Gene chip for detecting pathogenic fungi of poplar canker and application thereof

A technology for poplar canker and pathogenic fungi, which is applied in the field of gene chips and can solve problems such as population limitation and time-consuming technical regulations

Inactive Publication Date: 2014-11-05
严东辉
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the application of these technologies has greatly promoted the development of microbial ecology and greatly improved the ability to determine and analyze microbial populations,

Method used

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  • Gene chip for detecting pathogenic fungi of poplar canker and application thereof
  • Gene chip for detecting pathogenic fungi of poplar canker and application thereof
  • Gene chip for detecting pathogenic fungi of poplar canker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0075] Example 1 Construction of gene chip

[0076] A gene chip for detecting the pathogenic fungi of poplar canker disease, comprising a carrier, an oligonucleotide probe located on the carrier, a positive control probe for hybridization process and a blank dot coating; the oligonucleotide probe includes 28 fungal probes, the sequences of which are shown in SEQ ID NOs. 2, 3, 8 to 33; the positive control for the hybridization process is Biotin.

[0077] The carrier is a 1.5ml Eppendorf centrifuge tube, and the chip is dotted on the inner surface of the bottom of the cross section (ArrayTube chip).

[0078] The oligonucleotide probe, the positive control probe for the hybridization process and the blank dot coating are distributed on the carrier in an array.

[0079] The array type distribution layout on the carrier is as follows figure 1 The layout shown [Blank grids and shaded grids in the figure indicate each dot matrix arrangement. The dot matrix in the shaded area is used to indi...

Example Embodiment

[0081] Example 2 Evaluation of the effectiveness of gene chips

[0082] (1) Materials and methods of evaluation research

[0083] (1) Materials: The target strains used for the specific test strains include the target strains and related ascomycetes Ascochyta sp. Basidiomycetes Agaricus bisporus, Zygobacterium Cunninghamella bertholletiae, Oomycetes Phytophthora cactorum, see Table 2 for details.

[0084] Table 2 Test strains and sources

[0085]

[0086] (2) Instruments and equipment

[0087] All-round desktop high-speed refrigerated centrifuge ( Stratos), PCR machine (Biometra), ultra-fine nucleic acid protein analyzer (Nanodrop ND-1000), molecular hybridization constant temperature oscillator (Eppendorf Thermomixer comfort), HDL ultra-clean workbench, high-pressure steam sterilizer, constant temperature shaker, condensation Gel imaging system (Alpha imager EP), DYY-4 stabilized and steady flow electrophoresis instrument (Beijing Liuyi Instrument Factory), Eppendorf pipette, microwa...

Example Embodiment

[0117] Example 3 Multiplex PCR gene chip detection of poplar canker in Beijing area

[0118] (1) Materials and methods

[0119] (1) Material

[0120] The sampling location was poplar in Beijing area, sampling according to different areas, including Haidian District, Chaoyang District, Changping District, Yanqing District, Huairou District, Miyun District, Pinggu District, Shunyi District, Tongzhou District, Daxing District, Fangshan District, Mentougou District. Choose 3-8 points in each area to collect samples. The distribution of collection areas and the status of collection locations such as Figure 13 . One spot was selected for each region, susceptible and asymptomatic samples, a total of 24 samples were used for hybridization. Sample collection locations for chip hybridization environment, host and habitat conditions are shown in Table 4;

[0121] Table 4 Collection location, host and habitat of microarray hybridization samples

[0122]

[0123] (2) Method

[0124] DNA extract...

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Abstract

The invention discloses a gene chip for detecting pathogenic fungi of poplar canker. The gene chip comprises a carrier, and an oligonucleotide probe, a hybridization process positive control and a blank dot coating which are all located on the carrier, wherein the hybridization process positive control is Biotin; the oligonucleotide probe comprises 28 fungus probes which are as shown in SEQ ID NO. 2, 3, 8-33; or the oligonucleotide probe comprises 38 fungus probes which are as shown in SEQ ID NO. 1-35. The gene chip is applicable to detecting the pathogenic fungi of the poplar canker, covering 16 types under 5 categories. The technology for detecting and diagnosing the poplar canker by use of the chip flux of multiple PCR (Polymerase Chain Reaction) amplification environment samples is primarily applied to the detection of the poplar canker in the Beijing area. The gene chip for detecting the pathogenic fungi of the poplar canker is capable of serving for the establishment of the high-flux detection technology for other pests, and the application and study on the basis of related biological diversity.

Description

technical field [0001] The invention relates to a gene chip for detecting pathogenic fungi of poplar canker and an application thereof, belonging to the technical field of gene chips. Background technique [0002] There are various types of tree canker pathogens and a wide range of hosts. There is not a one-to-one correspondence between the sexual and anamorphic types. The anamorphic types are diverse, and their morphological characteristics have little difference. The sexual types are not often found in nature. The current detection and identification methods still rely on the isolation and culture of pathogenic bacteria, which limits the research on the occurrence, development and control of tree canker under ecological conditions. [0003] The identification and detection methods of pathogenic bacteria in the prior art mainly stay on cultivable technology, morphological biology, some biochemical techniques and conventional molecular methods. These technologies and methods...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C40B40/06C12R1/645
CPCC12Q1/6895
Inventor 严东辉张星耀赵文霞冯小慧李永贾秀贞
Owner 严东辉
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