First Report of Colletotrichum acutatum Causing Anthracnose on Olives in Albania

Abstract

Olive trees (Olea europaea L.) are among the most important fruit tree crops grown in Albania, covering an estimated 8% of the arable land of this country. The highest amount of olive production in Albania is concentrated in the coastal districts of Fier, Berat, Elbasan, Lezha, Tirana, Kruja and Vlora, all with Mediterranean climate conditions. Anthracnose is the main disease of olive fruit caused by different Colletotrichum spp. primarily belonging to two complexes, C. acutatum sensu lato (s.l.) and C. gloeosporioides s.l. (Cacciola et al. 2012; Schena et al. 2014.). On July 2018, field observations of about 50 olive trees in the Ishull Shengjin Lezhe Region (54% prevalence), 1 km away from the coast showed severe symptoms of mummified olive fruits in about 20 to 40% disease incidence and 30% disease severity of the Italian table olive cv. Frantoio trees resulting in premature fruit drop or mummification. The causal agent was isolated directly from infected fruits on potato dextrose agar and Rose Bengal nutrient media. Microscopic examinations of five single spore isolated fungal colonies showed acervuli with typical conidia of the genus Colletotrichum that were aseptate, straight, hyaline, subcylindrical with rounded ends and 12.3 to 22.1 µm long (mean = 17.4 µm) and 2.5 to 7.3 µm wide (mean = 5.9 µm) (n= 50 conidia) (Damm et al. 2012). To identify the fungal species, DNA from two single-spore isolates (Col-3-ALB and Col-9-ALB) isolated from fruits was extracted, and six genes were amplified (ITS, GAPDH, CHS-1, HIS3, ACT, and TUB2) using the primers reviewed in Damm et al. (2012). PCR products were sequenced, and BLAST analysis showed 100% identity to C. acutatum for both isolates (GenBank accession nos. for Col-9-ALB MT218337 [ITS1-5.8-ITS2], MT274748 [CHS-1], MT274749 [HIS3], MT274750 [GAPDH], MT274751 [TUB2], and MT274752 [ACT]). Phylogenetic analysis using the concatenated sequences of Col-9-ALB, the type species of C. acutatum (112996), a previously identified published Greek C. acutatum isolate (O9) and three C. godetiae strains confirmed the identification of Col-9-ALB as C. acutatum. Pathogenicity tests were performed in the laboratory to confirm the ability of C. acutatum isolates to cause disease on olive drupes. Fruits were surface disinfected with 0.1% NaClO for 3 min and rinsed with ddH2O. Artificial inoculations with the two above isolates were performed by spraying 24 olive fruits per isolate cv. Kalamon (eight olive fruits per replication) with a spore suspension (106 conidia/ml). Olive fruits sprayed with sterilized water served as untreated control. After inoculation, olive fruits were placed in closed sterile plastic boxes and kept at 26°C with a 12-h photoperiod. First rot symptoms and formation of acervuli by the pathogen were initiated 3 days after inoculation. Eight days post-inoculation, all treatments exhibited typical anthracnose symptoms similar to those observed in olive orchards (extensive fruit rot). To fulfil Koch’s postulates, C. acutatum was re-isolated from 10 random symptomatic olive fruits/isolate and their identity was confirmed from all samples. To our knowledge, this is the first report of C. acutatum causing fruit rot on olive trees in Albania. It is important to further study the epidemiology for the disease under local climate conditions and on different olive cultivars in order to develop effective management strategies for this very destructive disease of olive.

Susceptibility of Italian olive cultivars to various Colletotrichum species associated with fruit anthracnose

“…Interestingly, C . acutatum has recently been reported for the first time as causal agent of olive anthracnose in Albania and Greece (Cara et al, 2021; Iliadi et al, 2018); these were the countries from which it is very probable C . godetiae , the species responsible for the first epidemic outbreaks of OA in the 1950s, was inadvertently introduced into southern regions of Italy (Cacciola et al, 2012).…”