Molecular marker closely linked with major QTL (Quantitative Trait Loci) for resisting gray leaf spot of corn and application of molecular marker
A molecular marker, gray spot disease technology, applied in the fields of genetic engineering and molecular biology, can solve problems such as yield loss, and achieve the effects of high contribution rate, improved disease resistance traits, and convenient and fast detection methods
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Embodiment 1
[0026] Example 1: Acquisition of linked marker molecular markers for the main QTL of resistance to gray spot disease in maize
[0027] 1. Construction of fine positioning groups
[0028] In 2015, the main effect QTLqGLS1.02 was obtained from the RILs derived from the resistant parent Qi 319 and the susceptible parent Ye 478, such as figure 1 Shown, and select the resistant individual and the susceptible parent Ye 478 for backcrossing.
[0029] All maize plants were inoculated to ensure the concentration of bacteria and the accuracy of single plant phenotype, and the inoculation effect was identified 20 days after pollination.
[0030] 2. Take healthy leaves, about 16cm per plant 2 , Genomic DNA was extracted by the modified CTAB method.
[0031] 3. Primer Development and Synthesis
[0032] The primers used in the present invention were obtained from maizeGDB according to the reference genome of maize B73. And used to amplify the genomic DNA of the resistant parent Qi 319 ...
Embodiment 2
[0036] Example 2: Application of Molecular Marker COP-M1 in Improving Zhengdan 958 gray spot disease resistance
[0037] 1. Use the primers shown in SEQ ID NO.1 and SEQ ID NO.2 to amplify a band with a size of 185bp in the corn susceptible inbred lines Zheng 58 and Chang 7-2, and the nucleotide sequence is as shown in SEQ ID NO.2 ID NO.4 is shown.
[0038] 2. Using the molecular marker COP-M1, the main QTL for disease resistance from Qi 319 (male parent) was crossed with Zheng 58 (female parent) and Chang 7-2 (female parent) and backcrossed continuously. The leaf DNA of the offspring was extracted and amplified with the molecular marker COP-M1. Among the offspring, the offspring that amplified the 191bp band consistent with the Qi 319 band pattern were selected.
[0039] 3. The progeny selected according to the method in step 1 of Example 2 continued to serve as the female parent, and Qi 319 was used as the male parent for hybridization. Repeat step 1. The improved inbred l...
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