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Method for rapidly detecting citrus tristeza virus by electrospray ionization mass spectrometry

A kind of citrus Huanglongbing and ionizing technology, applied in the field of rapid detection of citrus Huanglongbing

Inactive Publication Date: 2019-05-31
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The real way to make a diagnosis is laboratory diagnosis, which requires section observation with the help of microscope technology, and there is a large possibility of missed detection; serology, nucleic acid hybridization and PCR and other methods can be detected when the bacterial concentration reaches a certain level, and during the incubation period of plant infection early stages of undetected

Method used

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  • Method for rapidly detecting citrus tristeza virus by electrospray ionization mass spectrometry
  • Method for rapidly detecting citrus tristeza virus by electrospray ionization mass spectrometry
  • Method for rapidly detecting citrus tristeza virus by electrospray ionization mass spectrometry

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Experimental program
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Embodiment 1

[0017] The method for rapid detection of citrus huanglongbing by electrospray extraction ionization mass spectrometry comprises the following steps:

[0018] (1) Collect fresh citrus leaves, accurately weigh 0.2000g sample, grind it into powder under liquid nitrogen, extract DNA with a plant DNA extraction kit, and the total volume of the PCR reaction system is 20 μl; (2) The PCR amplification program is: 94 Pre-denaturation at ℃ for 5min; denaturation at 94℃ for 30s, annealing at 55℃ for 30s, extension at 72℃ for 40s, cycle 35 times; extension at 72℃ for 10min;

[0019] (3) Agarose gel electrophoresis program: use 1% agarose to make the gel, the electrophoresis buffer is 1×TBE buffer, 120V constant voltage electrophoresis for 30min, and DNA Ladder is used as the DNA molecular weight standard; under the gel imaging system, observe and take pictures save;

[0020] (4) Accurately weigh 0.20 g of navel orange leaf fresh weight, grind it into powder under liquid nitrogen, add 2 m...

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Abstract

The invention provides a method for rapidly detecting citrus tristeza virus by electrospray ionization mass spectrometry. The electrospray ionization mass spectrometry is used to determine the changeof metabolites in citrus leaves, and the infected citrus nursery stocks at the early stage of the incubation period of tristeza virus can be identified prior to a routine detection method such as PCRbeing difficult to effectively detect the infected nursery stocks by combining a principal component analysis method. In a laboratory, samples are tested for 30 times, the detection result can reach 98%, during the field test, the samples are detected for 100 times, and the detection effect can reach 90%, so that the citrus tristeza virus can be effectively and rapidly detected.

Description

technical field [0001] The invention relates to a rapid detection method for citrus huanglongbing, belonging to the field of detection methods for plant diseases. Background technique [0002] Citrus huanglongbing is a devastating disease in citrus production, caused by bacteria that parasitize the phloem tissue of plants, including Phloembacterium asiatica, Phloembacterium africanica, and Phloembacterium americanum. After the plant is infected, the whole tree vigor is weakened, the growth is slow, the leaves are mottled and yellowed, the fruit is unevenly colored, and the symptoms of "red nose fruit" appear. -Withered within 8 years, seriously reducing the economic benefits of citrus. So far, there are no effective pesticides and resistant varieties against the disease, so early diagnosis of the disease is the basis and key to the prevention and control of citrus HLB. The real way to make a diagnosis is laboratory diagnosis, which requires section observation with the hel...

Claims

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Application Information

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IPC IPC(8): G01N27/68
Inventor 罗丽萍郝影宾陈焕文薛阿辉刘永泉崔萌郭夏丽吴栋
Owner NANCHANG UNIV
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