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

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

Active Publication Date: 2012-07-11
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 (DRB...

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 Containing D.radiodurans R1dap Gene (DRB0118) Recombinant Strain

[0038] 1. Experimental method

[0039] 1. The 2 recombinant Escherichia coli obtained in Example 1 were respectively inoculated in 20 mL of LB liquid medium (containing Amp antibiotics), and after the shake flask was cultured overnight (37° C.), they were then respectively transferred to 100 mL of LB liquid medium In the medium, try to keep the inoculum volume consistent, culture to OD 600 ≈0.5 is enough.

[0040] 2. After centrifuging 10mL of the bacterial solution, shock it in an equal volume of 4M NaCl salt solution for 2 hours, and immediately dilute each sample to 10 times with sterile deionized water. -4 , Take 10 μL and spot on the surface of LB solid medium, culture at 37°C for 16 hours, observe the colony formation and take pictures.

[0041] 2. Experimental results

[0042] image 3 It was shown that the growth state of the JM-dap strain containing the D...

Embodiment 3

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

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

[0047] According to the full-length coding sequence, 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), so as 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.

[0048] (2) Transforming Tobacco Using the Leaf Disk Method

[0049] Use a sterile t...

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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: 1799871) of Deinococcus radiodurans R1, and specifically relates to the application of the gene in improving the resistance of plants to salt stress. Background technique [0002] Deinococcus radiodurans R1 (D.radiodurans R1) has strong resistance to ionizing radiation, UV radiation, DNA damage reagents and osmotic stress, so it has become an ideal strain for studying the adaptation mechanism of microorganisms to abiotic stress, and can also be used as a research A model species for salt tolerance in higher plants (Battista et al., 2001). Derived from a large plasmid in the D. radiodurans R1 genome, the encoded protein is related to osmotic stress resistance. [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. Contents of the invention [0004] The purpose o...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/70C12N1/21C12N15/82A01H5/00C12R1/19
Inventor 张维江世杰王劲陈明滕超林敏
Owner LONGPING BIOTECHNOLOGY (HAINAN) CO LTD
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