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משרד החינוך והמזכירות הפדגוגית האגף לתכנון ופיתוח תכניות לימודים האוניברסיטה העברית בירושלים המרכז להוראת המדעים מנהלת מל"ם המרכז הישראלי לחינוך מדעי טכנולוגי ע"ש עמוס דה שליט המרכז הארצי למורי הביולוגיה המרכז הארצי של מורי הביולוגיה ומורי מדעי הסביבה בשיתוף עמותת בשער שימוש בביופחם והשפעתו על צמחים פרופ' יגאל אלעד ,המחלקה לפתולוגיה של צמחים וחקר העשבים ,מכון וולקני מתנדב בעמותת "בשער" 16.12.12 Climate Change Since the industrial revolution, atmospheric levels of greenhouse gases have been rising, mainly due to burning of fossil fuels. With them, mean global temperatures have been rising. Application of biochar to soil results in long term sequestration of fixed carbon •Biochar use agricultural plants Soil fertility 2006 Biochar applied once in 2003 Colombian Llanos (N=3) Major, J., M. Rondon, D. Molina, S.J. Riha and J. Lehmann, 2010. Maize yield and nutrition during 4 years after biochar application to a Colombian savanna oxisol. Plant and Soil 333, 117-128. Value of soil application of Biochar Increases soil fertility nutrient retention water holding capacity cation exchange capacity Increases beneficial soil microorganisms Reduces nutrient runoff, absorbs ammonia and ammonium Absorbs pesticides and soil toxins Reduces agricultural N2O and CH4 emissions 6 Pepper growth 0 3 5 240 200 Biochar (%) 0 1 a 3 a b b 160 120 65 70 75 80 85 Time after planting (days) Fruit weight (g/plant) Fruit weight (g/fruit) 440 ± 21 b 83 ± 7 b 565 ± 36 b 97 ± 5 a 732 ± 58 a 104 ± 4 a Graber et al. 2010, Plant and Soil Pepper canopy dry weight (g) BIOCHAR (%) Normalized pepper leaf area (%) Plant growth promotion under optimal fertigation a 19 17 b c 151 0 0 3 5 Biochar Content (%) Normalized tomato plant height (%) 600 a 400 a 200 b a a b b b Biochar (%) 0 1 3 35 45 55 65 Time after planting (days) Graber et al. 2010, Plant and Soil Normalized tomato leaflet area (%) Tomato growth 300 Biochar (%) 0 1 3 200 a a ab ab b b a ab b 100 45 55 65 Time after planting (days) נביטת חיטה Germination (%) 100 75 50 25 0 1 0 0 4 8 12 Time after planting (days) Height (cm) 60 40 20 1 % GHW 0 0 0 10 20 30 Time after planting (days) Biochar Increases soil fertility: nutrient retention, water holding capacity, cation exchange capacity Increases beneficial soil microorganisms Reduces nutrient runoff Absorbs pesticides and soil toxins Reduces agricultural N2O and CH4 emissions Biochar has a beneficial effect on plants also on the background of full fertilization What other effects? Disease severity (% leaf coverage) 20 a 10 0 0 a 0 3 2 4 b b 6 20 10 Effect of soil biochar application on leaf gray mold disease 8 Result: Suppression of gray mold on tomato and sweet pepper leaves a a a 0 b b b 3 0 0 2 4 6 Time after infection (days) Biochar AUDPC±SE (%) (%*days) 0 42.3±8.3 a 3 10.5±3.8 b Elad et al, Phytopathology 2010 Citrus wood biochar Effect of soil biochar application on tomato leaf powdery mildew 100 75 Upper leaves Biochar (%) Disease severity (% leaf coverage) 50 25 0 15 a b 30 45 60 0 1 3 AUDPC±SE (%*days) 269±19 a 57±10 10± 3 )Leveillula taurica( b b 100 75 a Middle leaves 50 25 0 15 100 b 30 45 a a b b 60 Lower leaves 75 0 1419±173 a 1 164± 39 b 3 106± 22 b 50 25 0 15 0 801±74 1 112±24 3 70±16 b 30 45 Citrus wood biochar 60 Time after infection (days) Elad et al, Phytopathology 2010 Effect of soil biochar application on pepper leaf powdery mildew b Leaf height Upper a b Middle a Lower b 0 Elad et al., 2010 Phytopathology a Biochar (%) 0 5 10 20 Disease severity (% leaf coverage) Result: Suppression of powdery mildew on tomato and sweet pepper leaves b 40 a b 20 b a 1 3 Biochar concentration in soil (%) Disease severity (% leaf coverage) Long term effect of soil application on leaf powdery mildew 80 a 0 60 a b 0 Biochar concentration in soil (%) 30 55 80 105 Time after infection (days) 3 b 1 b a 0 0 600 Disease severity Elad et al, Phytopathology 2010 1200 1800 AUDPC (%*days) Biochar concentration in soil (%) Broad Mite in Pepper 5 b b 3 0 a 0 5 10 15 Severity (%) 20 Control No biochar Elad et al, Phytopathology 2010 3% biochar )Polyphagotarsonemus latus( אקרית העיוותים Effect of parameters of biochar treatment : Biomass Biochar source concentration -------------Time after Pyrolysis Plants age infection by temperature Disease (time after pathogen assay biochar (days) treatment) Biochar concentration (%) Effect of biochar treatment parameters: Pyrolysis temp. C Greenhouse waste (GHW) 3.0 1.0 0.5 3.0 1.0 0.5 - b b c a GHW a 0 20 40 Disease severity )%( Elad et al, Phytopathologia Mediterranea (2012) 60 Plants age Time after infection (d.a.p) by Botrytis cinerea Assay (days) 350 AL 29 12 450 AL 72 14 Pyrolysis temp. C Biochar concentration (%) Olive pomice (OP) 3.0 1.0 0.5 0 b 20 3.0 1.0 0.5 0 40 OP a 10 20 b 10 DL 23 7 450 DL 16 10 450 AL 21 5 30 ab 5 350 60 b 3.0 1.0 0.5 0 OP a Plants age Time after infection (d.a.p) by Botrytis cinerea Assay (days) 15 Disease severity )%( Elad et al, Phytopathologia Mediterranea (2012) OP a Biochar concentration (%) Pyrolysis temp. C b 3.0 1.0 0.5 - c (EW) ab a Eucalyptus wood (EW) 3.0 1.0 0.5 - b 0 20 Plants age Time after infection (d.a.p) by Botrytis cinerea Assay (days) 350 DL 23 7 350 AL 47 10 a 40 60 Disease severity )%( Results: Suppression of foliar diseases in all cases Efficacy may differ with the biochar (temperature and biomass source) Three weeks of incubation are needed •Plant pathogens infect foliar plant parts •Biochar applied to soil (root zone) Induced resistance (systemic) What plant genes may be responsible for the effect of biochar on plant diseases? Systemic resistance pathways PRs WRKY (FaWRKY1)Transcription factors WRKY involved in plant responses to biotic and abiotic stresses PR1 PR1 (FaPR1) SA dependent related marker PR5 PR5 (Faolp2) Osmotin-Like (down regulated by ABA, induced by SA and wounding) PR10 Priming for defenses Necrotrphs Biotrphs (Fra a3) induced under stress and act as common allergens BIOCHAR ? LOX Lox (Falox) Lipoxygenase, central in JA signaling, involved in ISR QPCR מדידת כמות יחסית על ידי Control Extraction total RNA Reverse Transcriptase Treatment cDNA QPCR Normalisation with house keeping gene (GAPDH) Relative quantification/control plant Relative mRNA level (Fold Change) Effect of biochar amendments on expression of defense related genes in strawberry plants 210 Greenhouse waste (3 wks) A Biochar conc. (%) 0 1 3 180 150 15 160 110 15 10 5 5 FaPR1 a3 Falox PR1 Faolp2 PR5 Fra PR10 Lox FaWRKY1 WRKY SAR ISR Citrus wood 9 mths 135 10 0 B 0 a3 Falox FaPR1 PR1 Faolp2 PR5 Fra PR10 Lox FaWRKY1 WRKY SAR ISR Total RNA was isolated from leaves and subjected to qPCR analysis. Transcripts levels were normalized to FaGAPDH and 18-26S interspacer RNA genes and expressed relative to no biochar amendment plants using the Δ Δ Ct Ct method. Meller Harel et al, 2011 Enhanced resistance to Gray mold in strawberry plants grown on biochar Biochar, conc. (%) AUDPC ±SE 415.8± 157.1± 110.0± 117.8± Disease severity (rot area, mm2) 0 Greenhouse waste (GHW) 1 GHW 3 Citrus wood (CW) 3 50 ba 0 0 4 a b b 7 9 Time after inoculation (days) Meller Harel et al, 2011 a b b b a B. cinerea 100 50.5 33.0 32.0 24.3 after 9 days: Relative mRNA level (Fold Change) Effect of Botrytis cinerea infection on expression of defense related genes (3 weeks GHW biochar + 7 days post inoculation) 150 Botrytis cinerea + 40 30 100 20 Biochar conc. (%) 1 1 3 3 B. cinerea + + 50 10 0 FaPR1 Faolp2 a3 Falox PR1 PR5 Fra PR10 Lox FaWRKY1 WRKY SAR Meller Harel et al, 2011 ISR 0 FaPR1 Fra a3 Falox PR1 Faolp2 PR5 PR10 Lox FaWRKY1 WRKY SAR ISR Enhanced resistance to Powdery mildew in strawberry plants grown on biochar Biochar, conc. (%) AUDPC ±SE 0 Citrus wood (CW) 1 CW 3 - 80 Disease severity (% leaf coverage) 1373.4± 84.1 a 1129.7±181.2 a 720.4± 87.7 b - a P. aphanis a b 40 a a 0 10 b b b 0 20 No biochar 30 40 Time after infection (days) Meller Harel et al, 2011 after 28 days: 3% CW Relative mRNA level (Fold Change) Effect of Podosphaera aphanis infection on expression of defense related genes (9 momths with biochar + 28 days after inoculation) Podosphaera aphanis + 100 85 Biochar P. conc. (%) aphanis 160 135 1 1 3 3 110 60 + + 70 10 30 5 0 FaPR1 Fra a3 Falox PR1 Faolp2 PR5 PR10 Lox FaWRKY1 WRKY SAR ISR 0 FaPR1 PR1 Faolp2 Fra a3 Falox PR5 PR10 Lox FaWRKY1 WRKY SAR Meller Harel et al, 2011 ISR Systemic resistance pathways BIOCHAR •נקבוביות הביופחם משמשת מקלט למקרואורגניזמים ייחודיים •חומרים המצויים בביופחם מעודדים מקרואורגניזמים מסויימים ומעכבים אחרים Stimulation of beneficial soil microbial consortium General bacteria Rhizosphere Growth medium 1*107 1*108 3 1 0 1*109 Rhizosphere Biochar conc.)%( Bacillus spp. Growth medium 1 10 100 1,000 Rhizosphere Trichoderma spp. Growth medium 1 1*102 Population (CFU/g) 1*104 oBiochar improves plants growth. oBiochar induces in plants resistance towards pathogens. oChanges in microbial populations. oBiochar-borne chemicals and/or biochar-induced microbial populations are responsible for these activities. Research in cooperation with Institute of Plant Protection Yigal Elad Omer Frenkel Yael Meller-Harel Dalia Rav David Menahem Borenshtein Ran Shulchani Sergey Segal Amit Kumar Jaiswal Zeraye Mehari Haile Institute of Soil, Water and Environmental Sciences Ellen R. Graber Eddie Cytryn Max Kolton Avner Silber Larissa Kautsky Maya Ofek Dror Minz Hebrew University Faculty of Agriculture Inst. of Agricultural Engineering Zohar Pasternak Beni Lew http://www.agri.gov.il/pages/788.aspx Cited publications •Elad, Y., Rav David, D., Meller Harel, Y., Borenshtein, M., Ben Kalifa, H., Silber, A. and Graber, E.R. (2010) Induction of systemic resistance in plants by biochar, a soil-applied carbon sequestering agent. Phytopathology 100:913-921. •Graber, E.R., Meller Harel, Y., Kolton, M., Cytryn, E., Silber, A., Rav David, D., Tsechansky, L., Borenshtein, M., Elad, Y. (2010) Biochar impact on development and productivity of pepper and tomato grown in fertigated soilless media. Plant and Soil 337:481– 496. •Kolton, M., Meller Harel, Y., Pasternak, Z., Graber, E.R., Elad, Y. and Cytryn, E. (2011) Impact of biochar application to soil on the root-associated bacterial community structure of fully developed greenhouse pepper plants. Applied and Environmental Microbiology 77:4924-4930. •Elad, Y., Cytryn, E., Meller Harel, Y., Lew, B., and Graber, E.R. (2011) The Biochar Effect: plant resistance to biotic stresses. Phytopathologia Mediterrenea 50: 335−349.