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Application of OsMSH4 protein and coding gene thereof in regulating and controlling plant pollen fertility

The technology of encoding gene and transgenic plant is applied to the application field of rice OsMSH4 protein and its encoding gene in regulating rice pollen fertility, which can solve the problems of difficult identification, costing a lot of manpower and material resources, and few spontaneous male sterile plants, etc. The effect of improving rice quality and increasing yield

Active Publication Date: 2014-12-10
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although the use of male sterile lines plays a vital role in the preparation and breeding of hybrid rice, there are very few spontaneous male sterile plants in nature, and identification is difficult and requires a lot of manpower and material resources

Method used

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  • Application of OsMSH4 protein and coding gene thereof in regulating and controlling plant pollen fertility
  • Application of OsMSH4 protein and coding gene thereof in regulating and controlling plant pollen fertility
  • Application of OsMSH4 protein and coding gene thereof in regulating and controlling plant pollen fertility

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1. Map-based cloning of OsMSH4 gene and its tissue-specific expression analysis

[0059] 1. Phenotype analysis of mutants

[0060] The inventor of the present invention is 60 A sterile mutant was found in the rice variety Zhongxian 3037 induced by Co radiation. Compared with the wild-type Zhongxian 3037 plant, the mutant had no significant difference in vegetative growth and flowering time ( figure 1 Middle A). After heading, the mutants showed typical sterile phenotype and eventually did not set fruit ( figure 1 Middle B). The mature anthers of wild-type plants are yellow, and the anthers are full of pollen, and the pollen uses I 2 -KI staining can turn blue, while the mature anthers of the mutant appear white, and the pollen is stained with I 2 -KI staining will not turn blue, and presents typical sterile pollen characteristics ( figure 1 Middle C and D). The inventors of the present invention granted the mutant to wild-type pollen, and found that it st...

Embodiment 2

[0092] Example 2, Osmsh4 mutant cytological phenotype analysis

[0093] 1. Cytological phenotype analysis in wild type

[0094] In the pollen mother cells of the wild-type rice Zhongxian 3037, the chromosomes were very thin and intertwined threads at the leptotene stage. During ditene, homologous chromosomes begin to pair and synapse. At the pachytene stage, the synaptonemal complex has been assembled, covering all homologous chromosomes, and at this stage, crossovers also begin to appear ( Figure 4 Middle A). At the end of the stage of degeneration, the chromosomes were extremely condensed into dots, and 12 ring-shaped or rod-shaped bivalents could be clearly seen ( Figure 4 Medium B). In metaphase I, 12 bivalents are neatly arranged on the equatorial plate ( Figure 4 Middle C). At anaphase I, the homologous chromosomes separate from each other and move to the poles of the cell under the traction of the spindle filaments ( Figure 4 Middle D), eventually forming two...

Embodiment 3

[0098] Example 3, Obtaining and Functional Identification of OsMSH4 Gene RNAi Rice Plants

[0099] In order to further confirm that the mutation of the OsMSH4 gene is the cause of the sterility of the mutant, the inventors of the present invention have carried out RNA interference to OsMSH4 to reduce or completely eliminate its expression, and further observe whether the phenotype of the RNAi plant is consistent with the phenotype of the mutant Osmsh4 unanimous.

[0100] 1. Construction of RNAi expression vector

[0101] Design the following RNAi primer sequences according to sequence 2 in the sequence listing:

[0102] OsMSH4 RNAi-F: 5'-ACTGAACTGTGGGGGAACTA-3' (positions 698-716 of SEQ ID NO: 2);

[0103] OsMSH4 RNAi-R: 5'-CTGATAAATGTTGCGAAGA-3' (reverse complement of positions 1093-1111 of Sequence 2).

[0104] Using primers OsMSH4RNAi-F and OsMSH4RNAi-R to perform PCR amplification, the cDNA obtained by reverse transcription of total RNA extracted from 5 cm spikelets of ri...

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Abstract

The invention discloses application of OsMSH4 protein and coding gene thereof in regulating and controlling plant pollen fertility. The application of the invention means application of protein (OsMSH4 protein) constituted by an amino acid sequence as shown in sequence 1 in a sequence list or a coding engine (OsMSH4 gene) of the protein in regulating and controlling plant pollen fertility. Experiments prove that in a development process of paddy rice, OsMSH4 protein expression level in the paddy rice is regulated down through a ribonucleic acid (RNA) interference technology, thus resulting in decrease of pollen fertility of the paddy rice compared to wild rice seeds. In combination with a tissue specific suppressor gene expression method, the OsMSH4 gene is applicable to rice hybrid seed production and fertility control; therefore, the application disclosed by the invention is significant in rice breeding, and is capable of increasing rice output, improving rice quality and offering an important biological resource.

Description

technical field [0001] The invention belongs to the technical field of plant molecular biology and relates to the application of rice OsMSH4 protein and its coding gene in regulating rice pollen fertility. Background technique [0002] Rice is one of the most important food crops for human beings. Nearly half of the world's population, including most of the population in East Asia and Southeast Asia, feed on rice. Rice is the largest food crop in my country, so how to increase the yield of rice is a key issue in my country's agricultural production. Since the 1960s, with the popularization of semi-dwarf varieties, the yields of rice and wheat have achieved breakthrough growth. In the 11 countries that promoted the Green Revolution, rice yields increased by 63 percent in the late 1980s compared to the early 1970s. Therefore, this dwarf breeding is called a "green revolution" in modern grain production. In my country, the three-line hybrid rice developed by Yuan Longping in...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/84C12N1/15C12N1/19C12N1/21A01H5/00
CPCC07K14/415C12N15/8205C12N15/8206C12N15/8289
Inventor 程祝宽张蕾唐丁李亚非沈懿杜桂杰
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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