Molecular marker Caps7 in paddy rice cadmium accumulation related gene OsHMA3 and application thereof

A molecular marker and rice technology, applied in the field of molecular biology, can solve the problems of high cadmium pollution and aggravate the risk of excessive cadmium content in rice, and achieve the effect of speeding up the breeding process

Active Publication Date: 2018-07-17
INST OF SUBTROPICAL AGRI CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the main rice producing areas in southern my country, the single planting structure of indica rice, the acidic soil b

Method used

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  • Molecular marker Caps7 in paddy rice cadmium accumulation related gene OsHMA3 and application thereof
  • Molecular marker Caps7 in paddy rice cadmium accumulation related gene OsHMA3 and application thereof
  • Molecular marker Caps7 in paddy rice cadmium accumulation related gene OsHMA3 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Discovery of allelic functional differences of HMA3 in rice indica and japonica subpopulations

[0032] (1) Natural variation of gene HMA3

[0033] Aiming at the rice cadmium accumulation-related QTL gene OsHMA3, the sequences of the OsHMA3 gene in 105 germplasm resources (collected in our laboratory) were analyzed. Haplotype analysis was carried out by MEGA sequence analysis software and STRUCTURE software, and it was found that there were 9 genotypes of this gene in nature, which were recorded as HMA3-1~HMA3-9 in sequence; among them, HMA3-2~HMA3-6 belonged to indica rice genes type, while HMA3-1, HMA3-7~HMA3-9 belong to the japonica rice genotype, see figure 1 a. HMA3-1 and HMA3-2 are the two main allelic genes, which accounted for 83.8% of the total materials; further analysis found that most of HMA3-1 appeared in japonica varieties and a small part appeared in indica varieties, while HMA3-2 The haplotype analysis of OsHMA3 mainly occurs in indica rice...

Embodiment 2

[0036] Example 2: Development of molecular marker Caps7 and genotype identification method

[0037] (1) Development of molecular marker Caps7

[0038] Molecular marker Caps7 of the present invention, the principle of marker design is: use the BLAST tool in the bioinformatics website NCBI (https: / / www.ncbi.nlm.nih.gov / ) to sequence the sequences of HMA3-1 and HMA3-2 Compare and find the difference site, the difference site forms two genotypes, wherein the one derived from the japonica rice genotype HMA3-1 cannot be cut by the restriction endonuclease AscI, after being digested by the AscI enzyme after PCR amplification, passed A single band was detected by agarose electrophoresis, and the size of the PCR product was 337bp; the indica rice genotype HMA3-2 could be cut by the restriction endonuclease AscI, and two bands were detected by agarose electrophoresis, and the size of the PCR product was 196bp and 141bp ,See image 3 .

[0039] (2) Genotype identification method

[0...

Embodiment 3

[0051] (1) Identification and screening of allelic types of the rice cadmium accumulation-related gene OsHMA3 in 300 rice indica materials and its effectiveness test

[0052] Taking the overall average value as the standard for distinguishing the high and low cadmium content of rice, the molecular marker Caps7 of the present invention was used to identify the allelic type of OsHMA3 in 300 indica rice materials.

[0053] The rice cadmium accumulation phenotypes of 300 materials were collected in polluted field experiments. The field planting method was carried out in a dry-wet alternate manner to maximize the accumulation of cadmium in the rice grains (the concentration of cadmium in the field is heavily polluted, the concentration is 1.9mg / kg, pH value is 5.4). Extract its genomic DNA from 300 parts of indica rice materials in Table 1, utilize the intragenic marker Caps7 of rice cadmium accumulation-related gene OsHMA3 (its PCR amplification primer is shown in SEQ ID NO:4 and ...

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Abstract

The invention belongs to the technical field of molecular biology, and provides a molecular marker Caps7 in a paddy rice cadmium accumulation related gene OsHMA3 and an application thereof. A forwardprimer and a reverse primer are synthesized, paddy rice DNA is amplified in a PCR system, and then enzyme digestion is carried out for amplification products, finally agarose electrophoresis is used for detecting and determining that the paddy rice to be measured contains HMA3-1 or HMA3-2 allele, so that paddy rice cadmium accumulation trait is determined and seed selection of low cadmium accumulation variety of indica type rice is assisted. Paddy rice cadmium accumulation traits can be effectively determined, and a new technical means is provided for selecting low cadmium accumulation varieties of indica type rice with improved seed selection efficiency.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to an intragenic molecular marker Caps7 of a rice cadmium accumulation-related gene OsHMA3 and an application thereof. Background technique [0002] Cadmium (Cd) pollution in rice fields leads to excessive cadmium in rice, which seriously affects food safety and food security in my country. Therefore, in order to avoid the harm of cadmium pollution to the human body, it is urgent to reduce the cadmium accumulated in rice grains. Studies on the genetic characteristics of cadmium accumulation in rice cultivars found that indica cultivars tend to accumulate more cadmium in grains than japonica cultivars. However, in the main rice-producing areas in southern my country, the single indica rice planting structure, acidic soil background and high cadmium pollution degree have aggravated the risk of excessive cadmium content in indica rice varieties. Then, before the...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 孙亮陈彩艳
Owner INST OF SUBTROPICAL AGRI CHINESE ACAD OF SCI
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