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Rice male and female fertility-related protein, encoding gene and application thereof

A technology related to proteins and coding genes, applied to rice male and female fertility-related proteins, coding genes and their application fields, can solve undiscovered problems

Inactive Publication Date: 2014-07-02
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After literature search, no report on the functional analysis related to the gene MFS1 of the present invention has been found so far

Method used

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  • Rice male and female fertility-related protein, encoding gene and application thereof
  • Rice male and female fertility-related protein, encoding gene and application thereof
  • Rice male and female fertility-related protein, encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] The phenotype of embodiment 1MFS1 gene mutation

[0062] A sterile mutant in the rice material Zhonghua No. 9 (purchased from the Resource Center of the Rice Research Institute of Sichuan Agricultural University) was found to be sterile in both male and female organs through forward and reverse hybridization and cytological observation ( figure 1), and further using techniques such as paraffin section, it was found that the root cause of its sterility was the abnormal meiosis of mutant pollen mother cells ( figure 2 ) and abnormalities in embryo sac structure ( image 3 ).

Embodiment 2

[0063] Map-position cloning of embodiment 2MFS1 gene mutation

[0064] 1. Map-based cloning of MFS1 gene mutation

[0065] Using G630 etc. as the female parent to hybridize with the heterozygote containing the mutant gene, construct a segregated population, locate the gene, and realize fine-mapping interval marker encryption and sequence comparison analysis show that a fragment deletion of about 20 KB is generated in this region of the mutant, It resulted in the loss of expression products of 3 predicted genes (orf1, orf2 and orf3) ( Figure 4 ), thereby causing a sterile phenotype in rice. Through gene prediction and transgenic methods, introducing one of the normal wild-type genes MFS1 into the mutant can restore the normal fertile phenotype ( Figure 5 ), and the introduction of the other 2 genes will not restore the fertility.

[0066] The sequence of the wild-type MFS1 gene is shown in SEQ ID NO.1, and the encoded protein sequence is shown in SEQ ID NO.2. The full-len...

Embodiment 3

[0079] Embodiment 3 MFS1 gene rice transgenic test

[0080] In this example, the expression vector PHB (Mao et al., 2005, PNAS 102:12270-12275, commercially available) was used as the rice transgene vector. The vector encodes a bacterial origin of replication (ori), a kanamycin resistance gene (Kan r ), hygromycin resistance gene (Hyg r ), herbicide resistance gene (Bar r ), 2×35S promoter, NOS gene termination signal sequence and multiple cloning site (MCS) for connecting target fragments. The MFS1 gene or its fragment can be inserted into the restriction endonuclease site to construct a transgenic plasmid.

[0081] 1. Construction of MFS1 gene complementation expression vector

[0082] In this example, pBS-MFS1 and PHB were firstly digested with PstI and SacI restriction enzymes, and the about 1.8K full-length cDNA fragment digested by pBS-MFS1 was ligated into the PstI and SacI sites of PHB, and the recombinant Enzyme digestion and sequencing identification.

[0083] ...

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Abstract

The invention discloses a rice male and female fertility-related protein, an encoding gene and an application thereof. The rice male and female fertility-related protein has an amino acid sequence as shown in SEQ ID (sequence identity) NO. 2 or a derivative sequence which is formed by performing substitution, deletion or addition of one or multiple amino acid residues on the amino acid sequence as shown in the SEQ ID NO. 2 and has the same functions. The invention further provides a gene for encoding the protein. The protein and the encoding gene thereof, provided by the invention, can be used for fertility control or identification of rice male and female organs, as well as regulation of rice male and female fertility level.

Description

technical field [0001] The invention relates to plant genetic engineering, in particular to a protein related to male and female fertility of rice, its coding gene and application thereof. Background technique [0002] Male sterility is very common in the plant kingdom. According to a large number of literature searches, male sterility has been found in 43 families, 162 genera, 320 species and 297 interspecific hybrids, and the number is still increasing. Since male sterility is the basis of the utilization of heterosis, the exploration of the mechanism of male sterility has been a very active research field for decades, especially in recent years, the research on molecular biology of male sterility has been fruitful The article on the molecular mechanism of male sterility summarizes the research results in this area and puts forward some corresponding theories and hypotheses, which enriches the theoretical basis of heterosis utilization and improves people's understanding o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A01H5/00
Inventor 李双成李平高锋焱王世全邓其明郑爱萍朱军刘怀年王玲霞
Owner SICHUAN AGRI UNIV
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