The Strange Case of Ocellomma Rediuntum (Arthoniales: Roccellaceae) in Australia: a Remarkably Disjunct Lichen
Total Page：16
File Type：pdf, Size：1020Kb
The Lichenologist (2020), 52, 187–195 doi:10.1017/S0024282920000092 Standard Paper The strange case of Ocellomma rediuntum (Arthoniales: Roccellaceae) in Australia: a remarkably disjunct lichen Gintaras Kantvilas1 , Cécile Gueidan2 and Anders Tehler3 1Tasmanian Herbarium, Tasmanian Museum and Art Gallery, Box 5058, UTAS LPO, Sandy Bay, Tasmania 7005, Australia; 2Australian National Herbarium, National Research Collections Australia, National Collections and Marine Infrastructure, CSIRO, P.O. Box 1700, Canberra, ACT 2601, Australia and 3Swedish Museum of Natural History, P.O. Box 50007, S-104 05 Stockholm, Sweden Abstract The new combination Ocellomma rediuntum (Stizenb. ex Hasse) Kantvilas, Gueidan & Tehler is proposed, supported by morphological, anatomical and molecular data. Hitherto known only from the Californian coast, this species is here recorded for Kangaroo Island (South Australia), Victoria and Tasmania. It is described and illustrated from Australian collections. The ecology of the species and its remarkable distribution are discussed. Key words: distribution, Kangaroo Island, lichens, Schismatomma, Tasmania, taxonomy, Victoria (Accepted 20 December 2019) Introduction Whilst taxon sampling and support values were insufficient to cat- egorically confirm the placement of this species within Ocellomma, The order Arthoniales represents one of the largest groups of liche- they suggested a close affinity between it and O. picconianum.The nized Ascomycetes. It comprises fruticose, foliose and crustose taxa remarkable disjunction between North American and Australian and occurs in most ecosystems of the world, reaching its highest populations is discussed in the context of the ecology of the species. diversity in subtropical coastal habitats with a Mediterranean- or desert-type climate (Ertz & Tehler 2011). The group has had a che- quered taxonomic history but the most recent classification (Ertz & Materials and Methods Tehler 2011) recognized six families, including the Roccellaceae. This large family, estimated to comprise 28–42 genera and c. 300 Anatomy, morphology and chemistry species (Jaklitsch et al. 2016), includes taxa with (inter alia) a tren- The study is based chiefly on material collected by the first author in tepohlioid photobiont, rounded ascomata with a thalline margin South Australia (Kangaroo Island) and Tasmania, and on herbarium and reduced excipulum, cylindrical to clavate asci, and ascospores material housed in AD, BM, FH, HO, MEL and S. Anatomical and that lack a gelatinous sheath (Ertz et al. 2015). Features of the morphological observations were undertaken using light microscopy, Roccellaceae as currently circumscribed are its high degree of spe- with thin hand-cut sections mounted in water, 10% KOH, lactophe- cies endemism and the relatively large number of small or mono- nol cotton blue, Lugol’s iodine after pretreatment with dilute KOH, typic genera, one of which is Ocellomma Ertz & Tehler, based on and ammoniacal erythrosin. Ascospore measurements are presented the Mediterranean taxon O. picconianum (Bagl.) Ertz & Tehler. in the format: 5th percentile–average–95th percentile, with outlying Within the family, Ocellomma is characterized by a crustose, ecor- values given in brackets and n being the number of measurements. ticate thallus, erumpent, sessile, pruinose ascomata containing cal- Routine chemical analyses using thin-layer chromatography followed cium oxalate, and hyaline, 3-septate ascospores, and was standard methods (Elix 2014). Calcium oxalate was observed by elut- recognized as a distinct lineage on the basis of DNA sequence ing thin sections in 20% sulphuric acid, which induces the precipi- data (Ertz et al. 2015). In this paper, we record the North tation of clusters of needle-like crystals. Terminology of asci American species Schismatomma rediuntum (‘rediunta’) (Stizenb. follows Torrente & Egea (1989)asillustratedbyKantvilas(2004). ex Hasse) Tehler for southern Australia and Tasmania, and transfer it to the genus Ocellomma (O. rediuntum (Stizenb. ex Hasse) Comparative specimen of Ocellomma picconianum examined. Kantvilas et al. comb. nov.) on the basis of morphological and ana- Italy: Calabria, Crotone, near mouth of the River Neto, 39°13′N, tomical data. Molecular data were also obtained and indicated that 17°08′E, 5 m alt., on bark of Eucalyptus, 14 v 1989, D. Puntillo the widely separated Australian populations were conspecific. (Lichenotheca Graecensis 3, 57 (1996)) (BM). Author for correspondence: Gintaras Kantvilas. E-mail: [email protected]. gov.au DNA extraction, amplification and sequencing Cite this article: Kantvilas G, Gueidan C and Tehler A (2020) The strange case of Ocellomma rediuntum (Arthoniales: Roccellaceae) in Australia: a remarkably disjunct Four Australian specimens of Ocellomma rediuntum (three from lichen. Lichenologist 52, 187–195. https://doi.org/10.1017/S0024282920000092 Tasmania and one from Victoria) were newly sequenced © British Lichen Society 2020 Downloaded from https://www.cambridge.org/core. University of Athens, on 29 Sep 2021 at 08:22:52, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0024282920000092 188 Gintaras Kantvilas et al. (Table 1). Lichen material was detached from the substratum in PAUP* v.4.0a (build 163) (Swofford 2002). The most likely tree using clean tweezers and transferred to an Eppendorf tube. was computed with the sumt command in MrBayes and visualized Genomic DNA was obtained using a phenol-chloroform-based in PAUP*. Additional support values were obtained using a fast protocol modified from Zolan & Pukkila (1986), as described in bootstrap analysis of 1000 pseudoreplicates with RAxML, using Gueidan et al. (2007). Three markers were amplified: the region the four previously described partitions. including the 5.8S subunit of the nuclear ribosomal RNA gene Published and newly generated ITS sequences were also and the internal transcribed spacers 1 and 2 (ITS), the large sub- aligned using Mesquite but for a much smaller dataset due to unit of the nuclear ribosomal RNA gene (nrLSU), and the second the high variability of this locus across the Roccellaceae. This largest subunit of the RNA polymerase II gene (RPB2). small dataset was combined with RPB2 data with the aim of The primers used for amplification were: ITS1F and ITS4 for confirming the sister relationship between O. rediuntum and the ITS region (White et al. 1990; Gardes & Bruns 1993), LR0R O. picconianum. For the ITS-RPB2 dataset, two specimens of and LR5 for nrLSU (Vilgalys & Hester 1990) and RPB2-7cF Schismatomma pericleum were used as an outgroup and the and RPB2-11aR for RPB2 (Liu et al. 1999). For the nuclear ribo- ingroup included O. rediuntum (3 specimens), O. picconianum somal regions, 1 μl of genomic DNA was added to a master mix (5 specimens), Pseudoschismatomma rufescens (2 specimens), comprising 5 μl of MyFi buffer (Bioline, London, UK), 1 μlof two species of Dirina, and Roccella allorgei. A maximum like- each primer, 1 μl of MyFi polymerase and 16 μl of water. For lihood tree search and bootstrap analysis were performed using RPB2, the master mix was modified to include 2 μl of each primer. RAxML as described above. Both LSU-RPB2 and ITS-RPB2 Amplifications were carried out on a Mastercycler Pro S thermo- trees were visualized in PAUP* and edited with Illustrator cycler (Eppendorf, Hamburg, Germany) using standard PCR pro- (Adobe Systems, San Jose, CA, USA). The datasets were deposited grams. PCR products were sent to Macrogen (Seoul, Korea) for in TreeBASE (25482). purification and sequencing. Mixed and weak PCR products were cloned using a TOPO-TA cloning kit (Invitrogen, Thermo Results Fisher Scientific, Waltham, MA, USA) according to the manufac- turer’s instructions. Additional ITS, nrLSU and RPB2 sequences With the congruence test showing no conflict, the two markers were obtained for Ocellomma picconianum following previously nrLSU and RPB2 were concatenated into a combined dataset. described protocols (Tehler & Irestedt 2007;Tehleret al. 2009a, b, The resulting phylogeny is presented in Fig. 1, with posterior 2010). probabilities and bootstraps as support values. The recovered relationships were largely in agreement with the phylogeny of Ertz et al. (2015), with Psoronactis, Lecanactis, Isalonactis and Phylogenetic analysis Chiodecton forming early diverging groups. Roccellina clusters together with Crocellina, Vigneronia, Dendrographa and Sequences were assembled using Sequencher v.5.4.6 (Gene Codes ≤ Corporation, Ann Arbor, MI, USA). RPB2 and nrLSU sequences Syncesia (99% PP and 70% BS), and the lineage that includes were aligned manually in Mesquite v.3.51 (Maddison & Diromma, Dirina and Roccella is well supported (100% PP and Maddison 2017), together with previously published sequences BS). The four specimens of Ocellomma rediuntum form a mono- from various genera within Roccellaceae (mostly from Ertz et al. phyletic group (100% PP and BS). This species is resolved as sister to O. picconianum but the relationship is not supported (< 95% 2015) and some new sequences of Ocellomma picconianum ≤ (Table 1). For the combined nrLSU-RPB2 analysis, two species of PP and 50% BS). Similarly, Pseudoschismatomma is resolved Gyrographa were used as an outgroup. The ingroup included our as sister to the lineage that includes O. rediuntum and O. picconianum, but this relationship is not supported (< 95% PP four Australian specimens of O. rediuntum,aswellas39 ≤ Roccellaceae