SNP Molecular Marker of Maize Low Phosphorus Response Gene zmarf31 and Its Application
A molecular marker and corn technology, applied in the field of molecular genetics, can solve the time-consuming and labor-intensive problems of QTL positioning, and achieve the effect of improving low phosphorus tolerance and high application value
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Embodiment 1
[0031] Example 1 The acquisition of a SNP molecular marker that is significantly correlated with the total dry weight of the corn low phosphorus response gene ZmARF31 and the determination of its detection primers
[0032] A SNP molecular marker of the corn low phosphorus response gene ZmARF31 significantly correlated with the total dry weight of the present invention is obtained by the following method:
[0033] 1) A total of 331 maize inbred lines from China, the United States and Mexico were collected, and the related populations used for mapping were constructed. The population has rich genetic diversity, including 131 temperate maize inbred lines and 200 tropical / subtropical maize inbred lines.
[0034] 2) Identification of root phenotypes at the seedling stage of related populations. In 2010, Sichuan Agricultural University Duoying Farm Base and in 2012, Sichuan Agricultural University Ya'an Teaching and Experimental Greenhouse, 331 maize inbred lines were obtained in p...
Embodiment 2
[0041] Application test on the total dry weight of SNP marker corn of embodiment 2 of the present invention
[0042] 1) HRM detection of SNP markers in associated populations. The specific method is as follows: randomly select 16 maize inbred lines in the related population, use genomic DNA as a template, and use specific primers (SEQ ID NO. 2 and 3) of the flanking sequence of the site as primers. The reaction system for PCR amplification is: 2×Eva-GreenSuperMix 10 μl, both upstream and downstream primers are 0.25 pmol / μl, DNA template is 100 ng, and double distilled water is added to 20 μl. The amplification reaction was carried out on the CFX96TM real-timePCR (Bio-Rad, USA) detection system, the program was: 98°C pre-denaturation for 90s; 95°C denaturation for 5s, 57°C annealing for 5s, a total of 50 cycles; 70°C to 90°C for each cycle Rise by 0.5°C, read the plate for 1s to draw the melting curve. HRM test results ( figure 1 ) showed that the site-specific primers succe...
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