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

Method for improving antioxidant capacity and quality of seeds

An anti-oxidation ability and anti-oxidation technology, applied in the field of molecular biology and biology, can solve the problems of reduced seed viability, agricultural production loss, pest damage, etc., and achieve the effect of improving VE content and anti-oxidative stress ability

Inactive Publication Date: 2011-11-23
SHANDONG AGRICULTURAL UNIVERSITY
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the management is not good, it will cause serious damage to the pests, resulting in a reduction in the viability of the seeds, and in severe cases, the seeds will be moldy and rotten, causing great losses in agricultural production.

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
  • Method for improving antioxidant capacity and quality of seeds
  • Method for improving antioxidant capacity and quality of seeds
  • Method for improving antioxidant capacity and quality of seeds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Analysis of seed-specific expression of At2S3 promoter

[0019] 1. Extract the genomic DNA of Arabidopsis thaliana leaves, and use specific primers to obtain the full-length sequence of the At2S3 promoter, 1760bp, by PCR cloning.

[0020] 2. The At2S3 promoter was inserted into the upstream of the GUS gene of the expression vector pBI121, and transformed into Agrobacterium GV3101.

[0021] 3. Preparation of Agrobacterium infection solution: Agrobacterium grown to the static growth phase (OD600=0.8), centrifuged at 5000rpm for 5min, the supernatant was kept as a strain, the sediment was suspended with 5% sucrose solution, and 0.3 -0.5‰ Silwet L-77.

[0022] 4. Infection of Arabidopsis thaliana: Immerse the whole Arabidopsis inflorescence in the infecting solution, shake slightly for 6-8 sec, and then take it out quickly; The thaliana was kept in the dark for 12 hours, and then transferred to the long-day condition for growth; the infected Arabidopsis thalian...

Embodiment 2

[0027] Example 2: Construction of At2S3 promoter-driven antioxidant gene expression vector

[0028] 1. Extract total RNA from Arabidopsis thaliana leaves, reverse transcribe into cDNA, and obtain MSD1 (700bp), CAT1 (1504bp) and HPT1 (1197bp) fragments by PCR amplification with specific primers.

[0029] 2. The obtained fragments were respectively inserted downstream of the At2S3 promoter to construct expression vectors.

[0030] 3. The constructed expression vector was transformed into Agrobacterium GV3101.

Embodiment 3

[0031] Example 3: Acquisition of transgenic Arabidopsis and analysis of its ability to resist oxidative stress

[0032] 1. Agrobacteria containing different vectors were used to infect wild-type Arabidopsis inflorescences.

[0033] 2. The harvested Arabidopsis seeds transfected with different vectors were screened separately.

[0034] 3. The obtained transgenic plant seeds are continuously screened on the medium containing kanamycin until homozygous ones are obtained.

[0035] 4. RT-PCR and enzyme activity assay were used to analyze the expression of antioxidant genes at the transcription level and enzyme activity level, and then select the two At2S3:: MSD1(M7) and At2S3::CAT1(C9) were cross-pollinated to obtain a bivalent transgenic line M7 / C9.

[0036] 5. Measure the VE content in the seeds of each strain transformed with At2S3::HPT1 by high performance liquid chromatography, select a strain H3 with the highest content, and cross it with the bivalent transgenic strain M7 / C...

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 relates to a method for improving antioxidant capacity and quality of seeds. In the method, the recombination of a seed-specific promoter At2S3 and three antioxidant genes is applied to improve the antioxidant capacity of the seeds. The upstream 1760 bp promoter sequence of the At2S3 gene respectively fuses GUS (beta-glucuronidase) reporter gene, manganese-superoxide dismutase (MSD1) gene, catalase (CAT1) gene and a gene of a key enzyme (homogentisate plant-transferase; HPT1) during vitamin E biosynthesis to transform arabidopsis and carry out strain hybridization. The results show that the GUS reporter gene is specifically and efficiently expressed in the seeds; and the monovalent MSD1 (M7), bivalent MSD1 / CAT1 (M7 / C9) and trivalent MSD1 / CAT1 / HPT1 (M7 / C9 / H3) transgenic arabidopsis strains have significantly higher germination rate of seeds when being subjected to 10 mu M paraquat oxidative stress treatment and significantly higher survival rate of seedlings when subjected to 1 mu M paraquat oxidative stress treatment than those of wild-type strains. Therefore, the promoter is a seed-specific highly-efficient promoter, and the recombination of the promoter and three antioxidant genes has important economic and social benefits for prolonging storage period of the seeds and improving quality and stress resistance of the seeds.

Description

(1) Technical field [0001] The invention relates to the application of the promoter of Arabidopsis thaliana seed storage protein gene 2S albumin isoform 3 (At2S3) and three anti-oxidation genes in germplasm improvement such as improving the anti-oxidation ability of seeds, and belongs to the field of molecular biology and biotechnology. (2) Background technology [0002] Crop seeds are the most basic means of agricultural production, the internal cause of agricultural production increase, and the carrier of various technical measures. Cereals, such as corn seeds, have large embryos, strong breathing, and easy to generate heat. The embryo contains a lot of fat and is prone to oxidation and rancidity. Moreover, the embryo has a large amount of bacteria and is prone to mildew. Soybean seeds, an oil crop, have strong hygroscopicity, are prone to lose vitality, and damaged seeds are prone to mildew and deterioration, and have poor thermal conductivity. A large amount of protein...

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): C12N15/113C12N15/82A01H5/10
Inventor 郑成超郗冬梅吴长艾杨国栋黄金光
Owner SHANDONG AGRICULTURAL UNIVERSITY
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