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Alternaria alternate spore suspension liquid and application thereof

A technology of tobacco red spot bacteria and spore suspension, which is applied in the field of agricultural microorganisms, can solve the problems of difficult control of ultraviolet dose and time, low mycelial sporulation efficiency, and culture pollution, so as to achieve shortened culture period, large inoculum amount, and guaranteed The effect of cultivating quality

Active Publication Date: 2013-07-10
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, in the laboratory, the mycelium is generally cultured on a plate, and the spore suspension of Alternaria tabacum is prepared after natural spore production, but the mycelium usually takes more than 10 days to grow on the plate, and the mycelium is mature and the amount of spore production is sufficient to prepare The spore suspension needs to be further cultivated for 5-10 days. Due to its long cultivation time, it is easy to cause the medium to dry and the culture to be polluted, and the sporulation efficiency is low without induction of mycelia.
Although some studies have shown that sporulation can be accelerated through ultraviolet light induction, it is difficult to control the dose and time of ultraviolet light, which is easy to cause variation and contamination of cultures

Method used

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  • Alternaria alternate spore suspension liquid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0019] The medium described in step B is oat medium (OA), potato dextrose medium (PDA), potato sucrose medium (PSA), Czapek’s medium or Richard’s medium.

[0020] The solution for promoting spore germination described in step D is one or more combinations of 1-2% sterilized glucose, fructose, sucrose, and peptone solutions. Alternatively, the spore germination-promoting solution described in step D is 2-5% fresh mature tobacco leaf tissue mash liquid of susceptible varieties of tobacco, and the tissue mash liquid is sterilized by filtration. The susceptible varieties of tobacco are K326, Yunyan 85 or G140.

[0021] The tissue pounding described in step D adopts a mortar, pounder or grinder.

[0022] The filtering described in step D is filtering with absorbent cotton or 4-6 layers of absorbent gauze.

[0023] The application of the tobacco brown spot fungus spore suspension in the present invention in the determination of tobacco variety resistance to black spot disease is t...

Embodiment 1

[0029] Tobacco diseased leaves with typical symptoms were collected from diseased fields, and strains of Alternaria Alternaria were screened out through separation, purification, and pathogenicity measurement; the hyphae producing spores of the strains were spotted on the culture medium plate, and each plate was Inoculate at 3 points, under the condition of 18h dark and 6h alternating light, culture at 27°C for 4 days, select the culture with dark brown mycelium and denser mycelium for inducing sporulation; plate the culture medium that meets the conditions for inducing sporulation , placed in a refrigerator at 2°C in a sealed state for induction culture in the dark for 2 days, then transferred to an incubator for dark, gradient cooling culture, and cooled from 20°C to 15°C over 2 days (the cooling rate is about 1°C every 9.6h); induction After the cultivation is completed, use 1% sterilized glucose solution as the solution for promoting spore germination, wash off the spores a...

Embodiment 2

[0031] Tobacco diseased leaves with typical symptoms were collected from diseased fields, and strains of Alternaria Alternaria were screened out through separation, purification, and pathogenicity measurement; the hyphae producing spores of the strains were spotted on the culture medium plate, and each plate was Inoculate at 4 points, under the condition of 18h dark and 6h alternating light, culture at 29°C for 6 days, select the culture with dark brown mycelium and denser mycelium to induce sporulation; plate the culture medium that meets the conditions for sporulation induction , placed in a refrigerator at 4°C in a sealed state for induction culture in the dark for 3 days, and then transferred to an incubator for dark, gradient cooling culture, which took 3 days to cool down from 20°C to 15°C (cooling rate is about 1°C every 14.4h); induction After the cultivation is completed, use 2% sterilized fructose and sucrose solution as the solution to promote spore germination, use ...

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Abstract

The invention discloses an alternaria alternate spore suspension liquid and an application thereof. The alternaria alternate spore suspension liquid is prepared through germ separation, germ cultivation, spore production through induction, and preparation of spore suspension liquid. The alternaria alternate spore suspension liquid is used in measurement of tobacco brown spot, or in screening of drug resisting the tobacco brown spot, biocontrol bacterium or natural products, or in research of soil ecology of the alternaria alternate and germ infect circulation. The alternaria alternate spore suspension liquid adopts a multipoint inoculation method, has fast cultivation speed and short cultivation cycle; the alternaria alternate is promoted to generate large quantity of spores rapidly through the comprehensive action of methods such as low temperature high humidity stimulation, gradient cooling, dark induction and the like, and the quantity of the spore generated is 50-100 times that of conventional spore generation quantity. The alternaria alternate spore suspension liquid is light in contamination, the time for cultivating hypha and generating spore through induction is relatively fixed and concentrated, spore generation and growing are synchronous, and the spores prepared in the same batch and different batches are small in quality difference and more stable in quality.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and in particular relates to a spore suspension of Alternaria spp. tobacco and an application thereof. Background technique [0002] Tobacco brown spot (Tobacco Brown Spot) is a leaf disease that commonly occurs in the mature stage of tobacco leaves, and its pathogen is Alternaria ( Alternaria alternata ). At present, in the laboratory, the mycelium is generally cultured on a plate, and the spore suspension of Alternaria tabacum is prepared after natural spore production, but the mycelium usually takes more than 10 days to grow on the plate, and the mycelium is mature and the amount of spore production is sufficient to prepare The spore suspension still needs to be continuously cultivated for 5-10 days. Due to its long cultivation time, it is easy to cause drying of the medium and contamination of the culture, and the sporulation efficiency is low without induction of myceliu...

Claims

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Application Information

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IPC IPC(8): C12N3/00C12Q1/18C12Q1/02C12R1/645
Inventor 方敦煌刘勇李永平肖炳光
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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