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Application of OsDRAP1 genes of rice in enhancing plant drought resistance

A drought resistance, plant technology, applied in applications, plant products, genetic engineering, etc., can solve the problem of unknown gene function of EREBPs

Active Publication Date: 2013-12-25
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the mechanism of EREBPs transcription factors participating in abiotic stress response has been comprehensively studied in recent years, the functions of many EREBPs genes in rice are still unknown.

Method used

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  • Application of OsDRAP1 genes of rice in enhancing plant drought resistance
  • Application of OsDRAP1 genes of rice in enhancing plant drought resistance
  • Application of OsDRAP1 genes of rice in enhancing plant drought resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Spatiotemporal expression differences of OsDRAP1 gene in different rice varieties under different stress treatment conditions

[0027] Nipponbare, Oryza sativa ssp.japonica (publicly available from the Institute of Crop Science, Chinese Academy of Agricultural Sciences, non-patent literature that has recorded this material is: Yongqing Jiao, Yonghong Wang, Dawei Xue, Jing Wang, Meixian Yan, Guifu Liu , Guojun Dong, Dali Zeng, Zefu Lu, Xudong Zhu, Qian Qian and Jiayang Li(2010) Regulation of OsSPL14by OsmiR156defines ideal plant architecture in rice. Nature Genetics, 42, 541-544);

[0028] Typical drought-tolerant upland rice variety IRAT109 (the public can obtain from the Institute of Crop Science, Chinese Academy of Agricultural Sciences, the non-patent literature that has recorded this material is: San-Hong Liu, Bin-Ying Fu, Hua-Xue Xu, Ling-Hua Zhu, Hu -Qu Zhai, Zhi-Kang Li (2007) Cell death in response to osmotic and salt stresses in two rice (Oryza sativ...

Embodiment 2

[0045] Example 2, OsDRAP1 can bind GCC box and DRE cis elements to regulate the expression of downstream genes

[0046] 1. Preparation of prokaryotic expressed protein

[0047] The gene coding sequence of the AP2 conserved domain (67~193aa) of OsDRAP1 was amplified by PCR (i.e. the nucleotide sequence shown in the 703rd-1083rd position of Sequence 1 in the sequence listing), and constructed into the plasmid pET-32a (Novagen , USA, 69015-3), protein expression was carried out in Escherichia coli BL21(DE3)pLysS (TransGen Biotech, China, Cat.No: DR101), the product was purified by His-trap column (GE), concentrated by Miliproe ultrafiltration tube After desalting and desalting, the protein concentration was determined by the Bradford method. The specific method is as follows:

[0048](1) PCR amplification of the AP2 domain coding region of the target gene:

[0049] The primers used are: OsDRAP1-AP2-F: 5′-GCA CTGCAG CAACTCCACATGCAGCGG-3' (the underlined part is the Pst I rest...

Embodiment 3

[0080] Example 3. Overexpression of OsDRAP1 gene enhances drought resistance of transgenic rice

[0081] 1. Overexpression and RNAi expression vector construction

[0082] Primers were designed according to the full-length Nipponbare sequence, and the full-length OsDRAP1 gene was amplified using Nipponbare cDNA as a template. At the same time, Pst I and BamH I restriction sites were added to the 5' and 3' ends. The amplification primers were:

[0083] F: 5′-GCA CTGCAG ATGGCAGCTGCTATAGAAGG-3' (the underlined part is the Pst I restriction site);

[0084] R: 5′-TAA GGATCC GTTATTGTTGTTGAGCAGC-3' (the underlined part is the restriction site of BamH I), the amplified product is digested and purified by Pst I and BamH I, and the target DNA fragment (845bp) is recovered; at the same time, the expression vector is digested with Pst I and BamH I pCUbi1390 (contains Ubiquitin1 promoter) (publicly available from Institute of Crop Science, Chinese Academy of Agricultural Sciences, non...

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Abstract

The invention discloses application of OsDRAP1 genes of rice in enhancing plant drought resistance and provides application of proteins shown in the second sequence of a sequence list in enhancing the plant drought resistance and application of encoding genes of the proteins shown in the second sequence of the sequence list in enhancing the plant drought resistance. Experimental results show that the expression quantity of the OsDRAP1 genes increases rapidly under high salinity stress and drought stress, but has a small increase under low-temperature stress. OsDRAP1 transcription factors can be combined with a GCC box and a DRE in a downstream gene promoter area to regulate and control expression of downstream genes and participate in plant stress response reaction. The over-expression OsDRAP1 genes of the rice can improve the ability of drought-stress resisting of the rice. The application of the OsDRAP1 genes of the rice in enhancing the plant drought resistance has important theoretical and practical significance in improving and enhancing the stress resistance of the rice and accelerating the process of stress-resisting molecular breeding.

Description

technical field [0001] The invention relates to the application of rice AP2 / EREBP transcription factor family gene OsDRAP1 in enhancing plant drought resistance in the field of biotechnology. Background technique [0002] Various biotic and abiotic stresses are important factors affecting rice growth and yield. Discovering genes that can improve rice resistance to adversity stress will lay the foundation for genetic engineering to improve new rice varieties. During the long-term evolution, plants have developed various mechanisms to cope with stress at the molecular, cellular, physiological and biochemical levels, and transcription factors, as upstream regulatory genes, play an important role in the stress response pathway of plants. The AP2 / EREBP, bZIP, NAC, MYB and WRKY transcription factor families are mainly involved in the stress response in plants. Among them, AP2 / EREBP transcription factor family genes are widely involved in plant development, hormone signal transduc...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
Inventor 傅彬英黄立钰黎志康王文生
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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