Method and applications for fast detecting pathogenic bacteria molecules of bacterial soft rotting disease for banana

A bacterial and pathogenic technology, applied in the fields of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem that it is difficult for banana farmers to accurately judge the pathogen, effective methods, and strong destructiveness.

Inactive Publication Date: 2011-08-17
PLANT PROTECTION RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease has the characteristics of rapid spread and strong destructiveness, causing huge economic losses to banana production
[0003] The initial symptoms of banana bacterial soft rot are somewhat similar to those of banana wilt, so it is difficult for banana farmers to accurately determine the pathogen. Th...

Method used

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  • Method and applications for fast detecting pathogenic bacteria molecules of bacterial soft rotting disease for banana
  • Method and applications for fast detecting pathogenic bacteria molecules of bacterial soft rotting disease for banana
  • Method and applications for fast detecting pathogenic bacteria molecules of bacterial soft rotting disease for banana

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Experimental program
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Effect test

Embodiment 1

[0025] PCR technology detects the method for banana bacterial soft rot, and its steps are:

[0026] (1) Bacterial genomic DNA extraction: three strains of frozen banana bacterial soft rot pathogens (First report of a sort of banana in Mainland China Caused by a Dickeya sp. (Pectobacterium chrysanthemi). Plant Dis. 2010, 94 (5 ): 640-640.) were inoculated into NA slant medium (beef extract 3g, yeast extract 1g, peptone 5g, glucose 10g, agar 20g, distilled water to 1000ml) and cultured at 35°C for 48h. Pick a single colony and inoculate it in liquid NA medium, culture overnight at 27-35°C on a shaker at 180 rpm, and extract the genome of the pathogenic bacteria of banana bacterial soft rot with a bacterial genome DNA extraction kit (Beijing Kangwei Century Biotechnology Co., Ltd.) DNA is ready for use.

[0027] (2) Amplify the 16S-23S rDNA ITS sequence of banana bacterial soft rot: use the reported bacterial universal ITS primers (primer1: 5'-GAAGTCGTAACAAGG-3'; primer2: 5'-CAA...

Embodiment 2

[0042] (1) Sample collection and isolation and identification of pathogenic bacteria

[0043] September 2010, Panyu District and Nansha District, Guangzhou City, Guangdong Province, Dongguan City, Zhongshan City, Zhuhai City, Gaozhou City, Huazhou City, Donghai Island and Xuwen County of Zhanjiang City, Nanning City of Guangxi Province, Zhangzhou City of Fujian Province, Chengcheng City of Hainan Province In Mai County, Lingao County, Ledong County, Dongfang City and other banana planting areas (as shown in Table 1), samples of diseased plants were collected, soil and water samples from diseased areas were brought back and pathogenic bacteria were isolated.

[0044] (2) Detection of banana bacterial soft rot:

[0045] ① Sample treatment: Genomic DNA of pseudostem tissue of diseased plants was extracted by CTAB method; each 0.5g soil sample was directly soaked in 1ml sterile water for half an hour, centrifuged at 5000rpm for 5min, and the supernatant was taken for later use; wa...

Embodiment 3

[0068] The present invention carries out the above detection on banana tissue and soil samples not infected with banana bacterial soft rot, and no specific band amplification is found at the molecular weight of 171bp.

[0069] As can be known from the above results, use the primer LF / LR of the ITS sequence-specific region of banana bacterial soft rot to detect the banana plant tissue and soil samples tested, if the 171bp fragment can be amplified specifically, It can be determined that banana bacterial soft rot exists in banana tissue or soil samples.

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Abstract

The invention discloses a method and applications for fast detecting pathogenic bacteria molecules of bacterial soft rotting disease for banana. The method comprises the steps that: a specific primer is utilized to conduct PCR (polymerase chain reaction) on DNA contained in a sample, and the sample contains the pathogenic bacteria molecules of bacterial soft rotting disease for banana when the PCR product presents a 171bp condition. The method is simple to operate, takes less time and is large in flux. The method is used for conducting determined detection to banana seedlings, soil leach liquor of soil in a planting area, irrigation water and the like, thus realizing the purposes of construction and safety production in no disease banana seedling production base in China and simultaneously stopping of introduction of dangerous foreign disease banana.

Description

technical field [0001] The invention belongs to the fields of crop disease prevention and plant quarantine, and in particular relates to a method and application for rapid detection of pathogenic bacteria of banana bacterial soft rot disease, that is, using polymerase chain reaction (PCR) molecular biology technology to detect banana bacterial soft rot The high-sensitivity and rapid detection of disease pathogens can be used for banana seedling quarantine, early diagnosis of banana soft rot in the field, and monitoring and identification of pathogens. Background technique [0002] The pathogen of banana bacterial soft rot is Pectobacterium chrysanthemi, which belongs to the genus Erwinia of the family Enterobacteriaceae. The pathogenic bacteria is highly pathogenic and may be a highly pathogenic subspecies. This is a new dangerous disease that has invaded my country. It was first discovered in September 2009 in the banana planting area of ​​Panyu, Guangzhou. The disease is...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 林壁润李培谦沈会芳蒲小明潘群英
Owner PLANT PROTECTION RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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