Fruit germplasm culture method capable of spontaneously eliminating agriculture remanet

A breeding method and fruit technology, applied in the field of plant germplasm cultivation, can solve problems such as excessive pesticide residues, and achieve the effect of enhancing food safety

Inactive Publication Date: 2009-06-10
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Extensive application and abuse of pesticides have caused

Method used

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  • Fruit germplasm culture method capable of spontaneously eliminating agriculture remanet
  • Fruit germplasm culture method capable of spontaneously eliminating agriculture remanet
  • Fruit germplasm culture method capable of spontaneously eliminating agriculture remanet

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0034] Example 1: Tomato Germplasm Cultivation for Spontaneous Elimination of Pesticide Residues

[0035] (1) Clone the fruit-specific promoter according to the conventional method:

[0036] Genomic DNA was extracted from tomato leaves, and the 1.1KbE8 promoter was cloned by PCR (see figure 1 ), PCR primers were designed according to the E8 promoter sequence provided in Genbank ID: AF515784.1, synthesized by Shanghai Jierui Bioengineering Company, the sequence is:

[0037] P1: 5'-CCCAAGCTTAGGAATTTCACGAAATCG-3';

[0038] P2: 5'-CGCGGATCCTCTTTTGCACTGTGAATG-3';

[0039] The upstream primer introduces a HindIII restriction site, and the downstream primer introduces a BamHI restriction site. According to the instructions of the plasmid pGEM-T easy, connect the cloned product to the plasmid pGEM-T easy to transform E.coli TG1, obtain positive clones through blue-white screening, and send them to Beijing Sanbo Yuanzhi Biological Company for sequencing to obtain the correct E8 prom...

Example Embodiment

[0058] Example 2: Cucumber Germplasm Cultivation for Spontaneous Elimination of Pesticide Residues

[0059] (1) Clone the fruit-specific promoter according to the conventional method:

[0060] With embodiment 1;

[0061] (2) Synthetic gene of organophosphate hydrolase OPH:

[0062] With embodiment 1;

[0063] (3) Construct the fruit-specific OPH plant expression vector:

[0064] With embodiment 1;

[0065] (4) transformation of soil Agrobacterium and cucumber explants

[0066] The transformation soil agrobacterium is the same as embodiment 1;

[0067] Cucumber seeds Jinyan No. 4 were purchased from Tianjin Academy of Agricultural Sciences. Soak ripe cucumber seeds in warm water for 1 hour, soak them in 75% alcohol for 45 seconds after peeling, and then wash them with 0.1% HgCl 2Soak for 5 minutes, wash 5 times with sterile water, dry the water on sterilized paper and connect it to MS medium. When the seed germinates to a height of 5 cm and the cotyledon is fully stretch...

Example Embodiment

[0075] Example 3: Capsicum Germplasm Cultivation for Spontaneous Elimination of Pesticide Residues

[0076] (1) Clone the fruit-specific promoter according to the conventional method:

[0077] With embodiment 1;

[0078] (2) Synthetic gene of organophosphate hydrolase OPH:

[0079] With embodiment 1;

[0080] (3) Construct the fruit-specific OPH plant expression vector:

[0081] With embodiment 1;

[0082] (4) transformation of soil Agrobacterium and capsicum explants:

[0083] The transformation soil agrobacterium is the same as embodiment 1;

[0084] Pepper seeds Tianjiao No. 1 were purchased from Beijing Jintianli Company. Soak the surface of the seeds in 95% alcohol for 45 seconds, then soak them in 50% bleach for 10 minutes, wash them with sterile water for 5 times, dry them with sterilized paper, put them on MS medium, and culture them under light at 25°C. When stretching, the hypocotyls were excised and cultured on the pre-medium for 2 days. The Agrobacterium LB...

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Abstract

The invention discloses a breeding method capable of spontaneously eliminating fruit germplasm of pesticide residue, which comprises the steps of: (1) cloning a fruit-specific promoter according to the prior method; (2) artificially synthesizing a gene of organophosphorus hydrolase (OPH); (3) constructing a fruit-specific OPH plant expression vector, in which an E8 amplification product and an artificially synthesized OPH gene sequence are connected to pGEM-T easy to construct pGEM-E8 and pGEM-OPH respectively, then HindIII and BamHI are used to perform double enzyme digestion on pGEM-TE8 and pSH to construct E8 to pSE8, EcoRI is used to perform single enzyme digestion on the pGEM-OPH and the pSE8 to construct OPH to pSE8OP, and a forward connection vector is screened, thereby obtaining the plant expression vector driven by the fruit-specific promoter for standby; (4) transforming soil agrobacteria and a plant explant; and (5) performing the differentiation and transplantation of a transgenic strain. The method can reduce the pesticide residue in fruits and simultaneously can not affect the efficacy of field pesticides, thereby contributing to the improvement of the yield and the quality.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for cultivating plant germplasm capable of specifically degrading pesticide residues in fruits. Background technique [0002] Pesticide pollution is one of the global environmental problems, and organophosphorus pesticides are the most widely used and most toxic compounds. Some pesticides have "three causes" substances, and the incubation period of carcinogenic and mutagenic effects can reach more than several decades. Some pesticides are environmental hormones, which interfere with endocrine after entering animals and humans, and cause abnormal reproductive functions. In the "Environmental Hormone List" that has been identified so far, there are 67 organic compounds, of which 44 are pesticides, accounting for 65.7%. Millions of tons of pesticides are applied worldwide every year, but no more than 5% are used directly on target organisms, and the vast majority of pesticide ...

Claims

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Application Information

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IPC IPC(8): C12N15/55C12N15/82A01H1/00A01H5/08
Inventor 赵杰宏赵德刚潘晨韩洁
Owner GUIZHOU UNIV
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