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Gene for improving plant salt tolerance and drought resistance and use thereof

A technology of drought resistance and salt tolerance, applied to the application of drought stress resistance, improving the field of plants to salt, can solve the problems of D.radiodurans and so on.

Active Publication Date: 2009-04-29
LONGPING BIOTECHNOLOGY (HAINAN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, in the prior art, there is no relevant report that D.radiodurans can improve the resistance of plants to salt and drought stress

Method used

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  • Gene for improving plant salt tolerance and drought resistance and use thereof
  • Gene for improving plant salt tolerance and drought resistance and use thereof
  • Gene for improving plant salt tolerance and drought resistance and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 SEQ ID NO: 1 nucleotide sequence expression in Escherichia coli and analysis of salt tolerance and drought resistance performance

[0049] 1. Cloning of the nucleotide sequence SEQ ID NO: 1

[0050] The complete nucleotide sequence was amplified from D. radiodurans genomic DNA based on the published Deinococcus radiodurans genomic sequence involving a pair of PCR-specific primers.

[0051] 2. Construction of Escherichia coli expression vector and molecular verification

[0052] The above-mentioned cloned fragment was digested with NdeI and SacII double enzymes, connected with the vector pTtSacB containing the E.coli universal promoter groE, and replaced the SacB gene to generate an Escherichia coli expression vector of the gene. Transform E.coli JM109, smear it on LB solid medium containing Amp, pick white colonies, extract plasmids by alkali lysis method, screen different recombinants, digest with BglII and sequence verification, and obtain a strain expressi...

Embodiment 2

[0055] Embodiment 2 SEQ ID NO: artificial synthesis of 2 nucleotide sequence

[0056] According to the nucleotide sequence of SEQ ID NO: 1, firstly divide 7 segments into single-stranded oligonucleotide fragments with a length of 150-200bp and sticky ends according to the positive and secondary strand sequences respectively. Anneal the 7 complementary single-stranded oligonucleotide fragments in one-to-one correspondence between the positive strand and the auxiliary strand respectively to form 7 double-stranded oligonucleotide fragments with sticky ends. The mixed double-stranded oligonucleotide fragments are catalyzed by T4 DNA ligase and assembled into a complete gene. Both ends of the synthesized DNA fragment contain XbaI and SacI sites.

[0057] The above-mentioned artificially synthesized 5' and 3' end restriction sites are XbaI and SacI sites SEQ ID NO: 2, which are used for the construction of the following high salt tolerance and drought resistance gene plant expressi...

Embodiment 3

[0058] Example 3 Eukaryotic expression of the nucleotide sequence of SEQ ID NO: 2 in tobacco cells and identification of salt tolerance and drought resistance of transgenic plants

[0059] (1) Construction of expression vector containing target gene

[0060] According to the full-length coding sequence (SEQ ID NO: 2), design primers to amplify the complete coding reading frame, and introduce restriction endonuclease sites on the forward and reverse primers respectively (this can be determined by the vector selected), In order to construct the expression vector. Using the amplified product obtained in Example 1 as a template, after PCR amplification, the sequence cDNA was cloned into an intermediate vector (such as pBluescript), and further cloned into a binary expression vector (such as pBI121 and pCAMBIA2200). The expression vector identified under the premise was transformed into Agrobacterium, and the model plant Nicotiana tabacum was transformed by the leaf disc method. ...

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Abstract

The invention initially discovers a nucleotide coded sequence which can improve salt resistance and drought resistance of plants, and a nucleotide coded sequence which is artificially synthesized according to codons preferred by plants. The nucleotide coded sequences construct a recombining carrier containing genes and converts the genes into host cells of prokaryotic cells and eukaryotic cells respectively. Experiments prove that after the genes are expressed in the plant, the obtained transgenic plant has enhanced resistance to salt stress, drought stress and the like.

Description

technical field [0001] The present invention relates to a nucleotide coding sequence and another nucleotide coding sequence artificially synthesized according to plant preferred codons. The present invention also relates to the use of the nucleotide coding sequence in improving plant resistance to salt and drought stress Applications. Background technique [0002] Deinococcus radiodurans (D. radiodurans) is one of the most radiation-resistant organisms known to date. The bacterium has extreme resistance to lethal doses of ionizing radiation, UV radiation, and DNA-damaging reagents, and has the ability to completely restore hundreds of DNA double-strand breaks caused by ionizing radiation without mutation. Its extreme radiation resistance and its DNA loss repair molecular mechanism have aroused great interest in the scientific community. It is not only of great significance to the basic science of exploring the molecular mechanism of DNA repair, but also to promote the devel...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/63C07K14/415A01H1/00C12Q1/68
CPCC12Q2600/13C12N15/8273C12Q1/6895C07K14/195
Inventor 林敏陈明王劲潘婕周正富张维陆伟平淑珍
Owner LONGPING BIOTECHNOLOGY (HAINAN) CO LTD
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