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Transcription factor SiARDP for conditioning resilience resistance of rice and encoding gene and application of transcription factor SiARDP

A transcription factor and gene technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of induction, etc., and achieve the effect of enhancing plant resistance to abiotic stress and benefiting agricultural production

Inactive Publication Date: 2014-09-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most DREB transcription factors belong to ABA-independent signaling pathways, but many reports have shown that the transcription levels of some DREB transcription factors are induced by exogenous ABA

Method used

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  • Transcription factor SiARDP for conditioning resilience resistance of rice and encoding gene and application of transcription factor SiARDP
  • Transcription factor SiARDP for conditioning resilience resistance of rice and encoding gene and application of transcription factor SiARDP
  • Transcription factor SiARDP for conditioning resilience resistance of rice and encoding gene and application of transcription factor SiARDP

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Cloning of the gene SiARDP induced by abiotic stress

[0032] After mixing the millet materials subjected to drought, salt and low temperature stress, total RNA was extracted and a cDNA library was constructed. Using the DRE element sequence (ATACTACCGACATGAGC) in the promoter region of RD29A as bait, SiARDP was cloned from the abiotic stress cDNA library of millet by using the Clontech company yeast one-hybrid kit. The full-length sequence is shown in figure 1 . Northern and qRT-PCR analysis showed that the gene was induced by abiotic stresses such as ABA treatment and drought ( figure 2 ).

Embodiment 2

[0033] Example 2SiARDP has the function of transcription factor

[0034] In order to study the localization of SiARDP in cells, the plasmid 35S:SiARDP-GFP with GFP tag was constructed. In addition, using the known nuclear localization of AHL22 as a positive control, a plasmid 35S:AHL22-RFP with an RFP tag was constructed. The two plasmids were co-transformed into protoplasts of etiolated millet seedlings, and the localization of SiARDP protein was observed under confocal laser microscope after culturing in the dark for 16 hours. The results showed that SiARDP was localized in the same position as AHL22 and located in the nucleus ( image 3 ), which is consistent with the predicted properties of SiARDP as a transcription factor. The transcriptional activity of SiARDP was analyzed using the yeast transcriptional activation system. SiARDP was constructed into pBD-GAL4 plasmid and transformed into yeast YRG-2. Dilute the transformed positive yeast with sterile water to OD 600...

Embodiment 3

[0035] Example 3 Obtaining of SiARDP heterologously expressed Arabidopsis plants

[0036] In order to study the function of SiARDP, a 35S promoter-driven SiARDP expression vector p35S:SiARDP was constructed, and p35S:SiARDP was transformed into wild-type Arabidopsis plants by Agrobacterium-mediated transformation. A total of 48 positive plants were obtained by hygromycin screening. After two generations of screening, 3 plants were selected from the homozygous single-copy plants obtained in the T3 generation as materials for further research, and they were named L4, L15 and L22 respectively. The total RNA of wild-type and transgenic Arabidopsis thaliana was extracted for PCR detection and RT-PCR detection. The primers were RT-SiARDPS and RT-SiARDPR. The transgenic plants all had SiARDP inserted and expressed ( Figure 5 ).

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Abstract

The invention provides a transcription factor SiARDP for conditioning resilience resistance of rice and an encoding gene and an application of the transcription factor SiARDP. The amino acid sequence of the transcription factor is as shown in SEQ ID No.1, and the encoding gene is as shown in SEQ ID No.2. By adopting the transcription factor SiARDP for conditioning the resilience resistance of rice and the encoding gene of the transcription factor, the invention provides a novel method for improving the abiotic stress resistance of plants, thereby being beneficial for agriculture production.

Description

technical field [0001] The invention relates to plant genes, in particular to a transcription factor SiARDP regulating stress resistance of millet and its coding gene and application. Background technique [0002] The growth and development of plants in nature will be affected by many adverse natural disasters, among which drought, high temperature, low temperature, salinity, and pests and diseases are the main factors that limit crop yield. my country is in the process of rapid industrialization. With the intensification of industrialization, the pressure on the environment is increasing, and various extreme climates appear frequently, which seriously threatens my country's food security. Drought is one of the serious environmental threats facing our nation. According to statistics, about 1 / 2 of my country's land area is in arid and semi-arid state, and my country's water resources are unevenly distributed, with more in the south and less in the north, and more in the east...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00
CPCC07K14/415C12N15/8273
Inventor 于静娟李丛岳静
Owner CHINA AGRI UNIV
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