Università degli Studi di Firenze

Dipartimento di Scienze delle Produzioni Agroalimentari e dell’Ambiente (DISPAA)

Sezione di Patologia vegetale ed Entomologia

Action 16 & Action 17

Establishing a monitoring network to assess

lowland forest and urban plantation in Lombardy

and urban forest in Slovenia

Disease management in urban forests

Alessandro Ragazzi, Beatrice Ginetti, Salvatore Moricca

Phytopathology unit

www.pmnatura

Species: Acer pseudoplatanus, Quercus robur, Alnus cordata

Sampling years: 2005 , 2006

Sampling months: march…………october = 8

Number of sampled plants per species: 3 healthy and 3 declining = 18 (9 + 9)

Number of samples per plant: two plants every month = 288 (144 + 144)

Number of Petri dishes per sample: 5 = 1440 (720 + 720)

Number of Petri dishes evaluated (of the 5 dishes above): 3 = 864 (432 + 432)

Number of samples seeded per Petri dishe: 5 = 4320 (2160 + 2160)

Protocol of sampling and isolation

which pathogens have we found????

strakerenemy.blogspot.com

Biscogniauxia mediterranea (De Not) Kuntze

Agente di cancro carbonioso

Agent of charcoal canker

Protologo: Kuntze, O. 1891, Revisio generum plantarum: 398

Basionym: Sphaeria mediterranea De Not., 1851

Position in classification: Xylariaceae, Xylariales, Xylariomycetidae,

Sordariomycetes, Pezizomycotina, Ascomycota,

Biscogniauxia mediterranea (De Not) Kuntze

EPPO Alert list

Quercus spp.

Fraxinus excelsior

Fagus sylvatica

Castanea sativa From Marocco

present all the

our peninsula,

including the

islands

Foto Capretti-Ragazzi

Botryosphaeria dothidea (Moug. ex Fr.) Ces. et De Not.

Agente di cancro corticale

Agent of cortical canker

Protologo: Cesati et De Notaris 1863, Comm. Soc. crittog.

Ital. 1 (4): 212

Sanctioning author: Fr. (SM2 : 423, Sphaeria dothidea)

Anamorph: Fusicoccum aesculi

Position in classification: Botryosphaeriaceae,

Botryosphaeriales, Ascomycota

Botryosphaeria dothidea (Moug. Ex Fr.) Ces. Et De Not.

EPPO Alert list

Acer

pseudoplatanus

Quercus rubra

Q. robur

Q. suber

Ostrya spp.

Platanus spp.

present all the our peninsula,

including the islands

Locus: DQ168265 466 bp DNA linear PLN 04.10-2005

Definition: Botryosphaeria dothidea isolate B5 internal transcribed

spacer 1. Partial sequence; 5.8 ribosomal RNA gene and internal transcribed

spacer 2, complete sequence; and 28S ribosomal gene, partial sequence.

Accession: DQ168265

Version: DQ168265; GI: 76563848

Reference: 1

Journal: Plant Disease

Origin:

1 GCGGGCCGCG GTCCTCCGCG GCCGGCCCCC CTCCCCGGGG GGTGGCCAGC

51 GCCCGCCAGA GGACCATCAA ACTCCAGTCA GTAAACGATG CATCTGAAAA

101 AACATTTAAT TTTAAACTAA AACC ...

151

Molecular characterization of Botryosphaeria dothidea

(target regions of ribosomial DNA )

Botryosphaeria parva Pennycook & Samuels

Botryosphaeria obtusa (Schwein.) Shoemaker

Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers

& A.J.L. Phillips

Diplodia seriata De Not

Foto Capretti

……and a very high number of

endophytic fungi

(pathogens and other)

Plots Non thinned plot (2Ant)

State of healthy Symptomatic Asymptomatic

Endophytic fungi 2011 2012 2013 2011 2012 2013

Alternaria

alternata (Fr.: Fr.)

Keissl.

4.2 4.7 5.8 3.9 * *

Apiognomonia

quercina (Kleb)

Höhn

8.1a 3.1a 5.6a 4.4a 3.2a *

Aposphaereia sp. * * * - - -

Aureobasidium

pullulans (de Bary

et al.)

3.0 3.1 * 3.1 * *

Biscogniauxia

mediterranea (De

Not.) Kuntze

7.7a 3.1a 4.4a * * *

Botryosphaeria

dothidea (Moug et

al.)

51.6b 33.3b 41.4b 30.2b 17.6b 13.8a

Ceratocystis

coerulescens

(Münch) Bakshi

7.8a 4.3a 5.4a 4.9a 4.3a 4.1b

Chaetomium

indicum Corda

3.1 * * - -

Cladosporium

herbarum Pers.:

Fr.

- - - - - -

Curvularia lunata

(Wakker) Boedijn

8.1 * * 6.7 6.4 6.3

Diplodia mutila

(Fr.) Mont.

29.6c 17.4c 23.4c 17.1c 11.9c 10.4c

Tab. 3 – Endophytic fungi detected in twigs of symptomatic and asymptomatic tress, isolated from thinned (2At)

and non thinned (2Ant) plot in North Park (Lombardy, North Italy). The data show the average value of the

endophytic assemblage of seven trees present in both plots: Acer pseudoplatanus, Alnus cordata, Fraxinus

angustifolia, F. excelsior, F. ornus, Quercus cerris, Q. robur. The colonization frequency is expressed as percentage.

t = thinned; nt = non thinned

Tab. 3 – Endophytic fungi detected in twigs of symptomatic and

asymptomatic tress, isolated from thinned (2At) and non thinned

(2Ant) plot in North Park (Lombardy, North Italy). The data

show the average value of the endophytic assemblage of seven

trees present in both plots: Acer pseudoplatanus, Alnus cordata,

Fraxinus angustifolia, F. excelsior, F. ornus, Quercus cerris, Q.

robur. The colonization frequency is expressed as percentage.

t = thinned; nt = non thinned

Diplodina acerina

(Pass.) B. Sutton

6.8a 3.8a 4.9a 3.3a * *

Epicoccum nigrum

Link

6.4 6.6 5.9 7.1 7.6 7.3

Gliomastix

murorum (Corda)

S. Hughes

7.1 7.9 6.8 11.6 10.2 10.9

Glomerella

cingulata

(Stoneman et al.)

3.2 3.0 3.1 - - -

Gnomonia sp. * * * - - -

Nectria sp. 4.2 * * 3.6 4.2 4.1

Neofusicoccum

parvum

(Pennycook et

al.)

52.2b 30.3b 44.1b 28.7b 19.1b 17.6d

Phialocephala sp. 4.9 5.2 5.0 - - -

Phomopsis

quercina (Sacc.)

Höhn. ex Died.

8.2a 3.4a 5.1a 11.0d 4.1a 3.4b

Phomopsis

acerina Pirone &

J.C. Carter

4.2d 3.3a 4.1a 3.2a * 3.3b

Pseudovalsa

longipes (Tul.)

Sacc.

3.8d * * 4.3a 3.2a 3.9b

Ramichloridium

sp.

- - - - - -

Septoria alni

Sacc.

7.3a 3.3a 4.8a 4.2a 4.1a 3.2b

Trichoderma

atroviride P.

Karst.

18.8 16.4 16.6 20.9 18.8 17.4

Trichoderma

viride Pers.: Fries

19.1 17.7 17.6 20.6 18.8 18.9

Values in column followed by the same letter do not differ

significantly per P< 0,05. Duncan’s Multiple Range Test

performed according to the values of only fungal

(endophytic) pathogens.

*Isolation frequency below the 3% threshold

Outside the LIFE plots, we found

some important pathogens

a lot of Phytophthora species…

•Phytophthora lacustris

•Phytophthora taxon Pg chlamydo (1^ report in Italy)

•Phytophthora gonapodyides

•Phytophthora inundata

•Phytophthora taxon walnut (1^ report in Italy)

•Potential hybrid between P. lacustris et P. Pg chlamydo

The new pathogen Phytophthora acerina

(first description )

Phytophthora acerina B.Ginetti, T. Jung, D.E.L.

Cooke, S. Moricca sp. nov.

Phytophthora acerina B. Ginetti, T. Jung, D.E.L. Cooke , S. Moricca sp. nov.

Peronosporaceae, Incertae sedis, Oomycota, Chromista

not yet included in the EPPO lists

Acer

pseudoplatanus Lombardia

causal agent of widespread sycamore maple mortality

Anthostoma decipiens (DC.) Nitschke

Agent of canker

Basionym: Sphaeria decipiens DC., in Lamarck et de Candolle 1805

Sanctioning author: Fr.

Citations in published lists or literature: Saccardo's Syll. fung. I:

302; III: 263; XII: 21; XV: 45

Position in classification : Diatrypaceae, Xylariales, Xylariomycetidae,

Sordariomycetes, Pezizomycotina, Ascomycota

causal agent of widespread hornbeam mortality

>ITS Anthostoma decipiens

CAGGTCTCGTTGGTGACCAGCGGAGGGATCATTACAGAGTTATTTATCTCCTAACCTT

ATGTGAACTTACCTATGTTGCCTCGGCGAGGAAAGCCTACCCTGTAGTTACTTGGAG

GCGAGCTACCCTGTAGCCCGCTGCTGGCTAACCCGTCGATGGACCATTCTAACTCTTG

TTTTTCTGTGGCACATCTGAATCGTTTATACTTAATAAGTTAAAACTTTCAACAACGGA

TCTCTTGGTTCTGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATT

GCAGAATTCAGTGAATCATCGAATCTTTGAACGCACATTGCGCCCATTAGTATTCTAG

TGGGCATGCCTGTTCGAGCGTCATTTCGACCATCAAGTCTTATTTGCTTGGAGTTGGG

AATTTGCTTGCAAGTAATTCCTTAAAATTATTGGCGGAGTTGCGATAACCCCAAGCGT

AGTAATTATCTCTCGCTTTCGGTGTGTTAGCGCTGACATTTAGCCGTTAAACCCTCTAT

ATTTATGTAGGTTTGACCTCGGATCAGGTAGGAATACCCGCTGAACTTAGC

Anthostoma decipiens 18S rRNA gene (partial), ITS1, 5.8S rRNA

gene, ITS2 and 28S rRNA gene (partial), strain IPV-FW349

GenBank: AM399021.1

First Match: Anthostoma decipiens 18S rRNA gene (partial), ITS1, 5.8S rRNA gene, ITS2 and 28S

rRNA gene (partial), strain IPV-FW349

Score Expect Identities Gaps Strand Frame

1003 bits(543) 0.0() 559/566(99%) 3/566(0%) Plus/Plus

Query 8 CGTTGGTG-ACCAGCGGAGGGATCATTACAGAGTTATTTATCTCCT-AACCTTATGTGAA 65

||| |||| ||| ||||||||||||||||||||||||||| ||||| |||||||||||||

Sbjct 3 CGTAGGTGAACCTGCGGAGGGATCATTACAGAGTTATTTAACTCCTAAACCTTATGTGAA 62

Query 66 CTTACCTATGTTGCCTCGGCGAGGAAAGCCTACCCTGTAGTTACTTGGAGGCGAGCTACC 125

|||||||||||||||||||||||| |||||||||||||||||||||||||||||||||||

Sbjct 63 CTTACCTATGTTGCCTCGGCGAGG-AAGCCTACCCTGTAGTTACTTGGAGGCGAGCTACC 121

Query 126 CTGTAGCCCGCTGCTGGCTAACCCGTCGATGGACCATTCTAACTCTTGTTTTTCTGTGGC 185

||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||

Sbjct 122 CTGTAGCCCGCTGCTGGCTAACCCGTCGATGGACCATTCTAACTCTTGTTTTTCTGTGGC 181

Query 186 ACATCTGAATCGTTTATACTTAATAAGTTAAAACTTTCAACAACGGATCTCTTGGTTCTG 245

||||||||| ||||||||||||||||||||||||||||||||||||||||||||||||||

Sbjct 182 ACATCTGAACCGTTTATACTTAATAAGTTAAAACTTTCAACAACGGATCTCTTGGTTCTG 241

Query 246 GCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAAT 305

||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||

Sbjct 242 GCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAAT 301

Query 306 CATCGAATCTTTGAACGCACATTGCGCCCATTAGTATTCTAGTGGGCATGCCTGTTCGAG 365

||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||

Sbjct 302 CATCGAATCTTTGAACGCACATTGCGCCCATTAGTATTCTAGTGGGCATGCCTGTTCGAG 361

Query 366 CGTCATTTCGACCATCAAGTCTTATTTGCTTGGAGTTGGGAATTTGCTTGCAAGTAATTC 425

||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||

Sbjct 362 CGTCATTTCGACCATCAAGTCTTATTTGCTTGGAGTTGGGAATTTGCTTGCAAGTAATTC 421

Query 426 CTTAAAATTATTGGCGGAGTTGCGATAACCCCAAGCGTAGTAATTATCTCTCGCTTTCGG 485

||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||

Sbjct 422 CTTAAAATTATTGGCGGAGTTGCGATAACCCCAAGCGTAGTAATTATCTCTCGCTTTCGG 481

Query 486 TGTGTTAGCGCTGACATTTAGCCGTTAAACCCTCTATATTTATGTAGGTTTGACCTCGGA 545

||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||

Sbjct 482 TGTGTTAGCGCTGACATTTAGCCGTTAAACCCTCTATATTTATGTAGGTTTGACCTCGGA 541

Query 546 TCAGGTAGGAATACCCGCTGAACTTA 571

||||||||||||||||||||||||||

Sbjct 542 TCAGGTAGGAATACCCGCTGAACTTA 567

Goidanich, 1954

Prophylaxis and/or treatment

Eliminate in winter death or diseased plants

Avoid sawdust, or limit its spread, during cutting and pruning

Remove and destroy the cut material

Extirpate the stumps

Avoid any type of wound

Prune only during the winter months

Protect large cutting surface with polyvinylic paste added with a

chemical compound or with biological mastic

Periodically check the large cutting surface to control the development

of the scaring process

Importe propagating material only from areas where there are not

reported some pathogens

Respect the distance in new plantations

Use resistant clones

Observe the decrees of obligatory control

Educate the staff that manages the urban green

General criteria for urban forest

Inform the citizenship

Make the treatment in the early morning or evening ones

Never make treatment in presence of wind

Prohibit access to treated areas

Ensure the protection of the operator

Use of chemical compounds at doses indicated on the label

Before proceeding to a treatment in urban

areas need to follow specific precautions:

www.magieraansaloni.it

Prescriptions at European level to adopt criteria in urban

and suburban area for a phytosanitary pruning

Prevent cutting, keeping the plants in appropriate conditions (distance)

Prevent cutting of large twigs and branches working on small branches, on which

the compartimentation is more easy

Cutting after windbreakage or traumatic wounds: need to trim the injured part

and protect the exposed tissues

Cutting due to decay or necrotic processes: make the cut on the surface healthy (15-

20 cm below the dead tissue). Protect the exposed tissues.

Cutting of twigs or branches dead: the practice of "Dead-wooding" is needed to

secure the plant, thereby eliminating not only the dead branches, but even those

with necrotic phenomena in extension

Lightening of the crown: fundamental in areas with presence of foliar fungal

agents. The lowest humidity values of a crown "lightened" in fact hamper many

pathogns. Moreover a lightened crown, in high windy areas, allows the wind to

traverse the crown without resistance, thus avoiding injuries, potential sites of entry

of pathogenic agents

The cutting must always be carried out in the vicinity of buds or other branches,

in order to allow the plant to form the healing tissue

Criteria to control Biscogniauxia mediterranea (Franceschini, com. pers.)

Reduce the inoculum mass through the cutting of infected plants and

pruning of branches by way of desiccation or with cankers

Burn all the material or to carry it away, making sure to cover it with a cloth

during wood hauling

If the spread of the pathogen is not yet at epidemic level, we can cut as

simple coppice. The consequent "opening of the forest" create conditions less

favourable to the spread of the pathogen

If the pathogen reached an epidemic spreading is necessary to proceed as

coppice with standards or with a conversion to a high forest. In this case,

instead of preventing the further spread of the pathogen, is more important

to maintain soil moisture to avoid the plants come easily in water stress

resulting in colonization of its organs by the pathogen

Criteria to control Phytophthora spp.

.

.

Strict quarantine rules

Obligation to contact Regional plant protection services

Cortical treatments with phosphites and tacky at the base of the trunk,

repeating annually in the spring

Inspection of imported plants

In case we suspect the presence of Phytophthora in soil, it’s recommende soil

treatment, having uprooted and destroyed symptomatic plants, using

sunburn.

Criteria to control endophytic fungi

????????????????????????????????????????

Avoid thermic and water stress to the plants

Reduce the inoculum biomass by a phytosanitary pruning

Biological control in planta???

Pre-immunization??

The results of our work are perhaps the first to point

out that a thinning can lead to an increase in the incidence of some fungal pathogenic

endophytes in forest trees.

Increase the biomass of leaves, understood as “a

territory” by colonizing

The Thinning

humbleness

diagnostic

capacity

training

meticulousness

The monitoring The prophylaxis