Method for improving beet salt-resistance, draught-resistance and anti-herbicide chlorsulfuron characteristics by multi-gene transformation and application

A technology resistant to herbicides and chlorsulfuron, applied in the field of bioengineering breeding of crops, can solve problems such as undetermined technical routes and methods

Inactive Publication Date: 2006-05-10
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Research in this area is still in its infancy at home and abroad, and the best technical route and method have not yet been determined.

Method used

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  • Method for improving beet salt-resistance, draught-resistance and anti-herbicide chlorsulfuron characteristics by multi-gene transformation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Creation of sugar beet transgenic lines with excellent salt and drought tolerance and resistance to the herbicide chlorsulfuron through multi-gene transformation

[0036] 1. Acquisition of sugar beet transgenic plants

[0037] Take the top section of the young inflorescence about 1 cm long from the plant in the flower bud stage of the field, and wash it with 70% alcohol for 1min, 0.1% HgCl 2 After sterilizing for 5 minutes, cut them into 0.1-0.5cm sections, inoculate them on the medium supplemented with 1mg / L 6-BA in MS, and culture them in the dark at 26±2°C to induce clustered buds. The clustered buds were cut and then subcultured. Bud clusters stripped of young leaves are used for genetic transformation.

[0038] Agrobacterium tumefaciens (AGL1 or LBA4404) carrying a binary vector (Mini-Ti plasmid with herbicide resistance gene als and target gene betA, or with herbicide resistance gene bar and target genes NHX1 and PPase gene) In the YEP medium supplem...

Embodiment 2

[0054] Example 2: The sugar beet breeding material with excellent resistance to salt and drought resistance to the herbicide chlorsulfuron obtained by multi-gene transformation by sister crossing of plants with different target genes

[0055] 1. Obtaining of sugar beet transgenic plants Same as Example 1.

[0056] 2. Management and resistance detection of transgenic plants Same as Example 1.

[0057] 3. Multigene aggregation using sister plants transfected with different target genes

[0058] Roots of plants transfected with different target genes (different selection marker genes) from the same recipient genotype are planted in the same isolation area, open pollination, and produce seeds. The harvested seeds were screened with 2 different selection agents such as the herbicide chlorsulfuron and the herbicide glufosinate. Screening is generally carried out at the 4-leaf stage of the plants. First, the plants are treated with an aqueous solution of a selective agent (such as ...

Embodiment 3

[0060] Example 3: Using a plant expression vector with a double T-DNA region for transformation to obtain multigene-transformed sugar beet breeding materials with excellent salt and drought tolerance and resistance to the herbicide chlorsulfuron

[0061] 1. Acquisition of sugar beet transgenic plants

[0062] Take the top section of the young inflorescence about 1 cm long from the plant in the flower bud stage of the field, and wash it with 70% alcohol for 1min, 0.1% HgCl 2 After sterilizing for 5 minutes, cut them into 0.1-0.5cm sections, inoculate them on the medium supplemented with 1mg / L 6-BA in MS, and culture them in the dark at 26±2°C to induce clustered buds. The clustered buds were cut and then subcultured. Bud clusters stripped of young leaves are used for genetic transformation.

[0063] A binary vector (Mini-Ti plasmid with double T-DNA regions, the target gene NHX1 and PPase gene can be inserted in series in one of the T-DNA regions in the form of fusion genes, ...

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Abstract

The invention provides a method of improving the sugar beet property of salt and drought resistance and weedicide resistance of green-yellow ron through polygene inversion and the application; recombining gene NHX1 of sodium hydrogen inversing transferring protein from the plant racuolar membrane, gene Ppase of racuolar membrane pyrophosphatase , gene beta of choline dehydrogenase from bacillus coli and gene als of acetolactate synthetase resisting the accident mutation of weed killer of green-yellow ron from quasi-mustard in the plant expression carrier, transferring into the sugar beet cell for high effective expression and getting trans genetic plant; choosing trans plants of salt and drought resistance and weedicide resistance of green-yellow ron dramatically increased from the trans genetic plants and their generations; getting the trans genetic plants with 3-4 target genes by a second conversion or making the plants with different trans genes mate with each other; choosing individuals with outstanding properties of salt and drought resistance and weedicide resistance of green-yellow ron from generations of polygenetic inversed plants, and then producing the sugar beet of new strain with properties of salt and drought resistance and weedicide resistance of green-yellow ron.

Description

technical field [0001] The invention belongs to the field of bioengineering breeding of crops, and specifically relates to a method and application for improving sugar beet salt and drought tolerance and herbicide chlorsulfuron resistance through multi-gene transformation. Background technique [0002] Among all kinds of natural adversities, the impact of drought on the world's crop production ranks first, and its harm is equivalent to the sum of other natural disasters. Salinization is also a major environmental factor affecting crop yields. About half of my country's land area is in arid and semi-arid areas. Even major agricultural areas in non-arid regions are occasionally hit by drought. Among the 1.6 billion mu of arable land in my country, there are more than 100 million mu of saline-alkali land, and about 1 billion mu of medium and low-yield fields. Most of these low-yielding fields are also due to drought and salinity. In addition, there are 300 million mu of sal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H3/04A01H1/02
Inventor 杨爱芳张举仁谷晓峰张可炜尹小燕
Owner SHANDONG UNIV
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