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Antifungal compositions containing the endophyte fungus alternaria alternata and/or its metabolites, as antagonist agents of plasmopara viticola

A technology for downy mildew and alternaria of grapes, used in antifungal agents, chemicals for biological control, fungicides, etc.

Inactive Publication Date: 2009-07-15
乌迪内大学 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for grapes, there are no reports on the use of endophytic fungi (especially Alternaria alternata) as antagonist organisms and the metabolites produced by them.

Method used

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  • Antifungal compositions containing the endophyte fungus alternaria alternata and/or its metabolites, as antagonist agents of plasmopara viticola
  • Antifungal compositions containing the endophyte fungus alternaria alternata and/or its metabolites, as antagonist agents of plasmopara viticola
  • Antifungal compositions containing the endophyte fungus alternaria alternata and/or its metabolites, as antagonist agents of plasmopara viticola

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 126 endophyte microorganisms were cultured in liquid medium (nutrient broth=NB+sucrose) in 48-well sterile plates placed on an oscillating stirrer at 150 rpm at room temperature.

[0035] A suspension (suspension 1) containing 900 μl of NB and one microorganism was placed in each well. For each microorganism, 5 replicates were set.

[0036] Subsequently, small discs cut from healthy grape leaves were contacted with the antagonist suspension for 5 minutes, and then incubated at 20°C in a concentration of 4.25×10 5 spores / ml of downy mildew spores were inoculated overnight (suspension 2).

[0037] Then, the small discs were placed in a constant temperature humidity chamber at 20° C. for one week.

[0038] At the end, the presence of P. viticola spores was checked with the aid of a stereomicroscope. The inspection was repeated six times during 2002 and 2003.

[0039] Among the 126 microorganisms, 5 consistently inhibited the sporulation of P. viticola. Therefore, they...

Embodiment 2

[0044] The production of low-molecular-weight metabolites by the endophyte Alternaria alternans was carried out starting from the fungal culture broth. Culture flasks containing 250 ml of nutrient broth were inoculated with fractions of actively growing mycelium of Alternaria alternaria kept on agarized medium in Petri capsules. The broth was then incubated on a shaker at 20°C for 48 hours, then frozen (-40°C) and lyophilized. The lyophilized broth (20 g) was then dissolved in water and filtered; the aqueous solution was then extracted with n-butanol. By preparatory layer chromatography (PLC) on silica gel plates (Merck-Kieselgel 60 PF 254 ) on this organic phase is fractionated using 60:15:25 n-butanol / acetic acid / H 2 A mixture of Os was eluted until three bands, designated A, B, and C in order of increasing polarity, were obtained. Fractions A (54 mg) and B (48 mg) were further purified by thin layer chromatography (TLC) on silica gel plates using a mixture of dichloromet...

Embodiment 3

[0056] For the in-plant tests, rooted cuttings of the vine cultivar Pinot grigio grown under controlled conditions in a greenhouse (20°C, photoperiod 12 hours) were used; the cuttings were arranged on polystyrene plates using spikes , so that the lower blade surface can remain attached to the plate as much as possible.

[0057] The in planta test was carried out in two experiments:

[0058] 1) Evaluation of DKP efficacy after 2 hours and 24 hours after inoculation with Peronospora vine therapeutic experiment ;

[0059] 2) Evaluate the treatment effect before inoculating grape peronospora preventive experiment .

[0060] 1) therapeutic experiment

[0061] Each leaf was labeled with a number and letter based on the DKP type and concentration used; while on the lower leaf surface, small circles were drawn to highlight where the DKP-containing solution would be located. Then, using the 4.7×10 5 Leaves were sprayed with a suspension of downy mildew spores at a concentration...

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Abstract

The present invention relates to compositions containing the fungus Alternaria alternata and, or the secondary metabolites thereof, belonging to the family of the diketopiperazines (DKPs), for controlling the pathogen fungus Plasmopara viticola.

Description

technical field [0001] The present invention relates to compositions comprising the fungus Alternaria alternata and / or its metabolites belonging to the family of diketopiperazines (DKP) for the control of the pathogenic fungus Plasmopara viticola, which is Remedies for grape downy mildew. The main three metabolites are the following compounds with the following formulas: [0002] Cyclic (L-phenylalanine-trans-4-hydroxy-L-proline) (1) [0003] Cyclic (L-leucine-trans-4-hydroxy-L-proline) (2) [0004] Cyclo(L-alanine-trans-4-hydroxy-L-proline)(3) [0005] Background technique [0006] All varieties of grapes (Vitis vinifera) are susceptible to downy mildew, which is caused by the pathogen Peronospora viticola. Conversely, Vitis americana varieties from humid regions where the presence of parasites play a strong role in the selection of endemic populations are resistant. [0007] Until the 50's, chemical combating of grape downy mildews was based almost exclusively on t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N43/90A01N63/04A01P3/00A01N63/30
CPCA01N63/04A01N63/30A61P31/10A01N43/90A01N2300/00
Inventor 丽塔·穆塞蒂斯特凡诺·博尔塞利米凯莱·德安布罗西奥
Owner 乌迪内大学
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