Arabidopsis AtPGK2 gene for enhancing salt tolerance of plants and application of arabidopsis AtPGK2 gene

A technology of Arabidopsis thaliana and salt tolerance, applied in the field of plant genetic engineering, can solve problems such as unclear physiological functions, achieve wide application value, and improve salt tolerance

Inactive Publication Date: 2014-10-29
JIANGXI AGRICULTURAL UNIVERSITY
View PDF0 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Entering this century, although recent studies have shown that the content of PGK protein in plants increases significantly under salt stress conditions (Kosova

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
  • Arabidopsis AtPGK2 gene for enhancing salt tolerance of plants and application of arabidopsis AtPGK2 gene
  • Arabidopsis AtPGK2 gene for enhancing salt tolerance of plants and application of arabidopsis AtPGK2 gene
  • Arabidopsis AtPGK2 gene for enhancing salt tolerance of plants and application of arabidopsis AtPGK2 gene

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0028] Example 1: Arabidopsis thaliana encoding 3-phosphoglycerate kinase gene AtPGK2 Clone of

[0029] (1) The total RNA of Arabidopsis plants is extracted by the guanidine isosulfate-phenol method. The use, preparation and specific operation steps of the medicine refer to the method of Huang Peitang et al. (2002).

[0030] (2) According to known Arabidopsis AtPGK2 Design a specific primer for the CDS sequence of the gene and introduce it into the upstream primer Nco I restriction site, introduced in the downstream primer BstE II restriction site.

[0031] Upstream primer: 5’-CATG CCATGG CTTCCACCGCCGCAACTGCA-3’ (introducing Nco I restriction site)

[0032] Downstream primer: 5’-G GGTAACC TTAAACAGTGACTGGCGTTGCT-3’ (introduced BstE II restriction site)

[0033] (3) Take 1 mg RNA as a template for reverse transcription, and use P 2853 Use Promega's reverse transcriptase ImProm-II for primers TM Perform reverse transcription, P 2853 The primer sequence, reverse transcription pr...

Example Embodiment

[0049] Example 2: Use pCAMBIA1301 vector construction " CaMV 35S - AtPGK2 "Fusion Gene

[0050] (1) Extract vector pCAMBIA 1301 plasmid (purchased from CAMBIA company) from E. coli, use Nco I / BstE II Recover large vector fragments after double digestion.

[0051] (2) For the recovered in Example 1 AtPGK2 The CDS fragment of the gene is used Nco I / BstE II Double enzyme digestion, and recover the digested fragments by agarose gel electrophoresis (same as Example 1).

[0052] (3) Connect the above two fragments under the catalysis of ligase at 16°C overnight to complete the "pCAMBIA 1301" vector CaMV 35S - AtPGK2 "Fusion gene construction.

[0053] Connection system:

[0054] Reagent Adding amount (μl) AtPGK2 CDS fragment of gene (50 ng μl -1 ) 2.0 pCAMBIA1301 vector large fragment (50 ng μl -1 ) 3.0 Solution I ligase5.0

[0055] (4) Transform E. coli DH5α competent cells with the ligation mixture, the specific method is as follows:

[0056] According to conventional CaCl ...

Example Embodiment

[0061] Example 3: Preparation of Transgenic Arabidopsis Plants

[0062] (1) Constructed with Example 2 " CaMV 35S - AtPGK2 "The fusion gene was transformed into Arabidopsis thaliana, and the specific transformation method adopted the Agrobacterium-mediated Floral dip method (Clough and Bent, 1998). The obtained seeds were subjected to 50 mg l -1 Hygromycin resistance screening, normal-growing plants are transferred to soil for culture.

[0063] (2) Real-time quantitative RT-PCR detection of transgenic plants: the transgenic Arabidopsis thaliana identified by PCR in Example 2 was multiplied and passed 50 mg l -1 Hygromycin resistance screening to obtain T 3 Generation of homozygous transgenic lines, respectively obtain wild-type and transgenic Arabidopsis seedlings grown for 1 week, extract total RNA from plant tissues according to the method of Example 1, and reverse transcribed into mRNA, using the following primers, reaction procedures and reactions System for PCR reaction:

[00...

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 clones an arabidopsis 3-phosphoglycerate kinase gene AtPGK2 for enhancing salt tolerance of plants, and discloses an application of the arabidopsis 3-phosphoglycerate kinase gene AtPGK2. The nucleotide sequence of the gene is shown in SEQIDNO: 1, and the gene also comprises genes which have homologies of 90-100% with the SEQIDNO: 1 nucleotide sequence. Meanwhile, the invention also provides a construction and transgenosis method of a recombinant vector to apply the gene. The method can be used for cultivating new varieties of transgenic plants with stronger salt tolerance, and has wide application values.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular, it uses molecular biology technology to obtain a gene capable of enhancing the salt tolerance of plants. AtPGK2 and its application in transgenic plants. Background technique [0002] Soil salinization is very serious all over the world, and salt stress is one of the important abiotic stresses affecting global crop yields (Parvaiz and Satyawati, 2008). my country's saline soil has a large area, wide distribution, and many types, accounting for about 10% of the country's land area, and the salinization and secondary salinization are increasing every year, which threatens the sustainable development of agricultural production and seriously affects plants. The growth and development of economic crops and the yield and quality of commercial crops (Sun Jianchang et al., 2008). Therefore, it is a very urgent task to cultivate new varieties of transgenic crops with enhanced salt ...

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
IPC IPC(8): C12N15/54C12N15/10C12N15/84A01H5/00
Inventor 刘栋李卫春马利霞程建峰
Owner JIANGXI AGRICULTURAL UNIVERSITY
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