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EVERGREEN Project: Sustainable Plant Disease Control from Agricultural Waste

The EVERGREEN project aims to substitute pesticides with biomolecules from agricultural wastes for eco-friendly plant disease control. It will assess traditional pesticides' performance against plant diseases and monitor the environmental impact and benefits of the biomolecules. Through communication and dissemination actions, it promotes sustainable food production with minimal environmental risks. The project focuses on the biological activity of high-quality polyphenol extracts and soil quality assessments post-pesticide application. By investigating enzyme activities and heavy metal effects on soil systems, EVERGREEN seeks to enhance sustainable agriculture practices.

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EVERGREEN Project: Sustainable Plant Disease Control from Agricultural Waste

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  1. EU project LIFE13 ENV IT 000461 / “EVERGREEN” “Environmentally friendly biomolecules from agricultural wastes as substitutes of pesticides for plant diseases control” BENEFICCIARY; CSIC (CEBAS-CSIC) Carlos García Izquierdo. CEBAS-CSIC

  2. Facilities (CEBAS-CSIC) CEBAS Project LIFE13 ENV/IT/000461 PILOT PLANT CULTURE CHAMBERS CEBAS LABORATORIES EXOERIMENTAL FIELD

  3. EVERGREEN (LIFE) CEBAS Project LIFE13 ENV/IT/000461 CEBAS-CSIC (EVERGREEN) Dr. Carlos Garcia Dra. Teresa Hernández Dr. J. Moreno Lda. Mª Dolores Coll Lda. Blanca D. Manuel Campoy Dña. C. Montesinos

  4. CEBAS-CSIC GROUP: EVERGREEN Project LIFE13 ENV/IT/000461 WHAT CEBAS-CSIC GROUP WILL DO IN THE EVERGREEN PROJECT?? EVERGREEN:Thisprojectwilldemostratethesuccessful use of biomoleculesrecoveredfromagricultural vegetable wastes as substitutes of pesticidesfortheenvironmentalyfriendly and sustainable control of plantdiseases ACTIONS B1, B4 B1) Demostration of the performances of traditionalpesticidesforthe control of bacterial and nematodesdiseases of platsimportantforthe EU. B4) Demostration of thebiologicalactivity of thehighqualitypolyphenolsextractsrecoveredfromnotediblebiomass and wastes, againstplantpathogenic bacteria and nematode, in plant. ACTIONS C1, C2 and C4: C1: Monitoringontheenvironmentalimpact of cooper and nematicidesforthecropdefenceagainstphytopathogenic bacteria and nematodes C2.Monitoring of theabsences of sideeffectsforthehighqualitypolyfhenolicpreparationsoncommon targets of the living organisms at laboratorylevel. C4. Monitoring of the short-termenvironmentalbenefictsfromthe use of thehighqualitypolyphenolspreparations in plantdisease control at pilotscalelevel in fieldscreening ACTION D…:Communication and disseminationactions

  5. EVERGREEN (LIFE) Project LIFE13 ENV/IT/000461 B1)Demostration of the performances of traditionalpesticidesforthe control of bacterial and nematodesdiseases of platsimportantforthe EU. (FromOctober 2014 to March 2015: DONE) AGRICULTURE: WE NEED TO PRODUCE FOOD (9000 MILL IN 20150) 1) Use of agrochemical to combatplantdiseases; isituseful? YES 2) Are goodtheagrochemicalsfortheenvironment?? Whataboutsoil quality?? (Cu and nematicides) (OUR ACTYVITY B1) SUSTAINABLE PRODUCTION: WE MUST PRODUCE WITH NO RISK FOR THE ENVIRONMENT

  6. PHOTODEGRADAtION VOLATILIZACIÓN ABSORCION Y EXUDACION RUNOFF INFILTRACION ADSORPTION DESORPTION CHEMICAL DEGRADATION BIODEGRADATION LIXIVIACION CAPILARIDAD WATER TABLE TRANSFER ADSORTION-DESORPTION (Cu, organics) LEACHING VOLATILISATION (organicmetabolites) RUNOF ABSORTION BY PLANTS AND ORGANISMS (Cu, organicmetabolites) TRANSFORMATI0N CHEMICAL (Cu) PHOTOCHEMICAL BIOLOGICAL (Cu) DYNAMIC OF CONTAMINANTS IN SOIL-WATER SYSTEM Project LIFE13 ENV/IT/000461 HEAVY METALS “Cu” NEMATICIDES Agriculture Industrial Domestic Wastes SOIL

  7. EVERGREEN (LIFE) Project LIFE13 ENV/IT/000461 CEBAS WORKING-GROUP (EVERGREEN) EFFECT OF CuSO4 and NEMATICIDES ON SOIL SYSTEM OBJECTIVE: to demonstrate the environmentally negative impact of COOPER and NEMATICIDES on SOIL SYSTEMS ACTIVITIES B1 AND C1: FROM OCTOBRE 2014 TO MARCH 2015 (DONE!!!)

  8. EVERGREEN (LIFE) B1Act. Experimentpreparation: Controlledchamber 60 daysexperiment(sampling at 0, 15, 30 and 60 days) Cu salts and Neenazal, Mocap, Nemacur, Vidate

  9. EVERGREEN (LIFE) C1 Act. Project LIFE13 ENV/IT/000461 MONITORING OF SOIL QUALITY AND BIODIVERSITY AFTER CU salt AND NEMATICIDE ADDITION. ENVIRONMENT: TO LIMIT THE USE OF THESE PRODUCTS ANALYSIS ON SOIL + NEMATICIDES AND Cu SULPHATE General analysis: pH, CE, N, P, K, Ca, Mg, Mn, Na, Fe, Cu, Zn Biologicalanalysis: OM, TOC, soluble C, microbialrespiration. Biochemicalanalysis: Soilenzymeactivities (Urease, b-glucosidase, phosphatase). Microbiologicalanalysis: PFLA,s

  10. pH: No problemwithsoil pH after Cu salt and nematicideaddition (7-8). Itisindicativethatsomeelements (heavy metals) willnot be a problem forplants. EC: Thisparameterisindicative of salinity. Ourresultsindicatethatthe use of Cu salt and nematicidesisnot a problemforsoilsalinity.

  11. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461

  12. EVERGREEN (LIFE) C1 Enzyme activities Project LIFE13 ENV/IT/000461 Dheydrogenase enzymes catalyze the oxidation of organic compounds with the removal of two hydrogen atoms that are transferred to the molecule of NAD+ R-H2 + NAD+ NADH + R + H+ Indicator of global microbial metabolism b-Glucosidase enzymes catalyze the final limiting step of cellulose degradation Carbon cycle Phosphatase enzymes catalyze the hydrolysis of various organic phosphate esters Phosphorus cycle R-O-PO32- + H2O R-O-H + HO-PO32-

  13. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 B-Glucosidaseis a hydrolaseincluded in the C cycle. Itisimportantforsoilquality. Cu showed a negativeeffectonthisenzyme at 60 days. Hidrolases: Extracellularenzymes Transformmacrosustratesintosmallercompounds Responsibles of importantscycles : C, N, P cycles) Degradationpesticides and otherxenobiotics

  14. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 Phosphataseactivity: hydrolase of P cycle. It can hydrolizeorganic P to inorganic P, assimilablebyplants. A negativeeffect of Cu and somenematicides at 0 and 60 days has beenshowed.

  15. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 Ureaseactivity: hydrolase of N cycle. Ureaseactivitydecreasedwhen Cu and pesticideswereadded to thesoil. N Cycleisaffectedbythesepollutants (Mocap and Cu)

  16. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 Dehydrogenases are endocellularenzymes. Theyindicatethat catabolicactivity of soils. Neenazaal at highdoseincreased soilmetabolicactivity (substrateformicroorganisms???).

  17. EVERGREEN (LIFE) C1 Water soluble C is a labil C fraction. Forthisreason, thisparameterchangesalong of theexperiment. Water soluble C isinfluencedbyorganicmattermineralzation. In ourstudy, there are nottoovariationswithtreatments and time.

  18. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 Water soluble N: our results showed a negative effect on water soluble N when some pesticides were added to the soil.

  19. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 ICP-OES equipment Micro and macronutrients was analyzed by ICP-OES (Inductively coupled plasma-optical emision spectrometer) cuantitative determination, is an elemental analysis technic (IONOMIC SERVICE). The “Ionoma” of soil is important to know the equilibrium of the elements in the soil.

  20. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 Cu quantityincresed onsoil. Itisdue to use of Cu salt.

  21. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 SOIL BIODIVERSITY: FUNCTIONALITY SOIL RESPIRATION Thedegradation of organicmatteris a property of allheterotrophs, and itsrateiscommonlyused to indicatethelevel of microbialactivity

  22. MICROBIAL BIOMASS: PHOSPHOLIPID FATTY ACIDS Project LIFE13 ENV/IT/000461 Microbialbiomass C and N (Powlson et al., 1987) ATP, Ergosterol, etc. PhosphoLipidFattyAcids (PLFAs) Bligh & Dyer, 1959 Frostegard et al., 1993

  23. EVERGREEN (LIFE) C1 Bacterial. fungal. Gram+. Gram-. satured and monosatured PLFAs concentration in microcosm soils T0 The total PLFA and bacteria/fungi. Gram+/Gram- and monounsatured/satured ratios in microcoms soils T0

  24. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 INFLUENCE OF Cu Salt AND NEMATICIDES ON SOIL BIODIVERSITY Total PLFA decreased in thesoilwhennematicides and Cu wereadded. Itcould be due to thenegativeeffect of theseagrochemicalsonsoil-life. At 60 days, only Cu salt and Nemacurshowedthisnegativeeffect.

  25. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 INFLUENCE OF Cu Salt AND NEMATICIDED ON SOIL BIODIVERSITY ForFungi/Bact, differenceswereshowedwithtreatments and the time. At 0 days, Fungi/Bact ratio onlyincreasedforNeenazal. At 60 days, soilswithNemacur and Vidateincreasedthis ratio value.

  26. EVERGREEN (LIFE) C1 Project LIFE13 ENV/IT/000461 • CONCLUSIONS B1/C1 ACTIVITIES (CEBAS) • Cu salts and nematicides are used in agriculture (bacteria and • nematodes are combated in agriculturewiththeseproducts). • 2) The use in agriculture of Cu salts and nematicidesshould be • reduced. Theyhave a negativeeffectonsoilecosystem and • environment, particularlyonspecificmicrobialpopulationimplied • in the C, N and P cycles • 3) Ourworkshowed a negativeeffectonsoilbiodiversity • 4) The Cu is more toxicthatnematicidesdue to: • a) Nematicides are organicproducts, and theycouldbe • degradedbydifferentways. Cu is a heavy metal (inorganic • pollutant), and cannot be degradedbymicorganisms. • b) Cu could be absorbed byplants

  27. CEBAS-CSIC GROUP: EVERGREEN Project LIFE13 ENV/IT/000461 AGRICULTURAL WASTES (SPAIN) Capacity of someagriculturalwastes (horticultural wastes) as polyphenols “factory”. A regional study B1 Activity

  28. EVERGREEN: obtainingpolyphenols IMPORTANT FOR THE ENVIROMENT ORGANIC WASTES: A PROBLEM… * THERE IS AN INCREASE IN THEIR PRODUCTION WHICH IS NOT DESIDERABLE * A PROBLEM!!! (contamination, bad odours) Weneedgive a solutionforthesewastes ADD VALUES fororganicwastes?? POLYPHENOLS EXTRACTION Waterextraction AGRICULTURAL RESOURCES WASTES BYPRODUCTS

  29. EVERGREEN (LIFE) Production: differentaccording to climate, soilconditions,…

  30. EVERGREEN (LIFE)

  31. EVERGREEN (LIFE) Agriculturewastes: extraction of polyphenolswithwater and methanol

  32. EVERGREEN (LIFE) Project LIFE13 ENV/IT/000461 PRELIMINARY CONCLUSIONS Agriculturalwasteshavethepossibility to acting as “factory” of polyphenols and antioxidants Poliphenolsextraction, fractionation, and analysis. In addition, wastesafterextractioncould be usedforbiomethanization, and as organicamendments: EVERGREEN: “ZERO residues”

  33. EVERGREEN C2 ACTIVITY (FROM APRIL 2015) In preparation Monitoring of theabsences of sideeffectsforthehighqualitypolyfhenolicpreparationsoncommon targets of the living organisms at laboratorylevel. Project LIFE13 ENV/IT/000461 CEBAS EXPERIMENT “Effect of polyphenolsadditiononsomepathogenmicroorganisms and nematodes in soils”. BIOSTIMULANT AND BIOPESTICIDE EFFECT SOIL + POLYPHENOLS SOIL + Pathogen (bacteria and nematicide) SOIL + POLYPHENOLS + Pathogen CONTROL SOIL

  34. EVERGREEN (LIFE) B4 Act. Effects (biostimulant, biopesticide) of polyphenolsonplant B4 ACTIVITY (FromApril 2015) In preparation

  35. CEBAS-CSIC: PLANTS EXPERIMENT WITH OLIVE AND KIWI (BACTERIA INOCULATION) CEBAS

  36. EVERGREEN (LIFE) B4 PLANT KIWI AND OLIVE

  37. EVERGREEN (LIFE) B4 ACT PLANT KIWI AND OLIVE (PREPARATION)

  38. EVERGREEN (LIFE) B4 ACT. TOBACCO PLANTS (PREPARATION)

  39. EVERGREEN (LIFE) • ACTIVITY D (DISSEMINATION) • MATERIALS FOR DISSEMINATION • Notebooks for EVERGREEN (Done) • POSTER (Done) • NOTICE BOARD (Done) • PREPARATION FOR FAIR AND EVENT (in preparation) • Cap of coffeefor EVERGREEN (May, 2015) • EVENTS (Future): • Meeting: agriculturalstakeholder (September, 2015) • Meeting: environmentpolicies (December, 2015) • Sometechnical meeting from CSIC

  40. EVERGREEN (LIFE). D-ACT. NOTICE BOARD: EVERGREEN FROM CEBAS

  41. CEBAS CEBAS WORKING GROUP THANK YOU FOR YOUR ATTENTION

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