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Method for detecting sensitivity of phytophthora parasitica var nicotianae to bactericide

A technology of tobacco black shank and black shank bacteria, applied in the biological field, can solve the problems of not being able to reflect the growth dynamic characteristics of pathogenic bacteria, and achieve the effects of rapid detection, effective control, and increased growth rate

Inactive Publication Date: 2014-02-05
GUIZHOU TOBACCO SCI RES INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Gerd once used YBA culture medium to measure the comprehensive inhibitory effect of boscalid on spore germination and mycelium growth of Botrytis cinerea, but this method can only obtain the turbidity value of pathogen growth, and cannot reflect the specific dynamic characteristics of pathogen growth

Method used

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  • Method for detecting sensitivity of phytophthora parasitica var nicotianae to bactericide
  • Method for detecting sensitivity of phytophthora parasitica var nicotianae to bactericide
  • Method for detecting sensitivity of phytophthora parasitica var nicotianae to bactericide

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Embodiment 1

[0018] Utilize this method to measure the inhibitory effect of dimethomorph on the spore germination of 3 strains of tobacco black shank collected from the stems of flue-cured tobacco disease in Guizhou Province, comprising the following steps:

[0019] (1) Preparation of lima bean liquid medium: weigh 60g of lima beans and boil in 800mL sterile water for 1 hour, filter, dilute the filtrate to 1L, sterilize and set aside; Phenyl, the configuration mass concentration is the medicament of 100, 10, 1, 0.1, 0.05, 0.025, 0.013, 0.006mg / L;

[0020] (2) Cultivate tobacco black shank bacteria on a lima bean solid medium plate for 4 days, take 10 mycelium blocks with a diameter of 5 mm at 1 / 3 of the colony, put them into a petri dish filled with 20 mL of 10% V8 culture medium, and light After 4 days of culture, a large number of sporangia were formed on the surface of the mycelium; the black shank plate with sporangia formed was placed in a refrigerator at 4°C for 30 minutes, and then ...

Embodiment 2

[0029] Utilize this method to measure the inhibitory effect of dimethomorph on the hyphal growth of 3 strains of tobacco black shank on flue-cured tobacco collected from Guizhou Province, comprising the following steps:

[0030] (1) Preparation of lima bean liquid medium: weigh 60g of lima beans and boil in 800mL sterile water for 1 hour, filter, dilute the filtrate to 1L, sterilize and set aside; Phenyl, the configuration mass concentration is the medicament of 2, 1, 0.5, 0.25, 0.13, 0.063, 0.031mg / L;

[0031] (2) Cultivate tobacco black shank bacteria on a lima bean solid medium plate for 4 days, take 10 mycelium blocks with a diameter of 5 mm at 1 / 3 of the colony, put them into a petri dish filled with 20 mL of 10% V8 culture medium, and light After 4 days of culture, a large number of sporangia were formed on the surface of the mycelium; the black shank plate with sporangia formed was placed in a refrigerator at 4°C for 30 minutes, and then placed at room temperature for 2...

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Abstract

The invention discloses a method for detecting the sensitivity of phytophthora parasitica var nicotianae to a bactericide. The method comprises the following steps of diluting dimethomorph by using a lima bean liquid culture medium or an oat liquid culture medium to prepare agents with various mass concentrations; culturing the phytophthora parasitica var nicotianae on a lima bean solid culture medium plate, inducing by using a V8 liquid culture medium to obtain a zoospore suspension liquid, regulating the concentration of the zoospore suspension liquid to be 1+10<4> / mL; placing a 96-hole micropore plate in a monitoring culture box, culturing in the dark at a temperature of 28 DEG C for 72h, reading data of a phytophthora parasitica var nicotianae growth curve of 0-72h by adopting growth curve monitoring software; and when culturing for 0 and 72h, determining an OD value of the phytophthora parasitica var nicotianae in each pore of 590nm by adopting turbidity determination software, calculating an inhibition rate of phytophthora parasitica var nicotianae spore germination and mycelial growth of the bactericide according to a formula (1), and calculating an EC50 value of the bactericide to a strain. According to the method, the sensitivity of the phytophthora parasitica var nicotianae to the bactericide can be detected conveniently, simply, rapidly, efficiently and accurately.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting the susceptibility of tobacco blackleg bacteria to fungicides. [0002] Background technique [0003] Tobacco black shank ( Phytophthora parasitica var. nicotianae ) caused by tobacco black shank is one of the most serious diseases of tobacco in the world. It can infect multiple tissues such as tobacco roots, stems and leaves during the entire growth period of tobacco, usually leading to root necrosis and yellowing of leaves. , plant wilting, or even death. In the absence of preventive measures, the loss can reach 100%. At present, the prevention and treatment of tobacco black shank mainly relies on chemical pesticides, and the types of control agents are relatively single. The phenylamide fungicide metalaxyl has been used continuously on tobacco for many years. The problem of drug resistance is becoming more and more prominent, and the control effect is...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12R1/645
Inventor 汪汉成王茂胜陈庆园李文红陆宁夏海乾
Owner GUIZHOU TOBACCO SCI RES INST
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