which includes the following steps:
[0055] ...
Embodiment 2
[0067] This example provides the application of the capsicum fruit oriented molecular marker CaUP12 in the above-mentioned Example 1 in natural populations.
[0068] In this example, the obtained fruit orientation molecular marker primers were used to identify the genotypes of 113 GWAS pepper materials, and predict the fruit orientation phenotype through the identification results. The steps are as follows:
[0069] (1) Genomic DNA of pepper was used as a template at a concentration of 80-150ng / μl; molecular marker CaUP12 was used for PCR amplification, the forward primer sequence was SEQ ID NO.1, and the reverse primer sequence was SEQ ID NO.2. The total volume of the PCR reaction system is 20 μl, and the specific components are as follows: ddH 2 O 15.6μl, 10×PCR Buffer 2.0μl, d NTPs (10mM) 0.4μl, TaqDNA polymerase 0.2μl (5U / μl), forward primer (10mM) 0.4μl, reverse primer (10 10mM) 0.4μl and 1.0 μl DNA template;
[0070] (2) The PCR reaction was carried out on the S1000 PC...
Embodiment 3
[0081] This example provides the application of the capsicum fruit oriented molecular marker CaUP12 in the above-mentioned Example 1 in the isolation population.
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