InDel molecular marker relevant to heat resistance of rice during earing flowering period, primer and application
A technology of heading and flowering, molecular markers, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of limiting the speed and progress of genetic breeding and variety improvement
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Embodiment 1
[0040] Genome-wide association analysis (GWAS) of rice heat tolerance at heading and flowering stage
[0041] 1) Heat tolerance identification of rice heading and flowering stage
[0042] 338 rice resources with a wide range of sources and rich genetic diversity were selected from the Hunan Provincial Germplasm Resource Bank. In 2018, the sowing date was adjusted according to the growth period of each material, and sown in four phases from April to June in Beishan, Changsha County, Hunan Province. town. After 30 days, they were transplanted to the field, and the field management and pest control were carried out according to the local rice cultivation techniques. The temperature and humidity recorder (HOBE) was used to record the temperature change of the rice canopy. The temperature probes were distributed in different positions of the field and recorded every 30 minutes. Record the initial earing stage of all materials. When the daily maximum temperature is higher than 37°...
Embodiment 2
[0051] Identification method of heat-resistant molecular marker HT9 in rice
[0052] 1) PCR amplification: use the CTAB method to extract the genomic DNA of rice plants, and use the DNA as a template to add the designed HT9 forward primer and reverse primer into the PCR reaction system. The PCR reaction system is as follows: template DNA 10-100ng, 1 μL of 5 μM forward primer, 1 μL of 5 μM reverse primer, 10× PCR buffer (containing Mg 2+ ) 2.0 μL, 10 mM dNTPMix 1 μL, Taq DNA polymerase 1U, and deionized water to make up to 20 μL; the PCR program refers to the third edition of the "Molecular Cloning Experiment Guide", and is appropriately modified according to the conditions and requirements of the Hunan Rice Research Institute. The specific PCR reaction program was: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 30 s, and 35 cycles; extension at 72°C for 10 min.
[0053] 2) Identify the heat resistance of ri...
Embodiment 3
[0055] Identification of Heat Tolerance of Rice Germplasm Resources Using Molecular Marker HT9
[0056] Taking the overall average value as the standard for distinguishing the strength of rice heat tolerance, the molecular marker HT9 of the present invention was used to identify the allelic types of 205 rice germplasm resources.
[0057] The heat tolerance of rice at the heading and flowering stage was identified by staged sowing under field conditions. 205 rice germplasm resources that encountered high temperature during the heading and flowering stage were selected, and the seed setting rate under high temperature was used as the index of heat tolerance. Genomic DNA was extracted from 205 rice materials in Table 1, and genotype identification was carried out according to the method in Example 2 using the heat-resistant molecular marker HT9.
[0058] Table 1 HT9 genotypes of 205 rice germplasm resources and seed setting rate under high temperature
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