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A related protein tapao1 for inducing plant pollen abortion and its coding gene and application

A technology for pollen abortion and inducing plants, applied in the field of genetic engineering, can solve problems such as affecting pollen fertility

Active Publication Date: 2020-05-22
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Current reports on PaO include: clones obtained from maize lls1 (lethal leaf-spot 1) mutants and Arabidopsis acdl (accelerated cell death 1) mutants that exhibit necrotic spots (Gray et al., 1997; Yang et al. , 2004); Li Pengli et al. (2006) cloned the homologous gene of maize LLS1 from soybean, with a full length of 1817bp, and named it Gmlls1. However, there is no relevant report that wheat PaO affects pollen fertility

Method used

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  • A related protein tapao1 for inducing plant pollen abortion and its coding gene and application
  • A related protein tapao1 for inducing plant pollen abortion and its coding gene and application
  • A related protein tapao1 for inducing plant pollen abortion and its coding gene and application

Examples

Experimental program
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Embodiment 1

[0037] Example 1: cDNA cloning and semi-quantitative analysis of the gene TaPaO1 related to wheat-induced senescence and photothermo-sensitive male sterility

[0038] The anthers of the hybrid wheat variety BS366 were taken, and the total RNA of the anthers was extracted by Trizol method. Using barley PaO (GenBank: AK365037) as a probe to screen the TaPaO1 gene fragment from the wheat BS366 library.

[0039] Total RNA of anthers of hybrid wheat variety BS366 (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 primers P1 and P2. The sequences of primers P1 and P2 are as follows:

[0040] P1: 5'-GTTGCGTGTGGGTGGCTG-3'

[0041] P2: 5'-TCTCCCTAATCAATCAATGTC-3'

[0042]...

Embodiment 2

[0051] Embodiment 2: Cultivate male sterile transgenic plants with TaPaO1 gene

[0052] 1. Construction of recombinant expression vector

[0053] Construction of pCambia2300-RD29A-TaPaO1 Dicotyledon Recombinant Expression Vector

[0054] Using the cDNA obtained by reverse transcription of the total RNA of the hybrid wheat variety BS366 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 forwarded The recombinant vector pCambia2300-RD29A-TaPaO1 was obtained by inserting it between the SacI and SpeI restriction sites behind the promoter of the vector pCambia2300-RD29A.

[0055] The primer sequences are as follows:

[0056] TaPaO1[SacI]: 5’–TCCGAGCTC CCC TAA TCA ATC AAT GTC AGC-3’

[0057] TaPaO1[SpeI]: 5’–CGGACTAGT ACG CCA TGC GTG CGT TCT CCA G-3’

[0058] 2. Acquisition and identification of genetically modified tobacco

[0059] 1) Obtaining geneti...

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Abstract

The invention relates to the field of gene engineering, in particular to a plant pollen abortion inducing related protein TaPaO1, coding gene and application thereof. An amino acid sequence of the protein is as shown in SEQ ID NO.1, and a nucleotide sequence of the protein is as shown in SEQ ID NO.2 or 3. The protein and the coding gene thereof have certain effect on the research on the sterilitymechanism of photo-and thermo-sensitive male sterile wheat, and provide very important theoretical and practical significance on the research and development of a new approach of wheat heterosis utilization.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular, the invention relates to a related protein TaPaO1 for inducing plant pollen abortion, its coding gene and application. Background technique [0002] Pheophytin (methyl ester) acid oxygenase (PaO) is the most important key enzyme among the enzymes involved in chlorophyll degradation and metabolism discovered so far (Hortensteiner S et al., 1998; Curty C et al., 1995). [0003] Current reports on PaO include: clones obtained from maize lls1 (lethal leaf-spot 1) mutants and Arabidopsis acdl (accelerated cell death 1) mutants that exhibit necrotic spots (Gray et al., 1997; Yang et al. , 2004); Li Pengli et al. (2006) cloned the homologous gene of maize LLS1 from soybean, with a full length of 1817bp, and named it Gmlls1. However, there is no related report that wheat PaO affects pollen fertility. Contents of the invention [0004] The purpose of the present invention is to provide...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/82A01H5/02A01H6/46
CPCC12N9/0071C12N15/8289C12Y114/1202
Inventor 张立平苑国良赵昌平苑少华白建芳王娜高世庆张风廷单福华孙辉马锦绣
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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