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OsHAP2C (oryza sativa heme activator protein 2C) and application of encoding genes thereof to regulating rice bacterial leaf blight resistance

A disease-resistant, transgenic plant technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problem of lack of OsHAP2s

Active Publication Date: 2019-03-01
INST OF CROP SCI CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no report that OsHAP2s regulate rice disease resistance.

Method used

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  • OsHAP2C (oryza sativa heme activator protein 2C) and application of encoding genes thereof to regulating rice bacterial leaf blight resistance
  • OsHAP2C (oryza sativa heme activator protein 2C) and application of encoding genes thereof to regulating rice bacterial leaf blight resistance
  • OsHAP2C (oryza sativa heme activator protein 2C) and application of encoding genes thereof to regulating rice bacterial leaf blight resistance

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Experimental program
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Effect test

Embodiment 1

[0113] Example 1, OsHAP2C can regulate rice bacterial blight resistance

[0114] This embodiment provides a protein named OsHAP2C derived from 9804 rice, its sequence is the 240-448th position of sequence 3 in the sequence listing, the sequence of the coding region of the OsHAP2C gene in 9804 rice is sequence 2 of the sequence listing, the genome The sequence is sequence 1 in the sequence listing.

[0115] 1. Construction of recombinant vector

[0116] 1) OsHAP2C gene expression vector

[0117] Total RNA was extracted from 9804 rice leaves and reverse transcribed into cDNA. Using the cDNA obtained in the above steps as a template, PCR amplification was performed using primers attB-F: 5′-AAGCAGGCTTTGACTTTATGCAAAGTTTGTTTGCATCTA-3′ and primers attB-R: 5′-GGGTCTAGAGACTTTAGCGCCCTCTTTCGCACGAC-3′ to obtain amplified products. Through BP reaction, the amplified product obtained in the above steps was introduced into the vector pGWC to obtain the positive entry clone plasmid pGWC-Os...

Embodiment 2

[0164] Example 2, the expression level of OsHAP2C gene after inoculation of Xoo bacterial strain KS6-6

[0165] 1. Inoculation of rice line 9804 with Xoo strain KS6-6 and leaf sampling

[0166] The seeds of 9804 rice were sown in seedling trays filled with sterilized soil, cultivated in the greenhouse for 25 days, and then transplanted into the net room, and planted as a single plant. At the full tillering stage of rice plants, Xoo strain KS6-6 was used to artificially inoculate rice plants by leaf-cutting method, and each plant was inoculated with 5 leaves. After 0, 2, 4, 8, 12, 24, and 36 hours of artificial inoculation, the inoculated leaves were clipped and quickly frozen in liquid nitrogen, and three biological replicates were taken for each sample. All samples were frozen in liquid nitrogen and stored at -70°C.

[0167] 2. Real-time quantitative PCR analysis

[0168] RNA extraction: TRIZON reagent was used to extract the total RNA of the sample, and the purity of the ...

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Abstract

The invention discloses an OsHAP2C and application of encoding genes thereof to regulating rice bacterial leaf blight resistance. The OsHAP2C is shown as the followings of A1), A2) or A3), wherein theamino acid sequence of the A1) is the 240th to 448th proteins of a sequence 3; the A2) is proteins prepared by substituting and / or missing and / or adding of one or more amino acid residues passed by the amino sequence shown by the 240th to 448th proteins of the sequence 3; the A3) is fused proteins by connecting tags to the N terminals or / and the C terminals of the A1) or the A2). Experiments prove that, when OsHAP2C genes are knocked out, the disease resistance of plants can be improved, and the disease resistance of OsHAP2C transgenic plants is significantly reduced, so that the OsHAP2C genes can be applied to regulating the disease resistance of plants.

Description

technical field [0001] The invention relates to the application of OsHAP2C and its coding gene in regulating rice bacterial blight resistance in the field of biotechnology. Background technique [0002] Bacterial blight caused by Xanthomonas oryzae pv.oryzae (Xoo) is one of the most important bacterial diseases in rice cultivation in the world, and it is seriously harmful to rice fields in South China and Southeast Asia. Rice bacterial blight can generally lead to about 10% reduction in rice production, and in severe cases, it can reduce production by 50%-60%. Breeding disease-resistant varieties using resistance genes is currently the most economical and effective measure to control rice bacterial blight. So far, 42 rice bacterial blight resistance genes have been reported at home and abroad (http: / / www.shigen.nig.ac.jp / rice / oryzabase / gene / list). However, the resistance genes derived from wild rice are difficult to use; some resistance genes only have adult plant resistan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N5/10A01H5/00A01H6/46
CPCC07K14/415C12N15/8281
Inventor 周永力曾丹石英尧陈腾君黎志康
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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