Plant expression vector for controlling artificially synthesized antimicrobial peptide gene by using specific vascular promoter and method for culturing anti-greensickness cotton by using same

A technology of plant expression vector and specific promoter, applied in the field of plant genetic engineering, can solve the problems of reducing fiber quality, plant morbidity, death, etc., and achieves the effect of simple and easy method, remarkable effect and huge economic benefit.

Inactive Publication Date: 2011-09-07
SOUTHWEST UNIVERSITY
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Problems solved by technology

Cotton Verticillium dahliae mainly infects from the root of the plant, and then gradually expands to the upper part of the plant along with the vascular tissues of the roots, stems and leaves of the plant. Under suitable conditions, it can develop to the top of the plant and cause the disease of the entire plant or even death, causing severe yield loss and reduced fiber quality

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  • Plant expression vector for controlling artificially synthesized antimicrobial peptide gene by using specific vascular promoter and method for culturing anti-greensickness cotton by using same
  • Plant expression vector for controlling artificially synthesized antimicrobial peptide gene by using specific vascular promoter and method for culturing anti-greensickness cotton by using same
  • Plant expression vector for controlling artificially synthesized antimicrobial peptide gene by using specific vascular promoter and method for culturing anti-greensickness cotton by using same

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Embodiment Construction

[0051] Below in conjunction with accompanying drawing, the present invention will be described in further detail, but the following description does not limit the present invention, any deformation and change of the present invention, as long as they do not depart from the spirit of the present invention, all should belong to the definition of the appended claims of the present invention scope.

[0052] The pharmaceutical reagents in the implementation examples of the present invention are domestic conventional chemical reagents that are not specified, and the materials and methods that are not specified are referred to "Molecular Cloning Experiment Guide" (Sambrook and Russell, 2001).

[0053] 1. Extraction of DNA

[0054] 1.1 DNA extraction buffer:

[0055] (1) Plant DNA extraction buffer:

[0056] CTAB extract: 100 mmol / L Tris-HCl (pH8.0), 20 mmol / L EDTA (pH8.0), 1.5 mol / L NaCl, 2% CTAB (w / v), 4% PVP40 (w / v ) and 2% mercaptoethanol (v / v), PVP and mercaptoethanol were add...

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Abstract

The invention provides a plant expression vector for controlling an artificially synthesized antimicrobial peptide gene by using a specific vascular promoter and a method for culturing anti-greensickness cotton by using the same. In the method, an artificially synthesized antimicrobial peptide gene spCEMA (Signal Peptide modified Cecropin A-Melittin) under the control of a specific vascular promoter AAP2 is integrated to a cotton genome to realize the specific expression of the gene spCEMA in a cotton vascular tissue, so that the disease resistance of cotton to greensickness is obviously improved. By inoculating different pathogenic bacteria of different greensickness types to genetically modified cotton obtained with the method in a homozygosis plant infected field, a disease index is less than 15, which is reduced by more than 85% in contrast with that of non-genetically modified cotton.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering. In particular, it relates to a method for preparing an artificially synthesized antimicrobial peptide gene plant expression vector controlled by a vascular-specific promoter and a transgenic cotton containing the above-mentioned expression vector; in addition, the present invention also relates to a method for improving cotton resistance to Verticillium wilt by using genetic engineering technology . technical background [0002] Cotton is the most important natural fiber crop in the world, and it is also an important material related to the national economy and people's livelihood in my country. The cotton industry plays a pivotal role in the development of my country's textile industry and even the entire national economy. However, since cotton Verticillium wilt was introduced into my country in 1935, the harm to my country's cotton production has been increasing year by yea...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84C12N1/21C12N5/10A01H5/00C12R1/01C12R1/91
Inventor 李先碧裴炎罗明肖月华侯磊李德谋宋水清罗小英龙琴
Owner SOUTHWEST UNIVERSITY
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