A kind of rice male fertility regulation gene osstrl2 and its application

A technology for male fertility and gene regulation, applied in the fields of plant genetic engineering and plant breeding

Inactive Publication Date: 2020-06-30
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for a long time, the development of hybrid rice has been affected by the limitation of resources of hybrid varieties, and it is gradually reaching the bottleneck

Method used

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  • A kind of rice male fertility regulation gene osstrl2 and its application
  • A kind of rice male fertility regulation gene osstrl2 and its application
  • A kind of rice male fertility regulation gene osstrl2 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Cloning and functional analysis of rice fertility control protein coding gene OsSTRL2

[0048] 1. OsSTRL2 gene full-length cDNA cloning, expression vector construction and subcellular localization

[0049] The inventors used bioinformatics techniques well known to those skilled in the art to determine a complete rice male reproductive development control protein OsSTRL2 from the rice genome. After software analysis and cDNA cloning, its CDS is 1293bp, as shown in SEQ ID NO: 1, encoding a rice male reproductive development control protein with a full length of 430 amino acids, and its sequence is shown in SEQ ID NO: 2. The result of the whole nucleotide sequence analysis shows that: the full length of the OsSTRL2 gene is 2091bp (SEQ ID NO: 3, including the regulatory region and intron).

[0050] Using the RNA of the normal wild-type rice variety Zhonghua No. 9 as a template, the first-strand cDNA was synthesized, and the full-length cDNA of OsSTRL2 was passed...

Embodiment 2

[0053] Embodiment 2, creation of rice male sterile line

[0054] Create osstrl2 rice male sterile line by genetic engineering or other means

[0055] The sequence of the coding region of the OsSTRL2 gene in this example is shown in SEQ ID NO:2. The osstrl2 mutant material in this example is obtained from the conventional japonica rice variety Zhonghua 9 through sequence variation of the OsSTRL2 gene.

[0056] Specifically, the target site of the OsSTRL2 gene (SEQID NO: 9) was first designed using CRISPR / Cas9 technology, and then the VK005-01-OsSTRL2 knockout vector was constructed to identify the recombinant vector, wherein the expression of the target site was controlled by the OsU3 promoter drive. The knockout vector was introduced into the genome of wild-type rice Zhonghua No. 9 by the rice transgenic method in Example 1. After the obtained regenerated plants were transplanted and survived, the transformed plants were screened with hygromycin; the total DNA of the leaves...

Embodiment 3

[0060] Example 3, Restoration of the Fertility of the Rice Male Sterile Line osstrl2 That Is the Complementary Function of the Transgene

[0061] Using the DNA of the normal wild-type rice variety Zhonghua No. 9 as a template, the fragment SEQ ID NO was amplified by the primer pair OsSTRL2-COM-F (SEQ ID NO: 12) and OsSTRL2-COM-R (SEQ ID NO: 13) :14, the fragment was cloned into the vector pCMBIYA1300, and P1300-OsSTRL2-COM was successfully constructed. Similarly, the vector P1300-OsSTRL2-COM was introduced by the rice transgenic method in Example 1 into the young panicles that do not have hygromycin resistance but are male sterile phenotype in the BC1F2 offspring of the rice male sterile line osstrl2 in the callus tissue. The obtained regenerated plants were transplanted and survived, and the transformed plants were selected with hygromycin. The transgenic T0 generation plants of the positive plants were observed for rice fertility, and the findings ( Figure 4 ) showed tha...

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Abstract

The invention discloses a paddy rice male fertility regulating gene OsSTRL2 and application thereof and belongs to the technical field of plant biology. The nucleotide sequence of the paddy rice male fertility regulating gene OsSTRL2 is as shown in SEQ ID NO. 1, and the amino acid sequence of protein encoded by the paddy rice male fertility regulating gene OsSTRL2 is as shown in SEQ ID No. 2. The invention further provides a method for culturing male sterile plants. The paddy rice male fertility regulating gene OsSTRL2 and a male sterile line generated on the basis of the gene have the advantages that the male sterile line is stable in fertility, unaffected by environment conditions and capable of being recovered by wild transgenosis; the gene and the male sterile line provide necessary components for the building of novel cross breeding systems and are significant in production practice.

Description

technical field [0001] The invention belongs to the fields of plant genetic engineering and plant breeding, and in particular relates to a rice male fertility regulating gene OsSTRL2 and its application. Background technique [0002] Rice (Rice), as a kind of grass, is the most important crop for human nutrition and calorie intake, providing more than one-fifth of the calories consumed by humans worldwide. Rice is also the most widely consumed staple food for large segments of the world's population, especially in Asia. According to 2012 FAO statistics, rice is the third most produced agricultural commodity in the world, after sugarcane and maize. Crop breeding refers to the technology of improving the genetic characteristics of crops to breed high-yield and high-quality varieties, also known as crop variety improvement. It takes genetics as the theoretical basis and comprehensively applies the knowledge of various disciplines such as plant ecology, plant physiology, bioch...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/113C12N15/82A01H5/02A01H6/46
CPCC07K14/415C12N15/8218C12N15/8289
Inventor 李双成邹挺李平邓其明王世全王玲霞朱军梁越洋刘怀年郑爱萍
Owner SICHUAN AGRI UNIV
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