CN103205457B - Method for using microbial stress tolerance genes to increase plant salt tolerance - Google Patents
Method for using microbial stress tolerance genes to increase plant salt tolerance Download PDFInfo
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- CN103205457B CN103205457B CN201310086561.3A CN201310086561A CN103205457B CN 103205457 B CN103205457 B CN 103205457B CN 201310086561 A CN201310086561 A CN 201310086561A CN 103205457 B CN103205457 B CN 103205457B
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Abstract
The invention discloses a method for using microbial stress tolerance genes to increase plant salt tolerance. The stress tolerance genes carried by microbes can be used for increasing plant salt tolerance. The microbes are stenotrophomonas maltophilia, SMA, in Latin. By a section of stress tolerance genes of the SMA, salt tolerance of transgenic plants can be increased. The SMA can be achieved by other approaches in nature. The method is simple to apply and evident in effect.
Description
Technical field
The present invention relates to a kind of investigative technique that utilizes microorganism to strengthen plant salt endurance, relate in particular to a kind of method of utilizing microorganism adversity gene to increase plant salt endurance.
Background technology
The soil salinization is current global ecological problem and resource environment problem.How utilizing large-area saltings development agroforestry, is bio-science technology key subjects in the urgent need to address.Although adopt traditional genetic and breeding method cultivation salt-tolerant plant new variety simple and feasible, make slow progress.Along with the development of Protocols in Molecular Biology, people place hope on and utilize genetic engineering breeding technology to cultivate the new variety of plant that salt resistance ability significantly improves, and then increase the exploitation in saltings.Now, had much for the research of plant salt tolerance mechanism, but on the whole, Chinese scholars is confined to physiology aspect to the research of plant salt tolerance mechanism more, the molecular mechanism regulated and control network of plant salt tolerance is still unclear, has seriously limited salt-tolerant plant rearing new variety.Therefore, understand the molecular mechanism of plant under condition of salt stress in depth, Selection of Salt-Tolerant gene, has great importance to utilizing molecular breeding technology to cultivate salt-tolerant plant new variety.
Summary of the invention
The object of this invention is to provide a kind of method of utilizing microorganism adversity gene to increase plant salt endurance.
The object of the invention is to be achieved through the following technical solutions:
The method of utilizing microorganism adversity gene to increase plant salt endurance of the present invention, utilize adversity gene that microorganism is carried can increase the salt tolerance of plant, described microorganism is stenotrophomonas maltophilia, its Latin called after: Stenotrophomonas mahophilia, SMA.
As seen from the above technical solution provided by the invention, what the embodiment of the present invention provided utilizes microorganism adversity gene to increase the method for plant salt endurance, owing to having confirmed there is a kind of endophyte---stenotrophomonas maltophilia in plant, by one section adversity gene being transformed into the salt tolerance that has increased Arabidopis thaliana in arabidopsis thaliana, illustrate that it can give the characteristic of plant salt tolerance, application is simple, successful.
Brief description of the drawings
Fig. 1 is the Technology Roadmap in embodiment of the present invention research process.
Amino acid or nucleotide sequence explanation
Listed sequence is the DNA sequence dna that in the present invention, stenotrophomonas maltophilia is carried adversity gene.
Embodiment
To be described in further detail the embodiment of the present invention below.
The microorganism adversity gene that utilizes of the present invention increases the method for plant salt endurance, its preferably embodiment be:
Utilize adversity gene that microorganism is carried can increase the salt tolerance of plant, described microorganism is stenotrophomonas maltophilia, its Latin called after: Stenotrophomonas mahophilia, SMA.
Described stenotrophomonas maltophilia obtains from occurring in nature, and this stenotrophomonas maltophilia is carried adversity gene, and the DNA sequence dna of described adversity gene is in the registration of NCBI website, and its sequence number is KC129717.
Comprise step:
First,, after the DNA sequence dna of described adversity gene is connected with carrier pCLD04541, the plasmid that carries described adversity gene is transformed in Agrobacterium GV3101 by electric shock;
Then, described Agrobacterium GV3101 is shaken after bacterium cultivates, contaminate or the method for transformation of Ye Panfa is transformed into the DNA of described adversity gene in plant by flower, merge with Plant Genome, make the genome of plant obtain this adversity gene and express, can increase the salt tolerance of plant.
Described plant comprise following any: herbaceous plant, xylophyta.
The present invention passes through transgenic technology, by one section of DNA fragmentation that contains microorganism adversity gene (or gene cluster), this fragment gene is KC129717 at the sequence number of NCBI application, be transformed in Arabidopis thaliana, increased the salt tolerance of Arabidopis thaliana, thereby explanation the method microorganism adversity gene can increase the salt tolerance of plant.
In specific embodiment, in plant, there is a kind of endophyte---stenotrophomonas maltophilia by having studies confirm that, on it one section carries the characteristic that the DNA of adversity gene can give plant salt tolerance, makes a plant new approach of salt tolerant more, and the process of concrete research is:
Utilize DNA restriction endonuclease Bam HI to cut respectively pCLD04541 plasmid vector and DNA, be connected with DNA by plasmid vector, transform DH10B bacterial strain and build BIBAC library, and the Arabidopis thaliana inflorescence dip-dye method mediating by Agrobacterium GV3101 obtains the transfer-gen plant of arabidopsis thaliana transformation.In conjunction with transforming artificial chromosome library technology and sequencing technologies, a kind of endophyte of plant detected--a characteristic that large fragment DNA can be given plant salt tolerance in stenotrophomonas maltophilia.Transform the transgenic arabidopsis comparison of a DNA large fragment in endophyte stenotrophomonas maltophilia and do not had genetically modified wild-type Arabidopis thaliana to show the characteristic of salt tolerant.
Concrete technological line is as shown in Figure 1:
1, built BIBAC library;
2, extract BIBAC library plasmid;
3, plasmid is transformed in Agrobacterium GV3101 by electric shock;
4, Agrobacterium is shaken after bacterium cultivation, contaminate in Arabidopis thaliana by flower dip-dye method;
5, screening transformed plant, collects the plant seed after transforming.The transgenic arabidopsis of Selection of Salt-Tolerant on the flat board that contains 150mM NaCl;
6, the corresponding BIBAC of the salt tolerant transgenic arabidopsis library plasmid screening is checked order and bioinformatic analysis;
7, find that in this BIBAC library, this plasmid of BIBAC2A1F6 is the section of DNA of endophyte stenotrophomonas maltophilia.Analyze this DNA fragmentation and show, in this segment DNA, contain relevant adversity gene and caused transgenic arabidopsis salt tolerance to significantly improve.
The present invention has found a new way that makes plant salt tolerance: in plant, have a kind of endophyte---stenotrophomonas maltophilia, one section of adversity gene on it has given plant stronger salt tolerance.Can from other approach of nature, obtain this endophyte.
The above; only for preferably embodiment of the present invention, but protection scope of the present invention is not limited to this, is anyly familiar with in technical scope that those skilled in the art disclose in the present invention; the variation that can expect easily or replacement, within all should being encompassed in protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain of claims.
Claims (2)
- One kind utilize stenotrophomonas maltophilia ( stenotrophomonas mahophilia) adversity gene increases the method for plant salt endurance, it is characterized in that, comprise the steps:First,, after described adversity gene is connected with carrier, the plasmid that carries described adversity gene is transformed in Agrobacterium GV3101 by electric shock;Then, described Agrobacterium GV3101 is shaken to bacterium and cultivate, contaminate or the method for transformation of Ye Panfa is transformed into described adversity gene in plant by flower, merge with Plant Genome, make the genome of plant obtain this adversity gene and express, can increase the salt tolerance of plant;Wherein, the DNA sequence dna of described adversity gene is shown in SEQ ID No.1, and described plant is Arabidopis thaliana.
- 2. the method for increase plant salt endurance according to claim 1, is characterized in that, described stenotrophomonas maltophilia obtains from occurring in nature.
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| CN201310086561.3A CN103205457B (en) | 2013-03-18 | 2013-03-18 | Method for using microbial stress tolerance genes to increase plant salt tolerance |
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| CN103205457B true CN103205457B (en) | 2014-12-10 |
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| CN115011510B (en) * | 2022-05-20 | 2023-04-28 | 青岛农业大学 | Salt-tolerant growth-promoting stenotrophomonas maltophilia, microbial agent thereof and application thereof |
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| CN1266278C (en) * | 2004-10-01 | 2006-07-26 | 厦门大学 | Clone procaryotic biological salt-resistant related gene method |
| CN101712945B (en) * | 2009-06-30 | 2012-04-18 | 中国环境科学研究院 | Extreme halophilic bacteria, compound halophilic bacteria agent and application |
| CN101836548B (en) * | 2010-05-11 | 2011-07-27 | 沈阳化工学院 | Method for improving salt tolerance of peppermint |
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