Method for cultivating low-cadmium accumulated rice and use of related materials

A technology for rice accumulation, applied in chemical instruments and methods, botany equipment and methods, biochemical equipment and methods, etc., can solve the problems of no cadmium, cadmium reduction, etc.

Active Publication Date: 2019-04-30
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the widespread presence of cadmium in the environment, and there is no very effective method for in situ removal of cadmium in soil and groundwater, it is of great significance to reduce the accumulation of cadmium in rice grains through biotechnology.

Method used

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  • Method for cultivating low-cadmium accumulated rice and use of related materials
  • Method for cultivating low-cadmium accumulated rice and use of related materials
  • Method for cultivating low-cadmium accumulated rice and use of related materials

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1. OsCd1 / 2 is a cadmium accumulation-related protein and its application in regulating yeast cadmium accumulation

[0085] 1. Construction of yeast expression vector

[0086] A. Extract total RNA from rice Nipponbare and 9311 leaves respectively.

[0087] B. Reverse transcribe the total RNA extracted in step A into cDNA to obtain the cDNA of Nipponbare and the cDNA of 9311.

[0088] C. Using the cDNA of Nipponbare as a template, the primer pair composed of OsCd1S and OsCd1A was used for PCR amplification to obtain the PCR amplification product of OsCd1. OsCd1S: 5'-ATGGAGGTGTTCTACTACCT-3';

[0089] OsCd1A: 5'-TTAAGGATTCAGTGGCTCAT-3'.

[0090] Using the cDNA of 9311 as a template, PCR amplification was performed with the above-mentioned primer pair composed of OsCd1S and OsCd1A to obtain a PCR amplification product of OsCd2.

[0091] D. Using the PCR amplification product of OsCd1 as a template, a primer pair composed of OsCd1S1 and OsCd1A1 was used to obtain ...

Embodiment 2

[0107] Embodiment 2, cultivate low cadmium accumulation rice

[0108] 1. Creation of OsCd1 knockout rice based on CRISPR / Cas9 technology

[0109] 1. Construction of sgRNA expression vector

[0110] Design of single guide RNA (single guide RNA) target sequence According to the genome sequence of the rice cadmium transporter OsCd1, three sgRNAs targeting OsLCd1 were designed, and their names were CR-1, CR-2 and CR-3. The designed gRNA target sequence was compared with the rice genome sequence to exclude non-specific target sites. The nucleotide sequence of CR-1 is 5'-GCCUGGUCGCAAUUGUAUCC-3', targeting the 527-546th position (target site sequence) of SEQ ID No.1 of OsCd1 gene; the nucleotide sequence of CR-2 is 5'-CUUCUCGGCGUUCGAGUCA-3', targeting the 423-441th (target site sequence) of SEQ ID No.1 of the OsCd1 gene; the nucleotide sequence of CR-3 is 5'-CAAGCACUCCCCCCGAGUAC-3', targeting Positions 354-372 (target site sequence) of SEQ ID No. 1 of the OsCd1 gene.

[0111] Use...

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Abstract

The invention discloses a method for cultivating low-cadmium accumulated rice and use of related materials. The method for cultivating low cadmium accumulation rice comprises inhibiting or reducing the expression of OsCd1/2 gene in the target rice and obtaining low-cadmium accumulated rice, and the target rice is the rice containing the OsCd1/2 gene. The accumulation of cadmium in the low-cadmiumaccumulated rice is less than the accumulation of cadmium in the target rice; the OsCd1/2 gene encoding name is OsCd1/2 protein, OsCd1/2 is a) or b): a) protein having an amino acid sequence of SEQ IDNo. 2 or SEQ ID No. 4; and b) protein derived from a) and obtained by substituting and/or deleting and/or adding one or several amino acid residues to the amino acid sequences represented by SEQ ID No. 2 or SEQ ID No. 4 and having the biological cadmium accumulation activity regulation and control. The method can obtain the low-cadmium accumulated rice and reduce the food risk.

Description

technical field [0001] The invention relates to a method for cultivating rice with low cadmium accumulation and the use of related materials in the field of biotechnology. Background technique [0002] Cadmium is a toxic metalloid element widely distributed in nature. Soil cadmium pollution can lead to environmental degradation and decrease in crop yield and quality. Soil-crop-food migration is listed as the primary way for humans to absorb cadmium in the environment. Cadmium that enters crops enters the human body through the food chain, and then induces liver cancer, skin cancer or bladder cancer, etc., and also induces teratogenicity, thus endangering the human body healthy. [0003] Rice has a large area of ​​promotion and a high yield per mu. It is one of the three major staple foods in my country. In recent years, with the rapid development of industry and agriculture, the problem of cadmium pollution in rice has become increasingly serious in my country, and the num...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C07K14/415C12N15/29
CPCC07K14/415C12N15/8213C12N15/8218C12N15/8243
Inventor 何振艳闫慧莉许文秀高逸伟
Owner INST OF BOTANY CHINESE ACAD OF SCI
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