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LAMP primer group for rapidly detecting phytophthora. medicaginis at normal temperature

A root rot fungus and primer set technology, applied in the field of plant protection, to achieve the effect of reducing investment and simple detection instruments

Inactive Publication Date: 2011-05-11
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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Problems solved by technology

[0003] In 2007, my country issued and implemented the "List of Imported Plant Quarantine Pests of the People's Republic of China", which increased the number of quarantine pests from 84 to 435, and Phytophthora alfalfa root rot is one of the newly added diseases. So far, the quarantine of Phytophthora alfalfa root rot bacteria in various ports is still in a state of no basis, so it is necessary to establish detection methods and standards as soon as possible to prevent this dangerous pathogen from being introduced into my country

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  • LAMP primer group for rapidly detecting phytophthora. medicaginis at normal temperature
  • LAMP primer group for rapidly detecting phytophthora. medicaginis at normal temperature

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Embodiment 1

[0057] Embodiment 1, Phytophthora alfalfa LAMP primer detection method

[0058] DNA extraction: About 50 mg of mycelium, DNA was extracted with Qiagen Dneasy Plant Mini Kit kit.

[0059] Reaction system: the total volume of each reaction mixture is 25 μl

[0060] 10*ThermoPol Buffer 2.5μl

[0061] 10mM DNTP mix 1μl

[0062] 5M Betaine 1μl

[0063] 10 μM FIP 2 μl

[0064] 10 μl MBIP 2 μl

[0065] 10 μM F3 0.5 μl

[0066] 10 μM MB3 0.5 μl

[0067] 10 μL LF 1 μl

[0068] 10 μM LB 1 μl

[0069] 8U / μl Bst enzyme 1μl

[0070] DDH2O 12μl

[0071] Template DNA 0.5μl

[0072] 25μl

[0073] Reaction procedure: react at 65°C for 1h.

[0074] Result: No. 2 swimming lane is Phytophthora alfalfa root rot bacteria, and there is a ladder-like specific band, and all the other swimming lanes do not have a ladder-like band except the genomic DNA band. The results show that the specificity of the primer pair of the present invention is strong. Other fungi of the same genu...

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Abstract

The invention relates to an amplifying nucleic acid primer for rapidly detecting phytophthora. medicaginis at normal temperature. The LAMP amplifying nucleic acid primer comprises an FIP primer, an F3 primer, a BIP primer, a B3 primer, an LF primer and an LB primer. The LAMP detecting system of the invention has favorable specificity and can rapidly, conveniently and efficiently detect phytophthora. medicaginis under the condition of 63 DEG C and meet the rapid detecting requirement of a port on the phytophthora. medicaginis.

Description

technical field [0001] The invention relates to the field of plant protection, and provides a normal-temperature rapid detection method for Phytophthora alfalfa root rot, which is suitable for application in port inspection and quarantine institutions. Background technique [0002] Alfalfa is a perennial herbaceous plant of the leguminous family and the genus Alfalfa. It is the most planted forage species in the world. Because of its wide adaptability, high yield, and good quality, it is known as the "king of forage grass". Alfalfa root rot is one of the main diseases that cause the loss of alfalfa yield. Fungi, Pythium, Pythium, and Phytophthora can parasitize alfalfa, chickpea, kiwi fruit, carrots and other important economic crops, among which alfalfa and chickpea are the main hosts. Phytophthora alfalfa root rot is distributed in all continents, including: Japan, Pakistan, Turkey, Greece, Canada, the United States, Argentina, Australia, South Africa, etc. At present, ther...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张裕君崔铁军黄国明廖芳刘鹏王金成
Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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