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Protein TaPaO associated with plant senescence induction and male sterility, coding gene and application thereof

A technique for male sterility and inducing plants, applied in the field of genetic engineering

Inactive Publication Date: 2010-09-29
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This gene shows an obvious senescent up-regulation trend in natural aging or artificially induced aging leaf tissue, and it is related to the expression of receptor-like protein kinase, and it is expressed in the transgenic leaves of Gmlls1 gene after knocking out receptor-like protein kinase The Lls1 mutant has a typical leaf patchy chlorotic necrosis phenotype similar to maize; however, there is no report on PaO in wheat

Method used

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  • Protein TaPaO associated with plant senescence induction and male sterility, coding gene and application thereof
  • Protein TaPaO associated with plant senescence induction and male sterility, coding gene and application thereof
  • Protein TaPaO associated with plant senescence induction and male sterility, coding gene and application thereof

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Embodiment 1

[0107] Example 1: cDNA cloning and semi-quantitative analysis of the gene TaPaO related to wheat-induced senescence and photothermo-sensitive male sterility

[0108] The anthers of the hybrid wheat variety Jingmai No. 7 (Jingshenmai 2009003) were taken, and the total RNA of the anthers was extracted by Trizol method. 5'RACE kit (5'RACE System for Rapid Amplification of cDNA Ends Kit) (GIBCOBRL, CAT.NO.18374-058) and 3'RACE kit (3'RACE System for Rapid Amplification of cDNA Ends Kit) (GIBCOBRL, CAT.NO.18373-019) to obtain the TaPaO gene.

[0109] The total RNA of the anthers of hybrid wheat variety Jingmai No. 7 (Jingshenmai 2009003) was extracted by Trizol method, and cDNA was obtained by reverse transcription with superscript II (purchased from Invitrogen Company) reverse transcriptase. Primers P1 and P2 were designed according to the sequence of TaPaO gene coding region. Using the cDNA obtained by reverse transcription as a template, PCR amplification was performed with pr...

Embodiment 2

[0119] Embodiment 2: Cultivate male sterile transgenic plants with TaPaO gene

[0120] 1. Construction of recombinant expression vector

[0121] Construction of pBI29A-TaPaO Dicotyledon Recombinant Expression Vector

[0122] Using the cDNA obtained by reverse transcription of the total RNA of the hybrid wheat variety Jingmai No. 7 (Jingshenmai 2009003) as a template, PCR amplification was performed with specific primers containing SacI and SpeI linker sequences; then SacI and SpeI double-digested the PCR product , recovered, and inserted the digested product between the SacI and SpeI restriction sites behind the CaMV 29A promoter of the vector pBI121 in the forward direction to obtain the recombinant vector pBI29A-TaPaO.

[0123] The primer sequences are as follows:

[0124] TaPaO[SacI]: 5'-TCCGAGCTCTCAATCAATGTCAGCGTGCAC-3'

[0125] TaPaO[SpeI]: 5'-CGGACTAGT ATG CCG GTG CTG GCG ATG-3'

[0126] 2. Acquisition and identification of genetically modified tobacco

[0127] 1) O...

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Abstract

The invention relates to the field of gene engineering, in particular to a protein TaPaO associated with plant senescence induction and male sterility, a coding gene and application thereof. The protein has an amino acid sequence as shown in SEQ ID NO.1, and has a nucleotide sequence as shown in SEQ ID NO.2 or 3. The protein associated with senescence induction and male sterility and the coding gene thereof have certain effect on study into sterility mechanisms of photo-temperature sensitive male sterile wheat, and provide important theoretical and practical significance for studying and opening up novel ways of utilizing wheat heterosis.

Description

technical field [0001] The present invention relates to the field of genetic engineering, in particular, the present invention relates to a related protein TaPaO for inducing plant senescence and male sterility, its encoding gene and application. Background technique [0002] In modern agricultural production, people widely use heterosis to improve varieties. Heterosis refers to the phenomenon that the first generation of hybrids produced by the hybridization of two parents with different genetic compositions has advantages over their parents in vigor, growth potential, fecundity, stress resistance, yield and quality. Utilization of heterosis is considered to be the main way to greatly increase the wheat production level in the future. [0003] In wheat genetics and breeding, there are mainly three-line method and two-line method to utilize heterosis. The "three-line method" was the first to be discovered and became the main way to utilize heterosis in wheat for quite a lo...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A01H5/00A01H1/02
Inventor 张立平王灵云郑岑赵昌平单福华唐忠辉张凤廷苑少华高世庆唐益苗
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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