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A kind of breeding method of pest-resistant and low-phenol cotton variety

A cotton variety and insect-resistant cotton technology, which is applied in the field of genetic engineering, can solve the problems such as no reports of successful breeding of dominant low-phenol transgenic insect-resistant cotton, and achieves inhibition of programmed death, short time consumption, and reduced cotton production. The effect of phenolic content

Active Publication Date: 2022-01-11
邯郸市农业科学院
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AI Technical Summary

Problems solved by technology

Some researchers have also attempted to introduce exogenous Bt genes into low-phenol cotton to improve the insect resistance of low-phenol cotton, but so far, there have been no reports of successful breeding of transgenic insect-resistant cotton with dominant low-phenol traits

Method used

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  • A kind of breeding method of pest-resistant and low-phenol cotton variety
  • A kind of breeding method of pest-resistant and low-phenol cotton variety
  • A kind of breeding method of pest-resistant and low-phenol cotton variety

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Embodiment 1

[0046] A breeding method for insect-resistant and low-phenol cotton varieties, comprising:

[0047] Experimental materials: transgenic cotton MON531 seeds, cotton embryo cDNA, pEASY-T1 vector, screening marker pCAMBIA3301 vector for GUS gene, cotton embryo cDNA, pGL3-HBV synthesized in Shanghai Shenggong Company, Agrobacterium tumefaciens LBA4404.

[0048] 1. Cloning of cotton seed α-globulin promoter: Cotton embryo cDNA was used as template, forward primer: 5'-AAGTCTGCAGGCTTGCGCATATTTCTTACTATTTTAGCTCTC-3'; reverse primer: 5'-GGCCGTCGGATCGGGAACGATAATCTCTGTATGTTG-3', as primer pair α- The globulin promoter was amplified by PCR, the PCR product was detected by 1% agarose gel electrophoresis, the PCR product was recovered from the gel, and the recovered and purified PCR product was cloned into the pEASY-T1 vector, and the recombinant plasmid was named pEASY-T1-α- globulin promoter.

[0049] 2. Construction of plant binary expression vector of α-globin promoter + GUS reporter gen...

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Abstract

The invention provides a breeding method for an insect-resistant and low-phenol cotton variety, which belongs to the technical field of genetic engineering and includes cloning the CAD1-A gene to obtain a CAD1-A-1 plasmid; constructing an RNAi expression vector for the CAD1-A gene; The embryogenic callus of the Bt gene-transferred insect-resistant cotton is genetically transformed by the Agrobacterium carcinoma mediated method; wherein, the CAD1-A gene in the RNAi expression vector is driven by the α-globulin promoter, and the α-globulin promoter Modification with Enh I enhancer. The method can inhibit the methylation of the Bt gene promoter and the special promoter of the CAD1-A gene seed, thereby inhibiting the significant reduction of the transcription level of the Bt gene in compound transgenic crops and promoting the reduction of the transcription level of the CAD1-A gene; Toxic callus, inhibit the programmed death of transformed cells, and increase the frequency of transformation.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a method for breeding insect-resistant and low-phenol cotton varieties. Background technique [0002] Gossypol is toxic to bacteria, fungi, insects, etc., and is an important part of the defense mechanism of cotton. However, gossypol easily damages the gastric mucosa of humans and monogastric animals, causing digestive disorders, which has become an important reason for long-term obstacles to people's better development and utilization of cottonseed. Although gossypol can be removed by physical and chemical methods, the process is complicated, the investment is large, the energy consumption is high, and the cost is high. Moreover, the cottonseed cakes processed by these methods will lose their nutrients to varying degrees, and their food safety will also be affected. reduce. Therefore, breeding experts from all over the world have launched low-phenol cot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/52C12N15/84A01H5/00A01H6/60
CPCC12N15/8218C12N15/8286C12N15/8243C12N9/00
Inventor 米换房权月伟唐光雷翟雷霞李继军李文蕾
Owner 邯郸市农业科学院
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