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Wheat fertility related gene TaMS7 and application method thereof

A fertility restoration gene and gene technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of complex, large and limited wheat genome

Active Publication Date: 2018-05-22
BEIJING NEXT GENERATION HYBRID WHEAT BIOTECHNOLOGY CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The wheat genome is large and complex, so the research on the mechanism of wheat male sterility has so far accumulated very limited data

Method used

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  • Wheat fertility related gene TaMS7 and application method thereof
  • Wheat fertility related gene TaMS7 and application method thereof
  • Wheat fertility related gene TaMS7 and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1. Genome-wide expression profile analysis of wheat anthers at different developmental stages and acquisition of anther expression contig in early pollen development

[0080] Wheat anthers whose pollen was in meiosis, mononucleate, binucleate and trinucleate were collected, total RNA was extracted with Trizol (Invitrogen), treated with DNaseI (Promega), and then mRNA was purified (Ambion). The purified mRNA was subjected to reverse transcription (Invitrogen), ultrasonic fragmentation (Fisher), library preparation (illumina) and amplification (illumina), and finally a sequencing reaction on an illumina machine.

[0081] The results of high-throughput sequencing of the wheat transcriptome were first assembled by Trinity software, and the resulting spliced ​​sequences were further removed from redundancy and similarity clustering. For the expression change analysis of the spliced ​​transcript contig, the high-throughput sequencing sequence in each sample was first ...

Embodiment 2

[0083] Embodiment 2.RT-PCR verifies the tissue expression specificity of TaMS7 gene

[0084] Since wheat is an allohexaploid consisting of three sets of genomes A, B, and D, the average copy number of genes is 2.8, of which nearly half of the genes (46%) have 3-4 copies, and 12% of the genes have 1-2 copies, 42% of genes had ≥5 copies. Starting from the sequence of comp155942_c0_seq4, using the sequencing information of common wheat published by CerealsDB and IWGSC (International WheatGenome Sequencing Consortium), as well as the wheat ancestors Triticum urartu (Triticum urartu, A genome donor) and Goat grass ( The sequencing information of Aegilopstauschii, D genome donor) was electronically cloned, and three TaMS7 genes were obtained, named TaMS7-A, TaMS7-B and TaMS7-D respectively. The genomic DNA sequences of the three TaMS7 genes are respectively shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, and the homology between the three is 95%-97%; the CDS sequences are res...

Embodiment 3

[0094] Example 3. Obtaining of TaMS7 gene promoter sequence

[0095] Starting from the genomic DNA sequences of TaMS7-A, TaMS7-B and TaMS7-D genes, using the sequencing information of common wheat published by CerealsDB and IWGSC (International Wheat Genome Sequencing Consortium), and the wheat ancestor of wheat Uraltu published in Nature in 2013 (Triticum urartu, A genome donor) and Aegilops tauschii (Aegilops tauschii, D genome donor) were electronically cloned to obtain the promoters of TaMS7-A, TaMS7-B and TaMS7-D genes, which were named as TaMS7-A promoter, TaMS7-B promoter and TaMS7-D promoter, the promoters described in the present invention can also be called pTaMS7-A, pTaMS7-B and pTaMS7-D respectively, and their lengths are 2601bp and 2635bp respectively and 2821bp, the nucleotide sequences of which are shown in SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO: 18, respectively.

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Abstract

The invention discloses a wheat fertility related gene TaMS7 and an application method thereof, and belongs to the field of biotechnology. The wheat fertility related gene TaMS7 is obtained by analyzing a whole genome expression spectrum of wheat anthers in different developmental stages, and plant fertility is regulated by regulating the expression of the gene. The method is used for producing and maintaining a wheat male sterile line and preparing hybrid seeds. The gene and the method have important theoretical and practical significance for establishing efficient wheat hybrid seed production techniques, and studying wheat male sterility mechanism and heterosis.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a wheat hybrid breeding method, including wheat genetic male sterile line propagation and hybrid seed preparation, and more specifically relates to a wheat male sterile gene and its mutant and its application in hybrid breeding . technical background [0002] Wheat is a self-pollinated crop, and the core of heterosis utilization is to establish a technical system for efficient production of wheat hybrids. At present, there are mainly three ways to utilize heterosis in wheat production: one is the three-line method, that is, the male sterile line is used to produce hybrids through the three-line interaction of the sterile line, the maintainer line and the restorer line; the other is the three-line method. The first method is the chemical killing method, that is, the application of chemical andricides to induce male sterility in wheat for hybrid seed production; the third i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H1/02A01H5/02A01H6/46
CPCA01H1/02C07K14/415C12N15/8231C12N15/8289
Inventor 李健王峥马力耕邓兴旺
Owner BEIJING NEXT GENERATION HYBRID WHEAT BIOTECHNOLOGY CO LTD
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