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Breeding method for reducing rice seed cadmium content

A rice grain and content technology, applied in the field of plant ecology and agriculture, can solve the problems of unspecified gene silencing, unspecified site mutation, and inability to realize mutants, so as to improve cadmium accumulation traits, avoid safety hazards, and retain The effect of excellent characteristics

Active Publication Date: 2018-07-10
HUNAN UNIV OF ARTS & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the mutants obtained by these methods are random and uncontrollable, cannot silence specific genes, cannot mutate specific sites, and cannot achieve gene knockout mutants, so gene editing technology has practical significance for rice production major

Method used

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  • Breeding method for reducing rice seed cadmium content
  • Breeding method for reducing rice seed cadmium content
  • Breeding method for reducing rice seed cadmium content

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Embodiment 1

[0050] In the present embodiment, a kind of breeding method that reduces cadmium content of rice grain specifically comprises the following steps:

[0051] (1) Cloning the full length of the exon of OsNCX8 of the rice receptor material, and sequencing;

[0052] Design specific primer sequences to clone the full length of the exon of OsNCX8 of the rice receptor material and sequence; the primer sequence is: F1: 5'-ATGGCGCTCCTGCGCAGGCGG-3', which is shown in SEQ ID NO: 2; R1: 5' -CTATGCTGGCCACCAGGAGT-3', which is represented by SEQ ID NO:3. Cloning and sequencing obtained the full-length sequence of the NCX8 gene of different rice varieties such as Nipponbare, 9311, and Takata. Sequencing found that the gene sequence had no base difference, indicating that the gene was highly conserved during the evolution process. The full-length sequence of the gene is as follows. Shown as SEQ ID NO:1:

[0053] ATGGCGCTCCTGCGCAGGCGGGGGTACAGTGCCGCCGTCCCCAGCGCCTGCTGCTTCCTCCTCCTGCTGCTCGTCCTCTCC...

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Abstract

The invention discloses a breeding method for reducing the rice seed cadmium content. A CRISPER / Cas9 technology is used for directionally knocking out rice OsNCX8 genes to obtain transgenic plants with the knockout OsNCX8genes; homozygote plants with the completely inactivated OsNCX8 protein are selected from the plants; the plants without transgenic ingredients and the reduced rice cadmium content are finally obtained. The OsNCX8 protein is the first Na-Ca exchange protein family member identified in the rice, so that the seed cadmium content reduction by about 45 percent is caused after theOsNCX8 protein inactivation. In addition, the plant agronomic properties of the OsNCX8 protein inactivated plants are not obviously changed through being compared with that of normal plants. The homozygote plants with the completely inactivated OsNCX8 protein can be obtained in the T0 generation plants; the breeding time is greatly shortened; in addition, the plants containing no transgenic ingredients can be screened from the later generation; the potential safety danger due to transgene is avoided.

Description

technical field [0001] The invention relates to the fields of plant ecology and agricultural technology, in particular to a breeding method for reducing the cadmium content of rice grains. Background technique [0002] Rice is one of the main food crops in my country. With the development of industry and agriculture, heavy metals in the environment and soil, especially heavy metal cadmium, accumulate in various tissues of rice, affecting the yield and quality of rice. Cadmium cannot be degraded in the environment and can seriously endanger human health through the food chain. At present, the heavy metal pollution of rice in Hunan Province is serious, especially the heavy metal cadmium content in the grain (rice) exceeds the standard, which threatens human health. It is very important to reduce the cadmium content in the grain. The toxicity of cadmium to rice can be alleviated and the cadmium content in rice can be reduced by measures such as changing soil, optimizing water ...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00A01H6/46
CPCC12N15/8213C12N15/8243C07K14/415
Inventor 郝小花田连福李东屏陈良碧
Owner HUNAN UNIV OF ARTS & SCI
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