Paddy bacterial leaf streak resistance-related gene OsDRxoc6

A bacterial and streak disease technology, applied in genetic engineering, plant genetic improvement, plant peptides, etc., can solve problems such as yield and quality decline

Inactive Publication Date: 2013-03-20
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Rice is an important food crop in the world, but the impac

Method used

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  • Paddy bacterial leaf streak resistance-related gene OsDRxoc6
  • Paddy bacterial leaf streak resistance-related gene OsDRxoc6
  • Paddy bacterial leaf streak resistance-related gene OsDRxoc6

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 OsDRxoc6 Gene Structure Prediction and Analysis

[0039] Using the DNA coding sequence SEQ ID No.2 of the OsDRxoc6 gene as a template, the nucleotide database was searched by BLAST method, and it was found that the sequence of SEQ ID No.2 was located on the short arm of chromosome 5 of rice variety Nipponbare, which was consistent with the LOC_05g01380 gene predicted by Genebank At the same locus, it is located in the rice bacterial streak resistance QTL qBlsr5a. According to the coding sequence of SEQ ID No.2, the amino acid sequence of SEQ ID No.3 can be obtained, which consists of 309 amino acids.

Embodiment 2

[0040] Example 2 Analysis of the expression pattern of OsDRxoc6 gene in different rice varieties

[0041] 1. Background expression of OsDRxoc6 gene in resistant variety Acc8558 and susceptible variety Zhonghua 11

[0042] In order to detect whether there is a difference in the background expression level of the OsDRxoc6 gene in the disease-resistant and susceptible varieties, and to verify whether it is related to disease resistance, the present invention uses real-time quantitative RT-PCR technology to analyze the OsDRxoc6 gene in the rice disease-resistant variety Acc8558 and the susceptible variety ZH11. Differences in background expression in . Total RNA was extracted from leaves of rice plants, and 1-5 μg of total RNA was treated with DNaseI (Invitrogen, USA) for 30 minutes to remove genomic DNA contamination, and then oligo(dT)15 was used to oligomerize Primers and SuperScriptII reverse transcriptase (Invitrogen) were used for reverse transcription. Quantitative PCR an...

Embodiment 3

[0045] Example 3 Isolation of OsDRxoc6 Gene and Construction of Plant Expression Vector

[0046] 1. Isolation of OsDRxoc6 gene

[0047] Using the genomic DNA of the susceptible rice variety Zhonghua 11 as a template, by PCR technology, using primers OsDRxoc6F1 / OsDRxoc6R1 to amplify and obtain a partially suppressed expression DNA fragment of the OsDRxoc6 gene, its sequence is shown in the sequence table SEQ ID NO.4 (SEQ ID NO.2 sequence 11th to 580bp), 1% agarose gel electrophoresis detected its size 570bp ( Figure 5 ); Using the genomic DNA of the disease-resistant rice variety Acc8558 as a template, the OsDRxoc6F2 / OsDRxoc6R2 combination primers amplified the complete overexpression fragment, and the target band of 1151bp was detected by 1% agarose gel electrophoresis ( Image 6 ), whose sequence is shown in the sequence listing SEQ ID NO.1. Primer sequences are shown in Table 1

[0048] Table 1. Related primer sequences

[0049]

[0050]

[0051] 2. Construction o...

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Abstract

The invention relates to the technical field of plant genetic engineering and provides a paddy bacteria leaf streak resistance-related gene coding sequence of which a coded nucleotide sequence is shown as SEQ ID No.2 and a coded amino acid sequence is shown as SEQ ID No.3, a gene OsDRxoc6 at which the paddy bacteria leaf streak resistance-related gene coding sequence is located and of which the nucleotide sequence is shown as SEQ ID No.1, and a separating and cloning method as well as functional verification and application of the gene OsDRxoc6. The gene OsDRxoc6 is located in paddy bacteria leaf streak resistance QTL qB1sr5a, has no intron and is a positive control factor. The inventor also discovers that the resistance level of a transgenic plant capable of suppressing the gene OsDRxoc6 expression on the paddy bacteria leaf streak is remarkably reduced and the disease resistance of the transgenic plant capable of excessively expressing the gene OsDRxoc6 is remarkably improved.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering. It specifically relates to the isolation, cloning, functional verification and application of a rice resistance to bacterial stripe disease related gene OsDRxoc6. The OsDRxoc6 gene is located in the rice bacterial stripe resistance QTLqBlsr5a, the gene has no intron, and it is a positive regulator in rice disease resistance response. The resistance level of transgenic plants suppressing the expression of OsDRxoc6 gene to rice bacterial stripe disease was significantly reduced, while the disease resistance ability of transgenic plants overexpressing this gene was significantly improved. Background technique [0002] Plants are attacked by various pathogens during their growth. There are many types of plant pathogens, including viruses, bacteria, fungi, and nematodes. Pathogen invasion of plants leads to two results: (1) The pathogen successfully reproduces in the host plant, c...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415
Inventor 丁新华储昭辉常清乐
Owner SHANDONG AGRICULTURAL UNIVERSITY
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