Molecular marker of low-cadmium-accumulation control gene ZmCd1 of corn kernel and application

A molecular marker, low accumulation technology, applied in chemical instruments and methods, biochemical equipment and methods, and microbial assay/inspection, etc., can solve problems such as limited application, and achieve the effect of accelerating genetic improvement

Active Publication Date: 2020-06-09
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Functional molecular markers of key regulatory genes for low cadmium accumulation in maize have not yet been developed, which limits their application in molecular marker-assisted breeding of low cadmium accumulation maize varieties

Method used

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  • Molecular marker of low-cadmium-accumulation control gene ZmCd1 of corn kernel and application
  • Molecular marker of low-cadmium-accumulation control gene ZmCd1 of corn kernel and application
  • Molecular marker of low-cadmium-accumulation control gene ZmCd1 of corn kernel and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Using genome-wide association analysis to locate key regulatory genes for low grain cadmium accumulation

[0036] An associated group consisting of 436 maize inbred lines was planted in cadmium-contaminated farmland (cadmium content was 1.8±0.3mg / kg). The experiment was conducted in randomized block design with two independent repetitions. For each replicate, each maize inbred line was planted in a row with 10 plants in each row. Each row is 3m long and the row spacing is 60cm. Grain cadmium content was determined after maize was harvested. The average value of the cadmium content in the grains of the inbred lines corresponding to two replicates was used as the phenotype value of the inbred lines. Genotypes of associated populations were combined with reduced genome sequencing (GBS), high-density array technology, and deep RNA-sequencing data from 368 different maize inbred lines, identifying 1.25M SNPs with an allele frequency (MAF) greater than 0.05 ( Liu e...

Embodiment 2

[0039] Example 2 Candidate Gene Screening

[0040] According to the LD attenuation distance of the population in Example 1 (Liu et al., 2017, Molecular Plant, 10:414-426), the candidate genes regulating the cadmium content of maize grains were screened within 100 kb of the flanking sequence of the significant SNP site. According to gene function annotation, a candidate gene was locked and named ZmCd1. The gene is located within 100kb of the flanking sequence of the most significant SNP site (chr2.S_158408660).

[0041] Four high cadmium accumulating maize inbred lines Zheng 58 (0.26 mg / kg), Ye478 (0.21 mg / kg), P178 (0.44 mg / kg) and Mo17 (0.14 mg / kg) and four low cadmium accumulating maize inbred lines The ZmCd1 genes of cross-lines B73 (0.008mg / kg), Huangzaosi (0.009mg / kg), Jing 92 (0.014mg / kg) and Jing 724 (0.008mg / kg) were sequenced, and the sequences were compared and analyzed. , the four high cadmium accumulating maize inbred lines Zheng 58, Ye478, P178 and Mo17 all had ...

Embodiment 3

[0046] Example 3 Candidate gene ZmCd1 functional verification

[0047] Obtain the EMS mutant with termination mutation (EMS4-038f45) and missplicing (EMS4-038f36) in the ZmCd1 (Zm00001d005190) gene coding region from the corn mutant database MEMD (http: / / www.elabcaas.cn / memd / ) , each planted 5 rows in cadmium-contaminated farmland, and planted 10 plants in each row. After the corn was mature, mixed threshing was used to determine the cadmium content in the grain, and three biological replicates were measured. The measurement results showed that there was no significant difference in the cadmium content among the grains of the same mutant, and the cadmium content in the mutant grains was 0.26±0.16mg / kg and 0.24±0.07mg / kg, which were significantly higher than that of the wild-type material B73 (the cadmium content in the grain was 0.008mg / kg).

[0048] EMS4-038f45 and EMS4-038f36 were crossed with high cadmium accumulation maize inbred line Mo17 (with LTR retrotransposon inse...

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Abstract

The invention provides a molecular marker of low-cadmium-accumulation control gene ZmCd1 of a corn kernel and an application thereof. The invention comprises the following steps of: performing geneticanalysis on the cadmium content character of the corn kernels by taking a correlated group consisting of 436 parts of corn self-bred lines as basic materials, screening candidate genes for regulatingand controlling the cadmium content of the corn kernels in a range of 100kb of flanking sequences of significant SNP sites, and locking the candidate genes named ZmCd1; conducting sequence comparisonanalysis, wherein analysis results show that 7520bp deletion and 604bp of Gypsy type LTR retrotransposon insertion mutation exist in a promoter position of the ZmCd1 gene of the high-cadmium-accumulation corn self-bred lines compared with the low-cadmium-accumulation corn self-bred lines; and developing a functional molecular marker PCR-ZmCd1 based on PCR according to the difference sequence of the promoter position. The molecular marker can be used for early prediction, screening and breeding of low-cadmium-accumulation corn.

Description

technical field [0001] The invention relates to the technical field of crop molecular marker assisted breeding, in particular to the molecular marker and application of the gene ZmCd1 which controls the low accumulation of cadmium in corn grains. Background technique [0002] Cadmium is a heavy metal with significant biological toxicity, which will enter the human body along the food chain and seriously endanger people's health. Cadmium is listed by the World Health Organization (WHO) as a key food pollutant for research, and by my country as a key monitoring indicator for the implementation of total emission control. Corn is an important food and feed crop in my country, and its planting area and output rank first among all crops. Cadmium in soil is absorbed by corn roots and finally accumulates in edible parts, which seriously affects the quality and safety of corn and poses a threat to human and animal health. Therefore, improving the tolerance of maize to cadmium toxic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/6895C12N15/11
CPCC07K14/415C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 骆美洁赵久然汤彬赵衍鑫郭欢乐陈志辉张云霞孔梦思冯震宋伟张如养
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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