Method for detecting plasmodiophora brassicae in Chinese cabbages

A technology for Brassica rhizogenes and Chinese cabbage, which is applied in the field of microbial detection and achieves the effects of high repeatability, convenient operation and accurate detection.

Inactive Publication Date: 2013-05-22
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no relevant reports on the application of FT-IR to the detection of Plasmodium brassicae.

Method used

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  • Method for detecting plasmodiophora brassicae in Chinese cabbages
  • Method for detecting plasmodiophora brassicae in Chinese cabbages
  • Method for detecting plasmodiophora brassicae in Chinese cabbages

Examples

Experimental program
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Effect test

Embodiment 1

[0046] The preparation of embodiment 1 sample

[0047] Chinese cabbage samples were collected from Xinmin City under Shenyang City, Liaoning Province, China.

[0048] Chinese cabbage samples: roots of Chinese cabbage infected with Plasmodium brassicae;

[0049] Roots of healthy Chinese cabbage (means not infected with Plasmodium brassicae);

[0050] Leaves of Chinese cabbage infected with Plasmodium brassicae;

[0051] Leaves of healthy Chinese cabbage (means not infected with Plasmodium brassicae).

[0052] The basic identification and judgment of Chinese cabbage infected by Plasmodium brassicae is based on conventional symptom observation and the application of specific PCR experiment technology (R.Faggian S.Parsons.Arapid diagnostic test for clubroot[J].Horticulture Australia).

[0053] (1) Freeze-dry the Chinese cabbage sample at -70°C, put it in a mortar, and grind it into a powder with a particle size of less than 2 μm with a pestle;

[0054] (2) The potassium bromid...

Embodiment 2

[0056] Example 2 Collection and Data Analysis of Fourier Transform Infrared Spectroscopy (FTIR)

[0057] Scan the compressed tablet prepared in Example 1 with a Fourier transform infrared spectrometer to obtain the infrared spectrum of the Chinese cabbage sample, and compare and analyze the infrared spectrum of the Chinese cabbage sample infected by Plasmodium brassicae and the healthy Chinese cabbage sample. During the scanning process, dry air is continuously fed into the sample area in the Fourier transform infrared spectrometer to drive off water vapor.

[0058] The scanning conditions of the infrared spectrum are as follows:

[0059] Temperature: room temperature (25°C);

[0060] Scanning range: 4000~500cm -1 ;

[0061] Resolution: 4cm -1 ;

[0062] Number of scans: 100 times.

[0063] The analysis of infrared spectral data was obtained after analysis by Grams32 software (Galactic Industries, Salem, NH, USA), and the analytical spectral range was 4000~500cm -1 segm...

Embodiment 3

[0076] Example 3 Using polymerase chain reaction (PCR) and Fourier transform infrared spectroscopy (FTIR) to detect Plasmodium brassicae in Chinese cabbage

[0077] Root samples (D1) known to be infected with P. brassicae (P. Ltd) extracted Chinese cabbage tissue genomic DNA according to the method provided in the instructions, carried out PCR amplification with specific primers of Plasmodium brassicae, and analyzed the amplified products by 1% agarose gel electrophoresis.

[0078] Specific primers are:

[0079] NS7 (upstream primer): 5'-GAGGCAATAACAGGTCTGTGATGC-3';

[0080] Pb3R (downstream primer): 5'-ACCATACCCAGGGCGATTG-3'.

[0081] The PCR amplification system is shown in the table below.

[0082] Element

[0083] Reaction conditions:

[0084] Pre-denaturation at 95°C for 4min; denaturation at 95°C for 30s, annealing at 55°C for 30s, extension at 72°C for 30s, a total of 30 cycles; extension at 72°C for 10min.

[0085] Depend on Figure 5 It can be seen th...

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Abstract

The invention discloses a method for detecting plasmodiophora brassicae in Chinese cabbages. The method comprises the following steps of: making roots and leaves of the Chinese cabbages into tablets; acquiring infrared spectrograms of the tablets; and analyzing the infrared spectrograms, wherein if the tablets made from the roots have a characteristic peak in at least one part in the ranges of 1748.16+/-0.75cm<-1>, 2851.24+/-0.05cm<-1> and 3013.23+/-0.28cm<-1>, the roots of the Chinese cabbages are infected with the plasmodiophora brassicae; and if the tablets made from the leaves have a characteristic peak in at least one part in the ranges of 1252.54+/-0.50cm<-1> and 1051.98+/-4.23cm<-1>, the leaves of the Chinese cabbages are infected with the plasmodiophora brassicae. According to the method, the plasmodiophora brassicae in the Chinese cabbages can be rapidly and accurately detected in a lossless manner by virtue of a Fourier transform infrared spectroscopy technique, and the detection is convenient to implement, does not depend on skills of people and is high in security and repeatability.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, in particular to a method for detecting Plasmodium brassicae in Chinese cabbage. Background technique [0002] The Food and Agriculture Organization of the United Nations positions Chinese cabbage as one of the twenty most important vegetables and is an important source of food. [0003] Plasmodiophora brassicae is an obligate parasite in the flagellate subphylum (Mastigomycotina), which can cause diseases of cruciferous plants such as rapeseed, green vegetables, mustard greens, mustard mustard, etc. Bacteria can also infect Chinese cabbage and cause clubroot of Chinese cabbage. [0004] Clubroot is an important soil-borne disease of cruciferous crops, which can form tumors of different sizes and shapes on the main and lateral roots of plants. After the root is damaged, it can affect the growth of the aboveground part of the plant, making the leaf color light, growth retardation...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/35G01N21/3563
Inventor 李斌单长林黄洁琼葛梦禹石雨谢关林
Owner ZHEJIANG UNIV
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