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Identification and use of rice drought-inducible promoter Oshox24P

A promoter and drought technology, applied in the field of plant genetic engineering, can solve the problems of plant poisoning, death, burdening transgenic plants, etc.

Inactive Publication Date: 2010-09-15
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the overexpression in the reports usually uses a constitutive promoter. Although the overexpression effect of the constitutive promoter is high, it is usually accompanied by negative effects, such as toxicity or burden to the plant or lethality of the transgenic plant. (Shavindra et al., Transgenic approaches to increase dehydration-stress tolerance in plants. Mol Breed 5:493-503, 1999)
Although it has been reported in the literature that a stress-responsive promoter has been isolated from plants (Yamaguchi-Shinozaki et al., Characterization of the expression of a desiccation-responsive rd29 gene of Arabidopsis thaliana and analysis of its promoter in transgenic plants. Mol Gen Genet 23: 331-340 , 1993; Kasuga et al., A combination of the Arabidopsis DREB1A gene and stress-inducible rd29A promoter improved drought-and low-temperature stress tolerance in tobacco by gene transfer. Plant Cell Physiol 45: 346-350, 2004; Xiao et al., Over-expression of a LEA gene in rice improves drought resistance under the field conditions. Theor Appl Genet115:35-46, 2007), but there are few strong stress-inducible promoters used in plant genetic engineering in rice, an important food crop

Method used

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  • Identification and use of rice drought-inducible promoter Oshox24P
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  • Identification and use of rice drought-inducible promoter Oshox24P

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1, Oshox24P promoter isolation and identification

[0024] Through the analysis of the drought-induced gene expression profile of the rice variety "Zhonghan 5" (a commercial variety provided by the Shanghai Academy of Agricultural Sciences in China), a gene that was strongly induced by drought (the expression level increased by more than 30 times in the late stage of drought stress) was found. TIGR (http: / / rice.plantbiology.msu.edu / ) ID is LOC02g43330, named Oshox24 (Adamantia et al., Agenome-wide survey of HD-Zip genes in rice and analysis of drought-responsive family members, Plant Mol Biol 66: 87-103, 2008), the corresponding BAC clone number is AP004868, and the corresponding full-length cDNA number in the KOME database (http: / / cdna01.dna.affrc.go.jp / cDNA / ) is AK063685.

[0025] The next step is to isolate the promoter of the gene. The specific steps are as follows: Find the genome sequence (AP004868) of the japonica rice "Nipponbare" corresponding to the g...

Embodiment 2

[0026] Embodiment 2, detect the induced expression of rice endogenous gene Oshox24

[0027] Rice varieties "Zhonghan 5", "Zhonghua 11" and "Zhenshan 97" were used as materials, planted in sandy soil in greenhouses, and subjected to drought stress at the 3-leaf stage. Samples at time point d0 were taken before the stress, samples at time point d1 were taken when the leaves were slightly rolled up due to drought, samples at time point d2 were taken when the leaves were half-rolled, and samples at time point d3 were taken when the leaves were fully rolled. Total RNA was extracted using TRIZOL reagent (purchased from Invitrogen Company) (extraction method according to the above-mentioned TRIZOL reagent instruction manual), and reverse transcriptase SSⅢ (purchased from Invitrogen Company) was used to synthesize cDNA by reverse transcription (method according to Invitrogen Company reverse transcriptase reagent instructions). Using the above cDNA synthesized by reverse transcription...

Embodiment 3

[0028] Example 3, Oshox24P promoter stress-induced activity identification

[0029] The embodiment of the present invention is to construct the GUS gene expression vector of the Oshox24P promoter and transform it into the rice variety "Zhonghua 11" (commercial variety from the Crop Research Institute of the Chinese Academy of Agricultural Sciences), and quantitatively detect the adversity of the Oshox24P promoter including drought, high Induced expression activity of salt, ABA, cold stress. The specific operation is as follows:

[0030] First, the PCR product of the Oshox24P promoter isolated in Example 1 was ligated into the pGEM-T Easy vector (purchased from Promega), transformed into Escherichia coli DH5α (purchased from Promega) and positive clones were obtained. Oshox24P was recovered from the pGEM-T Easy positive clone by BamHI single digestion and then connected to the GUS expression vector p1391Z (from the vector publicly used by CAMBIA, the vector contains the GUS re...

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Abstract

The invention belongs to the technical field of plant genetic engineering and relates to the isolation, identification and use of a plant stress specifically inducible promoter Oshox24P. A rice drought-inducible promoter Oshox24P, which has a nucleotide sequence SEQ ID No.1 in a sequence table, is obtained by isolation and identification. The promoter of the invention can be used in plant genetic engineering for plant genetic improvement, in particular for plant drought resistance improvement.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering. It relates to the isolation, identification and application of a plant stress-specific inducible promoter. By isolating and identifying a rice drought-inducible gene promoter, it can be applied to plant genetic engineering to achieve the purpose of plant genetic improvement, especially improving plant drought resistance. Background technique [0002] Drought has always been one of the important reasons for crop yield reduction, and with the deterioration of the global environment, climate anomalies will increasingly endanger food production security. This is even more evident in China, a country with a shortage of fresh water resources. Improving plant drought resistance has always been a long-term and arduous task. On the one hand, transgenic has become an effective means to study functional genomics; on the other hand, the improvement of plant stress resistance through tran...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A01H5/00
Inventor 熊立仲杨梅
Owner HUAZHONG AGRI UNIV
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