Detection method of sweet potato black rot pathogen

A black spot fungus and detection method technology, applied in the field of bioengineering, can solve the problems of time-consuming and labor-intensive, incapable of rapid disease detection, and achieve the effect of preventing misdiagnosis and preventing spread

Active Publication Date: 2014-03-26
INST OF PLANT PROTECTION HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods cannot be used for early diagnosis of diseases, and the observation of the morphology of pathogens requires t

Method used

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  • Detection method of sweet potato black rot pathogen
  • Detection method of sweet potato black rot pathogen
  • Detection method of sweet potato black rot pathogen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Specific Amplification Experiment of Sweet Potato Black Spot Bacteria Detection Method

[0035] 1. Materials and methods

[0036] (1) Source of primers: According to GenBank, A. Ceratocystis fimbriata )gene sequence

[0037] (GenBank accession number: EF017220.1~EF017226.1) Design primers, the primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., and the size of the amplified target fragment was 392bp.

[0038] (2) Germ materials: Sweet potato black spot fungus, sweet potato soft rot fungus, sweet potato dry rot fungus (collected from the Plant Protection Institute of Henan Academy of Agricultural Sciences in May 2012, collected by Zhang Desheng), sweet potato black spot fungus (October 2011 month, from the Institute of Plant Protection, Hebei Academy of Agriculture and Forestry Sciences, provided by Chen Shulong).

[0039] (3) Nucleic acid extraction: use the total DNA extraction kit of Treasure Bioengineering (Dalian) Co., Ltd., and fo...

Embodiment 2

[0045] Example 2 Amplification experiment of different samples by detection method of sweet potato black spot bacterium

[0046] 1. Materials and methods

[0047] (1) Source of primers: According to GenBank, A. Ceratocystis fimbriata ) gene sequence (GenBank accession number: EF017220.1 ~ EF017226.1) to design primers, which were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. The size of the amplified target fragment is 392bp.

[0048] (2) Disease-like materials: air-dried mycelium of sweet potato black spot fungus, stems of various potato seedlings infected with black spot disease (collected from the Plant Protection Institute of Henan Academy of Agricultural Sciences in May 2012, collected by Zhang Desheng).

[0049] (3) Nucleic acid extraction: use the total DNA extraction kit of Treasure Bioengineering (Dalian) Co., Ltd., and operate according to the kit instructions to extract the total DNA of the sample to be tested.

[0050](4) PCR amplification reaction ...

Embodiment 3

[0055] Example 3 The detection method of sweet potato black spot fungus to the detection results of field diseased seedlings

[0056] 1. Materials and methods

[0057] (1) Source of primers: According to GenBank, A. Ceratocystis fimbriata ) gene sequence (GenBank accession number: EF017220.1 ~ EF017226.1) to design primers, which were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. The size of the amplified target fragment is 392bp.

[0058] (2) Disease-like materials: Potato seedlings and potato cubes (collected from the Nursery of Luoyang Academy of Agricultural Sciences, Henan Province in May 2012, collected by Zhang Desheng), the samples showed typical black spot symptoms.

[0059] (3) Nucleic acid extraction: use the total DNA extraction kit of Treasure Bioengineering (Dalian) Co., Ltd., and follow the instructions of the kit to extract the total DNA of the sample.

[0060] (4) PCR amplification reaction system 25 μL, composed of: 2.5 μL 10×PCR buffer, 0.5 μ...

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Abstract

The invention relates to a detection method of sweet potato black rot pathogen, and belongs to the technical field of bioengineering. The method specifically comprises steps as follows: a pair of sweet potato black rot pathogen specific primers SPCPF and SPCPR is designed and synthesized; total DNA (deoxyribonucleic acid) of a to-be-tested sample is extracted and taken as a template, and the primers SPCPF and SPCPR are used for performing PCR (polymerase chain reaction) amplification; and an amplification product is detected through agarose gel electrophoresis. The method is simple, convenient, fast and sensitive, not only can be used for detecting pathogen mycelia but also can be used for detecting sick potato seedlings and pieces; the detection method can be used for performing early detection on sweet potato black rot and further can be used for effectively distinguishing similar diseases, such as a black nevus disease, soft rot and dry rot, of sweet potatoes; and the method has significance on early warning and propagation prevention and control of the sweet potato black rot pathogen.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and relates to a detection method for pathogens, in particular to a detection method for sweet potato black spot pathogen. Background technique [0002] Sweet potato black spot disease is caused by the fungus C. Ceratocystis fimbriata Ellis and Halsted) is one of the three major diseases that seriously endanger sweet potato production. The annual yield loss due to this disease is generally 5% to 10%, and the loss caused by the disease is as high as 20% to 50%. or even higher. In addition, diseased potatoes can also produce toxic substances such as ipomeama rone and ipomeanine, which can cause poisoning and even death to humans and livestock after eating. The genus Ascomycota, Sclerotinia, Spheroidales, Sclerosaceae, Sclerosaceae. Infection of sweet potato mainly invades from wounds, and can also invade from bud eyes, lenticels, and root eyes on potato pieces. [0003] Sweet potato blac...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/645
CPCC12Q1/04C12Q1/686C12Q2531/113
Inventor 张德胜张振臣王爽秦艳红乔奇田雨婷王永江
Owner INST OF PLANT PROTECTION HENAN ACAD OF AGRI SCI
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