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XNDL2 gene, protein, overexpression vector, acquisition method for rice resistant to sheath blight, and application of XNDL2 gene

An overexpression vector and acquisition method technology, applied in the biological field, can solve the problems of lack of sheath blight resistant rice germplasm resources, and no rice varieties with high sheath blight resistance or immunity have been found.

Active Publication Date: 2020-03-24
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, due to the lack of rice germplasm resources resistant to sheath blight in my country, no rice varieties with high resistance or immunity to sheath blight have been found.

Method used

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  • XNDL2 gene, protein, overexpression vector, acquisition method for rice resistant to sheath blight, and application of XNDL2 gene
  • XNDL2 gene, protein, overexpression vector, acquisition method for rice resistant to sheath blight, and application of XNDL2 gene
  • XNDL2 gene, protein, overexpression vector, acquisition method for rice resistant to sheath blight, and application of XNDL2 gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Obtaining xndl2 transgenic rice plants edited by CRISPR / Cas9

[0047] 1) Acquisition of transgenic rice

[0048] Provide the OsXNDL2 gene sequence to Baige Gene Technology Co., Ltd., and the company will carry out target site sequence design, primer design, vector construction and transformation, transform rice variety Dongjin, and finally obtain T0 transgenic rice.

[0049] 2) Identification of transgenic rice

[0050] The obtained transgenic plants were further verified by sequencing analysis, such as figure 1 shown. Through the sequencing analysis of the positive plants, it was found that the base G was inserted after the 80th base of the OsXNDL2 coding gene sequence shown in Sequence 1, and the 81st to 84th bases were deleted, so that the OsXNDL2 gene function was lost ( figure 1 ), and the two gene-edited plants were named xndl2-1 and xndl2-2, respectively.

Embodiment 2

[0052] Obtaining XNDL2 Overexpression Transgenic Rice Plants

[0053] 1) Construction of a recombinant vector for overexpressing the XNDL2 gene

[0054] The nucleotide sequence of the OsXNDL2 (LOC_Os02g34970) gene is nucleotides 1-615 of SEQ ID No.1, the encoded protein is OsXNDL2, and the amino acid sequence of the protein is SEQ ID No.2.

[0055] The RNA of rice variety Dongjin was extracted and reverse transcribed into cDNA. Using this cDNA as a template, use SEQ ID Mo.3: 5'-AAGCTTATGGGAGGAGCTACAAACTT-3' and SEQ ID No.4: 5'-GGATCCCTATTTATTCGGCAAACTTAC-3' as primers to amplify, and use the cDNA obtained by reverse transcription as a template to amplify The OsXNDL2 gene of rice Dongjin was cloned by RT-PCR. The specific reaction system was as follows: 5×SFBuffer (with 10 mM MgSO4) 10 μL, dNTPMix (10 mM each) 1 μL, upstream primer (10 μm) 2 μL, downstream primer (10 μm) 2 μL, Phanta Super-Fidelity DNA Polymerase 1 μL, template cDNA 1 μL, ddH2O up to 50 μL; the reaction progr...

Embodiment 3

[0071] Phenotype observation of xndl2 transgenic rice and XNDL2-OX transgenic rice resistant to sheath blight after inoculation

[0072] Resistance identification was carried out by inoculating detached leaves of rice with sheath blight. The leaves of wild-type control, xndl2 transgenic rice and XNDL2-OX transgenic rice were inoculated with Rhizoctonia solani.

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Abstract

The invention provides an XNDL2 gene, a protein, an overexpression vector, an acquisition method for rice resistant to rice sheath blight disease and application of the XNDL2 gene, belonging to the technical field of biology. The nucleotide sequence of the XNDL2 gene is shown as SEQ ID No. 1. The XNDL2 gene provided by the invention has the effect of resisting rice sheath blight disease. When theXNDL2 gene is introduced into rice, the rice capable of resisting rice sheath blight disease can be obtained.

Description

technical field [0001] The invention belongs to the technical field of biotechnology, and in particular relates to an XNDL2 gene, a protein, an overexpression vector, an acquisition method and application of rice sheath blight-resistant rice. Background technique [0002] Rice sheath blight is a soil-borne fungal disease caused by Rhizoctonia solani. It is one of the three major diseases of rice, and it is also a rice disease that harms all over the world. The disease can occur in the whole growth period of rice, mainly damages leaf sheaths and leaves, has the characteristics of great harm, high prevalence, and wide host range, and seriously affects the high and stable yield of rice. In agricultural production, the prevention and control of rice sheath blight mainly adopts pesticide control and cultivation management, but long-term use of chemical pesticides not only causes environmental pollution, but also accelerates the mutation of pathogenic bacteria and produces drug re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8282
Inventor 孙倩玄元虎李天亚邱永春苑德鹏徐晓凤
Owner SHENYANG AGRI UNIV
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