Application of Arabidopsis thaliana AtDIQD gene in improving drought resistance and photosynthetic efficiency of plants
A technology of photosynthetic efficiency and Arabidopsis thaliana, applied in the application field of Arabidopsis AtDIQD gene in improving plant drought resistance and photosynthetic efficiency, to achieve the effects of enhancing drought resistance, increasing thousand-grain weight and yield
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0026] Example 1: Obtaining and identification of T-DNA insertion mutant atdiqd.
[0027] The acquisition of T-DNA insertion mutant atdiqd: on the Arabidopsis thaliana gene database website Tair website (https: / / www.arabidopsis.org / ), input the gene number of AtDIQD (AT3G49260), select and order the T-DNA insertion mutant ( SALK_047040C), T-DNA insertion mutant atdiqd material was obtained.
[0028] Identification of T-DNA insertion mutant atdiqd: Download the cDNA sequence of the gene, and design primers: AtDIQD-F: 5'-TACCAGACAGCATTCCAAT-3' and AtDIQD-R: 5'-TTCTTCGCATTCGTTTCC-3'. Real-time quantitative PCR was used to detect the expression of AtDIQD gene in T-DNA insertion mutants atdiqd and Col-0.
[0029] The specific steps are: (1) extract the RNA of the atdiqd mutant and Col-0 Arabidopsis leaves respectively; (2) reverse transcribe the RNA into cDNA, and use the obtained cDNA as a template for PCR reaction; (3) perform real-time quantification PCR reaction. The results...
Embodiment 2
[0037] Example 2: Construction and genetic transformation of AtDIQD overexpression vector.
[0038]In order to further study the role of AtDIQD gene in regulating the drought resistance of Arabidopsis, plants overexpressing AtDIQD gene were obtained through genetic transformation.
[0039] 1. Construction of AtDIQD overexpression vector
[0040] Download the CDS sequence (sequence as shown in SEQ ID NO:1) of this gene on Tair website, take AtDIQD gene CDS sequence as template (use Primer5 biological software to design primers (front and back primers add attB1 and attB2 respectively), primer is: AtDIQD -CDS-F: 5'-GGGGACAAGTTTGTACAAAAAAGCAGGCTTAATGGGGAAGAAAGGGAGTG-3', AtDIQD-CDS-R: 5'-GGGGACCACTTTGTACAAGAAAGCTGGGTAATGATCATGCCTCCAGCC-3'; PCR amplification of the AtDIQD target fragment, the amplified product is the sequence shown in SEQ ID NO: 1 of the present invention ( 1-1416bp), the PCR reaction conditions are: 95°C pre-denaturation for 5min; 95°C for 30sec, 56°C for 30sec, 7...
Embodiment 3
[0051] Example 3: Construction and genetic transformation of a vector for GUS expression driven by AtDIQD promoter
[0052] Take AtDIQD target gene coding region initiation codon ATG upstream 1576bp as its promoter (the sequence of the promoter is shown in SEQ ID NO: 3) amplification template, use Primer 5 software to design primers (before and after primers respectively add attB1 and attB2); the specific primer sequence is: AtDIQD-P-F: 5'-GGGGACAAGTTTGTACAAAAAGCAGGCTTAAAAAACCCGAACCAGAGG-3', AtDIQD-P-R: 5'-GGGGACCACTTTGTACAAGAAAGCTGGGTAATTTCCGGCAACACTCTT-3', the target fragment is amplified by PCR, and the amplified product is exactly SEQ ID NO of the present invention: 3 Sequences indicated (1-1576 bp). The PCR reaction conditions were: pre-denaturation at 95°C for 5 minutes; 32 cycles at 95°C for 30 sec, 58°C for 30 sec, and 72°C for 1 min; extension at 72°C for 5 min. Use the Gateway method to construct the vector, and use PEG8000 / 30mmol / L MgCl for the target fragment 2 A...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More