Method for preserving rice black-streaked dwarf virus indoor living bodies

A technology of rice black bar dwarfing and preservation method, which is applied in the field of agricultural science, can solve problems such as the difficulty of breeding disease-resistant varieties, and achieve the effects of extending the identification time of variety resistance, overcoming technical bottlenecks, and overcoming the difficulty of long-term preservation of plants

Active Publication Date: 2013-01-16
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

All the factors mentioned above have brought great difficulties to the selection and breeding of disease-resistant varieties around RBSDV and the work that requires RBSDV living materials to conduct various researches

Method used

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  • Method for preserving rice black-streaked dwarf virus indoor living bodies
  • Method for preserving rice black-streaked dwarf virus indoor living bodies
  • Method for preserving rice black-streaked dwarf virus indoor living bodies

Examples

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Effect test

Embodiment 1

[0019] Example 1. Indoor in vivo preservation method of rice black streaked dwarf virus

[0020] ① Fresh plants carrying rice black streaked dwarf virus were obtained.

[0021] In September 2010, rice with typical "green dwarf" symptoms was collected in a severely ill area in Ganyu County, Jiangsu. Refer to the Trizol (Reagent) manual to extract total RNA from rice. Design primers based on the published sequence of RBSDV-S5-ORF2 (primers were synthesized by Shanghai Handsome Biotechnology Company):

[0022] F: 5'-TAATCAGCGCGATGTCTAC-3'

[0023] R: 5'-CCCCCGCATCTAAGGAG-3'.

[0024] RT-PCR was used to determine whether the diseased strain carries rice black streaked dwarf virus. Reverse transcription was performed according to the instructions of M-MLV reverse transcriptase (Promega). The PCR amplification system is: cDNA template 3.0μL, Taq plus enzyme 0.5μL, dNTP (10mmol.L-1 each) 0.5μL, 10×PCR buffer (containing Mg2+) 2.5μL, 5'end primer (10pmol.L) -1) 1.0μL, 3'end primer (10pmol....

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Abstract

The invention provides a method for preserving rice black-streaked dwarf virus (RBSDV) indoor living bodies. According to the principle of the method, wheat serves as a virus source host, small brown planthopper imagoes carrying the RBSDV spread the virus on wheat seedlings, a descendant small brown planthopper nymphs obtain the virus from the wheat, and accordingly a large of small brown planthoppers with the virus are obtained at a short period under indoor feeding conditions. The method effectively overcomes the technical bottleneck in preserving a large number of RBSDV indoor living bodies in a long term, and a series of technical problems that an RBSDV virus source is difficult to preserve continuously through the living bodies, the conventional cycle of obtaining the virus source is long, virus feeding plants cannot be provided for a long term, and minimum of the instar of the virus feeding nymphs is difficult to operate, and the like are solved. The obtained small brown planthoppers with the virus can support rice and corn variety resistance identification at any time and can also serve as RBSDV purification and serum preparation materials, and the problems that materials are difficult to obtain and the virus content is low and the like in conventional purification are solved. By application of the method, research, development and utilization of the RBSDV are greatly accelerated.

Description

Technical field: [0001] The invention relates to an indoor living body preservation method of rice black streaked dwarf virus and its application in agriculture, and belongs to the field of agricultural science and technology. Background technique: [0002] Rice black-streaked dwarf virus (RBSDV) belongs to the Fijivirus genus of Reovirdae (Reovirdae). It is a virus that is transmitted by Laodelphax striatellus in a persistent, non-ovitic manner. Mainly infects maize to cause maize rough dwarf disease and infects rice to cause rice black-streak dwarf disease. The susceptible plants show that the growth of the plant is severely inhibited, the leaves are dark green, and wax drops may appear on the leaves, sheaths and stems. Shaped protrusions, black streaks can be formed in the upper and late stages of rice, and the back of corn leaves have obvious roughness. The seriously diseased plants do not head or loose powder, which seriously affects the yield of corn and rice. At present, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/033A01G7/00
Inventor 任春梅程兆榜周益军
Owner JIANGSU ACAD OF AGRI SCI
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