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Method for inducing grape elsinoe ampelina to produce spores

A cystic cavity and grape technology, which is applied in the field of inducing sporulation of Cystobacter scabbies, can solve the problems of slow growth and difficult sporulation, and achieves the effect of simple operation.

Active Publication Date: 2017-06-13
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the technical problem that P. viticola is the pathogenic bacterium of grape black pox, which grows slowly and is difficult to produce sporulation, the purpose of the present invention is to provide a method for inducing sporulation by P.

Method used

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  • Method for inducing grape elsinoe ampelina to produce spores
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  • Method for inducing grape elsinoe ampelina to produce spores

Examples

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Effect test

Embodiment 1

[0031] Embodiment 1: Isolation and identification of grape black pox bacteria

[0032] Collect 'Red Globe' grapes where black pox (with grape black pox pathogenic fungus) leaves are brought back to the laboratory, cut off the junction of disease and health with a scalpel, cut into small pieces of 3mm * 3mm with scissors, and Put the cut pieces into 2.5% NaClO solution for surface disinfection for 2 minutes, and then rinse them with sterile water for 3 times. After natural drying, place on the potato dextrose agar medium (PDA) containing 50ng / mL streptomycin. . Incubate in the dark at 25°C. After growing for 5 days, the bacterium colony that grew out of grape scab was transferred to a new PDA medium to continue culturing ( figure 1 A). The conidia are evenly coated on the PDA medium, and a single colony can grow out after 6 days of growth, and the bacterium colony of picking the single grape scab cyst cavity bacteria is inoculated on the new PDA medium ( figure 1 B). Util...

Embodiment 2

[0035] Embodiment 2: the influence of culture mode on the spore production of grape scab coelomyces

[0036] First pick the edge of the PDA culture dish grown on the PDA culture dish for 15 days, and inoculate it in a culture dish containing PDA medium and a culture bottle containing PDA medium ( figure 2 A and 2E), cultivated in the dark at 25°C, the colony morphology is as follows figure 2 As shown, after culturing for 25 days, place the petri dish and culture bottle at 21°C for 24 hours of induction, scrape the surface tissue of the colony with a sterile scalpel, extrude it into a powder, put it into sterile water, and shake it for 30 seconds , take the supernatant and carry out the spore counting of the coelomyces spp. with a hemocytometer to obtain the spores of the coelomyces spp.

[0037] The spore production of the colony of grape scab coelomyces cultivated in culture flasks can reach 5.3×10 6 individual / mL, while the spore yield of each colony of Grape scab in the...

Embodiment 3

[0038] Embodiment 3: the impact of cultivation time on the spore production of grape scab coelomyces

[0039] Due to the relatively slow growth of C. viticola, the color of the colony of C. viticola in the early stage was dark red, but the color of the colony changed to gray-green in the later stage. In order to explore the influence of culture time on the sporulation of C. Cultivate 20 days, 25 days, 30 days and 35 days in the culture bottle to induce sporulation. It is found that the spore production of the grape scab is the largest when it is cultivated on the culture bottle for 25 days, while the 35-day grape The spore production of sclerocystis was only 0.25×10 6 pcs / mL( image 3 D), it shows that the long culture time is not conducive to the spore production of grape scab coelomycetes. Find that by morphological observation, there are a large amount of aerial hyphae ( Figure 4 E), and a large number of oil cells were produced in the hyphae ( Figure 4 F), the sporul...

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Abstract

The invention discloses a method for inducing grape elsinoe ampelina to produce spores. The method comprises the following steps of selecting an outer ring mycelium of a grape anthrachose pathogenic fungus colony which grows for 15 days in a culture dish with a PDA (potato dextrose agar) culture medium, and inoculating the mycelium into a new sterile culture bottle with the PDA culture medium; culturing for 25 days at the temperature of 25 DEG C under the dark environment, and putting the culture bottle under the dark environment for 24h at the temperature of 21 DEG C; using a sterile scalpel to scrape colony surface tissues, extruding into powder, putting into sterile water, oscillating for 30s, obtaining supernatant liquid, and using a blood counting chamber to count the spores. The method has the advantages that the operation is simple, more conidiums can be stably obtained, the anthrachose-resisting grape variety can be conveniently sorted, and the guarantee is provided for the disease-resistant breeding and grape pathological studying.

Description

technical field [0001] The invention belongs to the technical field of fruit tree pathology, and in particular relates to a method for inducing spore production by coelomyces vine scab. Background technique [0002] There are many kinds of grape diseases, and the damage is serious. Among them, grape black pox is widely distributed and occurs in large numbers in the rainy areas in the north and south of my country. When the disease is severe, it can cause up to 50% fruit loss, seriously affecting the yield and quality of grapes, and has become a constraint on the health of the grape industry. One of the main factors of development. Grape black spot is a fungal disease caused by Elsinoe ampelina, which harms the shoots, vines, petioles, veins, fruit stalks and fruits of grapes. [0003] The asexual stage of Grape scab is Cystospora spp., and the sexual stage is Ascomycotina sclerosia spp. It grows slowly on the artificial medium. It has been reported that C. viticola grows we...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N3/00C12R1/645
CPCC12N1/14C12N3/00
Inventor 李智王西平
Owner NORTHWEST A & F UNIV
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