Protein influencing male fertility and coding gene and application thereof

A fertility and protein technology, applied in the direction of genetic engineering, plant gene improvement, application, etc., can solve the problems of undiscovered corn related reports

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 obtain new male sterile lines using the above technology, β-1,3-glucanase genes from different sources are required, such as corn, but no related reports have been found so far

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0077] The first embodiment, obtaining the sequence of ZmPRcallase

[0078] Using the known sequence of tobacco PR-Glucanase (pathogenesis-related Glucanase) (as shown in SEQ ID NO: 3 in the sequence listing), the nucleoside of β-1,3-glucanase (ZmPRcallase) of maize variety B73 was obtained The acid sequence is shown in SEQ ID NO:1 in the sequence listing, and the amino acid sequence is shown in SEQ ID NO:2 in the sequence listing.

[0079] The ZmPRcallase nucleotide sequence was synthesized by Nanjing GenScript Biotechnology Co., Ltd.; the 5' end of the synthesized ZmPRcallase nucleotide sequence (SEQ ID NO: 1) is also connected with a RsrII restriction site, and the ZmPRcallase The 3' end of the nucleotide sequence (SEQ ID NO: 1) is also connected with a SnaBI restriction site.

no. 2 example

[0080] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0081] 1. Construction of the recombinant cloning vector DBN01-T containing the nucleotide sequence of ZmPRcallase

[0082]The ZmPRcallase 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 the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector DBN01-T, which was constructed 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 initiation codon; SP6 is the promoter of SP6 RNA polymerase; T7 is the promoter of T7 RNA polymerase; ZmPRcallase is the nucleoside of ZmPRcallase acid sequence (SEQ ID NO: 1); MCS is a multiple cloning site).

[0083] Then, the recombinant cloning vecto...

no. 3 example

[0090] The third embodiment, the acquisition and verification of corn plants transferred to the ZmPRcallase nucleotide sequence

[0091] 1. Obtain the corn plant transferred to the ZmPRcallase nucleotide sequence

[0092] According to the commonly used Agrobacterium infection method, the immature embryos of the aseptically cultured maize variety Zong 31 (Z31) were co-cultured with the Agrobacterium described in 3 in the second example, so that the The T-DNA in the recombinant expression vector DBN100370 (including the ZmMAC2 promoter sequence, the ZmPRcallase nucleotide sequence, the promoter sequence of the maize Ubiquitin gene, the PMI gene and the Nos terminator sequence) was transferred into the maize genome, and the transferred The corn plant of the ZmPRcallase nucleotide sequence; meanwhile, the wild-type corn plant is used as a negative control.

[0093] For Agrobacterium-mediated transformation of maize, briefly, immature immature embryos are isolated from maize and t...

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Abstract

The invention relates to a protein influencing male fertility and a coding gene and application thereof. The protein influencing the male fertility comprises: (a) a protein constituted by an amino acid sequence shown as SEQ ID No:2; or (b) a protein which is constituted by performing substitution and / or deletion and / or addition of one or more amino acids on the amino acid sequence shown as (a), has male fertility influencing activity, and is derived from (a). The protein ZmPRcallase influencing the male fertility is beta-1,3-glucanase derived from of a corn variety B73 for the first time, and can be used for greatly reducing the amount of corn fertile pollen to obtain a completely male-infertile plant; the crossbreeding period can be shortened by adopting the produced corn male-infertile plant, a castration step in a hybrid seed production process is eliminated, mechanical damage and yield influence of mechanical castration on the corn plant are reduced, and the purity and yield of a crossbreed are ensured.

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 corn-derived protein that affects male fertility by degrading callose, 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 corn breeding and production are mainly obtained through conventional breeding methods, or introducing sterility traits from one variety to another through backcrossing. This m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/24C12N15/56C12N15/82A01H5/00A01H1/02
CPCA01H1/02C12N9/244C12N15/827C12N15/8289C12Y302/01006
Inventor 于彩虹李建勇李梅丁德荣张颖
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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