Method for rapidly identifying rice black-streaked dwarf virus and southern rice black-streaked dwarf virus

A technology of rice black-streaked dwarf and black-streaked dwarf virus, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microbial measurement/inspection, and can solve the problem that sequence homology is difficult to distinguish and has not yet been seen. and other issues to achieve the effect of specific identification methods

Inactive Publication Date: 2010-04-07
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since these two viruses have very similar characteristics in many aspects, traditional plant virus detection methods, such as electron microscope observation (similar in size and shape of virus particles), traditional biological inoculation identification (symptoms, mediators, host range similar), serological detection (sequences still have high homology) and other methods are difficult to distinguish
At present, molecular biology detection methods are mos

Method used

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  • Method for rapidly identifying rice black-streaked dwarf virus and southern rice black-streaked dwarf virus
  • Method for rapidly identifying rice black-streaked dwarf virus and southern rice black-streaked dwarf virus

Examples

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example 1

[0042] The mixed samples of rice that are known to be infected with rice black-streaked dwarf virus and southern rice black-streaked dwarf virus and rice samples infected with rice black-streaked dwarf virus and southern rice black-streaked dwarf virus respectively are used as the test objects. Healthy rice The sample is a negative control.

[0043] 1. Primer design:

[0044] (1) According to NCBI ( http: / / www.ncbi.nlm.nih.gov / sites / entrez? db=nucleotide The S9 nucleotide sequence of rice black streaked dwarf virus (AB011403, AF459812, AY050486, AY050487, AJ291706, AJ297429, AJ297430, NC_003731, AF536564, AF540976, AY039705) and southern rice black streaked dwarf virus (EU523359, EU784843) have been reported on ) S9 nucleotide sequence design degenerate primer, the primer sequence is as follows:

[0045] RB1_S9_F: 5`-GRTAGACAGGCAAAYMTAAGCGT-3`

[0046] RB2_S9_F: 5`-TTACAYCAAGCACTTTGCGAGG-3`

[0047] RB_S9_R: 5`-GGATTACAACAHACACAMCGAAA-3`

[0048] (2) The primer pairing and the size o...

example 2

[0079] The mixed samples of rice that are known to be infected with rice black-streaked dwarf virus and southern rice black-streaked dwarf virus and rice samples infected with rice black-streaked dwarf virus and southern rice black-streaked dwarf virus respectively are used as the test objects. Healthy rice The sample is a negative control.

[0080] 1. Primer design:

[0081] (1) According to NCBI ( http: / / www.ncbi.nlm.nih.gov / sites / entrez? db=nucleotide The S9 nucleotide sequence of rice black streaked dwarf virus (AB011403, AF459812, AY050486, AY050487, AJ291706, AJ297429, AJ297430, NC_003731, AF536564, AF540976, AY039705) and southern rice black streaked dwarf virus (EU523359, EU784843) have been reported on ) S9 nucleotide sequence design degenerate primer, the primer sequence is as follows:

[0082] RB1_S9_F: 5`-GRTAGACAGGCAAAYMTAAGCGT-3`

[0083] RB2_S9_F: 5`-TTACAYCAAGCACTTTGCGAGG-3`

[0084] RB_S9_R: 5`-GGATTACAACAHACACAMCGAAA-3`

[0085] (2) The primer pairing and the size o...

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Abstract

The invention provides a method for rapidly identifying the rice black-streaked dwarf virus and southern rice black-streaked dwarf virus, belonging to the field of plant protection. Three primers are designed according to the nucleotide sequence of the rice black-streaked dwarf virus and southern rice black-streaked dwarf virus, which are respectively [A] RB1_S9_F, [B] RB2_S9_F and [C] RB_S9_R, wherein the combination of A and C detects the rice black-streaked dwarf virus and the combination of B and C detects the southern rice black-streaked dwarf virus. After the total RNA is extracted fromthe samples, the two viruses of the rice black-streaked dwarf virus and the southern rice black-streaked dwarf virus can be detected by only one RT-PCR, thus realizing the identification of two viruses. Compared with the traditional method in which one RT-PCR can only detect one virus, the invention greatly reduces cost, reduces time, and is a detection method which is special, sensitive, economic and convenient.

Description

Technical field [0001] The invention relates to a method for quickly identifying rice black-streaked dwarf virus and southern rice black-streaked dwarf virus, belonging to the field of plant protection. Background technique [0002] Rice black-streaked dwarf virus (Rice black-streaked dwarf virus, RBSDV) is a serious virus disease on rice. It belongs to the Fijivirus of the plant reovirus family (Fijivirus). Laodelphax striatellus spreads. In the early stage, the symptoms of the field are mainly dwarfing and dark green leaves. In the later stage, wax drop-like protrusions appear on the leaves, sheaths and stems, and then black bars are formed, which have a great impact on the yield of rice. Can lead to no yield. In addition to infecting rice, the virus can also infect corn (maize rough dwarf disease), wheat (wheat green dwarf) and a variety of gramineous weeds. It was popular in Jiangsu, Shanghai, and Zhejiang of China in 1963 and 1966, and it broke out in many corn and rice pr...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/94
Inventor 季英华周益军程兆榜周彤熊如意
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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