Gene for resisting osmotic stress caused by salt stress of corn, molecular marker and application
A molecular marker, corn technology, applied in the field of genetic engineering, can solve problems such as slow research progress, and achieve the effect of accelerating the breeding process
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Embodiment 1
[0043] Example 1 Cloning and functional analysis of maize salt resistance gene
[0044] There were significant differences in salt tolerance among different inbred lines, showing different leaf water contents under 300mM NaCl treatment. Metabolomics analysis of 266 maize inbred lines under control and salt treatment conditions, combined with the data of population leaf water content, using bioinformatics methods to establish a salt-tolerant compound prediction model, obtained 37 compounds related to maize salt resistance The hallmark metabolites, named METO1 to METO37. Then, using the content of METO31 as a physiological index, genome-wide association analysis was carried out, and a locus with significant effect value was found at the position of 129,288,383-129,688,383kb on chromosome 9, and a candidate gene was identified in this QTL candidate region, and the locus corresponding to The gene named ZmSOT10 is a gene encoding cytochrome oxidase P450, such as figure 1 As shown...
Embodiment 2
[0049] Example 2 Obtaining the Deletion Fragment (ZmSOT10-Del) in the ZmSOT10 Intron of the Maize Salt Tolerance QTL Gene
[0050] According to the SNP corresponding to the highest effect value of METO31's GWAS, the candidate gene association analysis was carried out on the resequencing results of 200 inbred lines, and it was found that Del382 (a 50bp deletion / insertion) was significantly correlated with METO31 content, as shown in Figure 4 As shown in A. The q-PCR results showed that there were significant differences in the transcript levels of ZmSOT10 in inbred lines containing Del382 and without Del382, as Figure 4 As shown in B, it shows that Del382 leads to the change of the transcription level of ZmSOT10, which further affects the content of METO31 and the salt resistance of maize inbred lines.
[0051] According to the presence or absence of Del382, 200 materials were divided into two haplotypes Hap1 and Hap2.
[0052] Taking M1016 and Zheng 58 as examples, the ZmS...
Embodiment 3
[0056] Example 3 Deletion or insertion of ZmSOT10-Del in maize salt-resistant gene ZmSOT10 as a molecular marker
[0057] Because the ZmSOT10-Del deletion exists in maize inbred lines such as M1016, the ZmSOT10-Del deletion can be used as a molecular marker to determine whether an individual is salt-tolerant.
[0058] Primer pair I (ZmSOT10-F1 / ZmSOT10-R1) was composed of the primers ZmSOT10-F1 (forward) and ZmSOT10-R1 (reverse) designed based on the flanking sequence of ZmSOT10-Del deletion. Using primer pair Ⅰ as primers, the genomic DNA of maize inbred line Zheng 58 and maize inbred line M1016 were used as templates for PCR amplification. PCR system 20μl: 2×Super Multiplex PCR Mix 10μl, 10μM Primer ZmSOT10-F1 1μl, 10μM Primer ZmSOT10-R1 1μl, DNA 1μl, diH 2 O 7 μl. PCR program: pre-denaturation at 94°C for 2min, denaturation at 94°C for 30s, annealing at 56°C for 30s, extension at 72°C for 30s, 34 cycles from denaturation to extension, and finally extension at 72°C for 5min...
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