Biocontrol microbial agent for apple anthracnose leaf blight as well as preparation method and application thereof

A biocontrol agent and anthracnose leaf technology are applied in the field of microorganisms to achieve the effects of environmental friendliness, significant control effect and simple fermentation process

Active Publication Date: 2020-06-16
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, there is no report of using Pseudomonas syringae to prepare apple anthracnose leaf blight biocontrol agent for the control of apple anthracnose leaf blight

Method used

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  • Biocontrol microbial agent for apple anthracnose leaf blight as well as preparation method and application thereof
  • Biocontrol microbial agent for apple anthracnose leaf blight as well as preparation method and application thereof
  • Biocontrol microbial agent for apple anthracnose leaf blight as well as preparation method and application thereof

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Experimental program
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preparation example Construction

[0028] The preparation method of biocontrol agent provided by the present invention comprises the following steps:

[0029] (1) Streak inoculation of strain B-1 in NA medium, and activate and cultivate at 30°C for 24 hours to obtain a single colony A of activated strain B-1;

[0030] (2) Inoculate the single colony A of the activated bacterial strain B-1 in the NB medium with a bottled volume of 80mL / 250mL, 180r·min -1 , 30 ° C constant temperature shaking culture for 12 hours, to obtain the strain B-1 fermentation seed liquid B;

[0031] (3) The fermented seed liquid B was inoculated into NB culture medium at an inoculum size of 1:100 for expanded cultivation, and incubated at a constant temperature at 30° C. for 12 hours with shaking to obtain 10 7 cfu mL -1 Cell culture medium C of B-1 strain;

[0032] (4) put 10 7 cfu mL -1 Cell culture medium C of strain B-1, at 4°C, 12000r·min -1 Centrifuge for 15min, collect the cell pellet and resuspend with sterile water to obta...

Embodiment 1

[0037] Isolation Screening and Strain Identification of Pseudomonas syringae B-1

[0038] 1. Isolation and screening of strains

[0039]Mature healthy fruits were collected from the commercialized Fuji apple orchard in Yantai, Shandong. After surface disinfection, the fruit tissues were ground, added sterile water, and separated by conventional gradient dilution coating. They were cultured with NA medium at 28°C and colonies were picked. Colonies with obvious morphological differences were purified and preserved on NA medium, and the primary screening and multiple re-screening of antagonistic bacteria were carried out with Apple ringworm as the target pathogen, and finally a bacterial strain with strong antibacterial activity was obtained, named B -1.

[0040] 2. Identification of strains

[0041] (1) Morphological characteristics: After the strain B-1 was cultured on NA medium for 24 hours, the colony was milky white, round or nearly round, with a smooth surface and a raise...

Embodiment 3

[0047] Effects of Pseudomonas syringae B-1 on Mycelia Growth of Apple Anthracnose Leaf Blight

[0048] The seed fermentation liquid of Pseudomonas syringae (Pseudomonas syringae) B-1 was inoculated in NB medium, and cultured with constant temperature and shaking at 30°C for 12 hours to obtain the cell culture liquid of strain B-1, and the cells of strain B-1 were collected after centrifugation .

[0049] Configure the PDA medium, add the thalline of bacterial strain B-1 therein, make its final concentration be 10 4 、10 5 、10 6 and 10 7 cfu mL -1 , with the PDA medium without adding bacterial strain B-1 as a control, the activated apple anthracnose leaf blight was inoculated in the above PDA medium. Incubate in dark at constant temperature at 25°C for 3 days, and measure the colony diameter.

[0050] The above-mentioned PDA medium used has the following components and formula: weigh 200g after the potatoes are peeled, cut into small pieces and boil in water for 15-20min, ...

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Abstract

The invention discloses a biocontrol microbial agent for apple anthracnose leaf blight. The biocontrol microbial agent for apple anthracnose leaf blight is a bacterial suspension of a pseudomonas syringae strain B-1. The pseudomonas syringae B-1 disclosed by the invention has a remarkable prevention and treatment effect on the apple anthracnose leaf blight. The research of the embodiment shows that the strain B-1 can significantly inhibit the growth of apple anthracnose leaf blight hyphae, and the prevention and treatment effect of the bacterial suspension of the strain B-1 on apple anthracnose leaf blight is 60% or above. The biocontrol microbial agent for apple anthracnose leaf blight is environmentally friendly, the fermentation process of large-scale production is simple, and the production cost is low.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a biocontrol agent for apple anthracnose leaf blight and its preparation method and application. Background technique [0002] Apples are rich in carbohydrates, vitamins and inorganic salts, and are an important source of nutrition and material basis for people. As an important economic crop and export fruit in my country, apple is the first in the world in terms of cultivation area and output. Apple is one of the important pillar industries for farmers in northern my country to increase their income. However, diseases seriously restrict the healthy development of my country's apple industry. [0003] Anthracnose leaf blight (Glomerella leafspot, GLS) caused by Glomerella cingulata is a newly discovered disease on apples in my country in recent years. After the apple fruit is infected, a black lesion with a diameter of 1-2 mm will be formed on the fruit surface, which not...

Claims

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Application Information

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IPC IPC(8): A01N63/27A01N31/08A01N25/04A01P1/00C12N1/20C12R1/38
CPCA01N25/04A01N31/08C12N1/20
Inventor 王彩霞练森李保华朱德娟张清明
Owner QINGDAO AGRI UNIV
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