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Proteins Affecting Male Fertility, Their Encoding Genes and Their Uses

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

Active Publication Date: 2016-01-20
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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  • Proteins Affecting Male Fertility, Their Encoding Genes and Their Uses
  • Proteins Affecting Male Fertility, Their Encoding Genes and Their Uses
  • Proteins Affecting Male Fertility, Their Encoding Genes and Their Uses

Examples

Experimental program
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Effect test

no. 1 example

[0082] The first embodiment, obtaining the OsAmylase sequence

[0083]Using the known sequence of corn α-amylase (α-Amylase) (as shown in SEQ ID NO: 3 in the sequence listing), the nucleotide sequence of the α-amylase (OsAmylase) of the rice variety Nipponbare is obtained, as shown in SEQ ID NO in the sequence listing: 1, 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] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

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

[0086] The synthetic OsAmylase nucleotide sequence was ligated into the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector DBN01-T, which Build process such as figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ start codon; SP6 is the promoter of SP6 RNA polymerase; T7 is the promoter of T7 RNA polymerase; OsAmylase is the nucleoside of OsAmylase acid sequence (SEQ ID NO: 1); MCS is a multiple cloning site).

[0087] Then, the recombinant cloning vector DBN01-T was transform...

no. 3 example

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

[0095] 1. Obtaining transgenic rice plants

[0096] According to the routinely adopted Agrobacterium infection method, the calli of the aseptically cultured japonica rice 9522 variety mutant (obtained by the cooperation project of Shanghai Jiaotong University) with the homozygous recessive allele dpw was compared with the three institutes in the second embodiment. Agrobacterium co-cultivation as described above, so that the T-DNA in the recombinant expression vector DBN100259 constructed in 2 in the second embodiment (including gdpw fragment, PG47:OsAmylase:In2 fragment, Ubi:PAT:Nos fragment, Ubi:PMI:Nos Fragment) is transferred into the rice genome, and a rice plant with the OsAmylase nucleotide sequence transferred is obtained.

[0097] For Agrobacterium-mediated transformation of rice, briefly, rice seeds were inoculated in induction medium (N6 sa...

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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 coding gene and its use, in particular to a rice-derived protein that affects plant male fertility by degrading starch in advance, its coding gene and its use. Background technique [0002] The development of plant hybrid breeding technology has greatly improved the quality and quantity of crops. Various combinations of yield increases and desired traits such as disease resistance, insect resistance, heat tolerance, drought tolerance, and changes in plant composition are possible thanks to the application of hybridization techniques. Typically, the application of hybridization relies on the male parent providing pollen to the female parent, thereby obtaining hybrids. [0003] For a long time, male sterile lines used in rice breeding and production are mainly obtained through conventional breeding methods, or introducing sterility traits from one variety to another through back...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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