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Method for quickly detecting rice stripe virus and rice black-streaked dwarf virus in laodelphax striatellus

A technology for rice stripe virus and rice black stripe dwarfing, which is applied in the directions of biochemical equipment and methods, and the determination/inspection of microorganisms to achieve the effect of a specific identification method

Inactive Publication Date: 2014-11-12
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the molecular biology detection method based on RT-PCR technology is more widely used. It has been reported that RT-PCR method is used to detect RSV and RBSDV before, but no technical method that can detect RSV and RBSDV simultaneously has been reported. This method only Only one RT-PCR can realize two kinds of virus identification, on the premise of ensuring reliability and sensitivity, it simplifies the operation, saves cost and time

Method used

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  • Method for quickly detecting rice stripe virus and rice black-streaked dwarf virus in laodelphax striatellus
  • Method for quickly detecting rice stripe virus and rice black-streaked dwarf virus in laodelphax striatellus
  • Method for quickly detecting rice stripe virus and rice black-streaked dwarf virus in laodelphax striatellus

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0039] Example 1: Detection of virus-carrying (RSV) striatellus striatellus after feeding poisonous rice plants with black-streaked dwarf disease

[0040] 1. Toxic mediator

[0041] The mediator SBPH is screened and preserved in this laboratory, and the screening method refers to the method of Liu Haijian et al. (Each bottle has 3-4 rice seedlings at the two-leaf stage, the variety is Wuyujing No. 3), and each head of SBPH is marked, and it is taken out after 3-4 days after laying eggs, and the female parent is detected. The situation of carrying poisonous (RSV) of lice, its progeny continue to raise standby. Select a family with a higher hatch rate and a female parent that is infected with the virus and feed it for 3-4 generations, then detect the virus-carrying condition of its offspring to determine the virus-carrying rate of the family, and raise it for later use.

[0042] 2. Feed poison of SBPH

[0043] The diseased plants showing typical symptoms of rice black-streake...

example 2

[0074] Example 2: Simultaneous detection of the poisonous situation of the SBPH population in the field

[0075] In the spring of 2013, a checkerboard method was used to collect overwintering generations of SBPH in several wheat fields in Gaochun County, Nanjing City, Jiangsu Province. A total of 2,000 heads were collected at random. After freezing, 150 heads were randomly selected. RNA was extracted according to the method in Example 1, and RT-PCR was performed with 3 primers. Detection, it was found that 3 SBPHs carried RSV (a band of 1114bp size was detected), 2 SBPHs carried RBSDV (a band of 414bp size was detected), and no SBPH carried two viruses at the same time, calculated In 2013, the overwintering generation of SBPH in Gaochun area carried RSV: 2% and RBSDV: 1.3%, which provided data support for the prediction and prevention of viral diseases.

example 3

[0076] Example 3: Detection of rice disease samples

[0077] Rice samples infected with RSV and RBSDV respectively and mixed samples infected with RSV and RBSDV were used as detection objects, and healthy rice samples were used as negative controls. Extract RNA according to the method of example one, carry out RT-PCR detection with 3 primers, as a result the sample that infects RSV can detect the band of 1114bp size, the sample that infects RBSDV can detect the band of 414bp size, infects two kinds of viruses The mixed sample can detect bands of 1114bp and 414bp at the same time, and no band is detected in the healthy rice sample, which shows that this detection method is also applicable to the detection of rice samples. Of course, since the symptoms of rice samples infected with RSV and RBSDV are very easy to distinguish in the field, the detection of rice disease samples can be used for the early detection of rice disease before the symptoms are manifested, and it is not com...

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Abstract

The invention provides a method for quickly and synchronously detecting rice stripe virus (RSV) and rice black-streaked dwarf virus (RBSDV) in an insect vector laodelphax striatellus, and belongs to the field of plant protection. According to nucleotide sequences of the rice stripe virus RNA4 and the rice black-streaked dwarf virus S2, three primers are designed as follows: [A] RS-F, [B] RB-F, and [C] RSRB-R, wherein A and C are combined to detect the rice stripe virus; the B and C are combined to detect the rice black-streaked dwarf virus. After extracting total RNA of single laodelphax striatellus, only one RT-PCR is needed, so that the rice stripe virus and the rice black-streaked dwarf virus can be detected, and thus, and quick and synchronous detection for the two viruses in the laodelphax striatellus is realized. Compared with the conventional method for detecting one virus by virtue of one RT-PCR, the cost is greatly saved and the time is saved, and thus, the method is a quick, simple, flexible and specific detecting method. The method disclosed by the invention can be applied to monitoring work of a virus carried rate of a field laodelphax striatellus colony, and can also be applied to related research work of a virus and vector interaction mechanism.

Description

technical field [0001] The invention relates to a method for rapid and synchronous detection of rice stripe virus and rice black-streaked dwarf virus in the body of the vector insect SBPH, belonging to the field of plant protection. Background technique [0002] Rice stripe virus (RSV) is a typical member of the genus Tenuivirus, and the rice stripe blight caused by it is one of the important diseases of rice. The outbreak of the epidemic in the vast rice areas dominated by the region poses a great threat to rice production. For example, in 2004, the diseased area in Jiangsu Province reached 23.55 million mu, accounting for 79% of Jiangsu's rice planting area. Spread, because of its serious harm, was once called "cancer" on rice, which caused great social repercussions. [0003] Rice black-streaked dwarf virus (RBSDV), a serious virus disease on rice, belongs to the plant Reoviridae Fijivirus (Fijivirus), virion spherical, field symptoms In the early stage, the plants are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/686C12Q1/70C12Q2531/113C12Q2537/143
Inventor 李硕季英华王喜周益军
Owner JIANGSU ACAD OF AGRI SCI
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