Application of corn ZmAKIN beta gamma 1 gene in cultivation of lead stress resistant corn

A corn and gene technology, applied in the field of molecular biology, can solve problems such as lead-free resistance, and achieve the effect of huge economic benefit potential

Active Publication Date: 2020-05-15
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, maize ZmAKINβγ1 gene has been found to be related to maize disease, but there is no report related to lead tolerance

Method used

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  • Application of corn ZmAKIN beta gamma 1 gene in cultivation of lead stress resistant corn
  • Application of corn ZmAKIN beta gamma 1 gene in cultivation of lead stress resistant corn
  • Application of corn ZmAKIN beta gamma 1 gene in cultivation of lead stress resistant corn

Examples

Experimental program
Comparison scheme
Effect test

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 results of genome-wide association analysis 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.

...

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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Abstract

The invention discloses an application of a corn ZmAKIN beta gamma 1 gene in cultivating lead stress resistant corn, which determines the relationship between the corn ZmAKIN beta gamma 1 gene and thecorn lead stress resistance, and comprises the following steps: constructing an overexpression vector of the corn ZmAKIN beta gamma 1 gene, and transfecting a corn immature embryo by using agrobacterium tumefaciens; the expression of the ZmAKIN beta gamma 1 gene of the corn can be up-regulated, therefore the lead resistance of the corn is effectively improved, and the ZmAKIN beta gamma 1 gene hasa wide application prospect in the field of heavy metal pollution resistance of plants, particularly in the field of soil lead toxicity resistance of the corn, and has huge economic benefit potential.

Description

technical field [0001] The invention relates to the application of corn ZmAKINβγ1 gene in cultivating lead stress-tolerant corn, which belongs to the field of molecular biology. Background technique [0002] Corn is one of the most important food crops worldwide. However, with the continuous deterioration of the ecological environment, the content of heavy metals in the soil is increasing, and its growth and yield quality are seriously affected by abiotic stress including heavy metals, which seriously restricts crop yield. Excavating stress-related genes, studying their stress response mechanism, and cultivating new maize varieties with strong stress resistance are the most effective ways to cope with adversity stress and increase maize yield. At present, my country's maize stress-resistant germplasm resources are very scarce. It is not only a very difficult process to obtain stress-resistant germplasm resources through conventional breeding, but also the breeding cycle is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/12C12N15/82A01H5/00A01H6/46
CPCC12N9/1205C12N15/8213C12N15/8271C12Y207/01037
Inventor 沈亚欧张严玲陈珉侯凤霞李昭玲
Owner SICHUAN AGRI UNIV
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