Preparation method for plant mutant

A technology for mutants and plants, which is applied in the field of preparation of plant mutants, and can solve problems such as the inability to realize mass reproduction of sterile line seeds

Inactive Publication Date: 2018-12-11
QINGDAO YUANCE GRP CO LTD
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Problems solved by technology

The common features of this kind of sterile materials are stable sterility, safe hybrid seed production, and easy preparation of high-yield,

Method used

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  • Preparation method for plant mutant
  • Preparation method for plant mutant
  • Preparation method for plant mutant

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Embodiment Construction

[0046] The present invention is described in detail below in conjunction with embodiment. In order to make the purpose, technical solutions and advantages of the present invention clearer and clearer, the present invention will be further described in detail below, but the present invention is not limited to these examples.

[0047] In order to solve the above-mentioned technical problems, the present invention provides a method for preparing a plant mutant, comprising the following steps:

[0048] T1. Construct a CRISPR / cas9 gene knockout vector to knock out the CYP78A13 gene;

[0049] T2. Agrobacterium transformation;

[0050] T3. Gene knockout detection;

[0051] T4. Phenotype identification of transgenic plants.

[0052] The step T1. constructing the CRISPR / cas9 gene knockout vector, knocking out the CYP78A13 gene, the sgRNA sequence is as follows:

[0053] 5'-ggcaCACAATGGCTCCAGCATTCC-3'

[0054] 5'-AAACGGAATGCTGGAGCCATTGTG-3'.

[0055]In order to solve the above-men...

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Abstract

The invention discloses a preparation method for a plant mutant. The preparation method comprises the following steps that T1, a CRISPR/cas9 gene knockout vector is constructed, and a CYP78A13 gene isknocked out; T2, agrobacterium is transformed; T3, gene knockout detection is carried out; and T4, phenotype identification is carried out on a transgenic plant. According to the preparation method for the plant mutant, a rice cell nucleus fertility gene serves as a target gene, gene knockout is carried out by using CRISPR/cas9 gene silencing, the function is lost after gene silencing, a cell isdeveloped into a complete plant, but the complete plant does not have pollen which is developed to have normal functions, and the plant mutant can be taken as a research object and used for creating asterile line.

Description

technical field [0001] The invention relates to the technical field of molecular biological blood plant sterile mutant creation, in particular to a method for preparing a plant mutant. Background technique [0002] Male sterility is an important trait closely related to the utilization of heterosis in crops. Rice is one of the most successful crops using heterosis, and it is also one of the crops with the richest male sterility diversity found so far. The male sterile line has become the most effective weapon for the utilization of heterosis. Among them, the "three-line" method is the earliest and most common method of breeding and production of hybrid rice in my country. The "three-line method" mainly uses the nuclear-cytoplasmic interaction The male sterile system has completed the three-line matching of "restorer line", "maintainer line" and "breeder line". [0003] The sterility is determined by the genetic material of the cytoplasm and the genetic material of the nucle...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00A01H6/46
CPCC12N15/8213C07K14/415C12N15/8289C12N2810/10
Inventor 米铁柱张国栋刘佳音万吉丽王克响罗碧
Owner QINGDAO YUANCE GRP CO LTD
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