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

Method for testing drought tolerances of vigna unguiculata type I and vigna unguiculata type II by fluorogenic quantitative PCR (polymerase chain reaction), diagnostic genes and primers

A fluorescent quantitative and drought-tolerant technology, applied in the field of plant biology, can solve problems that have not yet been reported or developed, and achieve the effect of promoting drought-tolerant breeding of cowpea and improving efficiency

Active Publication Date: 2013-04-17
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology uses specific techniques such as fluorogenic qPCR or DNA sequencing to identify various forms of Cowpeas with varying degrees of resistance towards waterlogging damage caused by drilling them underground during rain events. By comparing these results against known sources of Drought Tolerancy (drill cuttings) data collected from previous years on other crops grown at risk of being affected), this method provides an efficient way to study how well drainage systems have been designed over time.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving crop production through reducing excessive rainfall caused by overwintered grassy weeds called vetchum persicae.

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 testing drought tolerances of vigna unguiculata type I and vigna unguiculata type II by fluorogenic quantitative PCR (polymerase chain reaction), diagnostic genes and primers
  • Method for testing drought tolerances of vigna unguiculata type I and vigna unguiculata type II by fluorogenic quantitative PCR (polymerase chain reaction), diagnostic genes and primers
  • Method for testing drought tolerances of vigna unguiculata type I and vigna unguiculata type II by fluorogenic quantitative PCR (polymerase chain reaction), diagnostic genes and primers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The method for screening diagnostic genes and primers for specifically distinguishing cowpea type I and type II drought tolerance comprises the following steps:

[0030] (1) Download and preprocessing of cowpea non-redundant gene sequence: from Harvest cowpea DNA database UP12 (http: / / www.harvest-web.org / hweb / bin / wc.dll?hwebProcess~hmain~&versid=68) Batch download of cowpea unigene sequences, a total of 29,728. Run the VecScreen program (http: / / www.ncbi.nlm.nih.gov / VecScreen / VecScreen.html) to scan and remove vector and adapter sequences contained on the downloaded sequences.

[0031] (2) Cowpea gene chip design: After scanning all 29,728 unigene sequences in the UP12 database, 4 oligonucleotide DNA sequences were designed as probes at different base positions of each sequence. These 4 sequences were in The weighted average of the signal values ​​in the microarray hybridization will define the signal value of a gene. The technical standards for design are: the GC cont...

Embodiment 2

[0037] The differential expression information of the diagnostic gene VuCETS1 in leaf tissue was analyzed by real-time quantitative PCR to identify type I and type II drought-tolerant materials.

[0038] (1) Material preparation: select a plump seed of 1 type I drought-tolerant cowpea variety and 1 type II drought-tolerant cowpea variety seeds and sow them in a nutrient bowl, and the substrate is vermiculite: nutrient soil = 3:1. After the cotyledons were flattened, the water was thoroughly watered, and then the watering was stopped, and the RNA of the leaves under normal growth and under stress conditions was extracted by the conventional Trizol method 14 days after the treatment.

[0039] (2) RAN extraction and reverse transcription: Take 0.1 g of leaf tissue and add liquid nitrogen to grind it into powder, and extract total RNA according to the kit instructions. Dilute the extracted RNA to 0.1 g / L with DEPC-treated water, and add 2 μL of anchor primer (AP) and 2 μL of RNA i...

Embodiment 3

[0042] Differential expression information of the diagnostic gene VuCETS1 in root tissue was analyzed by real-time quantitative PCR to identify type I and type II drought-tolerant materials.

[0043] (1) Material preparation: Select a plump seed of 1 type I drought-tolerant cowpea variety and 1 type II drought-tolerant cowpea variety seeds and sow them in a nutrient bowl, the substrate is vermiculite: nutrient soil = 3:1. After the cotyledons were flattened, the watering was stopped, and the roots of the plants were carefully pulled out 14 days after the treatment, washed with water quickly, and the RNA of the roots under normal growth and stress conditions was extracted by the conventional Trizol method.

[0044] (2) RAN extraction and reverse transcription: Take 0.1 g of root tissue and add liquid nitrogen to grind it into powder, and extract total RNA according to the kit instructions. Dilute the extracted RNA to 0.1 g / L with DEPC-treated water, and add 2 μL of anchor prime...

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 testing drought tolerances of vigna unguiculata type I and vigna unguiculata type II by fluorescence quantitative PCR, diagnostic genes and primers. According to the method, sequences of the tested diagnostic genes are shown as in SEQIDNO: 1; the fluorescence quantitative PCR primers have the sequences as follows: a forward primer: 5'-TGGATTGTGGTGGACATAC-3' and a downstream primer: 5'-GGTTCCTTCTGAGCATTGA-3'. The method, the diagnostic genes and the primers have the advantages that significant differentially expressed genes in materials of drought tolerances of the vigna unguiculata type I and the vigna unguiculata type II provide a diagnostic gene tool for distinguishing the drought tolerances of different types of vigna unguiculata specifically. In addition, direct technical support is provided for testing drought tolerance types of different breeding parent materials by real-time fluorescence quantitative PCR rapidly and conveniently based on the primer sequences of the tolerance diagnostic genes of the vigna unguiculata type I and the vigna unguiculata type II.

Description

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

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
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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