Method for promoting salt and drought tolerance of maize and wheat by combining betAíóNHX1íóPPase gene and transgene technology

A transgenic and drought-tolerant technology, 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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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Create a corn inbred line with excellent salt and drought tolerance

[0038] 1. Obtaining of transgenic maize plants

[0039] Corn backbone inbred line seeds are soaked in 70% ethanol for 10 minutes, then soaked in 0.1% mercury chloride for 10-12 minutes, and then washed 3-5 times with sterile water. After sterilization, the seeds were germinated in sterile Erlenmeyer flasks. After the seeds germinated (lubai), they were placed on the modified MS medium and continued to germinate under dark conditions. When the germ is extended to 3-5 cm, the coleoptile and 2-3 young leaves are peeled off to expose the growth cone at the top of the stem tip for transformation.

[0040] 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 supplemented with antibiotics, they were shaken and cult...

Embodiment 2

[0053] Example 2: Creating wheat breeding materials with excellent salt and drought tolerance

[0054] 1. Obtaining of wheat transgenic plants

[0055] Select excellent varieties such as Jinan 17, Wenqian No. 1, etc. for Agrobacterium-mediated genetic transformation. (1) Cut off all the fibrous roots of the sterile wheat etiolated seedlings, peel off the coleoptile and 1-2 young leaves with a dissecting needle and a scalpel, expose the shoot tip growth cone, and then stab the shoot tip growth cone with a dissecting needle . (2) Tilt a sterile petri dish with a diameter of 4.5 cm and pour it into the LBA4404 bacteria (Mini-Ti plasmid with herbicide resistance gene bar and target gene betA and NHX1, or with herbicide resistance) added with acetosyringone Gene bar and target gene PPase gene) solution (OD 600 =0.8), soak the sterile seeds with bare shoot tip growth cones in the bacterial solution for 6-8min, during this period, use 0.5×10 5 Treated at Pa atmospheric pressure t...

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PUM

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Abstract

This invention discloses a method for improving the salt resistance and drought resistance of the corn and wheat by aggregating the beta, NHX1, PPase genes with the transfer genes, the steps as follows: regrouping the solution film natrium hydrogen counter-operating protein gene NHX1, the solution film tar phosphatase gene PPase and the Cholinesterase dehydrogenase gene from the escherichia colim to the plant expression carrier, then adding the corn or wheet cells and make them express efficiently, then producing the transfer gene plant; screening out the transfer gene pure cell that has apparently higher salt resistance and drought resistance from the transfer gene plant and its offspring; getting the transfer gene plant with the three object genes through twice tranfering the transfer gene pure cell or making the plant with the different transfer genes mate mutully, then sceening out the ones that have better salt resistance and drought resistance in their offsprings, and making them mute and purify by themselves to creat the new corn and wheat seeds with the salt resistance and drought resistance; or making use of the transfer polymeric material to produce the mixing seeds directly.

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 the salt and drought tolerance of corn and wheat by transgenic polymerization of betA, NHX1 and PPase genes. 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 saline...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H1/02A01H1/04A01H4/00C12N15/29C12N15/31C12N15/53
Inventor 张举仁杨爱芳谷晓峰张可炜尹小燕
Owner SHANDONG UNIV
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