ZmGLK44 gene for regulating and controlling water utilization efficiency of corn under drought and application of ZmGLK44 gene

A drought and water technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve problems such as the inability to synthesize photosynthetic organs

Active Publication Date: 2021-09-10
HUAZHONG AGRI UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In Arabidopsis, glk1glk2 double mutants are pale green and fail to synthesize photosynthetic organs

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • ZmGLK44 gene for regulating and controlling water utilization efficiency of corn under drought and application of ZmGLK44 gene
  • ZmGLK44 gene for regulating and controlling water utilization efficiency of corn under drought and application of ZmGLK44 gene
  • ZmGLK44 gene for regulating and controlling water utilization efficiency of corn under drought and application of ZmGLK44 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Acquisition of ZmGLK44 Gene Regulating Water Use Efficiency in Maize under Drought

[0036] According to the cDNA sequence of ZmGLK44, a pair of homologous recombination primers were designed for PCR amplification. The primer sequences are as follows:

[0037] ZmGLK44-OE-F: 5'-GCACTAGTATCCCGGGAAGGCGCGCCATGGGGCTGGACGTCGG-3'; ZmGLK44-OE-R: 5'-CGTCGTATGGGTACATGGCCACGTATTTCCGGCTGTAGCC-3'.

[0038] The above PCR amplification uses Vazyme Phanta Max Super-Fidelity DNA polymerase for gene amplification, and its reaction system is as follows:

[0039] 2x phanta buffer 10ul dNTP 0.4ul ZmGLK44-OE-F 0.8ul ZmGLK44-OE-R 0.8ul Phantas max 0.4ul cDNA 100ng wxya 2 o

7.1ul

[0040] The reaction program is: 95°C for 3min; 95°C for 15s; 60°C for 15s; 72°C for 60s; 72°C for 5min; 35 cycles,

[0041] That is, the PCR product is obtained. After sequencing and detection, the sequence of the PCR product obtained i...

Embodiment 2

[0043] 1. Construction of ZZ0153-RD101p-3HA vector

[0044] 1) A primer pair was designed using the promoter sequence of the drought-induced expression gene ZmRD101 of the maize B73 inbred line as shown in SEQ ID NO.1 as a template, as follows: RD101-F: 5'-CCTGTCAAACACTGATA GTTT TTCACTTTTTTAGTCTGGCAAT-3', RD101-R: 5'-CCGGGATACTAGTGC GTTTAAAC GCTCACGGTTGCCTTG-3' (the underline is the PMEI restriction site);

[0045] 2) PCR was carried out using the sequence of the corn drought-induced expression gene ZmRD101 as a template, and the nucleotide sequence of the gene PCR product was sequenced as shown in SEQ ID No.3; the PCR amplification conditions were as follows:

[0046] reaction system:

[0047] 2x phanta buffer 10ul dNTP 0.4ul Forward primer RD101-F 0.8ul Reverse primer RD101-R 0.8ul Phantas max 0.4ul cDNA 1.5ul wxya 2 o

6.9ul

[0048] The PCR reaction conditions: pre-denaturation at 95°C for 3 min; denaturation at 95...

Embodiment 3

[0068] Example 3 Obtaining of Transgenic Maize Strains

[0069] 1. Send the carrier obtained in Example 2 to Jiangsu Weimi Biotechnology Co., Ltd. for transformation. The transformation background is corn inbred line C01, and the T1 generation is obtained from the T0 generation transgenic seeds obtained by the company;

[0070] 2. Screen T1-positive plants by smearing herbicides, harvest seeds, and cultivate T2 generation;

[0071] 3. Continue to perform PCR detection on the T2 generation plants to confirm that the target gene has not undergone genetic segregation loss, and finally obtain two positive homozygous transgenic maize families containing stable inheritance of ZmGLK44 and corresponding negative segregation materials; PCR detection primers are (RD101p-F / ZmGLK44-OE-R).

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a ZmGLK44 gene for regulating and controlling water utilization efficiency of corn under drought and application of the ZmGLK44 gene. A nucleotide sequence of the ZmGLK44 gene is as shown in SEQ ID No.1, the corn drought-resistant gene ZmGLK44 is introduced into a corn line C01, the drought-resistant phenotype and the drought-resistant physiological index of a transgenic plant are remarkably improved, and it is indicated that the ZmGLK44 gene can be used for improving the drought resistance of crops.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to a ZmGLK44 gene for regulating water use efficiency of maize under drought and application thereof. Background technique [0002] Maize (Zea may L), a cross-pollinated annual grass, is an important food crop, as well as an important feed and industrial raw material. In my country, the planting area of ​​corn has surpassed that of rice and has become the largest food crop in my country. Maize production is seriously threatened by drought, [0003] With the change of natural environment, the threat of drought to corn production will become more and more serious. In-depth research on the genetics and molecular mechanism of drought resistance in maize, identification and cloning of important drought resistance genes, and the combination of plant genetic engineering and traditional breeding can speed up the selection of varieties with excellent drought resistance. ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N1/21C12N15/11A01H5/00A01H6/46C12R1/19
CPCC07K14/415C12N15/8273
Inventor 代明球张斐吴锦峰
Owner HUAZHONG AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap