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A method for detecting bacterial leaf spot of areca catechu

A technology for bacterial leaf spot and bacterial leaf spot, which is applied in the field of molecular biological technology to detect bacteria, can solve the problems of low accuracy and time-consuming, and achieve the effects of high accuracy, short time and high specificity

Inactive Publication Date: 2019-01-11
HAINAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the deficiencies in the prior art, the object of the present invention is to provide a method for detecting betel nut bacterial leaf spot, aiming to solve the problems of time-consuming and low accuracy of conventional detection methods

Method used

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  • A method for detecting bacterial leaf spot of areca catechu
  • A method for detecting bacterial leaf spot of areca catechu
  • A method for detecting bacterial leaf spot of areca catechu

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment one: detection of betel nut bacterial leaf spot

[0024] The flow chart of the method for detecting betel nut bacterial leaf spot is as follows figure 1 As shown, the specific detection steps include:

[0025] S101. Pretreat the tested sample and extract the DNA of the tested sample; the specific operation is to wash and dry the tested betel nut sample with clean water, cut the tissue block at the junction of disease and health, and then follow the bacterial genome DNA of TIANGEN company The instructions of the extraction kit (DP302) extract the DNA of the betel nut sample to be tested.

[0026] S102. According to multiple sequence alignments, design specific primers Ba-F / Ba-R, the sequences of which are as follows:

[0027] Ba-F: 5'-ACCTCGCGCTATAGGGGCG-3';

[0028] Ba-R: 5'-CATCGATGTTCCTCCGCATA-3'.

[0029] Use Ba-F and Ba-R primers for PCR reaction; PCR reaction is as follows:

[0030] 20 μL reaction system: 10 μl of 2×TransTaq-T PCR SuperMix, 1 μl of ...

Embodiment 2

[0033] Example 2: Specificity, sensitivity and field application analysis of the detection method

[0034] Specificity analysis of PCR detection:

[0035] Take Burkholderia andropogonis, and Xanthomonas campestris pv.arecae, Colletotrichum gloeoporioides, Ganoderna lucidum, betel nut bud rot DNA of Phythophthorameadii and Thielaviopsis paradoxa were used as samples, and sterile ddH 2 O is a negative control to test the specificity of primers Ba-F and Ba-R.

[0036] Result shows: detect the product of each PCR tube through 2% agarose gel electrophoresis, only template is that Burkholderia andropogonis genomic DNA amplifies the single band that size is 513bp, shows to be positive, and all the other have no band, shows to be negative (see figure 2 ), it can be seen that the specificity of the PCR detection is very high.

[0037] Field applications of PCR detection:

[0038]PCR was performed with upstream and downstream primers Ba-F and Ba-R, and 5 suspected samples were rand...

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Abstract

The invention discloses a method for detecting bacterial leaf spot of areca catechu. The method comprises the steps of: preprocessing tested samples and extracting the DNA of the samples; Using specific primer Ba-F / Ba-R for PCR reaction; Electrophoretic detection. The method for detecting bacterial leaf spot of areca catechu overcomes the shortcomings of the conventional detection method, has theadvantages of high specificity, stable reaction, high accuracy, easy operation, short use time, and only 4-6h.

Description

technical field [0001] The invention belongs to the field of molecular biological technology detection of bacteria, in particular to a method for detection of betel nut bacterial leaf spot. Background technique [0002] Areca cathecu L., Palmaceae areca is a perennial evergreen tree with erect stems, arbor-like, more than 10 meters high, up to 30 meters high, native to the tropical Pacific region, Southeast Asia, South Asia and parts of East Africa. One of the important southern medicine resources. The main production areas of betel nuts in China are Hainan and Taiwan, among which the total output of areca nuts in Hainan ranks second in the world, and the unit output also ranks second in the world, reaching 3336kg / hm 2 . Betel nut is the largest economic crop in Hainan, and Hainan farmers planting betel nut is one of the ways to get rid of poverty and become rich. [0003] However, with the continuous expansion of the betel nut planting area, new diseases are also constan...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/04C12N15/11C12R1/01
CPCC12Q1/686C12Q1/6895C12Q2565/125
Inventor 施海涛韦运谢刘国银方娇
Owner HAINAN UNIVERSITY
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