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Method for detection of fungi belonging to the genus Geosmithia

A detection method and technology for fungi are applied in the detection field of fungi belonging to the genus Geosmithia, and can solve the problems of difficulty in specific and rapid detection, and the lack of elucidation of specific gene regions.

Inactive Publication Date: 2011-12-07
KAO CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above method does not elucidate specific gene regions of fungi belonging to the genus Geosmithia
Therefore, there is a problem that it is difficult to specifically and rapidly detect fungi belonging to the genus Geosmithia

Method used

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  • Method for detection of fungi belonging to the genus Geosmithia
  • Method for detection of fungi belonging to the genus Geosmithia
  • Method for detection of fungi belonging to the genus Geosmithia

Examples

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Effect test

Embodiment 1

[0076] Example 1 Analysis of specific nucleotide sequences of fungi belonging to the genus Geosmithia

[0077] The base sequences of various β-tubulin genes of the genus Geosmithia were determined by the following method. Use GenTorukun TM (manufactured by Takara Bio Co., Ltd.), DNA was extracted from cells of the genus Geosmithia cultured on a potato dextrose agar slant medium at 25° C. in the dark for 7 days. For PCR amplification of the target site, PuRe TaqTM Ready-To-GoPCR Beads (manufactured by GE Health Care UK LTD) were used, and Bt2a (5'-GGTAACCAAATCGGTGCTGCTTTC-3': SEQ ID NO: 6), Bt2b (5'-ACCCTCAGTGTAGTGACCCTTGGC -3': sequence number 7) (Glass and Donaldson, Appl Environ Microbiol 61: 1323-1330, 1995). The amplification conditions are as follows: denaturation temperature is 95°C, annealing temperature is 59°C, extension temperature is 72°C, and 35 cycles are implemented. The PCR product was purified using Auto SegTM G-50 (manufactured by Amersham Pharmacia Biotech...

Embodiment 2

[0079] Example 2 Detection of fungi belonging to the genus Geosmithia by the PCR method

[0080] (1) Preparation of primers

[0081]Among the specific nucleotide sequence regions (base sequence numbers 1 to 3) obtained above for fungi belonging to the genus Geosmithia, starting from a particularly high specificity region belonging to fungi belonging to the genus Geosmithia at the 3' end side, the following conditions are met: Partial regions of 4 conditions were studied: 1) the base sequence of the inherent gene is about 10 bases continuous, 2) the GC content of the oligonucleotide is about 30% to 80%, 3) the oligonucleotide The possibility of acid self-annealing is low. 4) The Tm value of the oligonucleotide is about 55-65°C. Based on the base sequence of this part of the region, 5 sets of primer pairs were designed, and the DNA extracted from various bacteria was used as a template to study the effectiveness of Geosmithia detection by PCR reaction. That is, studies were co...

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Abstract

A method for detecting a bacterium belonging to the genus Geosmithia, which comprises identifying the bacterium with a nucleic acid represented by the following nucleotide sequence (a) or (b): (a) a partial nucleotide sequence of a ss-tubulin gene depicted in any one sequence selected from SEQ ID NO:1 to SEQ ID NO:3, or a sequence complementary to the partial nucleotide sequence; or (b) a nucleotide sequence produced by deleting, substituting, inserting or adding one or several nucleotides in a nucleotide sequence depicted in any one sequence selected from SEQ ID NO:1 to SEQ ID NO:3, or a sequence complementary to the nucleotide sequence.

Description

technical field [0001] The present invention relates to a method for detecting fungi belonging to the genus Geosmithia. Background technique [0002] Fungi belonging to the genus Geosmithia are widely distributed in nature, multiply on crops such as vegetables and fruits, and contaminate foods and beverages made from these crops. However, since fungi belonging to the genus Geosmithia form chlamydospores with high heat resistance, even if the usual sterilization treatment process for other fungi is performed, such as heat sterilization of acidic beverages, etc., fungi belonging to the genus Geosmithia Fungi can also survive and reproduce, thus becoming the cause of mildew. Therefore, fungi belonging to the genus Geosmithia are being alerted as important harmful fungi that cause serious quality accidents. In order to prevent accidents caused by fungi belonging to the genus Geosmithia in foods and beverages and their raw materials, detection and identification of fungi belong...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12Q1/68
CPCC12Q1/6895
Inventor 细谷幸一中山素一德田一矢口贵志弘佑介
Owner KAO CORP
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