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Plant inducible promoter and application thereof

A promoter and inducible technology, applied in the field of plant inducible promoter sequence and its application in plant stress resistance regulation

Active Publication Date: 2016-04-20
BEIJING AGRO BIOTECH RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, previous studies were limited to morphology, anatomy, and physiology, and there were few studies on the molecular mechanism of salt tolerance and gene cloning of wild barley

Method used

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  • Plant inducible promoter and application thereof
  • Plant inducible promoter and application thereof
  • Plant inducible promoter and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] DNA molecules such as 1) or 2) or 3):

[0041] 1) sequence in the table The DNA molecule shown in SEQ ID NO.1;

[0042] 2) under high stringency conditions with the sequence in the table A DNA molecule that hybridizes to the nucleotide sequence shown in SEQ ID NO.1 and has promoter activity;

[0043] 3) A DNA molecule having more than 65% identity with the nucleotide sequence of 1) or 2) and having promoter activity.

[0044] A method for obtaining a DNA molecule shown in SEQ ID NO.1, comprising the steps of: 1) using the CTAB method to extract wild barley genomic DNA, 2) using the genome walking method to clone the wild barley HbCIPK2 promoter region; in step 2), two steps are carried out: A round of PCR amplification, using 2 reverse primers SP1 and SP2 respectively, the SP1 and SP2 primers are the sequence in the table DNA molecules shown in SEQ ID NO.2 and SEQ ID NO.3.

[0045] In this embodiment, the specific steps to obtain the DNA molecule shown in SEQIDN...

Embodiment 2

[0061] Application of 1), 2), 3) DNA molecules described in Example 1.

[0062] The biological material related to the DNA molecule described in Example 1 is any one of the following A1)-A6):

[0063] A1) an expression cassette containing said DNA molecule;

[0064] A2) a recombinant vector containing said DNA molecule, or A1) a recombinant vector of said expression cassette;

[0065] A3) a recombinant microorganism containing the DNA molecule, the expression cassette described in A1) or the recombinant vector described in A2);

[0066] A4) a recombinant cell line containing the DNA molecule, the expression cassette described in A1) or the recombinant vector described in A2);

[0067] A5) a transgenic plant tissue containing the DNA molecule, the expression cassette of A1) or the recombinant vector of A2);

[0068] A6) A transgenic plant organ containing the DNA molecule, the expression cassette of A1) or the recombinant vector of A2).

[0069] Use of said DNA molecule as ...

Embodiment 3

[0072] Method for obtaining stress-resistant transgenic plants using the DNA molecules described in Example 1.

[0073] A method for cultivating transgenic plants comprises introducing the DNA molecule and a target gene into plants to obtain a plant in which the target gene is induced by adversity stress and / or ABA-induced expression.

[0074] The method comprises the following steps: 1) Constructing the expression vector proHbCIPK2:GUS driven by the DNA molecule in Example 1; 2) Obtaining a transgenic line by using the expression vector proHbCIPK2:GUS.

[0075] The specific operation method of the step 1) is: use the primers proHbCIPK2-F, proHbCIPK2-R to amplify the DNA molecular sequence in Example 1 by PCR method and introduce restriction sites PstI and XhoI, insert the vector pYBA1121 after sequencing, and construct the obtained Expression vector proHbCIPK2:GUS; the sequences of the primers proHbCIPK2-F and proHbCIPK2-R are respectively the sequences in the table DNA mol...

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Abstract

The invention discloses a plant inducible promoter and an application thereof. A section of HbCIPK2 promoter sequence of 1,750bp is obtained from a halophyte wild barley genome through a chromosome primer walking method, an arabidopsis proHbCIPK2:GUS strain obtained through the promoter sequence can drive expression of GUS genes under stress of different concentrations and times, and the expression activity of GUS is enhanced along with increase of the stress times and concentrations. The inducible expression activity of the GUS genes driven by the HbCIPK2 promoter under PEG and ABA stress is relatively low. The identical transgenic lines are free of GUS gene expressions under stress-free treatment and low-temperature and high-temperature stress. The obtained DNA sequence of 1,750 bp is the novel stress inducible promoter, and has high promoter activity under high-salt stress, and proper expressions of transcription factors and other controlling genes can be induced.

Description

technical field [0001] The invention relates to the field of plant transgenic technology, in particular to a plant-inducible promoter sequence and its application in plant stress resistance regulation. Background technique [0002] Salinization and drought are important environmental factors restricting the sustainable development of agriculture. According to statistics, 103 countries in the world have salinized soil, covering an area of ​​380 million hectares, accounting for about 10% of the global arable land area; my country has about 140 million mu of saline cultivated land and 400 million mu of saline wasteland. What's more serious is that the area of ​​secondary salinized soil is still expanding at a rate of 3% per year due to the rise of global temperature, sea level rise, industrial pollution, agricultural irrigation, improper fertilization and other human factors; plus Water resources are becoming more and more tense, and droughts occur very frequently. Every year,...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/10C12N15/82C12N15/84A01H5/00
CPCC12N9/1205C12N15/1013C12N15/102C12N15/8273C12N15/8293C12Q2531/113
Inventor 李瑞芬陈亚娟魏建华王宏芝
Owner BEIJING AGRO BIOTECH RES CENT
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