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Separation method of tobacco root black rot bacteria

A black rot fungus and isolation method technology, applied in the field of germ isolation, can solve the problems of complicated operation process, easy to be covered by other fungi, slow growth rate, etc., and achieve the effect of safe test environment, saving test cost, less consumables and reagents

Inactive Publication Date: 2018-08-10
HENAN UNIV OF SCI & TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Petri dishes are required for disinfection with alcohol and sodium hypochlorite, and for cleaning with sterile water. The petri dishes need to be replaced every time they are disinfected and cleaned. One disease strain needs 5 pairs of petri dishes to complete the separation. The operation process is complicated, and More consumables and reagents are required, and the cost is higher
[0005] Moreover, since the tobacco root black rot fungus is a facultative parasitic fungus, and its growth rate is slower than other types of fungi such as Pythium, it is very easy to be covered by other fungi when isolated by conventional diseased tissue isolation methods, and it is difficult to obtain pure culture. pathogen

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A method for isolating tobacco root black rot fungus, the operation steps are:

[0030] Step 1: Collection of test samples Select the diseased plants with typical symptoms of tobacco root black rot and pull them out, gently peel off the soil above 2 mm from the root surface with a scalpel, shake off the rest of the soil and collect it as a rhizosphere soil sample, do not shake The fallen soil adhered to the roots is gently brushed off with a small brush, air-dried indoors, put into paper bags, numbered, and stored for later use.

[0031] Step 2: Induction of carrot discs The rhizosphere soil samples were crushed in a mortar and poured into sterilized Petri dishes. Disinfect the surface of fresh carrots with 75% alcohol, cut them into discs about 5 mm thick with a sterilized knife, dip the ground samples evenly, and place them on a Petri dish covered with absorbent cotton soaked in distilled water and filter paper cultured in a constant temperature incubator at 25 ℃, ob...

Embodiment 2

[0036] A method for isolating tobacco root black rot fungus, the operation steps are:

[0037] Step 1: Collection of test samples Select the diseased plants with typical symptoms of tobacco root black rot and pull them out, gently peel off the soil above 2 mm from the root surface with a scalpel, shake off the rest of the soil and collect it as a rhizosphere soil sample, do not shake The fallen soil adhered to the roots is gently brushed off with a small brush, air-dried indoors, put into paper bags, numbered, and stored for later use.

[0038] Step 2: Induction of carrot discs The rhizosphere soil samples were crushed in a mortar and poured into sterilized Petri dishes. Disinfect the surface of fresh carrots with 75% alcohol, cut them into discs about 6 mm thick with a sterilized knife, dip the ground samples evenly, and place them on a Petri dish covered with absorbent cotton soaked in distilled water and filter paper cultured in a constant temperature incubator at 25 ℃, du...

Embodiment 3

[0043] A method for isolating tobacco root black rot fungus, the operation steps are:

[0044] Step 1: Collection of test samples Select the diseased plants with typical symptoms of tobacco root black rot and pull them out, gently peel off the soil above 2 mm from the root surface with a scalpel, shake off the rest of the soil and collect it as a rhizosphere soil sample, do not shake The fallen soil adhered to the roots is gently brushed off with a small brush, air-dried indoors, put into paper bags, numbered, and stored for later use.

[0045] Step 2: Induction of carrot discs The rhizosphere soil samples were crushed in a mortar and poured into sterilized Petri dishes. Disinfect the surface of fresh carrots with 75% alcohol, cut them into discs about 4mm thick with a sterilized knife, dip the ground samples evenly, and place them in a Petri dish covered with absorbent cotton soaked in distilled water and filter paper , cultured in a constant temperature incubator at 25 °C, ...

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Abstract

The invention discloses a separation method of tobacco root black rot bacteria. The method includes the steps: acquiring rhizosphere soil of tobacco plants with tobacco root black rot symptoms; dryingand grinding the acquired rhizosphere soil; adhering the grinded rhizosphere soil onto carrot slices; placing the carrot slices adhered to the soil into a culture vessel to culture the carrot slicesfor 2-4 days; flushing the carrot slices by the aid of sterile water with 50 micro-liters / milliliters of penicillin and 50 micro-liters / milliliters streptomycin sulfate; continuing to culture the flushed carrot slices for 2-4 days; selecting mildew layers with tobacco root black rot typical symptoms on the carrot slices; purifying, culturing and storing the mildew layers. Growth of the tobacco root black rot bacteria is induced to restrain other bacteria from growing by the aid of different effects of components of carrot slices on the tobacco root black rot bacteria and the other fungi, and the separation method solves the problem that the other fungi easily covers the tobacco root black rot bacteria. The surfaces of treated carrot slices can rapidly grow hyphae, typical conidiophore stalks, conidiophores and chlamydospores, and the success rate reaches 96% or more.

Description

technical field [0001] The invention relates to a method for isolating pathogenic bacteria, in particular to a method for isolating tobacco root black rot pathogen. Background technique [0002] The isolation, purification and cultivation of plant pathogenic fungi are not only the basis for theoretical research such as morphological observation, pathogenicity determination, physiological ecology and molecular biology of pathogenic fungi, but also the identification and screening of disease resistance of varieties, pathogen resistance (anti- ) and other applied research foundations. The fungal mycelium in the diseased plant tissue can generally resume growth and reproduction if given suitable environmental conditions. However, under natural conditions, pathogenic bacteria are usually mixed with other miscellaneous bacteria, and pathogenic bacteria are isolated from diseased tissues or other substrates, which is called the isolation of plant pathogenic fungi. The isolation o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N1/02C12R1/645
CPCC12N1/02C12N1/14
Inventor 康业斌李淑君赵世民康蕾叶红朝周俊学董昆乐孔德辉王玉洁马君红李成军王海涛陈玉国白静科丘睿
Owner HENAN UNIV OF SCI & TECH
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