Applications of arabidopis thaliana JAV1 protein and coding gene of arabidopsis thaliana JAV1 protein in regulation of plant disease resistance and insect resistance

A technology encoding gene and protein is applied in the application field of Arabidopsis JAV1 protein and its encoding gene in regulating plant disease resistance and insect resistance, which can solve problems such as affecting resistance effect, and achieve broad application space and market prospect. Effect

Active Publication Date: 2014-02-19
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most of this technology is based on the overexpression of exogenous resistance gen

Method used

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  • Applications of arabidopis thaliana JAV1 protein and coding gene of arabidopsis thaliana JAV1 protein in regulation of plant disease resistance and insect resistance
  • Applications of arabidopis thaliana JAV1 protein and coding gene of arabidopsis thaliana JAV1 protein in regulation of plant disease resistance and insect resistance
  • Applications of arabidopis thaliana JAV1 protein and coding gene of arabidopsis thaliana JAV1 protein in regulation of plant disease resistance and insect resistance

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Experimental program
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Effect test

Embodiment 1

[0065] Embodiment 1, the acquisition of Arabidopsis JAV1 gene

[0066] Extract total RNA from Arabidopsis thaliana Columbia-0 ecotype (Arabidopsis Biological Resource Center (ABRC), seed number: CS6673) flowers, reverse transcribe to obtain cDNA, use the cDNA as a template, and use primer 1:5' -CGAATCAAAATTAAACATTGT-3' and primer 2: 5'-AAAACATTTTTATTTATTT-3' for PCR amplification. The obtained PCR product was sequenced, and the result is shown in sequence 2 of the sequence listing. Sequence Listing Sequence 2 is the full-length cDNA sequence of Arabidopsis thaliana JAV1 gene, wherein, the 260-838th position of the self-sequence listing sequence 2 is an open reading frame, encoding the JAV1 protein composed of 192 amino acids shown in the sequence listing sequence 1; The 1st to 259th positions at the 5' end of Listing Sequence 2 are 5'UTR regions; the 839th to 1129th positions at the 3' end of Sequence Listing 2 are 3'UTR regions.

[0067] Sequence 2 in the Sequence Listing c...

Embodiment 2

[0068] Example 2, Utilizing the RNAi vector of JAV1 to cultivate insect-resistant and disease-resistant transgenic Arabidopsis

[0069] 1. Acquisition of RNAi vector pBI121-JAV1-RNAi

[0070] Using the Arabidopsis cDNA of Example 1 as a template, use primer 3: 5'-GC TCTAGA ATGGCTAACCCCAACGAGTG-3' (the underlined base is the Xba I recognition sequence) and primer 4: 5'-TT GGTACC AGCAGAAGTAGTATTACCGG-3' (the underlined base is the Kpn I recognition sequence) was amplified by PCR to obtain a DNA fragment of about 300 bp, which was digested with Xba I and Kpn I, and combined with the The vector backbone fragments of vector pTCK303 were ligated to obtain intermediate vector 1.

[0071] Using the Arabidopsis cDNA of Example 1 as a template, use primer 5: 5'-CC ACTAGT AGCAGAAGTAGTATTACCGG-3' (the underlined base is the Spe I recognition sequence) and primer 6: 5'-TA GAGCTC ATGGCTAACCCCAACGAGTG-3' (the underlined base is the Sac I recognition sequence) was amplified by PCR to ...

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Abstract

The invention discloses applications of arabidopsis thaliana JAV1 protein and a coding gene of arabidopsis thaliana JAV1 protein in regulation of plant disease resistance and insect resistance. The invention also provides a method used for improving plant disease resistance and insect resistance, short hairpin RNA capable of reducing JAV1 gene expression material content, small interfering RNA, an interference vector, and interference segments. It is confirmed by experiments that, compared with wild type and null-transfection vector control groups, relative expression level of T3 generation pBI121-JAV1-RNAi transfected arabidopsis thaliana is reduced significantly, leaf infected area is 0.25 to 0.27cm<2> after inoculation by botrytis cinerea, and is obviously smaller than that of wild type and null-transfection vector control groups; the weight of beet armyworm larvae which are fed with leaves of 5 weeks for 6 days increases 1.58 to 2.03mg, and is obviously lower than that of wild type and null-transfection vector control groups; and plants are capable of growing normally in 12 days when bradysia odoriphaga larvae are put into soil with the plants of 18 days. According to the applications of arabidopsis thaliana JAV1 protein and the coding gene of arabidopsis thaliana JAV1 protein, a novel approach is provided for cultivation of disease-resistant and insect-resistant plants, and in the field of agricultural production, application range is wide, and market prospect is promising.

Description

technical field [0001] The invention relates to the application of Arabidopsis thaliana JAV1 protein and its coding gene in regulating plant disease resistance and insect resistance. Background technique [0002] In the process of evolution, plants will form a certain degree of ecological adaptability to avoid pests or pathogenic bacteria, to avoid pests, to tolerate pests, or to resist bacteria. Plant resistance genetic engineering based on plant resistance research is a biotechnology that utilizes the principles of molecular genetics to identify and isolate resistance genes, introduce resistance genes into recipients, and obtain regenerated individuals with stable expression and inheritance of resistance. Most of the resistance genes come from natural stress-resistant organisms, or the protective genes produced by plants in adversity. Transferring these genes to varieties with poor resistance can improve stress resistance. Most of this technology is based on the overexpr...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/63C12N5/10C12N1/21C12N7/01C12N15/84A01H5/00
Inventor 谢道昕胡珀周武
Owner TSINGHUA UNIV
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