The use of simultaneous P-solubilizingP and biocontrol activity of microorganisms for sustainable agriculture Svetlana Zhigletsova, PhD (Chem) State Research Center for Applied Microbiology and Biotechnology (SRCAMB) Obolensk, Moscow region, 142279 Russian Federation E-mail: sjigletsova@mail.ru
Provision of crops with available phosphorus: Deficiency of resources of natural phosphatic raw material; Low level of P assimilation by plants from chemical P-P fertilizers (10-25%); Ecological problems of manufacturing and application of chemical P-fertilizers; P High cost of chemical P-P fertilizers Challenges in plant cultivation: Injuriousness and problems of plant diseases with Fusarium head blight (FHB) as an example : Losses of a crop (20-50 %); Pollution of grain and forages by highly stable mycotoxins; Direct and cross resistance of phytopathogens to chemical fungicides; Absence of resistant cultivars to FHB
Structure of Research Search of antagonists for soilborne pathogens of agricultural crops among phosphate solubilising microorganisms Search of phosphate solubilising microorganisms among antagonists for soilborne pathogens of agricultural crops Assessment of availability of phosphoric ores for selected microbes Development of production techniques for P-biofertilizer with biocontrol activity
Search of antagonists for soilborne pathogens of agricultural crops among phosphate solubilising microorganisms In the center of a Petri dish, the agent of Fusarium head blight (Fusarium graminearum) was grown. Phosphate solubilising strains were grown by strokes.
Antagonistic properties of T. asperellum against F. graminearum T. asperellum F. graminearum Suppression of F. graminearum by T. asperellum
One of the modes of action of fungi-antagonists Mycoparasitism of Trichoderma sp. against Pythium on pea seed surface (Hubbard et al., 1983).. SEM image of phytopathogen Rhizoctonia solani surface after removing of Trichoderma s hypha. Cell wall erosion is a result of lytic enzymes action (Elad et al.; 1983)
Biocontrol activity of Bacillus subtilis IPM-215 and Pseudomonas fluorescens Р469 against different phytopathogens Phytopathogen Disease Zone of suppression, Ø mm by IPM-215 by Р469 Alternaria brassicae Dark spot 10-15 ++ Fusarium oxysporum Root rot 20-30 + Fusarium solani Root rot 20-30 + Fusarium culmorum Fusarium head blight 20-30 + Fusarium graminearum Fusarium head blight 20-30 ++ Colletotrichum atraventa Cereal anthracnose 20-30 ++ Phytophthora infestans Potato blight 15-20 + Rhizoctonia solani Bare patch 15-20 + Pseudomonas solanacearum 9049 Potato wilt 10-15 15-20 Pseudomonas sp. 37 Potato tuber rot 10-15 15-20 Pseudomonas sp. 115 Cabbage soft rot 10-15 15-20 Pseudomonas sp. 116 Potato soft rot 10-15 15-20 Erwinia carotovora pv. carotovora Potato rubber rot 15-20 5 Oospora lactis 5 Potato rubber rot 5 10-30 Phoma esigia 6 Phoma rot 5 10-30 Gaeumannomyces graminis var. tritici Root rot of wheat 10-15 5 Verticilium dahlia Cotton wilt 30-40 -
Antagonistic activity of the most prospective strains of genus Trichoderma against phytopathogens of genus Fusarium Phytopathogen Level of activity Gibberella pulicaris (Fr.) Sacc. (Fusarium oxysporum) Fusarium culmorum (W.G. Sm.) Sacc Fusarium graminearum var. graminearum Schwabe Pathogen index (number) in Trichoderma strain index collection 16 29 6 ++ + 1 ++ + 25 ++ + 4 ++ ++ 26 ++ + 34 +++ 35 +++ 36 +++ 31 +++ 32 +++ 33 +++ 28 +++ +++ 2 ++ Gibberella gordonii C. Booth (F.heterosporum Nees et T. Nees) F. sporotrichioides Sherb. (F. 3 +++ sporotrichiella Bilai) 24 ++ + Fusarium sp. 21 +++ ++
Content of 4-deoxynivalenol (DON) in wheat grain (wheat was infected by F. graminearum) 1- infected control 2-Trichoderma #16 (seed treatment) 3-Trichoderma #16 (seed treatment + spraing) 4 -Agat - chemical fungicide (seed treatment) 5- TMTD - chemical fungicide (seed treatment) DON, mg/kg 1,4 1,2 1 0,8 0,6 0,4 0,2 0 1 2 3 4 5 Treatment
Search of phosphate solubilising microorganisms among antagonists for soilborne pathogens of agricultural crops Solid medium with Ca 3 (PO 4 ) 2 or phosphate ore
Dynamics of phosphate releasing from Са 3 (РО 4 ) 2 by different microorganisms in liquid medium Р, mg/l 700 600 500 400 300 200 100 0 0 10 20 30 40 t, h 1 2 3 4 5 1- Pseudomonas fluorescens P469; 2 Bacillus subtilis IPM 215; 3 - Trichoderma longibrahiatum 29; 4 T. asperellum 16- GJS 03-35; 5 - Burkholderia cepacia E37.
Most common mechanism of phosphate releasing from mineral raw materials by microorganisms Carbon source (carbohydrates) Microorganism Organic acids Ca 3 (PO 4 ) 2 Phosphorites Apatites H 2 PO 4 -
Most effective mechanism of phosphate releasing from mineral raw materials by microorganisms H H H H H O H H H H H O H H H H H O O 2 O 2 HC---C---C---C---C---C HC---C---C---C---C---C HC---C---C---C---C---C OH OH OH OH OH H OH OH OH OH OH OH OH OH OH OH O OH All glucose (starch, cellulose) Gluconic acid 2-ketogluconic acid 4 C 6 H 12 O 7 + Ca 3 (PO 4 ) 2 = 2Ca(C 6 H 11 O 7 ) 2 + Ca(H 2 PO 4 ) 2 14C 6 H 12 O 7 +Ca 10 (PO 4 ) 6 F 2 =7Ca(C 6 H 11 O 7 ) 2 +3Ca(H 2 PO 4 ) 2 +2HF
How Complex Biofertilizer Works Organic acids Nutrients Soluble phosphates, antibiotics, enzymes Microorganism Phosphate ore
Most Important Results For the first time, the microorganisms combining high antagonistic activity to pathogens of agricultural crops with high values of phosphate solubilising activity are revealed. Among them there is the strain B. subtilis IPM-215 on the basis of which preparation Bactophyt against various diseases of cereal, fruit, vegetable and decorative cultures is manufactured in Russia. Most effective mechanism of phosphate solubilisation found out till now was revealed for some of the investigated microorganisms.
The goal of cooperation: Development of preparations against plant diseases raising at the same time quantity and quality of the foodstuffs, improving soil fertility and environmental conditions. Preparations can be created both on the basis of one strain with the complex of properties, and on the basis of several microorganisms with different type of activity
Tasks suggested for cooperation - scientific: Search of pathogens suppressors for the certain agricultural crops having additional properties, including phosphate solubilisation from mineral raw materials. Study of mechanisms of both antagonistic activity, and solubilisation of phosphates for the most active microorganisms. Finding-out of interrelation of suppressive properties and phosphate solubilising ability..
To implement scientific tasks, from the potential partner it is expected: Delivery of plant pathogens to be controlled, soil and ores samples for testing activity of our strains. Delivery of samples (or granting of an opportunity for sampling) from which isolation of new target microorganisms and also phytopathogens to be controlled will be carried out. Direct participation or granting to use equipment for analysis of methabolites causing suppressive properties and ability to release phosphates by microorganisms.
Tasks suggested for cooperation - practical: Development of formulations for P-fertilizers P with biocontrol activity: granules and/or composts. Development of P-fertilizers P with biocontrol activity for the certain crops and for the certain soil types.
To implement these tasks, from the potential partner it is expected: Direct participation or granting of an opportunity to use equipment for development of formulations (granules and/or composts). Carrying out (or assistance to carrying out) tests of designed formulations for certain crops on the certain soil types.
Thank you! SRCAMB 2009