Gene for enhancing draught-resistance of plant

A technology of plant stress resistance and genetics, applied in the field of plant genetic engineering, can solve the problems of slow effect, long cycle of stress-resistant varieties, etc., and achieve the effect of easy acquisition

Inactive Publication Date: 2009-01-07
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional breeding methods have a long period of time to cultivate new plant stress-resistant varieties, and the results are slow

Method used

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  • Gene for enhancing draught-resistance of plant
  • Gene for enhancing draught-resistance of plant
  • Gene for enhancing draught-resistance of plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: the cloning of the gene that improves plant stress resistance

[0019] 1. Reagents

[0020] Restriction enzymes, Taq DNA polymerase, T4 DNA ligase, PrimeStar hot start high-fidelity DNA polymerase, pMD18-T cloning vector, etc. were purchased from Dalian Bao Biological Engineering Company; Trizol reagent was purchased from Beijing Tianwei Times Technology Co., Ltd.; The reverse transcription kit was purchased from Japan ToYoBo Company; the plasmid extraction and DNA recovery kit was purchased from Anhui Youjing Bioengineering Co., Ltd.; the PCR primers were synthesized by Shanghai Handsome Biological Company; the rest of the reagents were imported or domestically produced analytically pure products .

[0021] 2. E. coli strains and plant material

[0022] The cloned strain of Escherichia coli was E.coli JM109, which was purchased from Clontech Company. The maize material was inbred line 616-1, provided by Sichuan Academy of Agricultural Sciences.

[002...

Embodiment 2

[0136] Embodiment 2: Real-time PCR (real-time quantitative PCR) analysis improves the expression pattern of the gene of plant stress resistance

[0137] 1. Experimental materials, reagents and instruments

[0138] Conventional reagents and corn materials are the same as in Example 1.

[0139] The Real-timePCR kit is SYBR Premix Ex Taq from Dalian Bao Biological Engineering Co., Ltd. TM (Perfect RealTime), the fluorescent quantitative PCR instrument is Baile Company's Fluorescent quantitative PCR instrument (Bio-Rad Laboratories, USA).

[0140] 2 methods

[0141] 2.1 Processing of corn material

[0142] Corn seeds were soaked in tap water at 37°C overnight. The next day, the seeds were placed in a petri dish covered with wet filter paper, and seedlings were germinated at 25°C under 16h light / 8h dark conditions, and water was added every day to keep the filter paper moist. After 10 days, when the corn seedlings had just grown the third leaf, the corn seedlings were treate...

Embodiment 3

[0169] Embodiment 3: the expression of the gene that improves plant stress resistance in tobacco and stress resistance test

[0170] 1. Materials

[0171] 1.1 Reagents and materials

[0172] The routine reagents are the same as in Example 1.

[0173] The Agrobacterium strain used for transgenesis was EHA105, purchased from Clontech Company; the tobacco variety used for transformation was 'Safflower Dajinyuan', provided by the Institute of Biology, Chengdu Branch of the Chinese Academy of Sciences; the plant expression vector was pHB, provided by the Shanghai Institute of Plant Physiology supply.

[0174] 1.2 Medium and solution

[0175] YEB medium: yeast extract 1g / L, beef extract 5g / L, egg white 5g / L, sucrose 5g / L, MgS0 4 .7H 2 O 0.5g / L. Adjust the pH to 7.0 with NaOH and autoclave.

[0176] YEP medium: yeast extract 10g / L, protein powder 10g / L, NaCl 5g / L. Adjust the pH to 7.0 with NaOH and autoclave.

[0177] Routine tissue culture medium:

[0178]

[0179]

...

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Abstract

The invention relates to a gene for improving the stress resistance of a plant the nucleotide sequence of which is expressed in the SEQ ID NO: 1. The nucleotide sequence expressed by the SEQ ID NO:1 is inserted into a vector to obtain the recombinant plasmid of the invention; the vector can select a known eukaryotic expression in the field. The recombinant plasmid is converted into tobaccos to obtain a transgenic plant. Test shows that based on an MS culture medium additionally provided with NaCl, mannitol and ABA, the transgenic plant grows better than a wild plant. After drying the transgenic plant and the wild plant for 18 days, the transgenic plant grows better than the wild plant. The detecting data for the content of malondialdehyde shows that the damage degree of the transgenic plant under stress conditions is lower than that of the wild plant. Therefore, the gene for improving the stress resistance of a plant can be applied in the fields of anti-drought of the plant and improving the salt improvement.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to a gene for improving stress resistance of plants, a recombinant plasmid and its application in improving plant tolerance to drought and salt Background technique [0002] Environmental stress (drought, high salinity, low temperature and high temperature, etc.) has a great negative impact on the growth and development of plants. Traditional breeding methods have a long period of time for cultivating new plant stress-resistant varieties, and the results are slow. The cycle of cultivating new stress-resistant plant varieties by transgenic methods is relatively short, and at the same time, it does not affect the existing good traits, so it has gradually become an important method for improving stress-resistant plant varieties. [0003] At present, there are two types of methods for improving plant stress resistance through transgenics: (1) Overexpression of transcription fac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/79C12N15/82A01H1/00
Inventor 刘永胜黄维藻牛向丽刘永生高永峰唐维罗笛
Owner SICHUAN UNIV
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