Molecular markers for assisted selection of peach varieties containing recessive single gene hd and their breeding methods and applications
A technology of assisted selection and molecular markers, applied in the fields of molecular biology and peach genetics and breeding, can solve the problems of relying on tree retention time or fruit hardness identification, time-consuming breeding efficiency, and long growth cycle, so as to improve breeding efficiency and reduce breeding costs , the effect of simple method
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Embodiment 1
[0060] A molecular marker for assisted selection of peach cultivars containing a recessive single gene hd,
[0061] 1. PpYUC11 locus non-variation characteristic primers (detection of no CACTA transposon insertion):
[0062] Design the forward primer WILD-F at 1593bp upstream of the ATG of peach PpYUC11 wild-type gene, design the reverse primer WILD-R at 2515bp upstream of ATG, if there is a site variation, a 3514bp band will be amplified, if there is no site variation , the expected band of 942bp will be amplified, and the primers used are as follows:
[0063] The forward primer WILD-F primer sequence is 5'-ACATCAAACGGCAATCAATA-3',
[0064] The reverse primer WILD-R primer sequence is 5'-TTAGCCACGAAGGTTACTCA-3';
[0065] 2. The PpYUC11 site contains a variant characteristic primer (to detect the insertion of a CACTA transposon, and the peach PpYUC11 mutant gene has a CACTA transposon insertion at 2142bp upstream of the start codon ATG):
[0066] Design the forward primer C...
Embodiment 2
[0075] One of the methods for breeding SH fleshy peach varieties with assisted selection of molecular markers containing a recessive single gene hd peach variety described in Example 1, which comprises the following steps:
[0076] (1) Select the natural population as the material, extract its DNA, then perform PCR amplification with the primer sequences shown in the primer pairs WILD-F and WILD-R and CACTA-INS-F and CACTA-INS-R respectively, and The amplified products were separated and analyzed by electrophoresis, and the band patterns of the amplified results were as follows: figure 2 , 3 shown;
[0077] From figure 2 It can be obtained that the varieties that amplified the N50 band with WILD-F and WILD-R as primers include 'Zhongyoutao 16', 'yumyeong', 'Qinwang', 'Jingyu', 'Huayu', 'Xia Crisp', the others all amplified the N20 band;
[0078] From image 3 It can be obtained that using CACTA-INS-F and CACTA-INS-R as the primer pair to amplify the L50 band, there are ...
Embodiment 3
[0098] The second method for breeding SH fleshy peach varieties using the molecular markers described in Example 1 to assist in the selection of a recessive single-gene hd peach variety, the method comprises the following steps:
[0099] (1) Select the natural population as the material, extract its DNA, then perform PCR amplification with the primer sequences shown in the primer pairs WILD-F and WILD-R and CACTA-INS-F and CACTA-INS-R respectively, and The amplified products were separated and analyzed by electrophoresis, and the band patterns of the amplified results were as follows: figure 2 , image 3 shown;
[0100] From figure 2 available in China, the varieties that amplified the N50 band with WILD-F and WILD-R primers include 'Zhongyoutao 16', 'yumyeong', 'Qinwang', 'Jingyu', 'Huayu', ' Xiacui', and the N20 band was amplified in the others;
[0101] From image 3 It can be obtained that the L50 strips amplified by the primer pair of CACTA-INS-F and CACTA-INS-R co...
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