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Tomato anti insect related gene, its coding protein and application

A tomato, gene technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem that the JA signal transduction pathway is poorly understood and so on

Inactive Publication Date: 2008-08-20
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, the biochemical pathway of JA synthesis has been basically understood, but the regulation of JA biosynthesis and the signal transduction pathway of JA are still poorly understood

Method used

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  • Tomato anti insect related gene, its coding protein and application
  • Tomato anti insect related gene, its coding protein and application
  • Tomato anti insect related gene, its coding protein and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1. Acquisition of Tomato Resistance Deletion Mutant JL1

[0030] The conventional tomato variety M82 was selected as the experimental material, and its seeds were soaked in 0.5% ethylmethyl sulfonate (EMS) for 12 hours for mutagenesis treatment, and the offspring materials were treated with protease inhibitor II after cultivation. No resistance testing, protease inhibitor II deficient material was self-propagated. Each generation was tested with the same method as above, and finally a single plant lacking resistance to protease inhibitors was selected for self-propagation. After 5 generations of selection, a stable tomato resistance-deficiency mutant material was obtained, named JL1 .

[0031] The most basic phenotypic characteristic of the mutant JL1 is the lack of expression of resistance-related proteins PIs after injury. The inoculation test was carried out on it, and the result was that the mutant lost resistance to insects. Further analysis showed that th...

Embodiment 2

[0032] Embodiment 2, the acquisition of LeAcx1

[0033] 1. Obtain tomato LeAcx1 by map-based cloning

[0034] Through the fine-mapping analysis of a large population containing 1200 individuals, the target gene was mapped to a region on chromosome 8 of tomato with a length of about 80 KB covered by BAC clones 166B24 and 232L13, such as figure 1As shown (the number represents the number of recombinants between the molecular marker and the target gene), the sequence analysis of the DNA fragment was carried out, and it was predicted that the BAC clone covered 8 genes, in which two acyl-coenzyme oxidases (Acyl-CoAOxidase, ACX) gene is considered to be an important candidate gene that causes JL1 resistance loss gene JL1 (the related genes that cause JL1 protease inhibitor resistance loss are collectively referred to as Jl1), and are named LeAcx1 and LeAcx2 respectively, wherein LeAcx1 has the The polynucleotide sequence of SEQ ID №: 1, SEQ ID № in the sequence listing: 1 consists ...

Embodiment 3

[0045] Embodiment 3, the verification experiment of LeAcx1 biological function

[0046] 1. Functional verification experiment of LeAcx1 in JA synthesis and fatty acid metabolism

[0047] Design primers to amplify LeAcx1 according to the cDNA sequence of the mutant LeAcx1 obtained in step 1 and the appropriate restriction site of the vector pET-23b. The primer sequences are as follows:

[0048] Primer 5: (upstream primer) 5'-C GAGCTC GGTAGAGGAGATGGAGGGTGTA-3' (the bases underlined are SacI recognition sites

[0049] Primer 6: (downstream primer) 5'-CCG CTCGAG CGGAACAGTTTGCTGCAGCTCTCG-3' (underlined bases are XhoI recognition sites)

[0050] Extract the total RNA of tomato conventional variety M82, use its reverse transcription product as a template, and carry out PCR amplification under the guidance of primer 5 and primer 6. After the reaction, the PCR product is purified, and the restriction endonuclease SacI and XhoI double digested the purified PCR product, then ligate...

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Abstract

The present invention discloses a tomato insect-resisting related gene, its coding protein and application. Said invention is aimed at providing a tomato insect-resisting related gene, its coding protein and application for breeding insect-resisting plant. The described tomato insect-resisting related gene has one of the following nucleotide sequences: (1), DNA sequence of SEQ ID No.1 in sequence table; (2), polynucleotide of SEQ ID No.2 protein sequence in coding sequence table; and (3), nucleotide sequence which can be hybridized with DNA sequence limited by SEQ ID No.1 in sequence table under the high stringent condition.

Description

technical field [0001] The invention relates to a plant gene, its encoded protein and its application, in particular to a tomato insect-resistance-related gene, its encoded protein and its application in cultivating insect-resistant plants. Background technique [0002] At present, in the world, the loss of crop yield due to insect infestation is very serious. The most common method of traditional control of insect infestation is the extensive application of chemical insecticides. According to statistics, 80% of the world's increased production through plant protection measures rely on pesticides. While the use of chemical pesticides has achieved certain economic benefits, it has also brought about serious problems such as pesticide residues, environmental pollution, ecological damage, pest resistance and re-emergence. Therefore, research and utilization of plant resistance genes to effectively control agricultural pests has become the goal that human beings are striving f...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N5/10C07H21/00
Inventor 李传友李常保蒋红玲吴晓燕
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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