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Sugarcane bacilliform virus detection method

A sugarcane baculovirus and detection method technology, applied in the field of plant protection, can solve the problems of expensive instruments, difficult detection, slow detection speed and the like, and achieve the effect of specific detection

Inactive Publication Date: 2010-12-08
SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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Problems solved by technology

Because the symptoms of SCBV are unstable, the diseased plants may be hidden under certain conditions, and it is difficult to transmit the virus through mechanical friction, so it is difficult to detect the symptoms of indicative plants; the immune electron microscope technology can directly observe the virus particles, and the diagnosis is relatively reliable. However, the equipment is expensive, the detection speed is slow, and the procedure is cumbersome; although the enzyme-linked immunoassay has high sensitivity, the preparation of high-purity antibodies is complicated and cumbersome

Method used

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Embodiment Construction

[0024] 1. Primer design:

[0025] According to the conserved sequence (sequence accession numbers AJ277091 and M89923) of the reported sugarcane baculovirus genome sequence in the GenBank database, the specific primer PC1 / PC2 for amplifying the sugarcane baculovirus was designed, and the amplified target fragment was 589bp in size. The sequence is as follows:

[0026] PC1 5′-ACCAGATCCGAGATTACAGAAG-3′

[0027] PC2 5′-TCACCTTGCCAACCTTCATA-3′

[0028] 2. Extraction of total DNA from sugarcane leaf tissue:

[0029] (1) Weigh 0.5g of sugarcane leaves and put them into a mortar, add an appropriate amount of liquid nitrogen to grind until powdery, then transfer to a 2ml centrifuge tube;

[0030] (2) Add 800ul of CTAB extract preheated at 65°C to the centrifuge tube containing the grinding material, mix it upside down and put it in a constant temperature bath at 65°C to incubate for 2 hours, and invert the centrifuge tube several times every 20 minutes;

[0031] (3) Take out the c...

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Abstract

The invention relates to a sugarcane bacilliform virus detection method. The method detects a sugarcane bacilliform virus by adopting a PCR (Polymerase Chain Reaction) amplification method after the total DNA (Deoxyribonucleic Acid) of sugarcane top tissues is extracted by using a specific primer PC1 / PC2 designed according to the genome sequence of the sugarcane bacilliform virus. The method overcomes the defects in the traditional detection method, and the rapid and sensitive method for specifically detecting the sugarcane bacilliform virus is provided.

Description

Technical field: [0001] The invention relates to the field of plant protection, specifically a method for detecting sugarcane baculovirus. Background technique: [0002] Sugarcane bacilliform virus (SCBV) is a new viral disease that spreads rapidly in recent years. It is mainly transmitted through the germplasm of Saccharicoccus sacchari and asexual reproductive materials. Due to the extensive exchange of sugarcane seedlings and breeding materials around the world, SCBV has spread to many sugarcane growing areas in the world such as Cuba, Australia, Madagascar, Maras, Mauritius, South America, the United States, Madrid, Morocco , Reunion, Papua New, Guinea and other countries, as well as my country's Taiwan, Guangdong, and Guangxi; although so far, there are few research reports on the impact of the virus on sugarcane production, the establishment of a rapid detection method for the virus is an important Important measure for the spread of this dangerous virus. [0003] Indi...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/94
Inventor 李文凤黄应昆王晓燕尹兴祥卢文洁罗志明
Owner SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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