Method for detecting sugarcane yellow leaf virus
A sugarcane yellow leaf virus and detection method technology, which is applied in the field of plant protection, can solve the problems that are not suitable for rapid detection of large quantities of samples in the field, the time required for detection is long, and sampling is complicated, etc., so as to achieve easy visual judgment, labor saving, and simple operation Effect
Inactive Publication Date: 2009-02-11
SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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Problems solved by technology
However, these methods have the disadvantages of complex sampling, cumbersome operation, long detection time, special expensive instruments and conditions, and high cost, and are not suitable for rapid detection of large quantities of samples in the field.
Method used
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Embodiment Construction
[0012] 1. Sample preparation and tissue imprinting: Take the lower midrib of the first leaf on the upper part of sugarcane, cut it transversely into a plane with a sharp blade, and press the cut plane vertically on the nitrocellulose membrane to obtain imprinted spots. The cellulose membrane was air-dried in a small box;
[0013] 2. Blocking: air-dry the blot, immerse the nitrocellulose membrane in blocking buffer, and incubate at 37°C for 45 minutes;
[0014] 3. Quickly wash the membrane with TBST buffer once: pour out the blocking buffer in the small box, immerse the nitrocellulose membrane in the TBST buffer, turn it gently a few times, and then discard the TBST buffer;
[0015] 4. Add antibody: immerse the nitrocellulose membrane in TBST buffer solution containing 0.01% sugarcane yellow leaf virus specific antiserum and 1% skimmed milk powder, incubate at 37°C for 2 hours, wash the membrane 3 times with TBST buffer solution, 3 minutes each time;
[0016] 5. Add enzyme-lab...
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The invention provides a method for detecting sugarcane yellow leaf virus. The method comprises the following steps of marking the tissue spot of a sample to be detected on nitrocellulose films; soaking the films into a sealed buffer solution containing 0.01% of specific antiserum of the sugarcane yellow leaf virus to incubate; transferring the films to a TBST buffer solution containing 0.1% of alkaline phosphatase labeled antibodies and 1% of skimmed milk powder; transferring two films into the primer / buffer solution (BCIP / NBT), adding 20ml double-distilled water to dissolve and developing for 5-10min; and determining the existence of the sugarcane yellow leaf virus in the sample if the tissue spot appears mauve. If no color is developed on the tissue spot, no sugarcane yellow leaf virus exists in the sample. The method can rapidly and accurately detect the sugarcane yellow leaf virus, and has the advantages that the detection result is easily judged by the visual observation, the operation is simple, the sampling amount is less, the labor consumption is reduced, the result is reliable, the contamination is prevented, no special instrument is in demand, and the method is suitable for detecting the large-scale field samples.
Description
Technical field: [0001] The invention relates to the field of plant protection, in particular to a method for detecting sugarcane yellow leaf virus that is fast, accurate and easy to operate. Background technique: [0002] Sugarcane yellow leaf disease is a worldwide sugarcane disease that seriously harms the growth of sugarcane. Since it was discovered in Hawaii, USA in 1989, it has successively appeared in more than 30 countries and regions around the world and has continued to spread. It is also quite common in various sugarcane areas in my country. Severe, the incidence rate is between 10%-100%. The occurrence of the disease leads to a decrease in sugarcane yield, poor quality and species degradation, and when it is severe, the sugarcane yield can be reduced by as much as 40%-60%. Establishing a rapid, accurate and operable virus detection method is the key to disease control and sugarcane detoxification. [0003] At present, the detection methods of sugarcane yellow le...
Claims
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Login to View More IPC IPC(8): G01N33/544G01N21/78
Inventor 李文凤黄应昆陈学宽蔡清卢文洁
Owner SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI