A molecular marker co-separated with the yellow seed coat of melon and its application
A melon, yellow technology, applied in the field of agricultural biology, can solve the problems of heavy workload, high cost, and easy to mix
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Embodiment 1
[0073] Example 1. Screening of Molecular Markers Related to Melon Seed Coat Color
[0074] 1. Construction and genetic analysis of isolated populations
[0075] From the collected melon varieties, a white thick-skinned melon variety Dongfangmi No. 1, and a yellow-thin-seeded thick-skinned melon variety Huangzi Jinyu ( figure 1 ). F 1 The seed coat is white. Using Dongfangmi No. 1 as the male parent and Huangzi Jinyu as the female parent to prepare a hybrid combination, the obtained F 1 The seed coat is yellow, indicating that the seed coat color exhibits a delayed genetic effect, and F 1 The seed coat develops from the maternal cells.
[0076] F obtained by crossing Dongfangmi No. 1 as the female parent and Huangzi Jinyu as the male parent 1 F obtained after selfing 2 A total of 470 seeds were planted in the field as individual plants, and each individual plant was self-crossed separately, and the seeds were harvested at maturity, namely F 3 After the seeds were dried, ...
Embodiment 2
[0085] Example 2, Molecular Marker YS Identification of the Seed Coat Color Genotype of Melon
[0086] Using the YS-labeled primer pair obtained in Example 1, the parents Dongfangmi No. 1, Huangzi Jinyu, F 1 (F 1 ) and the above 470 F 2 Generation single plant (358 strains of white seed coat single plant, 112 strains of yellow seed coat single plant) carries out genotype analysis, specifically as follows:
[0087] Genomic DNA of a single plant extracted by the CTAB method is used as a template, and the above-mentioned YS-labeled primer F and primer R are used for PCR amplification to obtain a PCR amplification product.
[0088]The above PCR amplification reaction system is as follows: 0.8 μL Buffer, 0.8 μL dNTPs (2 mmol / L each), 1 μL Primers (10 μmol / L, F+R (the final concentration of each primer is 0.5 μmol / L), 1 μL genome DNA (50ng / μL), 0.1μL DNA Polymerase for PAGE (purchased from Beijing Quanshijin Biotechnology Co., Ltd.) and 6.3 μL ddH 2 O.
[0089] The above PCR ...
Embodiment 3
[0120] Example 3. Application of Marker YS in Molecular Marker-Assisted Selection of Melon Yellow Seed Coat
[0121] In order to carry out the molecular marker-assisted selection breeding of the yellow seed coat gene, and verify the accuracy of the marker YS in the testa coat color, according to the method in Example 2, the YS marker was used to select other species except Dongfangmi No. 1 and Huangzijinyu. 12 melon varieties with different genetic backgrounds were actually tested, and a total of 14 melon varieties listed in Table 1 were identified. details as follows:
[0122] Table 1 is the information of 14 melon varieties
[0123]
[0124] The genomic DNA of each material was extracted by the CTAB method, and the 14 melon materials in Table 1 were subjected to PCR amplification using the marker YS according to the method in Example 2 to obtain PCR amplification products.
[0125] If the amplified product only contains 117bp, the genome of the muskmelon to be tested do...
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