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Improvement of biological seed treatments against damping-off in crop production

Contract nr: FAIR-CT96-1373
Project nr: 1373
Project type: SC
Starting date: 01/08/1997
Duration: 48 months
Total cost: 1,357,500 EUR
EC Contribution: 1,092,000 EUR
Scientific Officer: Arnaud BORCHARD
Research topic: Plant health
Acronym: IMPROBIOSEED

Background:
Seedling damping-off, caused by soil-borne fungal pathogens, results in persistent commercial losses in agriculture and horticulture. Seed treatment with antagonistic bacteria or fungi is an alternative to fungicides in the control of damping-off.
However, under commercial conditions, biological treatments frequently show more variable activity than fungicides.
The activity of biological seed treatments relies on efficient colonisation and metabolic activity of antagonists in the spermosphere and rhizosphere.
Failure to colonise and poor biocontrol activity was frequently linked to
(i) competition from the indigenous soil microflora and
(ii) susceptibility of biocontrol agents to environmental conditions (soil moisture, pH, temperature, soil types etc.) and
(iii) lack of appropriate seed coating technology for biologicals.


Objectives:
The main aim of the project is to use molecular approaches together with established soil microbiological methods to develop strategies which improve the efficacy of biological seed inocula. This aim will be achieved through academic and industrial research and subsequent field evaluation.
Academic research transforming bacterial antagonists with the lux AB reporter genes (Objective l.) will allow accurate study of the fate of seed inocula in the soil environment.
Use of lux marked strains of biocontrol agents will enable the determination of their environmental activity profiles (Objective 2.).
These profiles facilitate design of antagonist combinations, which extend biocontrol activity to a wider range of environmental conditions.
Lux-marked antagonists and Tn5 mutagenesis will be used to identify the physiological characteristics (eg. Catabolic activity range, motility, adhesion properties and siderophore and antimicrobial production) that contribute to rhizosphere competence (Objective 3.) of biocontrol agents. This information will be used to develop seed coat formulations and additives, which improve rhizosphere competence of bacterial seed inocula (Objective 4.).
Since individual antagonist treatments can rarely provide activity against all damping-off pathogens, it is proposed to identify combinations of antagonists with synergistic modes of action (Objective 5.) and to integrate biological with fungicide seed treatments (Objective 6).
Industrial research strategies to improved biocontrol activity developed under Objectives 2 to 6 (eg mixed inocula, addition of carbohydrates and other seed additives) require the development of appropriate fermentation, formulation and seed coating methods.
Two industrial partners (specialising in the production of microbial inocula and seed production/treatment) will therefore apply the fundamental data produced to improve and adapt
(i) fermentation technologies (Objective 7.) and
(ii) seed coating and formulation methods (Objective 8.) for the different bacterial antagonists.

Field trials throughout the EU (Objective 9.) are proposed to
(i) evaluate the activity of biological seed inocula under different climatic and soil conditions and cultural production methods and
(ii) to build confidence in both the scientific and user community in biological/integrated disease control.


Description:
Task 1.1.: Lux- marking of three bacterial antagonists
Objectives:
The transformation of bacterial antagonists with luxAB reporter genes.

Task 1.2: Determination of the relationship between luminescence and metabolic activity
Objectives: The transformation of bacterial antagonists with luxAB reporter genes.

Task 2.1: Effect of soil conditions on rhizosphere colonisation and activity of single antagonist inocula
Objectives: Identification of environmental activity profiles of bacterial biocontrol agents.

Task 2.2: Effect of soil conditions on rhizosphere colonisation and activity of mixed antagonist inocula
Objectives: Identification of environmental activity profiles of bacterial biocontrol agents.

Task 2.3: Dose response growth room trials
Objectives: Identification of environmental activity profiles of bacterial biocontrol agents.

Task 3.1: Metabolic profiling of antagonist strains
Objectives: Identification of physiological characteristics of biological control agents which are essential for spermosphere and rhizosphere competence.

Task 3.2: Identification of physiological characteristics important for spermosphere and rhizosphere competence using Tn5 mutants
Objectives: Identification of physiological characteristics of biological control agents which are essential for spermosphere and rhizosphere competence.

Task 4.1.: Effect of carbohydrate seed coat additives on rhizosphere competence
Objectives: 4. To test carbohydrate seed coat additives for their ability to improve spermosphere/rhizosphere competence by bacterial seed inocula.

Task 5.1.: Identification of interactions between fungal and bacteria antagonists in vitro
Objectives: To identify synergistic combinations of antagonists for combined seed and soil treatment.

Task 5.2: Identification of synergistic antagonist combinations for combined seed and soil treatments in vivo in glasshouse trials
Objectives: To identify synergistic combinations of antagonists for combined seed and soil treatment.

Task 5.3.: Identification of interactions between antagonist seed inocula and VA-mycorrhizal fungi
Objectives: To identify synergistic combinations of antagonists for combined seed and soil treatment.

Task 6.1.: Identification of compatible fungicide/antagonist combinations in vitro
Objectives: To identify compatible fungicide/antagonist combinations for integrated disease control.

Task 6.2.: Effect of fungicide addition on colonisation and activity of antagonists in vivo
Objectives: To identify compatible fungicide/antagonist combinations for integrated disease control.

Task 7.1.: Development of fermentation technologies for bacterial antagonists
Objectives To develop fermentation and inoculum formulation methods for bacterial antagonists.

Task 7.2.: Development of carrier based inoculum preparations
Objectives: To develop fermentation and inoculum formulation methods for bacterial antagonists.

Task 7.3.: Effect of inoculum storage on the survival and activity of antagonists
Objectives: To develop fermentation and inoculum formulation methods for bacterial antagonists.

Task 7.4.: Production and supply of bacterial inocula for field trials
Objectives: To develop fermentation and inoculum formulation methods for bacterial antagonists.

Task 8.1.: Integration of the application of bacterial antagonist into the pelleting process
Objectives: To develop seed coating methods for individual, mixed and integrated antagonist treatments

Task 8.2.: Optimisation of seed coat physical properties
Objectives: To develop seed coating methods for individual, mixed and integrated antagonist treatments.

Task 8.3.: Optimisation of seed coat composition and antagonist loading
Objectives: To develop seed coating methods for individual, mixed and integrated antagonist treatments.

Task 8.4.: Survival of antagonists during storage of pelleted seeds
Objectives: To develop seed coating methods for individual, mixed and integrated antagonist treatments.

Task 8.5.: Production of pelleted sugar beet seeds for all field trials
Objectives: To develop seed coating methods for individual, mixed and integrated antagonist treatments.

Task 9.1.: Sugar beet field trials with the most active antagonist and integrated seed treatments
Objectives: Field trials with the most active biological and integrated seed treatments developed in objective 2 to 8.

Current situation/results:
Results will be available after clearance by the exploitation committee, which will meet in July 2000.
Task 1.1., Task 1.2, Task 3.1, Task 5.1., Task 5.2., Task 6.1., Task 7.1., Task 7.2., Task 8.1., Task 8.2.: completed.
Task 2.1: started but behind schedule.
Task 2.2: start of work on task delayed until month 32.
Task 2.3: work in progress.
Task 3.2: work in progress and on schedule.
Task 4.1: work to start in project month 33
Task 5.3: ongoing (this has to be delayed until project months 30, start of cucumber cropping in the UK; Spring 2000).
Task 6.2: work is currently scheduled to start in project month 33.
Task 7.3: work on this task is ongoing and on schedule (scheduled to be completed in project month 42).
Task 7.4: work is scheduled to start in project month 33.
Task 8.3: work is ongoing and slightly ahead of schedule.
Task 8.4: ongoing.
Task 8.5: since a proportion of field trials was scheduled to start in year three of the project, work had to be re-scheduled for months 31.
Task 9.1: some field trials are now scheduled for year three (to start in project month 32).


Coordinator
Carlo LEIFERT
University of Aberdeen
St. Machar Drive
UK-AB24 3UU Aberdeen
Tel.: +44 1224 27 38 56
Fax: +44 1224 27 27 03
E-mail: c.leifert@abdn.ac.uk / bot154@abdn.ac.uk


Partners

  • Uwe FISCHER
    KWS Kleinwanzlebener Saatzucht ag Vorm. Rabbethgeand Giesecke
    Grimsehlstraße 31
    P.O. Box 1463
    D-37555 Einbeck
    Tel.: +49 5561 31 13 63
    Fax: +49 5561 31 15 40

  • Gerhard WOLF
    Georg-August Universität Göttingen
    Grisebachstraße 6
    D-37077 Göttingen
    Tel.: +49 551 39 37 23
    Fax: +49 551 39 41 87
    E-mail: cheppne@gwdg.de

  • Nicos E. MALATHRAKIS
    Technological Education Institute Heraklion
    Stauromenos
    P.O. Box 140
    GR-711 10 Heraklion
    Tel.: +30 81 25 41 03
    Fax: +30 81 25 05 48
    E-mail: ipro@lyttos.her.tei.gr

  • Adam BAHJAT
    Microbio Ltd
    Maxted road - Boundary way
    UK-HP2 7SU Hemel Hempset
    Tel.: +44 1442 39 91 28
    Fax: +44 1442 39 91 28
    E-mail: adam.hajjar@bbsrc.ac.uk
 
 
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