ACTIVITY OF CONSORTIUM INSTM Apr. 1, 2015 – Oct. 31, 2015 Technical Meeting Sergio Miele, Enrica Bargiacchi – EU project LIFE13 ENV IT / “EVERGREEN” “Environmentally friendly biomolecules from agricultural wastes as substitutes of pesticides for plant diseases control”

Evergreen: Targets and activities 1.Demonstrating the in vitro and in vivo efficacy and reliability of polyphenolic- based biomolecules extracted from agricultural non-food biomass and fresh vegetable waste from ready-to-eat produce processors as plant biostimulants and disease control products against phytopathogenic bacteria and nematodes, to replace current agrochemicals, included copper salts, in agriculture 2.Greater sustainability and lower energy-intensive crop production cycles 3.Lower agrochemical residue hazard in food and pollution in the environment, with less chance for resistance insurgence mechanisms 4.Reduction of point-source pollution from disposal of vegetable waste in soil 5.Projecting new process of non-food biomass and fresh vegetable waste where these materials undergo to a polyphenol extraction step, and then are exploited to produce new materials, and energy (i.e. anaerobic digestion) Sweet Chestnut (Castanea sativa Mill.) is an example of a new process cycle to exploit biomass potentials, according to INSTM research

Evergreen: Castanea sativa extracts WATER EXTRACTION REVERSE OSMOSIS & NANO- FILTRATION STANDARDIZED EXTRACTS PRE- TREATMENTS FEED FIBER (1% TANNINS) MULCHING TRACED SOURCE 1 Courtesy: Gruppo Mauro Saviola

Evergreen: Castanea sativa extracts uses 1) Microgranulated starter fertilizers for application at planting/transplanting with Castanea sativa and other polyphenols extracts: a plant biostimulant for boosting plant earliness and use of soil nutrients and water Courtesy: Color Glass S.p.A. 2) Castanea sativa and other polyphenols extracts in fertigation: a plant biostimulant to increase plant resistance to nematodes and pathogens Courtesy: Fattoria Autonoma Tabacchi

ACTIVITY FOR THE PROJECT Month 7-14 B1 / B7 – Demonstration of the performances of traditional pesticides for the control of bacterial and nematode diseases of plants important for the EU (2 nd CYCLE EXPERIMENT-June/October 2015) – Demonstration of the in vivo performances of the high quality polyphenolic bioactive preparations, recovered from vegetable not edible biomass and waste, at pilot scale level in field screenings (2 nd CYCLE EXPERIMENT-June/October 2015) B4 - Demonstration of the biological activity of the high quality polyphenolic extracts recovered from not edible biomass and waste, against plant pathogenic bacteria and nematode, in planta (Pot experiment in progress since Oct. 21, 2015) B7 – Preliminary activity: Microgranule formulations (April-May 2015) C4 – Monitoring of the short term environmental benefits from the use of the high quality standardizes polyphenolic preparations in plant disease control at pilot scale level in field screenings: Field soil sampling for residues and nematodes (October 2015) D7 - Articles and press releases D10 - International conferences and fairs

FIELD EXPERIMENT 2015: OXYMIL WAS UNAVAILABLE the increasing trend toward a concentration of R&D for top used agrochemicals may represent a threat for world agriculture, because less a.i. and production sites are available, while pest resistance and accidental events can occur worldwide; the necessity to develop alternative strategies for crop protection, less relying on bottle-neck agrochemicals, is therefore a must, to reduce market and food supply uncertainty. problems in Jan at the production site, the US manufacturer DuPont’s La Porte factory near Houston, Texas  no Oxymil worldwide

Tobacco Virginia Bright, field grown, single plot 500 m 2, June-Oct cv PVH 2310 (low tolerance to Meloidogyne arenaria, and resistant to Meloidogyne incognita race 1 and 3) Treatments B1 – Demonstration of the performances of traditional pesticides for the control of bacterial and nematode diseases of plants important for the EU (2 nd CYCLE EXPER.)

Conclusions – CHT: acts as a biostimulant, with minimal effects on nematode count; low evidence of nematode feeding on tobacco roots was found. – Better yield results for CHT, then the biological agent Flocter, then agrochemicals B1 – Demonstration of the performances of traditional pesticides for the control of bacterial and nematode diseases of plants important for the EU (2 nd CYCLE EXPER.)

B4 - Demonstration of the biological activity of the high quality polyphenolic extracts recovered from not edible biomass and waste, against plant pathogenic bacteria and nematode, in planta Tobacco Virginia Bright, greenhouse pot test, beginning Oct.21 st, 2015 Treatments – CHTNon-infested control – EtoprofosInfested control – CHT+Marc Meal (50%+50%)

B7 - Demonstration of the in vivo performances of the high quality poly-phenolic bioactive preparations, recovered from vegetable not edible biomass and waste, at pilot scale level in field screenings (preliminary activity: April-June 2015) Preliminary activity consisted in preparing at industrial pilot level* formulations for field application of Chestnut tannins, to be used in 2015 growing season: – A microgranulated fertilizer for application at sowing/transplanting along the row (pilot plant) – A fully and easily, non-foamy, wettable tannin extract powder for application in microirrigation (pilot plant) – A microgranulated composition of CHT + Marc

MAJOR MICROGRANULATED FERTILIZERS WITH TANNIN EXTRACTS %MICRO TANMICRO NP Tannin extract1812 Nitrogen (N)07 Phosphate (P 2 O 5 )030 Total Iron (Fe)32 Complex Tannin-Fe (Fe) Granule Size mm (> 80%) 100% mm 0.8 Authorization: Italian D.Lgs. 75/2010 as Iron-Tannin complex fertilizer and Nitrogen-Phosphate Fertilizer w/ Iron-Tannin complex

CORRECTIVES-VARIOUS CORRECTIVES * DRY TANNINS EXTRACT WP pHw3.0 – 3.5 ORGANIC CARBON (C)MIN. 50 % TANNIN CONTENTMIN. 73 % * Authorization: D.M , publ. on Italian Official Gazette n° 215 Sept. 15, 2011 (modifications of D.Lgs. 75/2010)

INSTM ACTIVITY FOR THE PROJECT Month 7-13 D2 – Notice boards & information displayed on INSTM website ( Link: at the Chestnut Tannin production site, and at the Tobacco experiment site; D3 Demonstration workshop in Italy  Visit to Tobacco experiments, and Ch Tannin microgranulate fertilizer production site (July 17, 2015) D8 – Networking: with Regione Umbria’s PSR funded Project QualiTaba (Traced quality of tobacco in a context of sustainable production in Central Italy) D10 - International conferences and fairs -Presentation at EXPO2015-Milano (June 25, 2015) of EVERGREEN -Oral presentation at CORESTA Agro-Phyto Meeting, Izmir (Turkey), Oct , 2015 (presentation on CORESTA WEB) D11 – Dissemination to institutions and policy makers  Event at Accademia dei Georgofili (Nov. 13, 2015)