AFTER-Cu AFTER-Cu LIFE (2014-2016) Carlos García Izquierdo. CEBAS-CSIC FAENZA, JANUARY 2015 PARTNER...

AFTER-Cu AFTER-Cu LIFE (2014-2016)

Carlos García Izquierdo. CEBAS-CSIC

FAENZA, JANUARY 2015FAENZA, JANUARY 2015PARTNER CSIC (CEBAS-CSIC)PARTNER CSIC (CEBAS-CSIC)

Project LIFE12 ENV/IT/000336“Anti-infective environmental friendly

molecules against plant pathogenic bacteria for reducing Cu" (AFTER-Cu)

CEBAS-CSIC (Murcia, CEBAS-CSIC (Murcia, Spain)Spain)

CSIC

Centro de Edafología y Biología Aplicada del Segura Centro de Edafología y Biología Aplicada del Segura

OUR INSTITUT: CEBAS-CSIC

CEBASCEBAS

CAMPUS UNIVERSITARIOCAMPUS UNIVERSITARIO

Dr. Carlos GarciaDr. Carlos GarciaDra. Teresa HernándezDr. J. MorenoLda. Mª Dolores CollIng. Carmen Chocano

CEBAS-CSIC (AFTER-Cu)CEBAS-CSIC (AFTER-Cu)

AFTER-Cu (LIFE)AFTER-Cu (LIFE)

EXPERIMENTAL GREENHOUSES (SANTOMERA-ABARÁNEXPERIMENTAL GREENHOUSES (SANTOMERA-ABARÁN

CEBAS WORKING-GROUPCEBAS WORKING-GROUP(AFTER-Cu)(AFTER-Cu)

““EFFECT OF CuSOEFFECT OF CuSO44 ON ON

SOIL-PLANT SYSTEMSOIL-PLANT SYSTEM””


DYNAMIC OF CONTAMINANTS IN SOIL-WATER DYNAMIC OF CONTAMINANTS IN SOIL-WATER

WATER TABLE

RUNOFF

PHOTODEGRADAtION

LIX

IVIA

CIO

N

ADSORPTION

VO

LA

TIL

IZA

CIÓ

N

CHEMICAL

DEGRADATION

BIODEGRADATIONDESORPTION

INF

ILT

RA

CIO

N

CAPILARIDADCAPILARIDAD

ABSORCION YEXUDACION

TRANSFERTRANSFER

ADSORTION-DESORPTION ADSORTION-DESORPTION (Cu)(Cu)LEACHINGLEACHING

VOLATILISATIONVOLATILISATION

RUNOFRUNOF

ABSORTION BY PLANTS AND ORGANISMS ABSORTION BY PLANTS AND ORGANISMS (Cu)(Cu)

TRANSFORMATI0NTRANSFORMATI0N

CHEMICAL CHEMICAL (Cu)(Cu)PHOTOCHEMICALPHOTOCHEMICAL

BIOLOGICAL BIOLOGICAL (Cu)(Cu)

AgricultureAgriculture

IndustrialIndustrial

DomesticDomestic

WastesWastes

HEAVY METALSHEAVY METALS““Cu”Cu”

CEBAS-CSIC EXPERIMENT (AFTER-Cu Project)CEBAS-CSIC EXPERIMENT (AFTER-Cu Project)

““EFFECT OF COPPER ON SOIL-PLANT SYSTEMS AT FIELD EFFECT OF COPPER ON SOIL-PLANT SYSTEMS AT FIELD LEVEL (DIFFERENT CROPS: kiwi, lemon, olive)LEVEL (DIFFERENT CROPS: kiwi, lemon, olive)

OBJECTIVEOBJECTIVE: to demonstrate the environmentally negative impact of to demonstrate the environmentally negative impact of COOPER on SOIL-PLANT SYSTEMSCOOPER on SOIL-PLANT SYSTEMS. It will be demonstrated at field level . It will be demonstrated at field level and on different crops and on different crops (kiwi, lemon and olive)(kiwi, lemon and olive)


CEBAS-CSIC EXPERIMENTCEBAS-CSIC EXPERIMENT

Greenhouse (kiwi, olive, lemon)Greenhouse (kiwi, olive, lemon)

Bacteria inoculation (Bacteria inoculation (pseudomona)pseudomona)

1) Cu addition (environmental problem)1) Cu addition (environmental problem)

2) Peptides use (clean agriculture)2) Peptides use (clean agriculture)

AFTER –Cu PROJECTAFTER –Cu PROJECT

CEBAS-CSICCEBAS-CSIC: PLANT EXPERIMENT WITH CITRUS, OLIVE AND KIWI: PLANT EXPERIMENT WITH CITRUS, OLIVE AND KIWI(BACTERIA INOCULATION: pseudomona)(BACTERIA INOCULATION: pseudomona)

http://www.google.es/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&docid=ED-h60Dg-vIKxM&tbnid=frVaMIjk7rtD2M:&ved=0CAUQjRw&url=http://www.guidaortogiardino.com/00116_innesti-attrezzatura.html&ei=Ep4EU9vvCIX80QXLw4GoDQ&bvm=bv.61535280,d.d2k&psig=AFQjCNExKbXC12o2vjKOqW2pP3yxp2Q55g&ust=1392897865415966

http://upload.wikimedia.org/wikipedia/commons/5/59/Pseudomonas_syringae_cultures.jpg


““In vitro” EFFECT OF BACTERIA INOCULATIONIn vitro” EFFECT OF BACTERIA INOCULATIONKiwi leaves infected by bacteriaKiwi leaves infected by bacteria

11 K Ps 5+ CuSO4-K Ps 5+ CuSO4- 2121 O Ps 6+ CuSO4+O Ps 6+ CuSO4+ 4141 K Ps 4+ CuSO4-K Ps 4+ CuSO4-

22 O Ps 10_ CONTROLO Ps 10_ CONTROL 2222 K Ps 6+ CuSO4-K Ps 6+ CuSO4- 4242 K Ps 8+ CuSO4-K Ps 8+ CuSO4-

33 O Ps 3+ CuSO4+O Ps 3+ CuSO4+ 2323 O Ps 8-CuSO4-O Ps 8-CuSO4- 4343 K Ps 2+ CuSO4-K Ps 2+ CuSO4-

44 K Ps 6+ CuSO4+K Ps 6+ CuSO4+ 2424 K Ps 8+ CuSO4+K Ps 8+ CuSO4+ 4444 O Ps 7-CuSO4-O Ps 7-CuSO4-

55 K Ps 7+ CuSO4+K Ps 7+ CuSO4+ 2525 K Ps 3+ CuSO4+K Ps 3+ CuSO4+ 4545 K Ps 10+ CuSO4+K Ps 10+ CuSO4+

66 K Ps 9+ CuSO4+K Ps 9+ CuSO4+ 2626 K Ps 5+ CuSO4+K Ps 5+ CuSO4+ 4646 K Ps 1+ CuSO4+K Ps 1+ CuSO4+

77 C Ps 4_ CONTROLC Ps 4_ CONTROL 2727 K Ps 4+ CuSO4+K Ps 4+ CuSO4+ 4747 K Ps 2+ CuSO4+K Ps 2+ CuSO4+

88 C Ps 6_ CONTROLC Ps 6_ CONTROL 2828 C Ps 2_CONTROLC Ps 2_CONTROL 4848 C Ps 1+ CuSO4-C Ps 1+ CuSO4-

99 C Ps 5_CONTROLC Ps 5_CONTROL 2929 C Ps9_CONTROLC Ps9_CONTROL 4949 C Ps 3+ CuSO4-C Ps 3+ CuSO4-

1010 O Ps 2_CONTROLO Ps 2_CONTROL 3030 O Ps 6+ CuSO4+O Ps 6+ CuSO4+ 5050 C Ps 10+ CuSO4-C Ps 10+ CuSO4-

1111 O Ps 1_CONTROLO Ps 1_CONTROL 3131 O Ps 5+ CuSO4+O Ps 5+ CuSO4+ 5151 C Ps 7+CuSO4-C Ps 7+CuSO4-

1212 O Ps 4_CONTROLO Ps 4_CONTROL 3232 O Ps 3+CuSO4-O Ps 3+CuSO4- 5252 C Ps 8+CuSO4-C Ps 8+CuSO4-

1313 O Ps 9_CONTROLO Ps 9_CONTROL 3333 K Ps 10_CONTROLK Ps 10_CONTROL 5353 K Ps 7_CONTROLK Ps 7_CONTROL

1414 KPs 1_ CONTROLKPs 1_ CONTROL 3434 K Ps 9_CONTROLK Ps 9_CONTROL 5454 K Ps 3_CONTROLK Ps 3_CONTROL

1515 O Ps 10+ CuSO4+O Ps 10+ CuSO4+ 3535 O Ps 5- CuSO4+O Ps 5- CuSO4+ 5555 O Ps 2+CuSO4-O Ps 2+CuSO4-

1616 O Ps 7+ CuSO4+O Ps 7+ CuSO4+ 3636 O Ps 9- CuSo4+O Ps 9- CuSo4+ 5656 O Ps 1+CuSO4-O Ps 1+CuSO4-

1717 O Ps 4+ CuSO4+O Ps 4+ CuSO4+ 3737 C Ps 7+ CuSO4+C Ps 7+ CuSO4+ 5757 C Ps 1+ CuSO4+C Ps 1+ CuSO4+

1818 O Ps 8+ CuSO4+O Ps 8+ CuSO4+ 3838 C Ps 2+ CuSO4+C Ps 2+ CuSO4+ 5858 C Ps 4+ CuSO4+C Ps 4+ CuSO4+

1919 C Ps 9+ CuSO4+C Ps 9+ CuSO4+ 3939 C Ps 3+ CuSO4+C Ps 3+ CuSO4+ 5959 C Ps 5+ CuSO4+C Ps 5+ CuSO4+

2020 C Ps 8+ CuSO4+C Ps 8+ CuSO4+ 4040 C Ps 6+ CuSO4+C Ps 6+ CuSO4+ 6060 C Ps 10+ CuSO4+C Ps 10+ CuSO4+

K= kiwifruit K= kiwifruit PsPs++= infected plant with pseudomone= infected plant with pseudomoneL= lemon L= lemon PsPs--= not infected plant with pseudomone = not infected plant with pseudomone O= Olive O= Olive CuSOCuSO4+4+= plant/soil treated with CuSO= plant/soil treated with CuSO4+4+ at 12k/Ha/year at 12k/Ha/yearControl= distilled water Control= distilled water CuSOCuSO4-4-= plant/soil not treated with CuSO= plant/soil not treated with CuSO4+4+ at 12k/Ha/year at 12k/Ha/year

AFTER-Cu (LIFE)AFTER-Cu (LIFE)GREENHOUSE PLANTS EXPERIMENT GREENHOUSE PLANTS EXPERIMENT

CONTROLCONTROL Ps+ CuSO4-Ps+ CuSO4- Ps+ CuSO4+Ps+ CuSO4+

P (g/100g)P (g/100g) 0,050a0,050a 0,051 a0,051 a 0,054 a0,054 a

K (g/100g)K (g/100g) 0,623 a0,623 a 0,643 a0,643 a 0,715 a0,715 a

Ca (g/100g)Ca (g/100g) 9,595 a9,595 a 11,280 a11,280 a 12,758 a12,758 a

S (g/100g)S (g/100g) 0,281 a0,281 a 0,311 a0,311 a 0,354 a0,354 a

Mg (g/100g)Mg (g/100g) 0,637 a0,637 a 0.716 a0.716 a 0,760 a0,760 a

Na (g/100g)Na (g/100g) 0,130 a0,130 a 0,146 a0,146 a 0,162 a0,162 a

Fe (mg/Kg)Fe (mg/Kg) 788,97 a788,97 a 9281,05 a9281,05 a 9484,84 a9484,84 a

Mn (mg/Kg)Mn (mg/Kg) 290,130 a290,130 a 325,20 a325,20 a 344,83 a344,83 a

Cu (mg/Kg)Cu (mg/Kg) 25.94 a25.94 a 24.,72 a24.,72 a 63.95 b63.95 b

Cd (mg/Kg)Cd (mg/Kg) 0,224 a0,224 a 0,246 a0,246 a 0,268 a0,268 a

Zn (mg/Kg)Zn (mg/Kg) 30,589 a30,589 a 34,331 a34,331 a 35,893 a35,893 a

Pb (mg/Kg)Pb (mg/Kg) 10,538 a10,538 a 12,256 ab12,256 ab 13,364 b13,364 b

Cr (mg/Kg)Cr (mg/Kg) 18,662 a18,662 a 21,485 a21,485 a 22,84422,844

Ni (mg/Kg)Ni (mg/Kg) 10,195 a10,195 a 12,184 a12,184 a 12,660 a12,660 a

For each parameter, different letters indicate significant differences between treatments according to Tukey test (p<0,05).For each parameter, different letters indicate significant differences between treatments according to Tukey test (p<0,05).

Kiwifruit_ T6 monthsKiwifruit_ T6 months

Concentrations of macro- and micronutrients and heavy metals in kiwi soils (dwt)Concentrations of macro- and micronutrients and heavy metals in kiwi soils (dwt)


Risk using CuSORisk using CuSO44 Soils showed > Cu concentration when CuSOSoils showed > Cu concentration when CuSO44 was used as bactericide. was used as bactericide. It could be negative on soil enzyme, It could be negative on soil enzyme, reducing the production of enzymes reducing the production of enzymes through its toxic effect on soil microflorathrough its toxic effect on soil microflora

CONTROL Ps+ CuSO4- Ps+ CuSO4+

P (g/100g)P (g/100g) 0,146 a0,146 a 0,220 a0,220 a 0,205 a0,205 a

K (g/100g)K (g/100g) 2,686 a2,686 a 3,535 a3,535 a 3,302 a3,302 a

Ca (g/100g)Ca (g/100g) 1,116 a1,116 a 2,130 a2,130 a 1,932 a1,932 a

S (g/100g)S (g/100g) 0,438 a0,438 a 0,630 a0,630 a 0,585 a0,585 a

Mg (g/100g)Mg (g/100g) 0,244 a0,244 a 0,422 b0,422 b 0,380 b0,380 b

Na (g/100g)Na (g/100g) 0,072 a0,072 a 0,195 a0,195 a 0,102 a0,102 a



Cu (mg/Kg)Cu (mg/Kg) 8,582 a8,582 a 13,182 a13,182 a 25.62 b25.62 b

Cd (mg/Kg)Cd (mg/Kg) <0,1 a<0,1 a <0,1 a<0,1 a <0,1 a<0,1 a


Pb (mg/Kg)Pb (mg/Kg) 6,190 a6,190 a 10,392 a10,392 a 14,237 a14,237 a

Cr (mg/Kg)Cr (mg/Kg) 1,116 a1,116 a 3,247 a3,247 a 3,987 a3,987 a



Concentrations of macro- and micronutrients and heavy metals in kiwi leaves (dwt)Concentrations of macro- and micronutrients and heavy metals in kiwi leaves (dwt)


Risk using CuSORisk using CuSO44. . Plants where CuSOPlants where CuSO44 was used as bactericide showed high was used as bactericide showed high quantity of Cu in leaves. It could be negative for fruits and quantity of Cu in leaves. It could be negative for fruits and alsoalso for human health for human health

CONTROLCONTROL Ps+ CuSO4-Ps+ CuSO4- Ps+ CuSO4+Ps+ CuSO4+P (g/100g)P (g/100g) 0,054 a0,054 a 0,052 a0,052 a 0,054 a0,054 a

K (g/100g)K (g/100g) 0,698 b0,698 b 0,550 a0,550 a 0,557 a0,557 aCa (g/100g)Ca (g/100g) 10,236 a10,236 a 14,514 a14,514 a 13,160 ab13,160 ab

S (g/100g)S (g/100g) 0,330 a0,330 a 0,278 a0,278 a 0,301 a0,301 aMg (g/100g)Mg (g/100g) 0,778 a0,778 a 0,780 a0,780 a 0,755 a0,755 a

Na (g/100g)Na (g/100g) 0,214 a0,214 a 0,130 a0,130 a 0,214 a0,214 aFe (mg/Kg)Fe (mg/Kg) 11502,810 a11502,810 a 9412,8322 a9412,8322 a 9150,808 a9150,808 a

Mn (mg/Kg)Mn (mg/Kg) 369,674 a369,674 a 350,010 a350,010 a 342,108 a342,108 aCu (mg/Kg)Cu (mg/Kg) 23,876 a23,876 a 24,900 a24,900 a 59.475 b59.475 b

Cd (mg/Kg)Cd (mg/Kg) 0,270 a0,270 a 0,294 a0,294 a 0,292 a0,292 aZn (mg/Kg)Zn (mg/Kg) 35,038 a35,038 a 37,232 a37,232 a 39,186 a39,186 a

Pb (mg/Kg)Pb (mg/Kg) 12,078 a12,078 a 11,866 a11,866 a 11,442 a11,442 aCr (mg/Kg)Cr (mg/Kg) 24,186 a24,186 a 23,296 a23,296 a 22,433 a22,433 a

Ni (mg/Kg)Ni (mg/Kg) 12,700 a12,700 a 13,858 a13,858 a 12,460 a12,460 aFor each parameter, different letters indicate significant differences between treatments according to Tukey test (p<0,05).For each parameter, different letters indicate significant differences between treatments according to Tukey test (p<0,05).

Lemon_T6 monthsLemon_T6 months

Concentrations of macro- and micronutrients and heavy metals in lemon soils (dwt)Concentrations of macro- and micronutrients and heavy metals in lemon soils (dwt)


Risk using CuSORisk using CuSO4 4

Our results obtained on lemon soils were similars to kiwi soil. Our results obtained on lemon soils were similars to kiwi soil. When CuSOWhen CuSO44 was used, high quantity of Cu in the soil was found. was used, high quantity of Cu in the soil was found. An increase of soil Ca content was observed. It could be due to An increase of soil Ca content was observed. It could be due to the bacteria inoculationthe bacteria inoculation


P (g/100g)P (g/100g) 0,094 a0,094 a 0,086 a0,086 a 0,155 a0,155 a

K (g/100g)K (g/100g) 2,270 a2,270 a 2,346 a2,346 a 2,880 a2,880 a

Ca (g/100g)Ca (g/100g) 1,368 a1,368 a 1,938 a1,938 a 1,266 a1,266 a

S (g/100g)S (g/100g) 0,420 a0,420 a 0,532 a0,532 a 0,445 a0,445 a

Mg (g/100g)Mg (g/100g) 0,218 a0,218 a 0,300 a0,300 a 0,225 a0,225 a

Na (g/100g)Na (g/100g) 0,028 a0,028 a 0,028 a0,028 a 0,080 a0,080 a



Cu (mg/Kg)Cu (mg/Kg) 7,076 a7,076 a 10,460 a10,460 a 19.16 a19.16 a

Cd (mg/Kg)Cd (mg/Kg) <0,1<0,1 <0,1<0,1 <0,1<0,1

Zn (mg/Kg)Zn (mg/Kg) 20,484 a20,484 a 31,588 a31,588 a 34,971a34,971a

Pb (mg/Kg)Pb (mg/Kg) 4,200 a4,200 a 5,772 ab5,772 ab 6,898 b6,898 b

Cr (mg/Kg)Cr (mg/Kg) 0,758 a0,758 a 1,424 b1,424 b 1,021 ab1,021 ab



Concentrations of macro- and micronutrients and heavy metals in lemon leaves(dwt)Concentrations of macro- and micronutrients and heavy metals in lemon leaves(dwt)



When CuSOWhen CuSO44 was used as bactericide on lemon crop, Cu was used as bactericide on lemon crop, Cu increased on lemon leaves. Cr and Ni also increased in leaves increased on lemon leaves. Cr and Ni also increased in leaves inoculated with bacteria. inoculated with bacteria. Heavy metal could be a risk to fruits Heavy metal could be a risk to fruits and the human healthand the human health


P (g/100g)P (g/100g) 0,050a0,050a 0,046 a0,046 a 0,047 a0,047 a

K (g/100g)K (g/100g) 0,632 a0,632 a 0,672 a0,672 a 0,716 a0,716 a

Ca (g/100g)Ca (g/100g) 8,178 a8,178 a 14,886 b14,886 b 12,684 ab12,684 ab

S (g/100g)S (g/100g) 0,280 a0,280 a 0,234 a0,234 a 0,271 a0,271 a

Mg (g/100g)Mg (g/100g) 0,624 a0,624 a 0,824 a0,824 a 0,747 a0,747 a

Na (g/100g)Na (g/100g) 0,128 a0,128 a 0,092 a0,092 a 0,097 a0,097 a



Cu (mg/Kg)Cu (mg/Kg) 19.97 a19.97 a 9.828 a9.828 a 59.552 b59.552 b

Cd (mg/Kg)Cd (mg/Kg) 0,252 a0,252 a 0,250 a0,250 a 0,262 a0,262 a




Ni (mg/Kg)Ni (mg/Kg) 10,648 a10,648 a 14,014 b14,014 b 12,798 ab12,798 ab


Olive_T6 monthsOlive_T6 months

Concentrations of macro- and micronutrients and heavy metals in olives soils (dwt)Concentrations of macro- and micronutrients and heavy metals in olives soils (dwt)AFTER-Cu (LIFE)AFTER-Cu (LIFE)


On soil olive crop, our results were similars to kiwi On soil olive crop, our results were similars to kiwi and lemon crops. When CuSOand lemon crops. When CuSO44 was used, > Cu in was used, > Cu in soil was found. Also Ca and Ni increased due to the soil was found. Also Ca and Ni increased due to the bacteria inoculation.bacteria inoculation.


P (g/100g)P (g/100g) 0,180 a0,180 a 0,173 a0,173 a 0,155 a0,155 a

K (g/100g)K (g/100g) 1,065 a1,065 a 1,156 a1,156 a 0,896 a0,896 a

Ca (g/100g)Ca (g/100g) 0,512 a0,512 a 0,620 a0,620 a 0,910 a0,910 a

S (g/100g)S (g/100g) 0,240 a0,240 a 0,230 a0,230 a 0,206 a0,206 a

Mg (g/100g)Mg (g/100g) 0,125 a0,125 a 0,140 a0,140 a 0,163 a0,163 a

Na (g/100g)Na (g/100g) 0,177 a0,177 a 0,153 a0,153 a 0,170 a0,170 a



Cu (mg/Kg)Cu (mg/Kg) 21,250 a21,250 a 14,510 a14,510 a 29.275 b29.275 b

Cd (mg/Kg)Cd (mg/Kg) <0,1<0,1 <0,1<0,1 <0,1<0,1






Concentrations of macro- and micronutrients and heavy metals in olive leaves(dwt)Concentrations of macro- and micronutrients and heavy metals in olive leaves(dwt)



When CuSOWhen CuSO44 was used as bactericide on olive crop, > Cu in was used as bactericide on olive crop, > Cu in Leaves was observed. Cr and Ni also increased in leaves Leaves was observed. Cr and Ni also increased in leaves inoculated with bacteria. inoculated with bacteria. Heavy metal could affect to fruits Heavy metal could affect to fruits and to the human healthand to the human health

Water soluble C and N Water soluble C and N is a labil fraction. For this reason, these parameters change along the is a labil fraction. For this reason, these parameters change along the experiment. Water soluble C and N are influenced by organic matter mineralzation There is experiment. Water soluble C and N are influenced by organic matter mineralzation There is not too differences between the different treatments. Only for kiwi, the CuSOnot too differences between the different treatments. Only for kiwi, the CuSO44 addition and addition and bacteria inoculation increased soluble N in soil.bacteria inoculation increased soluble N in soil.


0

100

200

300

400

500

Kiwi Lemon Olive

ControlPs-CuSO4-Ps-CuSO4+

0

10

20

30

40

50

Kiwi Lemon Olive


WATER SOLUBLE CWATER SOLUBLE C

WATER SOLUBLE NWATER SOLUBLE N

The degradation of organic matter is a property The degradation of organic matter is a property of all heterotrophs, and its rate is commonly of all heterotrophs, and its rate is commonly

used to indicate the level of microbial activityused to indicate the level of microbial activity

After 6 months of the experiment, no negative effect was After 6 months of the experiment, no negative effect was observed on observed on soil microbial respiration soil microbial respiration when Cu was added when Cu was added to the soil for lemon and kiwi. For olive, soil respiration to the soil for lemon and kiwi. For olive, soil respiration increased in control soil. A major negative effect could increased in control soil. A major negative effect could probably be observed with > Cu in soilprobably be observed with > Cu in soil


0

500

1000

1500

2000

2500

3000

3500

4000

Kiwi Lemon Olive


Microbial biomass: PhosphoLipid Fatty Acids (PLFAs)

Bligh & Dyer, 1959Frostegard et al., 1993

Bligh & Dyer, 1959Frostegard et al., 1993

PhosphoLipid Fatty Acids (PLFAs)

Microbial biomass C and N (Powlson et al., 1987)ATP, Ergosterol, etc.


Kiwi PLFA (nmol g-1)Kiwi PLFA (nmol g-1) Control T60Control T60 Ps+CuSO4-Ps+CuSO4- Ps+CuSO4+Ps+CuSO4+

BacteriaBacteria 9.349.34 9.229.22 9.919.91

Gram+ BacteriaGram+ Bacteria 6.696.69 5.695.69 6.596.59

Gram- BacteriaGram- Bacteria 2.652.65 3.533.53 3.313.31

FungiFungi 0.480.48 0.490.49 0.270.27

Total satured. PLFA Total satured. PLFA 17.8317.83 17.3417.34 16.7716.77

Total monounsat. Total monounsat. PLFA PLFA

1.361.36 1.641.64 2.032.03

ActinobacteriaActinobacteria 0.570.57 0.420.42 0.460.46

Bacterial. fungal. GramBacterial. fungal. Gram++. Gram. Gram--. satured and monosatured PLFAs concentration in kiwi soils T6 months. satured and monosatured PLFAs concentration in kiwi soils T6 months Bacterial. fungal. GramBacterial. fungal. Gram++. Gram. Gram--. satured and monosatured PLFAs concentration in kiwi soils T6 months. satured and monosatured PLFAs concentration in kiwi soils T6 months

Kiwi PLFA (nmol g-1)Kiwi PLFA (nmol g-1) Control T60Control T60 Ps+CuSO4-Ps+CuSO4- Ps+CuSO4+Ps+CuSO4+

Total PLFaTotal PLFa 19.6819.68 19.8819.88 19.1619.16

Fung/BacFung/Bac 0.050.05 0.050.05 0.030.03

Gram+/Gram-Gram+/Gram- 2.532.53 1.611.61 1.991.99

Sat PLFa/Mono PLFA Sat PLFa/Mono PLFA 13.113.1 10.5410.54 8.258.25

The total PLFA and bacteria/fungi. GramThe total PLFA and bacteria/fungi. Gram++/Gram/Gram-- and monounsatured/satured ratios in kiwi soils T6 months and monounsatured/satured ratios in kiwi soils T6 months .The total PLFA and bacteria/fungi. GramThe total PLFA and bacteria/fungi. Gram++/Gram/Gram-- and monounsatured/satured ratios in kiwi soils T6 months and monounsatured/satured ratios in kiwi soils T6 months .

PLFAs ON SOILS

Differences between control soil and soil with CuSODifferences between control soil and soil with CuSO44 addition were detected for gram- bacteria addition were detected for gram- bacteria and monounsaturated PLFA . Bacteria, fungi and actinobacteria did not show differencesand monounsaturated PLFA . Bacteria, fungi and actinobacteria did not show differences

KIWI CONTROL ( CuSO4- Ps-) Kiwi Ps+ CuSO4

- Kiwi Ps+ CuSO4+

T_6 months (27/10/2014)T_6 months (27/10/2014)T_6 months (27/10/2014)T_6 months (27/10/2014)

KIWI CONTROL KIWI CONTROL

( CuSO( CuSO44- - PsPs--))

Kiwi PsKiwi Ps+ + CuSOCuSO44-- Kiwi PsKiwi Ps+ +

CuSOCuSO44++

T_0 (07/05/2014T_0 (07/05/2014

n=5n=5 n=5n=5 n=10n=10T_6 months (27/10/2014)T_6 months (27/10/2014)

n=5n=5 n=2n=2 n=8n=8T9 monthsT9 months

n=3n=3 n=0n=0 n=2n=2

Survival kiwi plants Survival kiwi plants


Kiwi crop was affected by Kiwi crop was affected by bacteria inoculation.bacteria inoculation.CuSOCuSO4 4 seems to have aseems to have apossitive effect .possitive effect .

KIWIKIWI

Lemon CONTROL ( CuSO4-

Ps-)

Ps+ CuSO4- Ps+ CuSO4

+

KIWI CONTROL KIWI CONTROL

( CuSO( CuSO44- - PsPs--))

Kiwi PsKiwi Ps+ + CuSOCuSO44-- Kiwi PsKiwi Ps+ + CuSOCuSO44

++

T_0 (07/05/2014T_0 (07/05/2014 n=5n=5 n=5n=5 n=10n=10

T_6 months T_6 months (27/10/2014)(27/10/2014)

n=5n=5 n=5n=5 n=10n=10

T9 monthsT9 months n=5n=5 n=5n=5 n=10n=10

Survival lemon plants Lemons haven’t symptoms.

Survival lemon plants Lemons haven’t symptoms.



Lemon crop was not affected by Lemon crop was not affected by bacteria inoculation.bacteria inoculation.CuSOCuSO44 not seems to have any not seems to have any effect on lemon crop.effect on lemon crop.

LEMONLEMON

OLIVE CONTROL Olive CuSO4- Ps+ Olive CuSO4

+ Ps+

KIWI KIWI CONTROLCONTROL

( CuSO( CuSO44- -

PsPs--))

Kiwi PsKiwi Ps+ + CuSOCuSO44-- Kiwi PsKiwi Ps+ + CuSOCuSO44

++

T_0 (07/05/2014T_0 (07/05/2014 n=5n=5 n=5n=5 n=10n=10

T_6 months T_6 months (27/10/2014)(27/10/2014)

n=5n=5 n=3 with symptoms+ 2 n=3 with symptoms+ 2 without symptomswithout symptoms

n=6 with symptoms+ 4 without n=6 with symptoms+ 4 without symptomssymptoms

T9 months (13/ T9 months (13/ 01/2015)01/2015)

n=5n=5 n=3 with symptoms+ 2 n=3 with symptoms+ 2 without symptomswithout symptoms

n=6 with symptoms+ 4 without n=6 with symptoms+ 4 without symptomssymptoms

Survival olive plants Survival olive plants



Olive crop was affected by Olive crop was affected by bacteria inoculation.bacteria inoculation.CuSOCuSO4 4 have not a clearlyhave not a clearlypossitive effect .possitive effect .

OLIVEOLIVE


CONCLUSIONS TO THE EXPERIMENTCONCLUSIONS TO THE EXPERIMENT1)1) Cu is a heavy metal which can cause some problems on soil quality Cu is a heavy metal which can cause some problems on soil quality(soil enzyme activities, microbial population, soil contamination,…) (soil enzyme activities, microbial population, soil contamination,…) Soils where CuSOSoils where CuSO44 is used, Cu accumulation was found. is used, Cu accumulation was found.

2)2) Our study indicates that Cu could move from soil to plant (leaves). Our study indicates that Cu could move from soil to plant (leaves).

3)3) The use of CuSO The use of CuSO44 as bactericide should be considered as a risk to as bactericide should be considered as a risk to the environmentthe environment

4)4) Results obtained in the AFTER-Cu Project showed that kiwi is Results obtained in the AFTER-Cu Project showed that kiwi isthe crop more affected by bacteria, and where the use of CuSO4the crop more affected by bacteria, and where the use of CuSO4seems be more effective , seems be more effective ,

HIGH QUALITY ORGANIC HIGH QUALITY ORGANIC PRODUCTSPRODUCTS

(peptides, aminoacids)(peptides, aminoacids)

Antibiotics induction by organic productsCompetitive action between microorganismsEnzyme productions with biopesticide activitySystemic resistance induction for plants

Fungal supression to avoid root rooting and necrosis

Fusarium sp.Pythium sp.

Biopesticide effectBiopesticide effect

Without organic product

Organic product

NEW PRODUCTS FOR A CLEAN NEW PRODUCTS FOR A CLEAN AGRICULTURE: AGRICULTURE: BIOPESTICIDESBIOPESTICIDES

ReferencesReferences DescriptionDescription

Control Control Plant not treated. Plant not treated.

Bac Bac Plant treated with Pseudomone spp.Plant treated with Pseudomone spp.

P1_30P1_30 Plants treated with Pseudomone+ antimicrobial peptide P1 (AP17) 30 µMPlants treated with Pseudomone+ antimicrobial peptide P1 (AP17) 30 µM

P1_100P1_100 Plants treated with Pseudomone+ antimicrobial peptide P1 (Ap17) 100 µMPlants treated with Pseudomone+ antimicrobial peptide P1 (Ap17) 100 µM

P2_30P2_30 Plants treated with Pseudomone+ antimicrobial peptide P2 (Li 27) 30 µMPlants treated with Pseudomone+ antimicrobial peptide P2 (Li 27) 30 µM

P2_100P2_100 Plants treated with Pseudomone+ antimicrobial peptide P2 (Li 27)100 µMPlants treated with Pseudomone+ antimicrobial peptide P2 (Li 27)100 µM

Peptide assays T0Peptide assays T0We have performed a co-inoculation assay (Pseudomone spp.+peptide) in kiwi, lemon and olive plants in order to verify the antimicrobial capacity of two different peptides known that P1: AP17 and P2: Li 27 P1: AP17 and P2: Li 27 at two concentrations (30 and 100 µM).For this, were prepared the bacterial solutions in sterile physiological solution and were added the peptide in the corresponding quantity to the final concentration in plant 30 and 100 µM.

Each plant was infected with the corresponding Pseudomone sp:Each plant was infected with the corresponding Pseudomone sp:Kiwi: P. syringae pv. actinidaeLemon: P. syringae pv. syringaeOlive: P. savastanoi pv. Nerii

This final solution (bacteria+peptide) was sprayed on kiwi and lemon plants and in the olive case was incorporated to plant through stems wounds. The different references are showed in the following table:

Peptide assays T0Peptide assays T0We have performed a co-inoculation assay (Pseudomone spp.+peptide) in kiwi, lemon and olive plants in order to verify the antimicrobial capacity of two different peptides known that P1: AP17 and P2: Li 27 P1: AP17 and P2: Li 27 at two concentrations (30 and 100 µM).For this, were prepared the bacterial solutions in sterile physiological solution and were added the peptide in the corresponding quantity to the final concentration in plant 30 and 100 µM.

Each plant was infected with the corresponding Pseudomone sp:Each plant was infected with the corresponding Pseudomone sp:Kiwi: P. syringae pv. actinidaeLemon: P. syringae pv. syringaeOlive: P. savastanoi pv. Nerii

This final solution (bacteria+peptide) was sprayed on kiwi and lemon plants and in the olive case was incorporated to plant through stems wounds. The different references are showed in the following table:

T_0 (before treatment) T_1 (after symptom onset)ICP (soil) ICP (soil and leaves)

microrespiration microrespirationSoluble C and Nitrogen Soluble C and Nitrogen

PLFAs PLFAs



Bacteria culture: Bacteria culture: Kiwi: P. syringae pv. Actinidae. Lemon: P. syringae pv. syringaeOlive: P. savastanoi pv. NeriiPeptidesPeptides: P1: AP17 P1: AP17 and and P2: Li 27 , produced by P2: Li 27 , produced by Florence University (Dr. S.Tegli)Florence University (Dr. S.Tegli)

Kiwi control T0 Kiwi P1_30 T0 Kiwi P1_100 T0 Kiwi P2_30 T0 Kiwi P2 100 T_0

T1 (22/12/2014)T1 (22/12/2014)

AFTER-Cu (LIFE)AFTER-Cu (LIFE)LEMON AND KIWI TREE (Peptide assay)LEMON AND KIWI TREE (Peptide assay)

CONTROLCONTROL BacBac P1_30P1_30 P1_100P1_100 P2_30P2_30 P2_100P2_100

P (g/100g)P (g/100g) 0,073 a0,073 a 0,070 a0,070 a 0,075 a0,075 a 0,066 a0,066 a 0,072 a0,072 a 0,064 a0,064 a

K (g/100g)K (g/100g) 0,590 a0,590 a 0,650 a0,650 a 0,641 a0,641 a 0,605 a0,605 a 0,640 a0,640 a 0,586 a0,586 a

Ca (g/100g)Ca (g/100g) 15,593 a15,593 a 17,056 ab17,056 ab 17,368 ab17,368 ab 17,353 ab17,353 ab 17,590 ab17,590 ab 18,390 b18,390 b

S (g/100g)S (g/100g) 0,210 a0,210 a 0,232 a0,232 a 0,240 a0,240 a 0,205 a0,205 a 0,216 a0,216 a 0,188 a0,188 a

Mg (g/100g)Mg (g/100g) 2,123 a2,123 a 2,322 a2,322 a 2,338 a2,338 a 2,295 a2,295 a 2,322 a2,322 a 2,382 a2,382 a

Na (g/100g)Na (g/100g) 0,070 a0,070 a 0,062 a0,062 a 0,068 a0,068 a 0,060 a0,060 a 0,060 a0,060 a 0,062 a0,062 a

Fe (mg/Kg)Fe (mg/Kg) 7282,38 a7282,38 a 7561,67 a7561,67 a 7703,34 a7703,34 a 7343,85 a7343,85 a 7468,72 a7468,72 a 6926,93 a6926,93 a

Mn (mg/Kg)Mn (mg/Kg) 252,34 a252,34 a 269,17 a269,17 a 271,95 a271,95 a 258,38 a258,38 a 265,46 a265,46 a 254,20 a254,20 a

Cu (mg/Kg)Cu (mg/Kg) 19,903 a19,903 a 17,342 a17,342 a 18,126 a18,126 a 19,150 a19,150 a 17,250 a17,250 a 18,554 a18,554 a

Cd (mg/Kg)Cd (mg/Kg) 0,193 a0,193 a 0,220 a0,220 a 0,215 a0,215 a 0,255 a0,255 a 0,228 a0,228 a 0,224 a0,224 a

Zn (mg/Kg)Zn (mg/Kg) 30,380 a30,380 a 29,820 a29,820 a 30,775 a30,775 a 30,261 a30,261 a 30,830 a30,830 a 29,876 a29,876 a

Pb (mg/Kg)Pb (mg/Kg) 12,486 a12,486 a 13,000 a13,000 a 49,820 b49,820 b 12,851 a12,851 a 13,050 a13,050 a 13,060 a13,060 a

Cr (mg/Kg)Cr (mg/Kg) 19,660 a19,660 a 20,728 a20,728 a 20,901 a20,901 a 21,296 a21,296 a 20,826 a20,826 a 19,684 a19,684 a

Ni (mg/Kg)Ni (mg/Kg) 10,403 a10,403 a 10,726 a10,726 a 10,661 a10,661 a 10,666 a10,666 a 10,790 a10,790 a 10,828 a10,828 a



Concentrations of macro- and micronutrients and heavy metals in kiwi crop soil (dwt)Concentrations of macro- and micronutrients and heavy metals in kiwi crop soil (dwt)

Bacteria + peptide inoculation: > Ca in soil. It could be due to a possitiveBacteria + peptide inoculation: > Ca in soil. It could be due to a possitiveeffect on element absorptioneffect on element absorption

KIWIKIWI

Concentrations of macro- and micronutrients and heavy metals in LEMON peptide soil (dwt)Concentrations of macro- and micronutrients and heavy metals in LEMON peptide soil (dwt)

CONTROLCONTROL BacBac P1_30P1_30 P1_100P1_100 P2_30P2_30 P2_100P2_100

P (g/100g)P (g/100g) 0.076 a0.076 a 0.080 a0.080 a 0.082 a0.082 a 0.083 a0.083 a 0.076 a0.076 a 0.080 a0.080 a

K (g/100g)K (g/100g) 0.773 a0.773 a 0.714 a0.714 a 0.705 a0.705 a 0.710 a0.710 a 0.676 a0.676 a 0.713 a0.713 a

Ca (g/100g)Ca (g/100g) 16.37 a16.37 a 18.36 a18.36 a 18.23 a18.23 a 18.00 a18.00 a 17.33 a17.33 a 18.00 a18.00 a

S (g/100g)S (g/100g) 0.306 a0.306 a 0.272 a0.272 a 0.267 a0.267 a 0.271 a0.271 a 0.260 a0.260 a 0.278 a0.278 a

Mg (g/100g)Mg (g/100g) 2.343 a2.343 a 2.474 a2.474 a 2.522 a2.522 a 2.558 a2.558 a 2.410 a2.410 a 2.552 a2.552 a

Na (g/100g)Na (g/100g) 0.073 a0.073 a 0.060 a0.060 a 0.062 a0.062 a 0.060 a0.060 a 0.057 a0.057 a 0.067 a0.067 a

Fe (mg/Kg)Fe (mg/Kg) 10914.25 a10914.25 a 11367.98 a11367.98 a 10053.21 a10053.21 a 10052.50 a10052.50 a 9212.28 a9212.28 a 9548.01 a9548.01 a

Mn (mg/Kg)Mn (mg/Kg) 291.22 a291.22 a 288.27 a288.27 a 289.96 a289.96 a 300.75 a300.75 a 280.16 a280.16 a 290.36 a290.36 a

Cu (mg/Kg)Cu (mg/Kg) 23.40 a23.40 a 29.15 a29.15 a 29.54 a29.54 a 36.60 a36.60 a 22.78 a22.78 a 27.53 a27.53 a

Cd (mg/Kg)Cd (mg/Kg) 0.14 a0.14 a 0.20 a0.20 a 0.20 a0.20 a 0.19 a0.19 a 0.16 a0.16 a 0.16 a0.16 a

Zn (mg/Kg)Zn (mg/Kg) 35.17 a35.17 a 37.05 a37.05 a 34.20 a34.20 a 41.74 a41.74 a 32.99 a32.99 a 35.05 a35.05 a

Pb (mg/Kg)Pb (mg/Kg) 11.76 a11.76 a 10.83 a10.83 a 12.75 a12.75 a 13.57 a13.57 a 13.41 a13.41 a 14.26 a14.26 a

Cr (mg/Kg)Cr (mg/Kg) 23.02 a23.02 a 22.89 a22.89 a 21.90 a21.90 a 22.69 a22.69 a 21.48 a21.48 a 22.05 a22.05 a

Ni (mg/Kg)Ni (mg/Kg) 11.71 a11.71 a 11.53 a11.53 a 10.86 a10.86 a 10.93 a10.93 a 10.40 a10.40 a 10.49 a10.49 a



Concentrations of macro- and micronutrients and heavy metals in lemon crop soil (dwt)Concentrations of macro- and micronutrients and heavy metals in lemon crop soil (dwt)

Bacteria + peptide inoculation not showed any difference on control soilBacteria + peptide inoculation not showed any difference on control soil(lemon crop).(lemon crop).

LEMONLEMON

Concentrations of soluble Carbon and Nitrogen in kiwi peptide soil (dwt)

CONTROL Bac P1_30 P1_100 P2_30 P2_100

Cs (ppm) 305,36 a 257,38 a 309,80 a 315,38 a 297,96 a 284,10 a

Nt (ppm) 28,98 a 26,77 a 27,63 a 27,00 a 23,70 a 22,15 aFor each parameter, different letters indicate significant differences between treatments according to Tukey test (p<0,05).

Concentrations of soluble Carbon and Nitrogen in lemon peptide soil (dwt)

CONTROL Bac P1_30 P1_100 P2_30 P2_100

Cs (ppm) 388,81 b 209,45 a 210,60 a 245,03 239,92 a 251,22 a

Nt (ppm) 28,00 a 30,92 a 33,02 a 27,11a 20,38 a 28,34 aFor each parameter, different letters indicate significant differences between treatments according to Tukey test (p<0,05).


KIWIKIWI LEMONLEMON

Kiwi PLFAKiwi PLFA (nmol g-1)(nmol g-1)

Control T0Control T0 BacBac P1 30P1 30 P1 100P1 100 P2 30P2 30 P2 100P2 100

BacteriaBacteria 15.515.5 11.5711.57 11.0611.06 20.3520.35 8.388.38 12.2112.21Gram+ BacteriaGram+ Bacteria 8.588.58 6.776.77 6.276.27 9.479.47 4.244.24 7.157.15Gram- BacteriaGram- Bacteria 6.926.92 4.84.8 4.784.78 10.8710.87 4.144.14 5.065.06FungiFungi 1.51.5 0.190.19 0.730.73 1.331.33 0.340.34 1.251.25Total saturated Total saturated PLFAsPLFAs

27.1427.14 14.5314.53 14.5714.5729.5629.56 9.869.86 21.9321.93

Total monoin. Total monoin. PLFAsPLFAs

2.622.62 1.921.92 1.961.966.046.04 2.362.36 2.282.28

ActinobacteriaActinobacteria 0.570.57 0.410.41 0.440.44 0.620.62 0.240.24 0.440.44

Kiwi PLFAKiwi PLFA (nmol g-1)(nmol g-1)

Control T60Control T60 BacBac PP1 301 30 P1 100P1 100 P2 30P2 30 P2 100P2 100

Total PLFA Total PLFA 32.0132.01 17.0917.09 17.6717.67 37.5537.55 12.6912.69 26.326.3Fung/BacFung/Bac 0.10.1 0.020.02 0.070.07 0.070.07 0.040.04 0.10.1Gram+/Gram-Gram+/Gram- 1.241.24 1.411.41 1.311.31 0.870.87 1.021.02 1.411.41Sat PLFa/Mono Sat PLFa/Mono PLFAPLFA

10.1510.15 7.587.587.447.44 4.94.9 4.184.18 9.69.6

Bacterial. fungal. GramBacterial. fungal. Gram++. Gram. Gram--. satured and monosatured PLFAs concentration in kiwi soils T0 before treatments.. satured and monosatured PLFAs concentration in kiwi soils T0 before treatments.The total PLFA and fungi/bacteria. GramThe total PLFA and fungi/bacteria. Gram++/Gram/Gram-- and satured/monounsatured ratios in kiwi soils T0 before treatments. and satured/monounsatured ratios in kiwi soils T0 before treatments.

Bacterial. fungal. GramBacterial. fungal. Gram++. Gram. Gram--. satured and monosatured PLFAs concentration in kiwi soils T0 before treatments.. satured and monosatured PLFAs concentration in kiwi soils T0 before treatments.The total PLFA and fungi/bacteria. GramThe total PLFA and fungi/bacteria. Gram++/Gram/Gram-- and satured/monounsatured ratios in kiwi soils T0 before treatments. and satured/monounsatured ratios in kiwi soils T0 before treatments.

PLFAsPLFAs


Microbial biomass was affected by bacteria and peptides (also by peptide doses)Microbial biomass was affected by bacteria and peptides (also by peptide doses)

Peptide Li 21 assay two months laterPeptide Li 21 assay two months laterPeptide Li 21 assay two months laterPeptide Li 21 assay two months laterPeptide AP17 assay two months laterPeptide AP17 assay two months later


Kiwi leaves infected by bacteriaKiwi leaves infected by bacteria

Kiwi crop showed a negative effect with Kiwi crop showed a negative effect with bacteria inoculo; peptide (particularly Li21)bacteria inoculo; peptide (particularly Li21)could avoid this effectcould avoid this effect

Lemon Control T0

Lemon P1 30 T0 Lemon P1 100 T0 Lemon P2 T_0 Lemon P2 100 T_0


Lemon leaves: plants not showed infection sintoms by bacteria Lemon leaves: plants not showed infection sintoms by bacteria

LEMON CROPLEMON CROP.LEMON CROPLEMON CROP.

Olive Control Olive P1 30 Olive P1 100 Olive P2 30 Olive P2 100

OLIVE CROPOLIVE CROP.OLIVE CROPOLIVE CROP.


Olive crop with bacteria and peptidesOlive crop with bacteria and peptidesinoculation. At this moment, not negative inoculation. At this moment, not negative effects has been showedeffects has been showed

A NEW EXPERIMENT FROM CEBASA NEW EXPERIMENT FROM CEBAS

““Effect of peptides addition on some pathogen Effect of peptides addition on some pathogen microorganismsmicroorganismsin soils”. EFFECT ON SOIL BIODIVERSITYin soils”. EFFECT ON SOIL BIODIVERSITY

SOIL + PEPTIDES SOIL + PEPTIDES SOIL + PathogenSOIL + PathogenSOIL + PEPTIDES + PathogenSOIL + PEPTIDES + PathogenCONTROL SOIL CONTROL SOIL


AFTER-Cu (LIFE)AFTER-Cu (LIFE)NºNº DescriptionDescription NºNº DescriptionDescription

11 Control destilled waterControl destilled water 1818 Compost (10%) Control destilled waterCompost (10%) Control destilled water

22 Control SFS (0.85%)Control SFS (0.85%) 1919 Compost (10%) Control SFS (0.85%)Compost (10%) Control SFS (0.85%)

33 Bac 1Bac 1 2020 Compost (10%) Bac 1Compost (10%) Bac 1



66 Bac 1 P1 30Bac 1 P1 30 2323 Compost (10%) Bac 1 P1 30Compost (10%) Bac 1 P1 30












ParametersParameters C/N hidrosolubleC/N hidrosolublePLFAsPLFAsMicrobial respirationMicrobial respirationGlucoaminopeptidaseGlucoaminopeptidase

Β- GlucosidaseΒ- GlucosidaseFosfataseFosfatase

PEPTIDE MICROCOMS ASSAYPEPTIDE MICROCOMS ASSAYTable 1. Sample descriptionTable 1. Sample descriptionSampling: T0, T15, T30 and T60 daysSampling: T0, T15, T30 and T60 daysBac 1: Bac 1: P. syringae pv. actinidaeP. syringae pv. actinidaeBac 2: P. syringae pv. syringaeBac 2: P. syringae pv. syringaeBac:3Bac:3 P. savastanoi pv. nerii P. savastanoi pv. neriiBac (1,2,3) P1 30: Pseudomone+ antimicrobial peptide P1 (AP17) 30 Bac (1,2,3) P1 30: Pseudomone+ antimicrobial peptide P1 (AP17) 30 µµMMBac (1,2,3) P1 100: Pseudomone+ antimicrobial peptide P1 (Ap17) 100 Bac (1,2,3) P1 100: Pseudomone+ antimicrobial peptide P1 (Ap17) 100 µµMMBac (1,2,3) P2 30: Pseudomone+ antimicrobial peptide P2 (Li 27) 30 Bac (1,2,3) P2 30: Pseudomone+ antimicrobial peptide P2 (Li 27) 30 µµMMBac (1,2,3) P2 100: Pseudomone+ antimicrobial peptide P2 (Li 27)100 Bac (1,2,3) P2 100: Pseudomone+ antimicrobial peptide P2 (Li 27)100 µµMMSample (soil and soil 10% compost) were disposed in 100 c.c. Sample (soil and soil 10% compost) were disposed in 100 c.c.

Several containers, each one with one hundred g of soil or soli Several containers, each one with one hundred g of soil or soli amendment with 10% compost , were treated as show in Table 1, and amendment with 10% compost , were treated as show in Table 1, and placed in a growth chamber.Each treatment was replicated three times and placed placed in a growth chamber.Each treatment was replicated three times and placed in a random design into the growth chamber set at 16h photoperiod with a day/night in a random design into the growth chamber set at 16h photoperiod with a day/night temperature regime of 24/15temperature regime of 24/15ººC.C.

PEPTIDE MICROCOMS ASSAYPEPTIDE MICROCOMS ASSAYTable 1. Sample descriptionTable 1. Sample descriptionSampling: T0, T15, T30 and T60 daysSampling: T0, T15, T30 and T60 daysBac 1: Bac 1: P. syringae pv. actinidaeP. syringae pv. actinidaeBac 2: P. syringae pv. syringaeBac 2: P. syringae pv. syringaeBac:3Bac:3 P. savastanoi pv. nerii P. savastanoi pv. neriiBac (1,2,3) P1 30: Pseudomone+ antimicrobial peptide P1 (AP17) 30 Bac (1,2,3) P1 30: Pseudomone+ antimicrobial peptide P1 (AP17) 30 µµMMBac (1,2,3) P1 100: Pseudomone+ antimicrobial peptide P1 (Ap17) 100 Bac (1,2,3) P1 100: Pseudomone+ antimicrobial peptide P1 (Ap17) 100 µµMMBac (1,2,3) P2 30: Pseudomone+ antimicrobial peptide P2 (Li 27) 30 Bac (1,2,3) P2 30: Pseudomone+ antimicrobial peptide P2 (Li 27) 30 µµMMBac (1,2,3) P2 100: Pseudomone+ antimicrobial peptide P2 (Li 27)100 Bac (1,2,3) P2 100: Pseudomone+ antimicrobial peptide P2 (Li 27)100 µµMMSample (soil and soil 10% compost) were disposed in 100 c.c. Sample (soil and soil 10% compost) were disposed in 100 c.c.

Several containers, each one with one hundred g of soil or soli Several containers, each one with one hundred g of soil or soli amendment with 10% compost , were treated as show in Table 1, and amendment with 10% compost , were treated as show in Table 1, and placed in a growth chamber.Each treatment was replicated three times and placed placed in a growth chamber.Each treatment was replicated three times and placed in a random design into the growth chamber set at 16h photoperiod with a day/night in a random design into the growth chamber set at 16h photoperiod with a day/night temperature regime of 24/15temperature regime of 24/15ººC.C.

CEBAS WORKING GROUPCEBAS WORKING GROUP

THANK YOU FOR YOUR ATTENTIONTHANK YOU FOR YOUR ATTENTION

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AFTER-Cu AFTER-Cu LIFE (2014-2016) Carlos García Izquierdo. CEBAS-CSIC FAENZA, JANUARY 2015 PARTNER...

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