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Application of dap gene in Deinococcus radiodurans R1 in breeding of salt tolerant plants

A plant and gene technology is applied in the application field of the gene in improving the resistance of plants to salt stress, and can solve problems such as unseen

Active Publication Date: 2014-05-14
LONGPING BIOTECHNOLOGY (HAINAN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, there is no research report on the function of the dap gene (DRB0118, GeneID: 1799871) in Deinococcus radiodurans R1 in improving plant salt tolerance

Method used

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  • Application of dap gene in Deinococcus radiodurans R1 in breeding of salt tolerant plants
  • Application of dap gene in Deinococcus radiodurans R1 in breeding of salt tolerant plants
  • Application of dap gene in Deinococcus radiodurans R1 in breeding of salt tolerant plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0037] Example 2 Salt tolerance experiment of recombinant strain containing D. radiodurans R1dap gene (DRB0118)

[0038] 1. Experimental method

[0039] 1. Inoculate the two recombinant Escherichia coli obtained in Example 1 into 20 mL LB liquid medium (containing Amp antibiotics), shake the flask overnight (37°C), and then transfer to 100 mL LB liquid medium. Try to keep the same amount of inoculum and cultivate to OD 600 ≈0.5 is fine.

[0040] 2. After centrifugation, 10 mL of the bacterial solution is shocked in an equal volume of 4M NaCl salt solution for 2 hours, and each sample is immediately diluted to 10 times with sterile deionized water. -4 , Take 10μL spot on the surface of LB solid medium, incubate at 37℃ for 16h, observe the colony formation and take pictures.

[0041] 2. Experimental results

[0042] image 3 It was shown that the growth status of the JM-dap strain containing the D. radiodurans R1dap gene (DRB0118) before the impact of the 4M NaCl salt solution was basica...

Embodiment 3

[0045] Example 3 Expression of dap gene (DRB0118) in tobacco and identification of salt tolerance of transgenic plants

[0046] (1) Construction of expression vector containing target gene

[0047] Based on the full-length coding sequence, design primers that amplify the complete coding reading frame, and introduce restriction endonuclease sites on the positive and negative primers (this may depend on the selected vector) to construct an 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). Under the premise, the identified expression vector is transferred to Agrobacterium, and the leaf disc method is used to transform the model plant tobacco.

[0048] (2) Using leaf disc method to transform tobacco

[0049] Pick the positive colonies on the YEB selection plate with a...

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Abstract

The invention discovers that dap gene (DRB0118) in Deinococcus radiodurans R1 improves the resistance of prokaryotes and plants. The invention constructs a recombinant vector comprising the gene; and the recombinant vector is transferred into prokaryotic and eukaryotic host cells. Experiments prove that: after the dap gene (DRB0118) is expressed in prokaryotic host cells and tobacco, the salt resistance of the prokaryotic host cells and the tobacco can be improved.

Description

Technical field [0001] The invention relates to the new function of the Dap gene (DRB0118, GeneID: 1798871) of Deinococcus radiodurans R1 (Deinococcus radiodurans R1), and specifically relates to the application of the gene in improving the resistance of plants to salt stress. Background technique [0002] D. radiodurans R1 (D.radiodurans R1) has strong resistance to ionizing radiation, UV radiation, DNA damage reagents, and osmotic stress, and has become an ideal strain for studying the adaptation mechanism of microbial abiotic stress. It can also be used as a research A model species for salt tolerance in higher plants (Battista et al., 2001). It is derived from a large plasmid in the D. radiodurans R1 genome, and the protein encoded by it is related to osmotic stress resistance. [0003] However, there is no research report on the function of the dap gene (DRB0118, GeneID: 1798871) in Deinococcus radiodurans R1 in improving plant salt tolerance. Summary of the invention [0004...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12R1/19
Inventor 张维江世杰王劲陈明滕超林敏
Owner LONGPING BIOTECHNOLOGY (HAINAN) CO LTD
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