Application of Maize zmakinβγ1 Gene in Breeding Maize Tolerant to Lead Stress
A corn and gene technology, applied in the field of molecular biology, can solve the problems of no lead tolerance and achieve great potential for economic benefits
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Embodiment 1
[0021] Example 1. Expression pattern analysis of maize ZmAKINβγ1 gene under lead stress
[0022] 1-1. Lead stress treatment of corn inbred line materials
[0023] The seeds of the plump corn backbone inbred line 178 were sterilized with 75% ethanol for 1 min and 10℅ of H 2 o 2 The solution was soaked for 15 minutes and shaken continuously during disinfection, and then rinsed with deionized water for 3 to 5 times until the residual H 2 o 2 Rinse well, then soak in deionized water for 4 hours, and finally germinate with filter paper under dark conditions at 28°C. After about 2-3 days, transfer the germinated corn to a floating plate, and culture it with Hoagland nutrient solution under the conditions of light (16h) / darkness (8h) at 28°C for hydroponics, change the culture medium twice a week, and wait until the seedlings grow to Stress treatment was carried out at the three-leaf stage.
[0024] 1-2. Lead stress treatment:
[0025] Select 30 seedlings with consistent growth...
Embodiment 2
[0028] Example 2. Candidate genes identified by genome-wide association analysis.
[0029] Using 312 maize inbred lines, the processing method and phenotypic identification were the same as 1-2, combined with 56110 SNP genotype data, the genome-wide association analysis results were as follows figure 2 As shown, the place marked in the figure is the location of the ZmAKINβγ1 gene.
[0030] Four SNP loci were detected on chromosome 1 (PUT-163a-60399874-3004, SYN7988, SYN7984, PZE-101256211, respectively located on chromosome 1 298967209bp, 298967713bp, 298969383bp, 299548320bp, Maize Gen_v3 Ref) Surface area, root dry weight, secondary root length, aboveground dry weight, biomass, and total root length were significantly associated. Among them, the results of correlation analysis are shown in Table 1. The ZmAKINβγ1 (299461477-299469668bp) gene falls within the LD (500kb) segment of these four SNPs. The importance of this gene in maize lead stress was further verified.
[0...
Embodiment 3
[0033] Example 3. Analysis of lead tolerance of Arabidopsis homologous gene mutant plants
[0034]The Arabidopsis homologue SALK_074210 was ordered in the TAIR Arabidopsis mutant library, in which image 3 and Figure 4 Labeled as Mutant. This mutant is a T-DNA insertion mutant in which the homologous gene of ZmAKINβγ1 in Arabidopsis has lost function. First, use 75% absolute ethanol to sterilize the mutant and Columbia wild-type seeds for 1 min, then sterilize with 1% NaClO for 15 min, and finally wash with sterile water for 3 to 5 times, sow and culture them on 1 / 2 MS medium for 7 days, then The wild-type plants and mutant plants were transplanted in 0g / L and 0.15g / L Pb(NO 3 ) 2 Continue to cultivate on the 1 / 2MS medium, observe the plant growth status after 10 days, the results are as follows: image 3 and Figure 4 as shown, image 3 The right figure in the figure shows the growth of Columbia wild type and Arabidopsis mutant under normal conditions respectively (Ara...
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