Unlock instant, AI-driven research and patent intelligence for your innovation.

Quick testing method for rice dwarf virus

A dwarf virus and detection method technology, applied in the field of plant virology, can solve problems such as expensive equipment, complicated technology, and unsuitable for large-scale detection

Active Publication Date: 2012-09-05
云南省农业科学院生物技术与种质资源研究所
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Electron microscope observation technology, expensive equipment, complicated technology, requires professional technicians, not suitable for mass inspection

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0016] Embodiment technical scheme of the present invention is as follows:

[0017] 1. Primer design:

[0018] Upstream primer: 5-GCAAAAATCGCCACCTGCCACTAT-3, downstream primer: 5-CGGGCATTTTCCTCCACGGCTAC-3, used to amplify S8 fragment 1370bp.

[0019] 2. Immunocapture of Virions

[0020] Add RDV rabbit polyantiserum diluted to 1:5000 into the PCR tube, coat at 4°C overnight, and wash 3 times with PBST. Rice disease samples were ground with PBST, centrifuged at 1000 g for 1 min, and 100 μl of the supernatant was added to a PCR tube, incubated for 3 h; washed 3 times with PBST, H 2 O washed once, centrifuged briefly, and sucked off the residual liquid.

[0021] 3. High temperature denaturation

[0022] Add 25 μl of DEPC-treated ultrapure water, denature at 97°C for 5 minutes, immediately insert into ice × pre-cool for 3-5 minutes.

[0023] 4.one step RT-PCR

[0024] The RT-PCR reaction was carried out using the one step RT-PCR kit from Dalian Bao Biological Company. RT-PCR...

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

PropertyMeasurementUnit
diameteraaaaaaaaaa
thicknessaaaaaaaaaa
Login to View More

Abstract

The invention provides a quick testing method for rice dwarf virus. The method includes the following steps: specifically capturing the rice dwarf virus by means of rice dwarf virus antibody, obtaining virus RNA as template by way of high-temperature denaturation and utilizing rice dwarf virus specific primer to carry out one-step reverse transcription polymerase chain reaction (one-step RT-PCR) and testing the target fragment by way of agarose gel electrophoresis. The method utilizes the high titer and specificity of the antibody to specifically and quickly capture the target fragment, consequently, a quick and specific method is provided for genome sequence amplification carried out for the research on the molecular variation of the virus, and thereby the immnocapture RT-PCR technology is perfected.

Description

technical field [0001] The invention relates to the technical field of plant virology, in particular to a rapid detection method for rice dwarf virus. Background technique [0002] Rice dwarf disease caused by rice dwarf virus is widely distributed in Japan, North Korea, Nepal, South Korea, Philippines, as well as my country's Zhejiang, Fujian, Yunnan, Guangxi, Jiangsu, Anhui, Jiangxi, Hunan, Hubei, Guangdong, Chongqing, Sichuan And Guizhou and other rice areas. When an outbreak occurs, it can cause a 30%-50% yield loss, or even no harvest. [0003] The pathogen of rice dwarf disease is rice dwarf virus, the English name is Rice dwarf virus, abbreviated as RDV, which belongs to the plant reovirus genus of the Reoviridae family. The RDV genome encodes 6-7 structural proteins, including P1, P2, P3, P5, P7, P8, and P9; and at least six nonstructural proteins, including Pns4, Pns6, Pns10, Pns11, and Pns12. Among them, P8 is the main coat protein of RDV. The RDV virion is a sp...

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): C12Q1/70C12Q1/68
Inventor 张仲凯董家红尹跃艳丁铭李婷婷方琦彭潞波张丽珍苏晓霞
Owner 云南省农业科学院生物技术与种质资源研究所