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Promoter of gene Me094 related to bacterial-blight resistance of Oryza meyeriana

A technology for resisting bacterial blight and verrucous wild rice, which is applied in the field of molecular biology, can solve the problem of weakened disease resistance of transgenic rice and achieve the effect of improving resistance

Inactive Publication Date: 2015-03-11
云南省农业科学院生物技术与种质资源研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through the research, it was found that the expression of Me094 in transgenic rice can improve the rice's resistance to bacterial blight, and the disease resistance of transgenic rice was significantly weakened after RNAi interference technology was used to interfere with the expression of Me094 gene

Method used

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  • Promoter of gene Me094 related to bacterial-blight resistance of Oryza meyeriana
  • Promoter of gene Me094 related to bacterial-blight resistance of Oryza meyeriana
  • Promoter of gene Me094 related to bacterial-blight resistance of Oryza meyeriana

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Cloning and analysis of the promoter of the bacterial blight resistance-related gene Me094 in wild rice

[0049] (1) Design primers to amplify the Me094 promoter

[0050] Me094 gene is a metallothionein gene screened and cloned by the applicant from the SSH library induced by Xanthomonas oryzae pv. oryzae in the early stage. It has been functionally verified, but its upstream promoter sequence is unknown, which limits it. Research on the regulation of the gene expression. According to the full-length cDNA sequence information of the Me094 gene, combined with the principle of TAIL-PCR technology, primers designed by the biological software Primer5.0 were used to amplify the promoter of the Me094 gene. The nucleotide sequence of the full-length cDNA of the Me094 gene is shown as SEQ in the sequence table. ID NO: 12, the designed primer for amplifying the promoter of the present invention (ie, the promoter of the bacterial blight resistance-related gene Me094 of Oryz...

Embodiment 2

[0081] Example 2 Construction of a high-efficiency expression vector for the Me094 promoter of the present invention and transformation of Arabidopsis thaliana to study the function of the promoter

[0082] 1. Construction of Me094 promoter expression vector

[0083] The promoter expression vector was constructed on the basis of obtaining the Me094 promoter sequence, that is, the Me094 promoter was used to replace the 35S promoter on the vector pBI121 (vector pBI121 is commercially available). According to the restriction sites on both sides of the 35S promoter on pBI121 and the sequence analysis results of the Me094 promoter, the pBI121 and Me094 promoter cloning vectors were digested with HindⅢ and Xba Ⅰ. The restriction system is:

[0084]

[0085] Reaction at 37°C for 3 hours, then 1% agarose gel electrophoresis, gel cutting and recovery of the target bands. For the method, see the gel recovery kit instructions (gel recovery kit purchased from Shanghai Shenggong). Then use T 4 L...

Embodiment 3

[0109] Example 3: Verification and expression analysis of the expressed sequence tag of the bacterial blight-resistant gene Me094 of wild rice

[0110] (1) Cultivation and processing of materials

[0111] Jinghong wart grain wild rice (Oryza meyeriana) is planted in a plastic greenhouse until the flowering period.

[0112] Activate bacterial blight pathogen Y8 with NA medium, culture at 28°C for 2-3 days, wash the lawn with sterile distilled water, and prepare OD 560 Bacteria liquid with a treatment value of 0.6. After 15:00 in the afternoon (this period is the time when the pathogenic bacteria of bacterial blight has the strongest infectivity), the leaves of wild rice verrucosa in the greenhouse were inoculated by the method of leaf cutting. The control group was inoculated with sterile water as a mock inoculation. 48h, 72h, 96h, 120h were sampled in equal amounts. The control, that is, the wild rice verrucosa with sterile water cut leaves, was also sampled in equal amounts. Immedi...

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Abstract

The invention discloses a promoter of a gene Me094 related to bacterial-blight resistance of Oryza meyeriana. The promoter belongs to tissue specificity expression promoters and can drive specific expression of a GUS gene in leaves of transgenic Arabidopsis. The promoter can be applied to seed breeding of rice resistant to bacterial blight, and can improve resistance of the rice to bacterial blight by driving concentrated specific expression of other exogenous genes especially genes resistant to bacterial blight in leaves of the transgenic rice. Additionally, the promoter has great theoretical and practical significance in prevention of other foliage diseases of the rice.

Description

Technical field [0001] The invention relates to a promoter of bacterial blight-resistant related gene Me094 cloned from wild rice verrucosa, belonging to the technical field of molecular biology. Background technique [0002] Gene expression and regulation is one of the important contents of plant genetic engineering research. As an important regulatory element, promoters regulate the expression of foreign genes in specific tissues, specific developmental stages and certain environmental conditions of transgenic plants. The use of genetic engineering technology to introduce relevant resistance genes into plants, so that plants can withstand various biotic or abiotic stresses, is a better way to improve plant resistance. The configuration of promoters and the on-demand expression of foreign genes are It seems very important. At present, the most widely used promoter in plant transgenic research is the 35S promoter derived from cauliflower mosaic virus (CaMV) (Odell JT, et al, Ide...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/11C12N15/84A01H5/00
Inventor 李定琴程在全钟巧芳肖素勤柯学李维蛟余腾琼张敦宇付坚王玲仙陈玲陈越蒋聪罗红梅曾民王波黄兴奇
Owner 云南省农业科学院生物技术与种质资源研究所
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