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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  • 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 fe

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

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Example Embodiment

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

[0059] 1. Mutant phenotype analysis

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

Example Embodiment

[0092] Example 2. Cytological phenotype analysis of Osmsh4 mutant

[0093] 1. Cytological phenotype analysis in wild type

[0094] In the pollen mother cells of wild-type rice Zhongxian 3037, the chromosomes appear to be very thin and intertwined threads at the thin line stage. In the even-line phase, homologous chromosomes begin to pair and join together. At the thick line stage, the synaptic complex has been assembled, covering all homologous chromosomes, and at this stage, the crossover also begins to appear ( Figure 4 In A). At the end of the terminal metamorphosis stage, the chromosomes are extremely concentrated into punctate, and 12 bivalents in the form of rings or rods can be clearly seen ( Figure 4 In B). In mid-stage I, 12 bivalent bodies are neatly arranged on the equatorial plate ( Figure 4 Medium C). At the later stage I, the homologous chromosomes separate from each other and move to the cell poles respectively under the traction of the spindle filaments ( Fig...

Example Embodiment

[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 infertility of the mutant, the inventors of the present invention performed RNA interference on OsMSH4 to reduce or completely eliminate its expression, and further observed whether the phenotype of the RNAi plant is the same as that of the mutant Osmsh4 Consistent.

[0100] 1. Construction of RNAi expression vector

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

[0102] OsMSH4 RNAi-F: 5'-ACTGAACTGTGGGGAACTA-3' (positions 698-716 of sequence 2);

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

[0104] Using the cDNA obtained by reverse transcription of total RNA extracted from 5cm spikelets of rice variety Nipponbare as a template, PCR amplification was carried out with primers OsMSH4RNAi-F and O...

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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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