Anth thracnos ose of of oliv ol ive tr trees s and d fr - - PowerPoint PPT Presentation

TUESDAY 16 FEBRUARY 2016 - KLEIN JOOSTENBERG, MULDERSVLEI

SA OLIVE ANNUAL GENERAL MEETING

Dr Antonia Carlucci

University of Foggia, Italy

Anth thracnos

• se of
• f
• l
• liv

ive tr trees s and d fr fruit it

Plan ant Path atholog

• gy

y and nd Diag agnos

• sis,

s, Uni niversi rsity ty of Fogg

• ggia

Apulia is the most region of Italy with the biggest area cultivated with olive trees, with more of 377 thousands hectares (32%), 12 milions of quintals

• f
• live

drupes (35%). The 34,4% of Italian olive oil is produced in Apulia region.

Region Hectares %

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s, Uni niversi rsity ty of Fogg

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Italian Olive production

Anthracnose of olive is caused from Colletotrichum spp. and also Fusarium lateritium

Doctoral thesis during 2008-2011

Morphological and molecular characterization of Colletotrichum spp. as causal agent of olive Anthracnose in southern Italy (Apulia): preliminary control means

Plan ant Path atholog

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y and nd Diag agnos

• sis,

s, Uni niversi rsity ty of Fogg

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OLIVE DISEASES Anthracnose - Colletotricum spp.,

The disease is favored from warm and wet conditions; the rains increase the infection cycles. The disease cause high oleic acid values, so the

• il is not high quality.

Plan ant Path atholog

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y and nd Diag agnos

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s, Uni niversi rsity ty of Fogg

• ggia

Plan ant Path atholog

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y and nd Diag agnos

• sis,

s, Uni niversi rsity ty of Fogg

• ggia

The Anthracnose infections of olive It is possible to think that the infections start from the stalk of the fruits. But secondary infections can start from mesocarp of olive fruits. The Colletotrichum infections can occur also on leaves, indeed olive plants often show stunting and leaf falls (strong defoliation). We ascertained that Colletotrichum spp. are able to infect leaves. Olive drupes infected from Colletotrichum

• spp. give oil called “red oil”. It is enough

5-10% of olive fruits infected to have an

• il with high acidity.

OLIVE DISEASES Anthracnose - Colletotricum spp.,

Plan ant Path atholog

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y and nd Diag agnos

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s, Uni niversi rsity ty of Fogg

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Survey has been carried out in more of 300 olive orchards located in Apulia region from February to December for three years (2008-2011)

Apulian provinces

• n. of towns
• n. of olive
• rchards

examinated % Disease Incidence 2008/2009 2009/2010 2010/2011 Foggia 27 45 >1 Bat 7 28 2 2 8 Bari 19 39 11 9 17 Taranto 15 44 65 52 68 Brindisi 32 66 67 65 77 Lecce 44 109 82 78 86 Plan ant Path atholog

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y and nd Diag agnos

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s, Uni niversi rsity ty of Fogg

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*Means calculated on the hottest months of the year.

• Medit. sea

*RH=68 RH=71 RH=77

Fungal species isolated Stem Leaf stalk Drupe Acremonium curvulum 1,1 3,4 4,5 Alternaria alternata 2,3 0,3 0,7 5,7 9 Aspergillus niger 0,2 0,2 0,4 Aureobasidium oleae 0,9 2,8 3,7 Botryosphaeriaceae 11,7 4,3 11,3 27,3 Cadophora malorum 3,4 0,4 1,2 0,7 5,7 Cladosporium tenuissimum 4,6 0,3 4,9 Colletotrichum acutatum complex 6,5 1,5 18,9 42,7

69,6

Colletotrichum gloeosporioides 2,2 0,8 3,4 12,3

18,7

Cytospora oleina 7,3 1,8 9,1 Eutypa lata 8,5 0,4 8,9 Fusarium lateritium var. lateritium 0,9 9,6 10,5 Glomerella cingulata 1,3 2,2 3,5 Lecythophora lignicola 6,8 0.2 6,8 Microsphaeropsis olivacea 5,9 0,8 1,2 7,9 Monochaetia karstenii 4,6 4,6 Phaeoacremonium angustius 2,3 0,6 2,9 Phaeoacremonium inflatipes 1,2 1,2 Phaeoacremonium mortoniae 1,2 1,2 Phaeoacremonium rubrigenum 3,2 0,6 3,8 Phaeoacremonium aleophilum 7,2 1,6 8,8 Phoma incompta 2,4 2,4 Phomopsis spp. 5,6 0,7 0,6 0,8 7,7 Pleurophoma cava 1,2 1,2 Pleurostomophora richardsiae 1,4 1,4 Pseudocercospora cladosporioides 0,9 0,5 1,4 Septoria olea 1,5 1,1 1,1 3,7 Spilocaea oleaginea 1,5 0,2 0,2 1,9 Stemphylium botryosum 1,2 2,3 3,5

• thers

4,5 91,1 65,3 2,9 100% 100% 100% 100%

Isolation frequency (%) of fungi from olive tissues related to Lecce province

Plan ant Path atholog

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s, Uni niversi rsity ty of Fogg

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Colletotrichum acutatum

c g h i

Conidia are hyaline elliptical- fusiform, pointed at both ends

Plan ant Path atholog

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y and nd Diag agnos

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s, Uni niversi rsity ty of Fogg

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Colletotrichum gloeosporioides

d

The conidia are sub-cylindrical, rounded at both ends

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a e f g h i c d

10 EF543858 Musicillium EF543859 Musicillium DQ286219 DQ286221 99 AY376522 AY376521 DQ286160 100 87 AF411765 A2 397 AJ749683 A2 PT135 Olivo AY376507 A2 STE U4459 Protea AJ301964 A2 BBA 71292 Lupino 225 226 229 AF272782 A2 HV83C Anemone 79 DQ286117 C lupini AR2820 AJ301959 A1 BBA 71249 var.lupini AJ301933 A1 BBA 70358 var. setosum DQ286119 C lupini AR2826 64 AF521205 A8 TOM9 AF521210 A8 TOM12 AF521196 A8 TOM21 61 72 69 AJ749694 A5 PT227 Olivo AY376513 A5 STE U4466 HAKEA AY376498 STE U164 PINUS AY376510 G acutata STE U 5292 AY376502 A5 STE U4460 PROTEA 445 372 C359 393 266 435 438 704 391 DQ286132 G acutata ATCC56816 58 DQ286121 MELO G acutata ATCCMY AJ749686 A3 PT170 AY376509 STE U5287 MALUS 314 310 DQ286124 G acutata VACCINO MEP 76 88 97 AJ749700 A6 PT250 AF411719 A6 S2 99 AF411766 A4 NI90 AJ749688 A4 CBS193 32 272 284 C222 91 7 66 89 567 6 C368 365 357 335 325 215 703 8 61 349 628 572 518 510 507 245 244 418 324 100 401 219 218 10 C L 98 AJ301925 A7 BBA 65797 DQ286140 F phormii MEP1334 319 808 79 73 100 AB105957 C higginsianum IFO618 AB105956 C higginsianum MAFF23 AB105966 C fuscum MAFF238340 97 AB057437 C linicola 313 98 99 DQ286152 C trichellum MEP1535 AJ301985 C truncatum BBA71528 DQ286154 C dematium AR3563 LIR AJ301954 C dematium BBA 99 58 AB105970 C coccodes MAFF712102 1 2 AY376528 C coccodes STE U5301 100 327 AJ301955 C circinans BBA67846 100 68 58 56 75 DQ286205 C crassipes CBS15975 DQ286156 C capsici AR4028 AY376531 C dematium STE U5299 AY376543 G truncata STEU5294 75 100 66 DQ286190 G cingulata AR2801 AY376532 C gloeosporioides STE AY376534 C gloeosporioides STE 220 223 99 100 A7 A4 A6 A3 A5 A8 A1 A2

• C. boninense
• C. agavensis

Outgroup

Glomerella acutata/Colletotrichum acutatum complex

Colletotrichum gloeosporioides Colletotrichum circinans Colletotrichum coccodes Colletotrichum dematium Colletotrichum linicola Colletotrichum higginsianum

Plan ant Path atholog

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y and nd Diag agnos

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s, Uni niversi rsity ty of Fogg

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Phylogenetic tree based on Neighborg Joining (NJ) analysis of ITS sequences

Species identified

• A2= C. simmondsii
• A3= C. fiorinae
• A4= C. clavatum
• A5= C. acutatum sensu strictu
• A7= C. phormii
• C. gleosporioides
• C. clavatum

Anthracnose of olive is caused from Colletotrichum spp.

Plan ant Path atholog

• gy

y and nd Diag agnos

• sis,

s, Uni niversi rsity ty of Fogg

• ggia

Colletotrichum acutatum Complex

• A2= C. simmondsii

(IF, 4.5%)

• A3= C. fiorinae

(IF, 7.4%)

• A4= C. clavatum

(IF, 44.6%)

• A5= C. acutatum sensu strictu (IF, 25.8%)
• A7= C. phormii

(IF, 11,3%)

• C. gleosporioides

(IF, 6.4%) Highlight

• C. clavatum was isolated with highest Isolation frequency

then C. gleosporioides. The first hypothesis was that C. clavatum is the really responsible of Anthracnose of olive in Apulia.

Anthracnose of olive is caused from Colletotrichum spp.

Plan ant Path atholog

• gy

y and nd Diag agnos

• sis,

s, Uni niversi rsity ty of Fogg

• ggia

So, the subsequent aims of this study were:

• Assessment of pathogenicity of Colletotrichum spp.

in vivo and in vitro;

• Assessment of control trials against Anthracnose in
• pen field.

(a)C. gloeosporioides (b) C. simmondsii (c) C. fiorinae (d) C. clavatum (e) C. phormii (f) C. acutatum (g)C. coccodes (h) C. circinans (i) C. destructivum

a b c d e f i h g

Plan ant Path atholog

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y and nd Diag agnos

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s, Uni niversi rsity ty of Fogg

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Pathogenicity Test in vitro conditions

(a)C.gloeosporioides,

• C. acutatum sensu strictum

Anthracnose of olive is caused from Colletotrichum spp. artificially inoculated

Plan ant Path atholog

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y and nd Diag agnos

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s, Uni niversi rsity ty of Fogg

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Anova, Duncan test P<0,5

Severity Disease

Fungal species

• C. gleosporioides
• C. simmondsii
• C. acutatum sensu strictum
• C. fiorinae
• C. clavatum
• C. coccoides
• C. destructivum
• C. circinans

Control

Plan ant Path atholog

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s, Uni niversi rsity ty of Fogg

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All Colletotrichum species inoculated were able to infect the

• live drupes

although values of Severity Disease index were different. The most aggressive fungus was C. simmondsii, which never has been isolated from olives, this is the first time.

• C. acutatum sensu strictum was the second fungus able to

infect olive drupes more than C. clavatum, although this last fungus was the most isolated. Moreover, C. coccoides, C. circinans e C. destructivum resulted to be able to infect olive drupes but with values of Severity Disease index very low.

Results

Experimental trial was carried out from September 2008 to October 2008 in two olive

• rchards (two cultivar: cv Ogliarola salentina and Picholine).
• Extensive orchard (cv Ogliarola), 10 x 10 mt, >100 years old, Disease Diffusion

2007 was 88%

• Intensive orchard (cv Picholine ), 3 x 3 mt, <35 years old, Disease Diffusion 2007

was 85%. The trial included 10 olive trees for each treatments, and replicated 5 times. The treatments were carried out in September and October (before and during ripening stage), and were the following;

• 50 trees as Control not treated ;
• 50 Trees treated with Difenconazolo/Copper;
• 50 Trees treated with Dodina;
• 50 Trees treated with Boscalid/Pyraclostrobin;
• 50 Trees treated with Kresoxim-methyl;

Chemical control trials

Plan ant Path atholog

• gy

y and nd Diag agnos

• sis,

s, Uni niversi rsity ty of Fogg

• ggia

Plan ant Path atholog

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y and nd Diag agnos

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s, Uni niversi rsity ty of Fogg

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Results

Treatment % drupes infected from Colletotrichum species by isolation in cultures

Control not treated

81.2

Difenconazolo/Copper

43.5

Dodina

34.9

Boscalid/Pyraclostrobin

33.7

Kresoxim-methyl

11.5

No fungicide was quite able to control olive Anthracnose during the field trial. It was possible to observed that the fungicide more efficacious was Kresoxim-methyl. This result is very important because was obtained from a trial that included just two treatments.

Plan ant Path atholog

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y and nd Diag agnos

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s, Uni niversi rsity ty of Fogg

• ggia

CONCLUSIONS

• 1. Preliminary study allowed to detected five species belonging to C.

acutatum complex such as C. simmondsii, C. fiorinae, C. clavatum, C. phormii, C. acutatum sensu strictum.

• 2. The most aggressive species resulted to be C. acutatum s.s., while the

most frequent in Apulia region was C. clavatum.

• 3. C. gleosporioides resulted to be isolated occasionally, and moreover, it

was not be able to infect olive drupes in pathogenicity tests.

• 4. Chemical control carried out in open field showed that no fungicide was

really able to protect olive drupes from Colletotrichum anthracnose, except Kresoxim-methyl resulted to be more efficacious. .

• 5. Maybe, the treatments should be started before September, and the

number of treatments should increase.

Plan ant Path atholog

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s, Uni niversi rsity ty of Fogg

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A B C D E

F

G H I J K

F E

H G J A I K D C B J D K I B H C

F

A E G

Control of Anthracnose of olive fruit

Each trial consists of at least 10 plants. A= Pyraclostrobin B= Tebuconazolo C= Mancozeb D= Trifloxystrobin E= Boscalin F= Control G= Cuprolivo (Copper) H= Difeconazolo I= Dodina J= Kendal TE (for organic agriculture) K= Agroallium (extracts from onion and garlic)

Trial dates

• 20 August (lignification seed)
• 15 September (darkening of olive

drupes)

• 10 October (ripening of olive drupes)

Harvesting: from 10 November

Randomized blocks scheme Therefore, on the base of results obtained next August, a control trial will be started with 7 fungicides and two organic products

Plan ant Path atholog

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y and nd Diag agnos

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s, Uni niversi rsity ty of Fogg

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Th Thank ank yo you u fo for r yo your ur at attent entio ion