Molecular marker for rapidly identifying density of spines of cucumber line, identification method and application
A molecular marker, cucumber technology, applied in the direction of biochemical equipment and methods, determination/inspection of microorganisms, DNA/RNA fragments, etc.
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Embodiment 1
[0023] Example 1 Construction of genetic segregation population
[0024]The North China type cucumber inbred line 9930 was used as the female parent, and the North China type cucumber inbred line Shannong CNS5 was used as the male parent. The fruit phenotype of the female cucumber inbred line 9930 is dense thorn tumor; the male cucumber inbred line Shannong CNS5 is a natural mutant isolated from the North China type cucumber Nongcheng No. 2 inbred line Shannong CNS2. The phenotype of its fruit is sparse spiny tumor.
[0025] Using these two parents to prepare a cross combination, get F 1 The prickly tumor density of the fruit was between the two parents, F 1 F 2 78 strains were randomly selected for genetic segregation analysis.
[0026] In 78 F 2 Among individuals, the fruit prickly tumor density of each plant was counted, and the chi-square analysis was used to analyze the segregation ratio of dense prickly / sparse prickly traits. Finally, it was determined that the pric...
Embodiment 2
[0027] Example 2 Whole genome resequencing of cucumber
[0028] Using population segregation analysis (BSA) from F 2 In the isolated population, 20 individual plants of S. spp. and S. spp. were randomly selected, and the gene pools of S. spp. (fs1 fs1) and S. spp. (FS1_) were established.
[0029] The constructed gene pools of cucumber with sparse tumor (fs1 fs1) and cucumber with dense tumor (FS1_) were sent to Shanghai Meiji Biomedical Technology Co., Ltd. for whole genome resequencing of cucumber.
[0030] The association analysis between the SNP markers obtained by sequencing and the phenotypic traits of tumor density showed that the fs1 gene fs1, which controls the thin thorn phenotype of cucumber, was mapped to the end of the long arm of chromosome 6.
Embodiment 3
[0031] Example 3 Development of Molecular Markers
[0032] Analyze the SNP marker results obtained by resequencing the whole cucumber genome, design marker primers according to the position of the sparse thorn gene fs1, use the designed CAPS marker primers and SNP marker primers to perform PCR amplification on the two parents, and use 1.2% agarose Gel electrophoresis detection (the 1.2% agarose gel means that 100 ml of TBE buffer contains 1.2 g of agarose powder).
[0033] The following 6 markers were screened from the designed CAPS markers and SNP markers to carry out PCR amplification between the two parents. The results of the amplification primers, restriction enzymes and amplified fragments are shown in Table 1.
[0034] Table 1 Marker primer sequences and amplified fragment results of 6 kinds of molecular markers
[0035]
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