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A technology of insect resistance and homology, applied in the biological field, can solve the problems of pest resistance and re-rampant, unreasonable, and human food safety threats.
Active Publication Date: 2019-06-21
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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Problems solved by technology
Pests and diseases seriously affect agricultural production, affect the yield and quality of crops, and restrict the stable development of agricultural economy
For a long time, people have generally used chemical pesticides to control pests. However, with the passage of time, especially the large and unreasonable use of chemical pesticides has brought about problems such as pesticide residues, pest resistance and re-emergence, and killing natural enemies. Serious problems such as destruction of natural ecological balance, environmental pollution, and threats to human food safety
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Embodiment 1
[0073] Embodiment 1, the acquisition of the recombinant vector that suppresses SlMTB1, SlMTB2, SlMTB3 gene expression
[0074] The total RNA of tomato (Solanum lycopersicon) variety M82 was extracted, and the cDNA obtained by reverse transcription was used as a template. Primer 1 and primer 2 were used to amplify SlMTB1 by PCR, primer 3 and primer 4 were used to amplify SlMTB2 by PCR, and primer 5 and primer 4 were used to amplify SlMTB2 by PCR. Primer 6 was used to amplify SlMTB3 by PCR; then the amplified product of the previous three steps was used as a template to amplify SlMTB1 / 2 / 3 by PCR using primer 1 and primer 6.
[0075] Primer 1:
[0076] 5′-GGGGACAAGTTTGTACA AAAAAGCAGGCT CCTCTTAGGAGATGCCAT-3' (upstream primer, the base underlined is the attB1 site);
[0077] Primer 2: 5'-CCCTGATTTCAGCGGAGCTCACCCTTACAA-3';
[0078] Primer 3: 5'-CCGCTGAAATCAGGGGTTGTGGAGCTT-3';
[0079] Primer 4: 5'-GCAGAAACTAAACTATCCACCTTCGGAGAA-3';
[0080] Primer 5: 5'-TAGTTTAGTTTCTGCTGAATGTT...
Embodiment 2
[0086] Example 2, SlMTB1, SlMTB2, SlMTB3 gene silencing transgenic tomato acquisition and functional identification
[0089] The recombinant vector pHELLSGATE2-SlMTB1 / 2 / 3-RNAi obtained by Example 1 for silencing SlMTB1, SlMTB2, and SlMTB3 gene expression was transformed into Agrobacterium AGLO (recorded by Li C, Liu G, Xu C, Lee GI, Bauer P, Ling HQ, Ganal MW, Howe GA. The tomato suppressor of prosystemin-mediated responses2 gene encodes a fatty acid desaturase required for the biosynthesis of jasmonic acid and the production of a systemic wound signal for defense gene expression. Plant Cell. 2003.15(7):1646-1661. The public can obtain it from the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences), pick a single clone, and shake it overnight at 28°C in LB liquid medium containing spectinomyci...
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Abstract
The invention discloses a method for improving insect resistance of tomato and provides application of protein or related biomaterials thereof in regulating plant insect resistance. The related biomaterials are nucleic acid molecules capable of expressing protein or an expression box, a recombinant vector, recombinant bacteria or a transgenic cell line containing the nucleic acid molecules, wherein the protein is SlMTB1 protein, SlMTB2 protein and SlMTB3 protein. Experiments prove that transgenic tomato with SlMTB1, SlMTB2 and SlMTB3 genes being silent can express more PI-II than wild tomato when being infringed by cotton bollworm, and resistance of the transgenic tomato to cotton bollworm is improved remarkably. Therefore, SlMTB1, SlMTB2 and SlMTB3 protein and part of coding genes thereofcan be used for cultivating and identifying insect-resistant plant varieties needed for agriculture and animal husbandry and ecological environment management and are of important significance in thefield of plant insect-resistant breeding.
Description
technical field [0001] The invention belongs to the field of biotechnology and relates to a method for improving insect resistance of tomato. Background technique [0002] Tomato is a very important economic crop. It is both a fruit and a vegetable. It has rich nutritional value and is deeply loved by consumers. Tomato is one of the most common fruits and vegetables cultivated in my country. At present, China is the country with the largest tomato cultivation area and the largest production volume in the world. With the continuous improvement of agricultural technology, the tomato planting area is expanding day by day, and the pests and diseases are becoming more and more serious. Pests and diseases seriously affect agricultural production, affect the yield and quality of crops, and restrict the stable development of agricultural economy. For a long time, people have generally used chemical pesticides to control pests. However, with the passage of time, especially the lar...
Claims
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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82
CPCY02A40/146
Inventor 李传友刘圆圆翟庆哲邓磊蒋红玲
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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