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Nucleotide sequences mediating male fertility in plants and methods of using same

A nucleotide sequence, male sterility technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, plant peptides, etc.

Active Publication Date: 2015-12-30
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The deficiencies that must be overcome include segregation of genes causing male sterility, production of 100% male sterile offspring, achievement of complete female fertility and subsequent restoration of pollen fertility in F1 hybrids

Method used

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  • Nucleotide sequences mediating male fertility in plants and methods of using same
  • Nucleotide sequences mediating male fertility in plants and methods of using same
  • Nucleotide sequences mediating male fertility in plants and methods of using same

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0123] The first embodiment, obtaining the sequence of ZmMYB103

[0124] From the known sequence of Arabidopsis AtMYB103 (as shown in SEQIDNO:3 in the sequence listing), the nucleotide sequence of the maize variety B73 transcription factor ZmMYB103 is shown in SEQIDNO:1 in the sequence listing, and its amino acid sequence is shown in the sequence listing Shown in SEQ ID NO:2.

[0125] To regulate male sterility by disrupting ZmMYB103 function, a dominant ZmMYB103 repressor needs to be established. The 12 amino acid EAR sequence was shown to function as a dominant repressor motif when fused to other transcription factors, such as LLDDLELRLGFA. The short nucleotide sequence encoding the EAR sequence is fused to the 3' end of the ZmMYB103 open reading frame to generate a ZmMYB103+G+EAR fusion, the nucleotide sequence of which is shown in SEQ ID NO:4 in the sequence listing.

[0126] The ZmMYB103+G+EAR nucleotide sequence was synthesized by Nanjing KingScript Biotechnology Co., ...

no. 2 example

[0127] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0128] 1. Construction of recombinant cloning vector DBN01-T containing ZmMYB103+G+EAR nucleotide sequence

[0129] Ligate the ZmMYB103+G+EAR nucleotide sequence into the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and follow the instructions of the pGEM-T vector produced by Promega to obtain the recombinant cloning vector DBN01-T , its construction process is as follows 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 SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; ZmMYB103+G+EAR It is the nucleotide sequence of ZmMYB103+G+EAR (SEQ ID NO: 4); MCS is the multiple cloning site).

[0130] Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 competent ce...

no. 3 example

[0137] The third embodiment, the acquisition and verification of maize plants transferred to the ZmMYB103+G+EAR nucleotide sequence

[0138]1. Obtaining corn plants transferred to the ZmMYB103+G+EAR nucleotide sequence

[0139] 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 DBN100369 (including the ZmMAC2 promoter sequence, ZmMYB103+G+EAR nucleotide sequence, the promoter sequence of the maize Ubiquitin gene, the PMI gene and the Nos terminator sequence) was transferred into the maize genome, A maize plant transformed with the ZmMYB103+G+EAR nucleotide sequence was obtained; at the same time, a wild-type maize plant was used as a control.

[0140] For Agrobacterium-mediated transformation of maize, briefly, immature embryos were isolated from maize a...

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Abstract

The invention relates to a nucleotide sequence capable of mediating plant male fertility and a use method of the nucleotide sequence. An MYB transcription factor comprises: (a) protein consisting of an amino acid sequence shown in SEQ ID NO: 2; or (b) protein which is derived from (a) by carrying out substitution and / or lacking and / or addition of one or more amino acids on the amino acid sequence in the (a) and has the function of controlling the development of pollens; or (c) a protein consisting of an amino acid sequence shown in SEQ ID NO: 2. According to the nucleotide sequence and the method, ZmMYB 103 is a first transcription factor which is separated from maize in the variety of B73 and is connected with an EAR sequence, so that the amount of maize fertile pollens can be greatly lowered, and a plant with complete male sterile is obtained; by adopting the produced maize male sterile plant, the crossbreeding period can be shortened, the castration step in the hybrid seed production process is omitted, the mechanical injury and yield influence on the maize plant from mechanical castration are lowered, and the purity and the yield of hybridization germplasm are ensured.

Description

technical field [0001] The present invention relates to a nucleotide sequence that mediates male fertility in plants and methods of using the same, in particular to a gene for pollen formation and a genetic method for producing male sterile plants by using a construct that disrupts the expression of the gene . Background technique [0002] Heterosis refers to the performance of F1 hybrids that are superior to those of the F1-producing parents. Heterosis in crop plants is often manifested in production as larger plants, stress tolerance, disease resistance, uniformity and increased yield, collectively referred to as hybrid vigor. [0003] Heterosis has been observed and documented in many important crop species, and the development of hybrids has become routine by plant breeders. However, hybrids can only be used in such plants when an effective and economical method of pollination control exists in the plant to ensure cross-pollination and prevent self-pollination. Mechan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8218C12N15/8289
Inventor 李建勇于彩虹张颖陶青鲍晓明卓壮
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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