The Third Report of Colletotrichum japonicum Worldwide

is paper reports the first finding of Colletotrichum japonicum (Hemmi) Bedlan on Berberis aquifolium Pursh [= Mahonia aquifolium (Pursh) Nutt.] in Poland. e fungus was collected in the Botanical Garden of Maria Curie-Skłodowska University (UMCS) in Lublin. is is the second locality of the species in Europe and the third worldwide. In this paper, morphological characteristics, microphotographs, and distribution comments are provided.


Introduction
Colletotrichum is traditionally recognized as an asexual genus of fungi, with a number of species linked to sexual morphs assigned to the Glomerella genus (Sordariomycetes, Glomerellaceae, Ascomycota). Over the years, Colletotrichum species were considered to be host specific, which led to the description of a large number of taxa Sharma & Shenoy, 2016). Species from this genus are economically one of the most important fungi, and cause anthracnose and other diseases in a wide range of plant species Hyde et al., 2009;Sutton, 1980). e comprehensive monographic study conducted by von Arx (1957) based on morphological characteristics with little emphasis placed on pathological features led to drastic reduction from approximately 750 to 11 species. Molecular studies of Colletotrichum species, which have become an integrated element in taxonomic research, have been developing intensively since the end of the twentieth century. e first applications of DNA sequence data to distinguish between species within this genus used the internal transcribed spacer (ITS1) region of nrDNA. Later, multilocus analyses became a common approach based on the use of ITS2 and large ribosomal subunit (LSU) as well as partial sequences of genes encoding, e.g., histone H3 (HIS), translation elongation factor 1 alpha (TEF-1α), tubulin beta chain (TUB), and actin (ACT) . Scientists working on Colletotrichum do not agree on the taxonomic approach and characters that should be employed to identify and describe a new Colletotrichum species. Hence, a reliable secondary barcode marker is indispensable for the accurate identification of Colletotrichum species (Sharma & Shenoy, 2016).
Identification of Colletotrichum species based on morphology has always been problematic; therefore, the current progress in molecular phylogenetic methods facilitates identification of stable and well-separated clades within Colletotrichum. However, the taxonomy of this genus is still unsatisfactory and there is a need for a polyphasic approach for identification, which reflects the natural classification of species and subspecific taxa within the genus .
Berberis aquifolium (Berberidaceae) is a plant native to western North America. is species was introduced to Europe as an ornamental plant in the 1820s. Table 1 Features of Colletotrichum species reported to be associated with Berberis aquifolium leaves.

C. nymphaeae (Pass.) Aa
Leaf spots Up to 10 mm 1-5 mm 1-1.5 mm 0. At present, it grows in many parts of Europe (Sorokopudov et al., 2017). It was introduced in Poland in 1839, and the status of this species has changed from a crop plant to a locally established and invasive species (Tokarska-Guzik et al., 2012).

According to A preliminary checklist of micromycetes in Poland, 23
Colletotrichum species were reported in Poland up until 2008 (Mułenko et al., 2008), while 26 species are currently known (Farr & Rossman, 2020;Jayawardena et al., 2016;Okorski et al., 2018;Pszczółkowska et al., 2016Pszczółkowska et al., , 2017. e present paper provides information about a new Colletotrichum species for Polish funga (C. japonicum on B. aquifolium); it also reports the second locality of the fungus in Europe and third worldwide.

Material and Methods
e infected leaves of B. aquifolium were collected in the Botanical Garden of Maria Curie-Skłodowska University (UMCS) in Lublin in 2018. e material sample was stained with cotton blue in lactic acid and gently heated. Observations were made using an Olympus SZX10 stereoscopic microscope and an Olympus CX31 light microscope. Photographic documentation was taken with an Olympus XC50 microscope camera. Measurements were made at ×600 and/or ×300 magnifications. Structure size ranges were specified based on 10 measurements of acervuli, 15 setae, and 30 conidia. e publications mentioned in Table 1 were used for identification of the fungus. e name of the host plant and its synonym is accepted following e Plant List (http://www.theplantlist.org/). e examined specimen is deposited in the herbarium of Maria Curie-Skłodowska University in Lublin (LBL).

Comments
e analyzed Polish sample exhibits the features of C. japonicum provided in the literature from Austria (Table 1), i.e., the only locality worldwide where the fungus has been noted on the same host (B. aquifolium) (Bedlan, 2012).