Application of rice osdrap1 gene in enhancing plant drought resistance
A technology of drought resistance and rice, applied in application, plant products, genetic engineering, etc., can solve problems such as unknown EREBPs gene function
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Embodiment 1
[0026] Example 1. Spatiotemporal expression differences of OsDRAP1 gene in different rice varieties under different stress treatment conditions
[0027] Nipponbare (Nipponbare, Oryza sativa ssp.japonica) (the public can obtain from the Crop Science Research Institute of the Chinese Academy of Agricultural Sciences, the non-patent literature that has recorded this material is: Yongqing Jiao, Yonghong Wang, DaweiXue, Jing Wang, Meixian Yan, Guifu Liu, Guojun Dong, Dali Zeng, Zefu Lu, Xudong Zhu, Qian Qian and Jiayang Li(2010) Regulation of OsSPL14by OsmiR156defines idealplant 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 -QuZhai, Zhi-Kang Li (2007) Cell death in response to osmotic and salt stres...
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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