Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Malic dehydrogenase related to high photosynthetic capacity and resisting reversal of wheat, coded genes and method for breeding plants of resisting reversal

A technology of malate dehydrogenase and encoding, which is applied in botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., and can solve the problems of low photosynthetic efficiency

Inactive Publication Date: 2009-07-01
INST OF BOTANY CHINESE ACAD OF SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, wheat belongs to C3 plants, and its photosynthetic efficiency is lower than that of C4 plants, such as corn. At the same time, in production practice, the losses caused by environmental adversities such as drought and salinity are very huge.

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
  • Malic dehydrogenase related to high photosynthetic capacity and resisting reversal of wheat, coded genes and method for breeding plants of resisting reversal
  • Malic dehydrogenase related to high photosynthetic capacity and resisting reversal of wheat, coded genes and method for breeding plants of resisting reversal
  • Malic dehydrogenase related to high photosynthetic capacity and resisting reversal of wheat, coded genes and method for breeding plants of resisting reversal

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Isolation and sequence analysis of wheat malate dehydrogenase gene probe

[0029] Wheat (Triticum aestivum L) was planted in a greenhouse, watered and fertilized normally, until it grew to 2-3 internodes, and roots, stems, and leaf tissues were collected and tested with TRI reagent (Molecular Research Center, Inc, Cincinnati, USA) Total RNA was extracted with PolyAT mRNA Isolation kit (Promega company, Madison, USA) separates Poly(A) + RNA, the content and purity of RNA were analyzed by UV spectrophotometer.

[0030] The conditions for reverse transcription to synthesize the first strand of cDNA are: Poly(A) + RNA 1ug, 5μmol / L Primer 5'-GACTCGAGTCGACATCGA(T) 17 -3', 0.5mmol / L dNTP, 50 units of RNasin, incubate at 65°C for 10 minutes, after cooling on ice, add 200U SuperScript TM IIRNase H-Reverse Transcriptase (Gibco), incubated at 42°C for 1 hour, added EDTA to a final concentration of 10 mM, incubated at 95°C for 10 minutes, and cooled on ice.

[00...

Embodiment 2

[0034] Example 2. Screening and sequence analysis of wheat malate dehydrogenase gene

[0035] With Wheat Stem Poly(A) + Using RNA as a template, cDNA was synthesized with a ZAP vector (Stratagene, LaJolla, CA, USA), and packaged in vitro to construct a cDNA library of wheat stems. Take 50ng probe DNA, add 50uCi 32 P-dCTP was labeled, incubated at 37°C for 1 hour, and EDTA was added to a final concentration of 10mM to terminate the reaction. After the probe passed through the Sephadex G-50 column, it was boiled at 100° C. for 10 minutes, and immediately cooled on ice for hybridization.

[0036] Take 50000pfu to plate, and transfer it to nitrocellulose membrane, denature in 0.5M NaOH plus 1.5M NaCl for 2 minutes, 1.5M NaCl plus 0.5M Tris-HCl (pH8.0) for 5 minutes, 0.2M Tris- Rinse with HCl (pH7.5) plus 2 x SSC for 30 seconds, sandwich the membrane between two layers of Whatman filter paper, bake in vacuum at 80°C for 2 hours, then add hybridization solution (6 x SSC, 5 x Denh...

Embodiment 3

[0039] Embodiment three, the translation of wheat malate dehydrogenase gene coded protein

[0040] Artificially synthesize a pair of primers:

[0041] 5'-Primer: 5'-CGGGATCCATGGCTGCGAAGGAACCGATG-3'

[0042] 3'-primer:

[0043] 5'-ATAAGAATGCGGCCGCTTACGCCAGGCATGAGTAGG-3'

[0044] BamHI and NotI restriction sites were introduced at both ends of the primers, and TaMDH gene was used as a template for PCR amplification. Amplification conditions: 10ng DNA, 1μmol / L primers, 0.4mmol / LdNTP, 2.5U Taq DNA polymerase (Gibco Company, Grand Island, NY, USA). Denaturation at 95°C for 5 minutes, followed by 30 cycles (1 minute at 95°C, 1 minute at 55°C, 1.5 minutes at 72°C), and a final extension at 72°C for 10 minutes.

[0045] PCR products were subjected to 1.0% agarose gel electrophoresis, using DNA Isolation Kit (Gibco Company, Grand Island, NY, USA) purified the electrophoresis product, digested with BamHI and NotI double enzymes, connected to the vector pET-28a (Novagen Company, U...

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 provides a malate dehydrogenase related to high light efficiency and stress resistance of wheat, a gene encoding the enzyme and a plant expression vector containing the gene. The invention also provides a method for cultivating transgenic plants with high light efficiency and stress resistance by using the gene of the invention.

Description

field of invention [0001] The invention relates to malate dehydrogenase related to high light efficiency and stress resistance of wheat, a gene encoding the enzyme and an expression vector containing the gene. Background of the invention [0002] Malate dehydrogenase is responsible for catalyzing the conversion between oxaloacetate and malate. It is ubiquitous in various animals, plants and most bacteria, and plays an important role in various metabolic processes. In plant cells, five types can usually be distinguished according to the specificity of its coenzyme and the distribution in the cell (Gietl, C., 1992, Biochim.Biophys.Acta, 1100:217-234): (1) NADP-dependent Chloroplast MDH (EC 1.1.1.82), this type of enzyme is part of the malate shuttle cycle, which plays an important role in maintaining the balance of reducing forces between the cytoplasm and grana. In C4 plants it is also involved in the enrichment of CO into the leaf sheath cell chloroplast 2 . (2) NAD-depen...

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 Patents(China)
IPC IPC(8): C12N15/53C12N9/04C12N15/63C12N15/82
Inventor 马庆虎丁郁
Owner INST OF BOTANY CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com