Protein influencing male fertility and encoding gene and application thereof

A fertility and protein technology, applied in genetic engineering, plant genetic improvement, application, etc., can solve undiscovered problems

Active Publication Date: 2014-03-26
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to use the above technology to obtain new male sterile lines, thereby reducing the risk of pollen-mediated gene drift, α-amylase genes from different sources are needed, such as rice, and no related reports have been found so far

Method used

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  • Protein influencing male fertility and encoding gene and application thereof
  • Protein influencing male fertility and encoding gene and application thereof
  • Protein influencing male fertility and encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0082] The first embodiment, obtain OsAmylase sequence

[0083] Using the known sequence of corn α-amylase (α-Amylase) (as shown in SEQ ID NO: 3 in the sequence list), the nucleotide sequence of the α-amylase (OsAmylase) of the rice variety Nipponbare is obtained as shown in the sequence table SEQ ID NO: 1 is shown, and its amino acid sequence is shown in SEQ ID NO: 2 in the sequence listing. The OsAmylase nucleotide sequence was synthesized by Nanjing GenScript Biotechnology Co., Ltd.

no. 2 example

[0084] Second embodiment, construction of recombinant expression vector and transformation of Agrobacterium with recombinant expression vector

[0085] 1. Construction of the recombinant cloning vector DBN01-T containing the nucleotide sequence of OsAmylase

[0086] The synthesized OsAmylase nucleotide sequence was connected to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of Promega product pGEM-T vector to obtain the recombinant cloning vector DBN01-T. The build process is like figure 1 As shown (where Amp represents the ampicillin resistance gene; f1 represents the origin of replication of phage f1; LacZ is the LacZ start codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; OsAmylase is the OsAmylase nucleoside Acid sequence (SEQ ID NO:1); MCS is the multiple cloning site).

[0087] Then the recombinant cloning vector DBN01-T was transformed into E. coli T1 comp...

no. 3 example

[0094] The third embodiment, the acquisition and verification of rice plants transformed into OsAmylase nucleotide sequence

[0095] 1. Obtain genetically modified rice plants

[0096] According to the conventional Agrobacterium infection method, the callus of the aseptically cultivated japonica rice 9522 mutant with the homozygous recessive allele dpw (obtained from the cooperation project of Shanghai Jiaotong University) was compared with the 3 in the second example The aforementioned Agrobacterium was co-cultured to combine the T-DNA (including gdpw fragment, PG47:OsAmylase:In2 fragment, Ubi:PAT:Nos fragment, Ubi:PMI:Nos) in the recombinant expression vector DBN100259 constructed in the second embodiment 2 Fragment) was transformed into the rice genome, and the rice plant with the nucleotide sequence of OsAmylase was obtained.

[0097] For Agrobacterium-mediated rice transformation, briefly inoculate rice seeds in an induction medium (N6 salt 3.1g / L, N6 vitamins, casein 300mg / L, ...

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Abstract

The invention relates to a protein influencing male fertility and an encoding gene and application thereof. The protein influencing the male fertility comprises: (1) a protein constituted by an amino acid sequence shown as SEQ ID NO:2, or (b) a protein obtained by performing substitution and / or deletion and / or addition of one or more amino acids on the amino acid sequence shown in (a), has male fertility influencing activity and is derived from (a). The protein OsAmylase influencing the male fertility, disclosed by the invention is alpha-amylase which is firstly separated from a rice variety named Nipponbare, can degrade starch in pollen grains, causes pollen sterility of rice transgene, is high in accuracy, and effectively prevents transgenic crops from spreading transgenic elements to other wild animal varieties via pollen; the protein can be used for keeping the homozygous recessive states of male sterile plants; meanwhile, the step of castration in a hybrid seed production process is eliminated, the influences on the investment of labor force and yield are lowered, and higher cost effectiveness is achieved.

Description

Technical field [0001] The present invention relates to a protein that affects male fertility, its encoding gene and its use, in particular to a protein from rice that affects plant male fertility by degrading starch in advance, its encoding gene and its use. Background technique [0002] The development of plant hybrid breeding technology has improved the quality and quantity of crops to a considerable extent. Due to the application of hybridization technology, various combinations of increased yield and expected characteristics (such as disease resistance, insect resistance, heat tolerance, drought tolerance, and changes in plant composition) are possible. Under normal circumstances, the application of hybridization technology relies on the male parent who provides pollen to the female parent to obtain hybrids. [0003] For a long time, male sterile lines used in rice breeding and production are mainly obtained through conventional breeding methods, or sterile traits are introdu...

Claims

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Application Information

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IPC IPC(8): C12N9/32C12N15/56C12N15/82A01H5/00A01H1/02
CPCA01H1/02C12N9/2422C12N15/827C12N15/8289C12Y302/01001
Inventor 于彩虹李建勇陶青张颖陈淑阳
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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