Cucumber CsDIR16 gene and application of cucumber CsDIR16 gene in lowering plant propamocarb hydrochloride pesticide residues

A technology for pesticide residues and propamocarb, applied in the field of genetic engineering applications, can solve problems such as unreported and reduced propamocarb residues, and achieve the effects of reducing pesticide residues, reducing pesticide usage, and reducing production costs

Active Publication Date: 2017-07-07
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] DIR Transporter genes play an important role in plant detoxification and metabolism, and participate in various stress responses, but for Cs DIR16 There is no report on reducing propamocarb residues in plants

Method used

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  • Cucumber CsDIR16 gene and application of cucumber CsDIR16 gene in lowering plant propamocarb hydrochloride pesticide residues
  • Cucumber CsDIR16 gene and application of cucumber CsDIR16 gene in lowering plant propamocarb hydrochloride pesticide residues
  • Cucumber CsDIR16 gene and application of cucumber CsDIR16 gene in lowering plant propamocarb hydrochloride pesticide residues

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Experimental program
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Effect test

Embodiment 1

[0025] CsDIR16 Gene isolation and cloning and expression analysis

[0026] In the previous study, we identified a significantly up-regulated expression induced by propamocarb by analyzing the expression profile of cucumber downy mildew stress. CsDIR16 gene, in order to obtain its full-length sequence information, we searched in the cucumber genome database (http: / / www.icugi.org / ) CsDIR16 The full-length sequence was designed using Primer Premier 5.0 to design specific primers for PCR amplification. The primer sequences are as follows:

[0027] Forward primer: 5′-ATGGCTGGAATCTCTCCAAT-3′

[0028] Reverse primer: 5′-TCAATAATGAAGCACGTAAATGTTA-3′

[0029] Start with Trizol (Invetrigen TM) method to extract cucumber fruit, stem, leaf and total RNA (the kit was purchased from Invetrigen, and the operation method was in accordance with the instructions). Then take 1 ul of the RNA sample, and reverse transcribe it into the first strand of cDNA according to the instructions of the...

Embodiment 2

[0040] CsDIR16 Three-dimensional molecular structure diagram of protein

[0041] Use CPHmodels 3.2 (http: / / www.cbs.dtu.dk / services / CPHmodels / ) to generate atomic coordinates (PDB format), and then use Rasmol windows software to output the three-dimensional molecular structure diagram of CsDIR protein, analyze and predict the protein structure ( Figure 4 ), the protein contains 7 β-sheets (yellow) and 1 α-helix (pink).

Embodiment 3

[0043] CsDIR Family protein and phylogenetic analysis

[0044] Arabidopsis AtDIR The protein is used as the query sequence, and all annotated protein sequences of BLAST cucumber have an e-value value of 1e-20. The results showed that there are a total of 23 in cucumber CsDIR Family genes, sorted according to the distribution of genes on chromosomes, named CsDIR1 , CsDIR2 ... CsDIR23 , as shown in Table 1, 23 CsDIR The genes are distributed on 7 chromosomes of cucumber, among which chromosome 4 is the most. will be 23 CsDIR Compare the proteins of the family genes to construct a gene phylogenetic tree, such as Figure 5 As shown, these CsDIR genes were divided into four subfamilies a, b, d, and e according to the phylogenetic analysis, none of which belonged to the c subfamily. DIRa , DIRb , DIRd with DIRe The differentiation in genetic distance suggests that the possible biological functions of members of this subfamily are not limited to disease resist...

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Abstract

The invention discloses a cucumber CsDIR16 gene and application of the cucumber CsDIR16 gene in lowering plant propamocarb hydrochloride pesticide residues. A nucleotide sequence of the cucumber CsDIR16 gene is shown as SEQ ID NO:1, the total length of the CsDIR16 is 588bp, an amino acid sequence of protein encoded by the CsDIR16 is shown as SEQ ID NO:2, 195 amino acids are encoded, and the CsDIR16 belongs to a Dirigent like protein family. An agrobacterium tumefaciens-mediated genetic transformation method is utilized to transform cucumbers, and an obtained transgenic plant is subjected to biological function verification, shown that the cloned CsDIR16 gene of the invention or the protein encoded by the CsDIR16 gene has the function of lowering the plant propamocarb hydrochloride pesticide residues, so that investment of a pesticide is reduced, the agricultural production cost is lowered, efficient development of agricultural industry is facilitated, and a new genetic resource is supplied to implement green agriculture and organic agriculture.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering application and relates to cucumber CsDIR16 Gene and its application in reducing plant propamocarb pesticide residues. Background technique [0002] As an important cultivated crop of vegetable production in the world, cucumber is loved by people because of its delicate taste and crisp taste, and occupies an important position in market consumption. However, cucumbers are often harmed by various diseases and insect pests in the production process, seriously affecting the yield or quality, among which cucumber downy mildew is the most serious. Propamocarb (PM) is a fungicide that can effectively control cucumber downy mildew. It is mainly controlled by spraying. When the disease is controlled, propamocarb will also produce residues in cucumbers. In recent years, with the improvement of the quality of life, people pay more and more attention to food safety issues, especially pesticide ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00C12N15/11
CPCC07K14/415C12N15/8205C12N15/8271
Inventor 辛明秦智伟刘春红周秀艳
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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