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Application of Rice osrhd1-1 Gene in the Breeding of Rice Male Male Sterile Line

A technology of male sterile line, rice, applied in application, genetic engineering, plant genetic improvement and other directions

Active Publication Date: 2022-03-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Through sequence comparison analysis, it was found that there are 4 homologous genes in rice with similar functions to AtRHD1, encoding UDP-glucose / galactose epimerase, but there is no report on the function of these genes regulating rice pollen development

Method used

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  • Application of Rice osrhd1-1 Gene in the Breeding of Rice Male Male Sterile Line
  • Application of Rice osrhd1-1 Gene in the Breeding of Rice Male Male Sterile Line
  • Application of Rice osrhd1-1 Gene in the Breeding of Rice Male Male Sterile Line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Construction of Rice OsRHD1-1 Gene Knockout Vector and Rice Genetic Transformation

[0023] The base sequence of the rice OsRHD1-1 gene is shown in SEQ ID NO.1.

[0024] (1) According to the "http: / / skl.scau.edu.cn / targetdesign / " website, design the specific knockout target of OsRHD1-1 gene, the sequence is as follows:

[0025] Knockdown target: TCCTCCAGCTTCTCCAACT.

[0026] According to the knockout target sequence, the expression sequence that expresses the corresponding sgRNA sequence is designed.

[0027] (2) Use NEB's BsaI endonuclease to double-digest (two positions with the same enzyme) pYLCRISPR / Cas9Pubi-H vector

[0028] The system is as follows:

[0029] buffer 2 μL; vector 1 μL; BsaI endonuclease 1 μL; ddH 2 O was added to a total volume of 10 μL.

[0030] The digestion conditions are: 37°C for 30 minutes.

[0031] (3) Knockout target synthesis

[0032] The target sequences of the synthesized primers are as follows:

[0033] 595100-aFP: 5...

Embodiment 2

[0050] Example 2: Gene editing of rice OsRHD1-1 gene knockout lines

[0051] In order to detect the pollen development in the florets of the rice OsRHD1-1 gene knockout line, the DNA of the gene-edited seedlings was extracted, and the amplified fragment was sent to Hangzhou Shangya Company for sequencing.

[0052] Such as figure 1 As shown in the results of sequencing, it was found that the two DNA single strands of the three sequenced strains were mutated.

Embodiment 3

[0053] Example 3: Observation of anther traits in florets of rice OsRHD1-1 gene knockout lines

[0054] The anthers in the florets of rice OsRHD1-1 gene knockout lines became smaller, lighter in color and stunted.

[0055] Observation of the floret traits of the rice OsRHD1-1 gene knockout line obtained in Example 2, such as figure 2 As shown, the anthers in the florets of the rice OsRHD1-1 gene knockout line became smaller, lighter in color, and stunted.

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Abstract

The invention discloses the application of the rice OsRHD1‑1 gene in the cultivation of rice male sterility lines. The present invention constructs a gene knockout vector by designing a knockout target of the OsRHD1-1 gene, and performs genetic transformation in rice, and finds that the loss of OsRHD1-1 gene function can lead to pollen sterility, which is helpful for a new type of male sterile line The development of hybrid breeding is facilitated.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the application of the rice OsRHD1-1 gene in the cultivation of rice male sterility lines. Background technique [0002] Rice is one of the most important food crops in the world. How to increase rice production to meet the growing food demand is an urgent problem to be solved today. Due to the limited arable land area and fresh water resources in our country, the method of increasing rice yield per unit area is the main way to increase rice yield. There are many factors that affect the yield of rice. The intrinsic genetic characteristics that affect the number of tillers per plant, the number of grains per panicle and the grain weight of rice are the determinants of rice yield; external factors include abiotic stresses such as light, temperature, water and salinity and The biological stress of diseases and insect pests has an important impact on the yield; in addition, h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/61C12N15/84C12N15/55A01H5/02A01H6/46
CPCC12N9/90C12N15/8218C12N15/8289C12N15/8205C12N9/22C12Y501/03002
Inventor 甘银波杨帅淇吴君宇黄琳丽刘一华刘伯涵
Owner ZHEJIANG UNIV
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