Application of CbDREB2AL gene in preparation of salt-tolerant transgenic plant

A transgenic and genetic technology, applied in the fields of applications, plant peptides, plant products, etc., can solve the problem of undiscovered CbDREB2AL gene transgenic plants, etc.

Pending Publication Date: 2021-07-02
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The gene has the characteristics of drought resistance and frost resistance, and thus obtained the Chinese invention patents with the authorized patent numbers "CN108018291B" and "CN108018293B". The inventor did not find that the CbDREB2AL gene has not been used to prepare transgenic plants with salt-tolerant characteristics through searching

Method used

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  • Application of CbDREB2AL gene in preparation of salt-tolerant transgenic plant
  • Application of CbDREB2AL gene in preparation of salt-tolerant transgenic plant
  • Application of CbDREB2AL gene in preparation of salt-tolerant transgenic plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Cloning of the sequence of CbDREB2AL coding gene of Ion mustard:

[0023] Utilize RNA extraction and separation reagent (Trizol, Invitrogen) to isolate the total RNA of the regenerated seedlings of Alpine mustard, and the specific method is: collect 100 mg of the regenerated seedlings of Alpine mustard, immediately place it in liquid nitrogen and grind it into powder, then add 1ml of Trizol reagent, and mix well , inhale into a 1.5ml centrifuge tube; place at room temperature for 5min; add 0.2ml of fresh chloroform, shake vigorously for 15s, and let stand at room temperature for 3min; centrifuge at 12000g for 15min at 4°C; transfer the supernatant to a new 1.5ml centrifuge tube , add 0.5ml of isopropanol and mix well, and centrifuge at 12000g for 10min at 4°C to precipitate RNA; wash the RNA pellet with 1ml of 75% ethanol, dissolve it in an appropriate amount of DEPC-treated water, and store it at -70°C for later use.

[0024] The full-length sequence of the gene was ob...

Embodiment 2

[0032] Obtaining of CbDREB2AL Transgenic Plants and Determination of Salt Tolerance

[0033] The construction of the plant overexpression vector of the CbDREB2AL gene of Ion mustard alpine: the fragment obtained in Example 1 obtained by sequencing verification by using the Gateway technology of Invitrogen Company through the BP reaction (BP II Enzyme mix, Invitrogen No.11789020) was recombined into the pDONR / Zeocin vector (Invitrogen No.12535-035), transformed into Escherichia coli DH5α competent cells, and the entry clone was obtained by screening with 20mg / L Zeocin, and then the plasmid was extracted and passed through Gateway The LR reaction (LR II Enzyme mix, Invitrogen No.11791100) the CbDREB2AL gene was recombined onto the PMDC32 carrier, transformed in Escherichia coli DH5α competent cells, and successfully recombined overexpression vector PMDC32-CbDREB2AL (such as image 3 ).

[0034] Agrobacterium-mediated transformation: transform the constructed overexpression pla...

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Abstract

The invention provides novel application of a CbDREB2AL gene. The gene can be applied to breeding of salt-tolerant transgenic crops to improve the salt tolerance of the crops and has wide application prospects.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of CbDREB2AL gene of mustard mustard in preparation of salt-tolerant transgenic plants. Background technique: [0002] Salt stress is one of the important abiotic stresses. High salt will directly or indirectly affect the growth and development of crops at the molecular and physiological levels, and ultimately affect the yield of crops (Abogadallah 2014). Salt stress is also a major threat to modern agriculture, and according to statistics, the land area affected by salt stress is increasing (Wang, Vinocur et al.2003). With the increase of population and the decrease of land resources, people pay more and more attention to the development and utilization of saline-alkali land (Chang, Yang et al.2014). Under salt stress conditions, some plants can re-form ion and osmotic balance by regulating the expression of related genes in cells to reduce or avoid dam...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/20A01H6/82
CPCC07K14/415C12N15/8273
Inventor 岳修乐安黎哲张国艳赵智星
Owner LANZHOU UNIVERSITY
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