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

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Anth thracnos ose of of oliv ol ive tr trees s and d fr fruit it Dr Antonia Carlucci University of Foggia, Italy SA OLIVE ANNUAL GENERAL MEETING TUESDAY 16 FEBRUARY 2016 - KLEIN JOOSTENBERG, MULDERSVLEI

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Italian Olive production Region Hectares % 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 of olive drupes (35%). The 34,4% of Italian olive oil is produced in Apulia region.

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Anthracnose of olive is caused from Colletotrichum spp. and also Fusarium lateritium Morphological and molecular characterization of Colletotrichum spp. as causal agent of olive Anthracnose in southern Italy (Apulia): preliminary control means Doctoral thesis during 2008-2011

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia 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 oil is not high quality.

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia 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 oil with high acidity.

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia OLIVE DISEASES Anthracnose - Colletotricum spp.,

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Survey has been carried out in more of 300 olive orchards located in Apulia region from February to December for three years (2008-2011) Medit. sea % Disease Incidence n. of olive Apulian orchards provinces n. of towns examinated 2008/2009 2009/2010 2010/2011 Foggia 27 45 0 0 >1 Bat 7 28 2 2 8 *RH=68 RH=71 RH=77 Bari 19 39 11 9 17 Taranto 15 44 65 52 68 Brindisi 32 66 67 65 77 Lecce 44 109 82 78 86 *Means calculated on the hottest months of the year.

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Isolation frequency (%) of fungi from olive tissues related to Lecce province Fungal species isolated Stem Leaf stalk Drupe Acremonium curvulum 4,5 0 0 1,1 3,4 Alternaria alternata 2,3 0,3 0,7 5,7 9 Aspergillus niger 0,4 0,2 0 0,2 0 Aureobasidium oleae 0 0 0,9 2,8 3,7 Botryosphaeriaceae 27,3 11,7 0 4,3 11,3 Cadophora malorum 3,4 0,4 1,2 0,7 5,7 Cladosporium tenuissimum 4,9 4,6 0 0 0,3 Colletotrichum acutatum complex 69,6 6,5 1,5 18,9 42,7 Colletotrichum gloeosporioides 18,7 2,2 0,8 3,4 12,3 Cytospora oleina 9,1 7,3 1,8 0 0 Eutypa lata 8,9 8,5 0 0,4 0 Fusarium lateritium var. lateritium 10,5 0,9 0 0 9,6 Glomerella cingulata 3,5 1,3 0 0 2,2 Lecythophora lignicola 6,8 6,8 0 0.2 0 Microsphaeropsis olivacea 7,9 5,9 0,8 0 1,2 Monochaetia karstenii 4,6 4,6 0 0 0 Phaeoacremonium angustius 2,9 2,3 0 0,6 0 Phaeoacremonium inflatipes 1,2 0 0 0 1,2 Phaeoacremonium mortoniae 1,2 1,2 0 0 0 Phaeoacremonium rubrigenum 3,8 3,2 0 0,6 0 Phaeoacremonium aleophilum 8,8 7,2 0 1,6 0 Phoma incompta 2,4 2,4 0 0 0 Phomopsis spp. 7,7 5,6 0,7 0,6 0,8 Pleurophoma cava 1,2 1,2 0 0 0 Pleurostomophora richardsiae 1,4 1,4 0 0 0 Pseudocercospora cladosporioides 1,4 0,9 0 0 0,5 Septoria olea 3,7 1,5 1,1 0 1,1 Spilocaea oleaginea 1,9 0 1,5 0,2 0,2 Stemphylium botryosum 3,5 1,2 0 0 2,3 others 4,5 91,1 65,3 2,9 100% 100% 100% 100%

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia  Conidia are hyaline elliptical- Colletotrichum acutatum fusiform, pointed at both ends c g h i

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Colletotrichum gloeosporioides  The conidia are sub-cylindrical, rounded at both ends c d a d g e f h i

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

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Anthracnose of olive is caused from Colletotrichum spp. 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.

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Anthracnose of olive is caused from Colletotrichum spp. 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 open field.

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Pathogenicity Test in vitro conditions (a) C.gloeosporioides , (a) C. gloeosporioides (b) C. simmondsii (c) C. fiorinae (d) C. clavatum (e) C. phormii a b c (f) C. acutatum (g) C. coccodes (h) C. circinans (i) C. destructivum C. acutatum sensu strictum d e f g h i

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Anthracnose of olive is caused from Colletotrichum spp. artificially inoculated Severity Disease Fungal species C. gleosporioides C. simmondsii C. acutatum sensu strictum C. fiorinae C. clavatum C. coccoides C. destructivum C. circinans Control Anova, Duncan test P<0,5

Plan ant Path atholog ogy y and nd Diag agnos osis, s, Uni niversi rsity ty of Fogg oggia Results All Colletotrichum species inoculated were able to infect the olive 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.