Pseudomonas syringae and application thereof to prevention and treatment of apple ring rot

A technology of pseudomonas and cloves, which is applied in the field of microorganisms and achieves the effects of low production cost, environmental friendliness and stable control effect.

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 (Pseudomonas syringae) for the control of apple ring spot

Method used

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  • Pseudomonas syringae and application thereof to prevention and treatment of apple ring rot
  • Pseudomonas syringae and application thereof to prevention and treatment of apple ring rot
  • Pseudomonas syringae and application thereof to prevention and treatment of apple ring rot

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preparation example Construction

[0029] The preparation method of biocontrol agent provided by the present invention comprises the following steps:

[0030] (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;

[0031] (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;

[0032] (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 24 hours to obtain 10 9 cfu mL -1 Cell culture medium C of B-1 strain;

[0033] (4) put 10 9 cfu mL -1 Cell culture medium C of strain B-1, at 4°C, 12000r·min -1 Centrifuge for 15 minutes, collect the supernatant and filter it through a 0.22 μm microporous membra...

Embodiment 1

[0038] Isolation Screening and Strain Identification of Pseudomonas syringae B-1

[0039] 1. Isolation and screening of strains

[0040]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.

[0041] 2. Identification of strains

[0042] (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 2

[0048] Effect of Pseudomonas syringae B-1 on Mycelia Growth of Apple Ringworm

[0049] Configure PDA medium, add bacterial suspension of bacterial strain B-1 to it, make its final concentration be 10 5 、10 6 、10 7 and 10 8 cfu mL -1 , with the PDA without adding bacterial strain B-1 as a control, the activated good apple ringworm was inoculated in the above-mentioned PDA medium. Incubate in dark at constant temperature at 25°C for 5 days, and measure the colony diameter.

[0050] The above-mentioned PDA medium used has the following components and formula: weigh 200g after peeling the potatoes, cut into small pieces and boil in water for 15-20min, add 20g of glucose and 15g of agar powder after filtering through four layers of gauze, and dilute to 1000mL. The pH value is natural, and it is sterilized by high-pressure steam at 121°C for 20 minutes.

[0051] The results showed that adding different concentrations of bacteria strain B-1 had a significant inhibitory effect o...

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Abstract

The invention discloses a pseudomonas syringae strain B-1. The pseudomonas syringae strain B-1 is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number of thepseudomonas syringae strain B-1 is CCTCC NO: M2015813. The pseudomonas syringae strain B-1 disclosed by the invention has a remarkable prevention and treatment effect on apple ring rot; research of embodiments indicates, the strain B-1 can obviously inhibit the growth of physalospora piricola hyphae; the antibacterial spectrum is wide, and besides the apple ring spot bacteria, the obvious antagonism effect on various fruit and vegetable pathogenic fungi such as fruit and vegetable postharvest botrytis cinerea, fruit and vegetable postharvest penicillium bacteria, apple tree valsa mali, pear tree valsa mali, apple anthracnose leaf blight bacteria, apple mold heart bacteria and peach brown rot bacteria is also achieved; bacterial suspension and fermented supernatant of the bacterial strain B-1 have prevention and treatment effects on apple branch and trunk and fruit ring rot of 70% or above, the bacterial strain B-1 has the characteristics of being stable in prevention and treatment effect, environmentally friendly and the like, 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 microbes, in particular to a strain of Pseudomonas syringae and its application. Background technique [0002] Apple ring spot is one of the important diseases of apple branches and fruits, which is caused by Botryosphaeria dothidea. Apple ring spot bacteria can infect the branches of apple trees throughout the year, forming ring spot tumors, dry rot spots or saddle-shaped spots, seriously weakening the tree vigor, causing dead branches or even dead trees; the fungus infects fruits, causing Rotten fruit. Since Botrytis can invade young apple fruits from May, it will occur in large numbers near the ripening stage, and even cause a large number of rotten fruits during the storage and transportation of apples. [0003] At present, nationwide surveys have found that the incidence rate of apple branch ring disease is as high as 77.6%; the incidence rate of fruit ring disease is 5% to 20% in normal years, and...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/27A01P3/00C12R1/38
CPCA01N63/00C12N1/20C12N1/205C12R2001/38
Inventor 王彩霞李保华练森赵圆秀晏朦张清明
Owner QINGDAO AGRI UNIV
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