Print
A regional study of the genus Phyllopsora (Ramalinaceae) in Asia and Melanesia
expand article infoSonja Kistenich, Mika Bendiksby§, Charles S. Vairappan|, Gothamie Weerakoon, Siril Wijesundara#, Patricia A. Wolseley, Einar Timdal
‡ University of Oslo, Oslo, Norway
§ Norwegian University of Science and Technology, Trondheim, Norway
| Universiti Malaysia Sabah, Kota Kinabalu, Malaysia
¶ The Natural History Museum, London, United Kingdom
# National Institute of Fundamental Studies, Peradeniya, Sri Lanka
Open Access

Abstract

Phyllopsora is a crustose to squamulose lichen genus inhabiting the bark of trees in moist tropical forests and rainforests. Species identification is generally challenging and is mainly based on ascospore morphology, thallus morphology and anatomy, vegetative dispersal units, and on secondary chemistry. While regional treatments of the genus have been conducted for Africa, South America and Australia, there exists no study focusing on the Asian and Melanesian species. Previously, 24 species of Phyllopsora s. str. have been reported from major national studies and checklists representing 13 countries. We have studied herbarium material of 625 Phyllopsora specimens from 18 countries using morphology, anatomy, secondary chemistry, and molecular data to investigate the diversity of Phyllopsora species in Asia and Melanesia. We report the occurrence of 28 species of Phyllopsora including the following three species described as new to science: P. sabahana from Malaysia, P. siamensis from Thailand and P. pseudocorallina from Asia and Africa. Eight species are reported as new to Asia. A key to the Asian and Melanesian species of Phyllopsora is provided.

Keywords

Malaysia, Sri Lanka, Thailand, rainforest, TLC, phylogeny, identification key

Introduction

The genus Phyllopsora Müll. Arg. consists of 54 crustose or squamulose species (Kistenich et al. in press). They grow mostly on bark of trees in (sub-)tropical rainforests or moist woodlands. The genus was described in 1894 from New Zealand (Müller 1894), but the first modern revision of the pantropical genus was conducted 87 years later by Swinscow and Krog (1981) focusing on the East African species. Ten years later, Brako (1991) monographed the Neotropical species, while Elix (2009) summarized the Australian species and their occurrence. Additional reports and regional studies of the genus and its species, distribution can be found from Eastern Africa (Timdal and Krog 2001), Peru (Timdal 2008b) and the West Indies (Timdal 2011). From Asia, however, only a few reports exist for selected countries. Upreti et al. (2002) listed five Phyllopsora species from India. Later, Mishra et al. (2011) described two species and one variety from India as new to science. Recently, Kondratyuk et al. (2016) described a new species from South Korea. Phyllopsoroid specimens have been reported in additional checklists and geographical studies from, for example, Bangladesh (Aptroot and Iqbal 2011), Northeast India (Logesh et al. 2017), Sri Lanka (Weerakoon and Aptroot 2014), South Korea (Joshi et al. 2011) and Thailand (Aptroot et al. 2007). A general Asian, transnational study focusing on Phyllopsora has to date not been published. So far, 24 of the 54 accepted Phyllopsora species have been reported to occur in Asia and Melanesia (Table 1). An additional nine species reported from Asia represent either synonyms or have recently been excluded from the genus (Kistenich et al. in press; 2018b; Table 1).

Species of Phyllopsora are generally challenging to identify by morphology only. In a molecular phylogeny of the lichen family Ramalinaceae C. Agardh, Kistenich et al. (2018b) showed the genus Phyllopsora to be polyphyletic. Consequently, they excluded ten species from the genus. Three of the excluded species most likely belong in the family Malmideaceae Kalb, Rivas Plata & Lumbsch. An additional three of the excluded species were transferred to the new genus Parallopsora Kistenich, Timdal & Bendiksby. The species of Parallopsora grouped together in a poorly resolved clade with a number of tropical genera, such as Eschatogonia Trevis., Krogia Timdal and Physcidia Tuck. (Kistenich et al. 2018b). Little is known about these genera in Asia, which are generally very similar to Phyllopsora in their macromorphology. They often differ, however, from Phyllopsora in ascospore size and arrangement, presence of prothallus, thallus construction and chemistry (Kalb and Elix 1995; Kistenich et al. 2018b; Timdal 2008a; 2009). Recently, Kistenich et al. (2018a) described three new species of Krogia from Asia and Oceania, which were all tentatively identified as Phyllopsora sp., indicating the morphological similarity between these two genera.

The scope of the present study is to revise Asian and Melanesian Phyllopsora specimens mainly collected between 1990 and 2017 by the authors. Herein, we provide an overview of the species of Phyllopsora occurring in the Asian countries with an updated taxonomy based on multiple sources of evidence, including DNA sequence data. We describe three new species and provide a key to the Asian and Melanesian species of Phyllopsora.

Species of Phyllopsora reported from Asia and Melanesia.

Species Authorship Cambodia China Fiji India Indonesia Japan Malaysia Nepal New Caledonia Papua New Guinea Philippines Solomon South Korea Sri Lanka Taiwan Thailand Vanuatu Vietnam
Accepted species
P. africana Timdal & Krog ** ** ** * ** ** ** ** *
P. breviuscula (Nyl.) Müll. Arg. * [10] ** ** ** [17] *
P. buettneri (Müll. Arg.) Zahlbr. * [16] ** * [1, 5, 15] [5] * [3] ** [6] [4]
P. castaneocincta (Hue) Kistenich & Timdal * * [16] * T ** ** * * * T * ** [17] * **
P. chodatinica Elix ** ** * [5] *
P. cinchonarum (Fée) Timdal ** [12] * T ** [6]
P. confusa Swinscow & Krog [10] * * ** ** [1, 15] ** [17] * **
P. cuyabensis (Malme) Zahlbr. **
P. dolichospora Timdal & Krog * ** ** [17]
P. foliata (Stirt.) Zahlbr. ** ** [17]
P. furfuracea (Pers.) Zahlbr. [10] [5] [8] * [2, 7] T * [17] [3] ** [6, 19] [4]
P. gossypina (Sw.) Kistenich et al. [12] [13] [8] [14, 15] ** [17] [6] [4]
P. halei (Tuck.) Zahlbr. [1, 2] ** [3]
P. himalayensis G.K. Mishra et al. T
P. isidiosa Kistenich & Timdal ** ** ** ** **
P. kalbii Brako [10] **
P. loekoesii S.Y. Kondr. et al. ** ** * T
P. longiuscula (Nyl.) Zahlbr. ** ** *
P. mediocris Swinscow & Krog *
P. neofoliata Elix *
P. parvifolia (Pers.) Müll. Arg. [18] [16] [7, 8] [14] *
P. parvifoliella (Nyl.) Müll. Arg. ** * * **
P. porphyromelaena (Vain.) Zahlbr. * * [10] * ** ** * * * T * ** * T **
P. pseudocorallina Kistenich & Timdal ** ** **
P. sabahana Kistenich & Timdal **
P. santensis (Tuck.) Swinscow & Krog [12] [2] [5] *
P. siamensis Kistenich & Timdal **
P. subhispidula (Nyl.) Kalb & Elix **
Reported, not confirmed species
P. chlorophaea (Müll. Arg.) Zahlbr. [10] [3]
P. corallina (Eschw.) Müll. Arg. [9, 16] [11] [17] [3] [4]
P. isidiotyla (Vain.) Riddle [10]
P. mauritiana (Taylor) Swinscow & Krog [10]
P. nemoralis Timdal & Krog [10]
P. pyxinoides (Nyl.) Kistenich et al. [6, 19]
P. swinscowii Timdal & Krog [10]
Excluded species
P. catervisorediata G.K. Mishra et al. T
P. densiflorae (Vain.) Gotth. Schneid. T
P. griseocastanea (Vain.) Gotth. Schneid. T
P. manipurensis (Müll. Arg.) Müll. Arg. T
P. subcrustacea (Malme) Brako [10]
P. viridis Paulson T
P. borbonica Timdal & Krog [17]
P. sorediata (Aptroot & Sparrius) Timdal [6]
P. soralifera Timdal [9]

Materials and methods

The specimens

We investigated material from 18 different countries in Asia and Melanesia (Table 1) based on herbarium collections made mainly between 1990 and 2017. Older material of Phyllopsora is generally not suitable for DNA sequencing (Kistenich et al. in press). In addition to material from our own herbaria directly available to us (BM, BORH, O, PDA), we received loans from the institutional herbaria B, E, H, TNS, and UPS, as well as from the private herbarium of P. Diederich. In total, we investigated 908 specimens of Phyllopsora and related genera. Author names for the studied species are provided in Tables 1 and 2.

The definition of Melanesia follows the United Nations geoscheme for Oceania as devised by the United Nations Statistics Division based on the M49 coding classification (https://unstats.un.org/unsd/methodology/m49/). Accordingly, it includes the five countries Fiji, New Caledonia, Papua New Guinea, Solomon Islands, and Vanuatu.

Morphology and secondary chemistry

All specimens were studied morphologically and when necessary, also anatomically. Microscope sections were prepared using a freezing microtome and mounted in water, 10% KOH (K), lactophenol cotton blue, and a modified Lugol’s solution in which water was replaced by 50% lactic acid. The types of upper cortex referred to in this paper (types 1 and 2) are those described by Swinscow and Krog (1981). Amyloid reactions in the apothecium were observed in the modified Lugol’s solution after pretreatment in K, and crystals of lichen substances were observed using polarized light. Ascospore measurements are given as X ± 1.5 × SD rounded to 0.5 µm, where X is the arithmetic mean and SD the standard deviation.

We performed thin-layer chromatography (TLC) as routine investigation for identification of lichen substances in accordance with the methods of Culberson (1972), modified by Menlove (1974) and Culberson and Johnson (1982). Generally, we examined the acetone-extracts in solvent system B’; fatty acids were not examined. In difficult cases, we additionally used solvent systems A and C for lichen substance identification.

Molecular methods and phylogenetic analysis

For DNA extraction, PCR amplification and DNA sequencing of the mitochondrial ribosomal small subunit (mtSSU) and the nuclear ribosomal internal transcribed spacer region (ITS: ITS1, 5.8S, ITS2), we followed the protocols outlined in Kistenich et al. (2018a). For sequence assembly and preliminary alignment, we used Geneious R9 (Kearse et al. 2012).

As many of the specimens, from which we generated sequences, had not been previously identified, we needed to find out, which specimens belonged in Phyllopsora s. str. and consequently, which sequences to use in the final phylogenetic analyses. Hence, we phylogenetically analysed a combined alignment of our Ramalinaceae dataset (Kistenich et al. 2018b) and the newly generated sequences using standard RAxML (i.e., applying the GTR substitution model for each pre-defined partition [mtSSU, ITS1, 5.8S and ITS2] with 100 rapid bootstrap inferences and the GAMMA model for evaluating and optimizing the likelihood of the final tree; Stamatakis 2014). Based on these RAxML trees, we selected those specimens falling into Phyllopsora s. str. and incorporated them into our Phyllopsora dataset (Kistenich et al. in press). This dataset was analysed phylogenetically in more detail (see below) to provide evidence for undescribed species.

Each marker was aligned separately using MAFFT v.7.408 (Katoh and Standley 2013) with the E-INS-i algorithm and the nucleotide scoring matrix set to 1PAM / κ=2. We trimmed the ends of the ITS alignment to comprise only the ITS-region and deleted the residual 18S and 28S sequence information. Each dataset was initially analysed by IQ-TREE v.1.6.7 (Nguyen et al. 2015) to infer a maximum likelihood tree using 1000 ultrafast bootstrap repetitions (Hoang et al. 2018). We checked for gene-tree incongruence using compat.py (Kauff and Lutzoni 2002) with a cut-off of 90. As we did not find any strongly supported incongruences, which would affect the circumscription of the new species, we concatenated the mtSSU and ITS alignments. We ran a detailed IQ-TREE analysis to find the best-fitting nucleotide substitution models and partitioning schemes (Chernomor et al. 2016; Kalyaanamoorthy et al. 2017) among models implemented in MrBayes (i.e., 1-, 2-, and 6-rate models) and to infer a maximum likelihood tree using 1000 standard non-parametric bootstrap repetitions (BS). We defined four subsets, one for mtSSU and three for ITS corresponding to the ITS1, 5.8S and ITS2 regions, and analysed those with the TESTMERGE function resembling PartitionFinder2. In addition, we analysed the dataset with MrBayes v.3.2.6 (Altekar et al. 2004; Ronquist and Huelsenbeck 2003) as described in Kistenich et al. (2018b). The temperature increment parameter was set to 0.05. We projected the BS values from the IQ-TREE analysis onto the MrBayes consensus tree with posterior probabilities (PP) and collapsed branches with BS < 50 and PP < 0.7. The resulting trees were edited in TreeGraph2 (Stöver and Müller 2010) and FigTree v.1.4.4 (http://tree.bio.ed.ac.uk/software/figtree).

Results

Morphology and secondary chemistry

Morphological identification of many specimens was challenging, but with data obtained by TLC, many specimens could be identified to species level. Of the 908 studied specimens, we found 625 specimens to belong in Phyllopsora, while 283 specimens were found to belong in other genera of the Malmideaceae and Ramalinaceae (not treated in this study). Of the 625 Phyllopsora specimens, 480 were identified to species level in Phyllopsora (Table 2, Suppl. material 2: Table S1), while 141 specimens (23%) were left unidentified (not included in Suppl. material 2: Table S1), most of which were not sequenced and did not contain lichen substances. The morphology and anatomy of the Phyllopsora species have been described in detail by Swinscow and Krog (1981) and Brako (1991), and are not repeated here. We often found the distinction between cortex type 1 and type 2 useful for species identification; however, in many species the cortex type is intermediate (type 1–2). The chemistry of the 54 accepted Phyllopsora species is summarized in Kistenich et al. (in press).

Information about all Phyllopsora species may also be found on our Phyllopsora website: http://nhm2.uio.no/lichens/Phyllopsora.

Specimens used in this study with voucher information and GenBank accession numbers. New sequences are indicated by accession numbers in bold. – indicates missing data.

Species Extract # mtSSU ITS Country Year Voucher Herbarium
Biatora beckhausii (Körb.) Tuck. MG925858 AF282071 Norway 1995 Holien, H. 6744 TRH
B. vacciniicola (Tønsberg) Printzen MG925861 MG925960 Norway 2013 Klepsland, J. JK13-L330 O
Crocynia molliuscula (Nyl.) Nyl. 7359 MK352275 La Réunion 1996 Krog, H. & Timdal, E. RE18/03 O
7360 MK352276 Mauritius 1991 Krog, H. & Timdal, E. MAU58/02 O
Phyllopsora africana Timdal & Krog ch1 470 MK412413 MK412480 Thailand 1993 Aguirre, James & Wolseley 2475a BM
471 MK412414 MK412481 Thailand 1992 Aguirre–Hudson, B. & Wolseley, P.A. 1327 BM
509 MK352138 MK352317 La Réunion 1996 Krog, H. & Timdal, E. RE08/13 O
1436 MK352175 MK352348 La Réunion 1996 Krog, H. & Timdal, E. RE22/09 O
4037 MK352199 MK352370 Thailand 2012 v.d. Boom, P. 46982 hb. v.d. Boom
7224 MK412469 MK412512 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-517 PDA
P. africana ch1? 1012 MK412425 Indonesia 2000 Wolseley, P. T15 BM
P. africana ch2 477 MK352122 MK352301 Japan 1995 Thor, G. 13199 UPS
6770 MK412461 MK412504 Sri Lanka 2017 Weerakoon, G. Ri056 PDA
P. africana ch3 472 MK412415 Solomon Islands 1965 Hill, D.J. 9242 BM
1416 MK412435 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. D.8.04.oQ BORH
1427 MK412443 Indonesia 2000 Wolseley, P. T22 OQ BM
6348 MK352231 MK352401 Philippines 1994 Diederich, P. 13345 hb. Diederich
6351 MK412447 Philippines 1994 Diederich, P. 13213 hb. Diederich
6352 MK412448 Philippines 1994 Diederich, P. 13119 hb. Diederich
6772 MK412462 MK412505 Sri Lanka 2017 Weerakoon, G. Im015 PDA
7205 MK412463 MK412506 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-543 PDA
P. amazonica Kistenich & Timdal 3619 MK352194 MK352365 Brazil 2014 Barbosa, R.S., Haugan, R. & Timdal, E. 90 O
4155 MK352208 MK352379 Brazil 2015 Kistenich, S. & Timdal, E. SK1-85 MPEG
P. breviuscula (Nyl.) Müll. Arg. 528 MG925892 MG925990 La Réunion 1996 Krog, H. & Timdal, E. RE36/18 O
1305 MG925893 MG925991 Brazil 1980 Kalb, K. & Marcelli, M. in: Kalb, Lich. Neotropici 515 GZU
1432 MK412445 Sri Lanka 2007 Jayalal, U. A4-5-8-5 PDA
2100 MK352355 Philippines 1992 Tan, B.C. 92-187 B
6752 MK352245 MK352412 New Caledonia 2016 Rikkinen, J. 35509 H
6754 MK412456 MK412499 New Caledonia 2016 Rikkinen, J. 35503 H
6760 MK412457 MK412500 Sri Lanka 2017 Weerakoon, G. Im042 PDA
P. breviuscula 6764 MK412458 MK412501 Sri Lanka 2017 Weerakoon, G. Mn093 PDA
6765 MK412459 MK412502 Sri Lanka 2017 Weerakoon, G. Mo81 PDA
7212 MK352256 MK352422 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-642 PDA
7213 MK412465 MK412508 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-601 PDA
7217 MK412466 MK412509 Sri Lanka 2017 Weerakoon, G. 982 PDA
7218 MK412467 MK412510 Sri Lanka 2017 Weerakoon, G. 1013 PDA
7229 MK412470 MK412513 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-649 PDA
7234 MK412516 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-648 PDA
7235 MK412472 MK412517 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-640 PDA
P. buettneri (Müll. Arg.) Zahlbr. ch1 428 MK352103 MK352283 Thailand 1994 Wolseley, P. & Kanajriavanit, S. s.n. BM:734816
995 MK352146 MK352322 Thailand 1993 James, P.W. & Wolseley, P.A. 2466a BM
1041 MK352160 MK352335 Kenya 2007 Divakar, Lumbsch & Mangold 19553D hb. Pérez-Ortega
P. buettneri ch2 6464 MK352239 MK352406 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. AM-37 O
7177 MK352252 Venezuela 1984 Brako, L. 8110 GZU
P. buettneri ch3 429 MK352104 MK352284 Thailand 1993 Aguirre, B., James, P.W. & Wolseley, P. 2736 BM
493 MK352131 MK352311 Thailand 1994 Wolseley, P. & Kanajriavanit, S. s.n. BM:1104011
6462 MK352238 Japan 1995 Thor, G. 13183 UPS
P. byssiseda (Nyl.) Zahlbr. 4737 MK352211 MK352382 Venezuela 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-220 VEN
4739 MK352212 MK352383 Venezuela 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-229 VEN
P. canoumbrina (Vain.) Brako 3627 MK352195 MK352366 Brazil 2014 Barbosa, R.S., Haugan, R. & Timdal, E. 166 O
P. castaneocincta (Hue) Kistenich & Timdal 460 MK352116 MK352295 Tanzania 2008 Timdal, E. 10912 O
461 MK412412 MK412479 Thailand 1993 Aguirre, James & Wolseley 2482B BM
998 MK412420 Thailand 1991 Wolseley, P.A. & Aguirre–Hudson, B. 5564 BM
999 MK412421 MK412486 Thailand 1993 Wolseley, P.A. & David, F. 3314 BM
1022 MK412427 MK412490 Thailand 1992 Wolseley, P.A. & Aguirre–Hudson, B. 5583 BM
P. castaneocincta 1032 MK412429 Nepal 2007 Sharma, L.R., Olley, L., Cross L7.1 E
1045 MK412431 Thailand 1993 James, P.W. & Wolseley, P.A. 2466b BM
1264 MK412433 MK412492 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. M.3.10.1 BORH
1420 MK412439 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. M.3.10.2a BORH
1421 MK412440 MK412493 Malaysia 2012 Thüs, H., Wolseley, P. & Vairappan, C. M110 BORH
3560 MK352186 MK352358 South Africa 2014 Burrows, J. & Timdal, E. 14280 O
4032 MK352196 MK352367 Thailand 2012 v.d. Boom, P. 47239 hb. v.d. Boom
6743 MK352243 MK352410 Kenya 2013 Kirika, P., Mugambi, G. & Lumbsch, H.T. 3011 O
7232 MK412515 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-594 PDA
7255 MK352270 MK352434 Australia 1992 Elix, J.A. 32834 CANB
P. chlorophaea (Müll. Arg.) Zahlbr. 529 MK352145 MK352321 La Réunion 1996 Krog, H. & Timdal, E. RE36/17 O
1051 MK352165 MK352340 Kenya 2002 Killmann, D. & Fischer, E. s.n. hb. Killmann
1309 MK352172 Venezuela 1986 Brako, L. & Berry, P.E. 8685 GZU
SE382 MG925894 MG925992 La Réunion 1996 Krog, H. & Timdal, E. RE08/10 O
P. chodatinica Elix 513 MK352139 Australia 1986 Elix, J.A. & Streimann, H. 21023 O
1539 MK352177 MK352350 New Caledonia 2005 Elvebakk, A. 05:691 O
6456 MK352237 MK352405 Malaysia 2014 Paukov, A. 2232 B
P. cinchonarum (Fée) Timdal 439 MK352105 Thailand 2002 Sipman, H. 48664 B
440 MK352106 MK352285 Japan 2006 Thor, G. 21521 UPS
4168 MK352210 MK352381 Venezuela 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-201 VEN
6063 MK352227 Guatemala 2004 v.d. Boom, P. 33395 hb. v.d. Boom
P. concinna Kistenich & Timdal 4041 MK352202 MK352373 Panama 2010 v.d. Boom, P. 43947 hb. v.d. Boom
4776 MK352224 MK352395 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-445 O
6455 MK352236 MK352404 Venezuela 2015 M.S. Dahl, J.E. Hernández M., S. Kistenich, E. Timdal & A.K. Toreskaas SK1-225 O
7176 MK352251 MK352418 Guatemala 2002 Andersohn, C. s.n. B
P. confusa Swinscow & Krog 514 MK352140 MK352318 Kenya 1972 Krog, H. & Swinscow, T.D.V. K48/177 O
1018 MK412426 MK412489 Thailand 1991 Wolseley, P.A. 1049 BM
P. confusa 1024 MK352150 MK352325 Cuba 2007 Tønsberg, T. 37813 BG
1300 MK352169 MK352343 Venezuela 1969 Oberwinkler, B., Oberwinkler, F. & Poelt, J. s.n. GZU
1417 MK412436 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. M.3.10.6 BORH
3571 MK352190 MK352362 Ecuador 2014 Prieto, M. s.n. HUTPL
4741 MK352214 MK352385 Venezuela 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-237 VEN
6360 MK412451 Papua New Guinea 1992 Diederich, P. 11056 hb. Diederich
6361 MK412452 Papua New Guinea 1992 Diederich, P. 10319 hb. Diederich
6766 MK412460 MK412503 Sri Lanka 2017 Weerakoon, G. Ri030 PDA
7185 MK352253 MK352419 Cameroon 1999 Frisch, A. & Tamnjong Idi 99/Ka1213 hb. Frisch
7220 MK412468 MK412511 Sri Lanka 2017 Weerakoon, G. 176 PDA
7236 MK352260 MK352426 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-609 PDA
7239 MK412473 MK412518 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-567 PDA
7240 MK412474 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-532 PDA
P. corallina (Eschw.) Müll. Arg. 1316 MK352173 MK352346 Venezuela 1986 Brako, L. & Berry, P.E. 8659 GZU
4164 MK352209 MK352380 Venezuela 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-185 VEN
4762 MK352220 MK352391 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-377 O
4775 MK352223 MK352394 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-430 O
P. cuyabensis (Malme) Zahlbr. 449 MK352107 MK352286 Peru 2006 Timdal, E. 10258 O
450 MK352108 MK352287 Thailand 1993 Aguirre, B., James, P.W. & Wolseley, P. 2467a BM
1290 MK352166 MK352341 Venezuela 1996 Hafellner, J. 53910 GZU
1291 MK352167 MK352342 Guatemala 1979 Kalb, K. & Plöbst, G. s.n. GZU
2048 MK352180 MK352352 Bolivia 2008 Flakus, A. & Rodriguez, P. 12792 O
P. dolichospora Timdal & Krog 515 MK352141 MK352319 Mauritius 1991 Krog, H. & Timdal, E. MAU65/22 O
6357 MK352233 Papua New Guinea 1992 Diederich, P. 10847 hb. Diederich
P. dolichospora 6359 MK412450 Papua New Guinea 1992 Diederich, P. 10846 hb. Diederich
6763 MK352247 MK352414 Sri Lanka 2017 Weerakoon, G. Hg40 PDA
6767 MK352248 MK352415 Sri Lanka 2017 Weerakoon, G. Si113B PDA
7258 MK352271 MK352435 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-643 PDA
P. fendleri (Tuck. & Mont.) Müll. Arg. 2098 MK352183 MK352354 Costa Rica 1985 H. Sipman & A. Chaverri 20806 B
7473 MK352277 MK352437 Venezuela 1979 Sipman, H. 10688 B
P. foliata (Stirt.) Zahlbr. 1035 MK352157 MK352332 Japan 2004 Kashawadani, H. 46389 TNS
7238 MK352261 MK352427 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-627 PDA
7247 MK352265 MK352431 Australia 2006 Elix, J.A. 38235 CANB
P. foliatella Elix 7243 MK352262 MK352428 Australia 1986 Elix, J.A. & Streimann, H. 20241 CANB
7246 MK352264 MK352430 Australia 1986 Elix, J.A. & Streimann, H. 20203 CANB
7253 MK352268 Australia 2005 Elix, J.A. 37286 CANB
7254 MK352269 Australia 1998 Streimann, H. 61609 CANB
P. furfuracea (Pers.) Zahlbr. 451 MK412411 MK412478 Thailand 1993 Aguirre, James & Wolseley 2918 BM
452 MK352109 MK352288 La Réunion 1996 Krog, H. & Timdal, E. RE36/22 O
453 MK352110 MK352289 Trinidad And Tobago 2008 Rui, S. & Timdal, E. 10799 O
455 MK352111 MK352290 Peru 2006 Timdal, E. 10183 O
P. furfurella Kistenich & Timdal 3570 MK352189 MK352361 Ecuador 2014 Prieto, M. s.n. HUTPL
4036 MK352198 MK352369 Dominican Republic 2008 v.d. Boom, P. 39069 hb. v.d. Boom
P. glaucella (Vain.) Timdal 1000 MK352147 MK352323 Dominican Republic 1987 Harris, R.C. 20779 BM
2125 MK352184 MK352356 Argentina 2013 Ferraro, L.I., Aptroot, A. & Cáceres, M.E.S. 10761 O
4766 MK352221 MK352392 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-393 O
4780 MK352225 MK352396 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. AM-44 O
P. gossypina (Sw.) Kistenich et al. AY584615 Costa Rica 2002 Lücking, R. 16052 DUKE
P. gossypina ch1 3575 MK352192 MK352363 Brazil 2014 Barbosa, R.S., Haugan, R. & Timdal, E. 141 O
3576 MK352193 MK352364 Brazil 2014 Barbosa, R.S., Haugan, R. & Timdal, E. 34 O
4160 MG925867 MG925967 Brazil 2015 Kistenich, S. & Timdal, E. SK1-108 O
P. gossypina ch1 4746 MG925868 MG925968 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-287 O
7201 MK352254 MK352420 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-584 PDA
P. gossypina ch2 4750 MK352219 MK352390 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-297 O
P. halei (Tuck.) Zahlbr. ch2 457 MK352113 MK352292 Tanzania 2008 Timdal, E. 10931 O
1044 MK352161 MK352336 Kenya 2007 Divakar, Lumbsch & Mangold 19574K hb. Pérez–Ortega
P. halei ch3 7221 MK352257 MK352423 Sri Lanka 2017 Weerakoon, G. 1008 PDA
P. hispaniolae Timdal 1545 MK352178 Ecuador 1999 Palice, Z. 3875 hb. Palice
3569 MK352188 MK352360 Ecuador 2014 Prieto, M. s.n. HUTPL
4039 MK352201 MK352372 Panama 2010 v.d. Boom, P. 44158 hb. v.d. Boom
P. imshaugii Timdal 3558 MK352185 MK352357 Ecuador 2014 Prieto, M. s.n. HUTPL
4043 MK352204 MK352375 Guatemala 2004 v.d. Boom, P. 33433 hb. v.d. Boom
4744 MK352217 MK352388 Venezuela 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-253 VEN
P. isidiosa Kistenich & Timdal 430 MK412409 MK412476 Thailand 1991 Wolseley, P.A. & Aguirre–Hudson, B. 5552 BM
1027 MK352153 MK352328 USA 2006 Lendemer, J.C. 7765 dupl. BG
1030 MK352155 MK352330 Nepal 2007 Sharma, L.R., Olley, L., Cross, A., Joshi, M. & Regmi, B. M16 E
1031 MK412428 Nepal 2007 Sharma, L.R., Olley, L., Cross L25-2 E
1259 MK412432 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. S.P.5 BORH
2099 MK412446 MK412494 Indonesia 2003 L. Sudirman & H. Sipman 51474 B
4035 MK352197 MK352368 Dominican Republic 2008 v.d. Boom, P. 39012 hb. v.d. Boom
4781 MG925907 MG926004 Brazil 2007 Lücking, R & Rivas Plata, E. 23302 SP
6349 MK352232 Philippines 1994 Diederich, P. 13210 hb. Diederich
7251 MK352267 MK352433 Australia 2006 Elix, J.A. 38478 CANB
P. isidiotyla (Vain.) Riddle 1315 MG925906 MG926003 Brazil 1979 Kalb, K. & Plöbst, G. in: Kalb, Lich. Neotrop. 343 GZU
P. kalbii Brako 456 MK352112 MK352291 Thailand 1993 Aguirre, B., James, P.W. & Wolseley, P. 2695 BM
458 MK352114 MK352293 Tanzania 2008 Timdal, E. 10913 O
459 MK352115 MK352294 Venezuela 1989 Kalb, K. & A. s.n. O
1028 MK352154 MK352329 USA 2010 Lendemer, J.C. 25770 BG
2052 MK352182 Bolivia 2010 Flakus, A. & Quisbert, J. 19221 O
P. loekoesii S.Y. Kondr. et al. 1033 MK352156 MK352331 Nepal 2007 Sharma, L.R., Olley, L., Cross A. C5 E
7478 MK352279 MK352439 Japan 1994 Thor, G. 12574 TNS
P. longiuscula (Nyl.) Zahlbr. 454 MG925899 MG925996 Peru 2006 Timdal, E. 10433 O
467 MK352117 MK352296 Trinidad And Tobago 2008 Rui, S. & Timdal, E. 10730 O
1011 MK412424 MK412488 Thailand 1992 Wolseley, P.A. & Aguirre–Hudson, B. 5580 p.p. BM
1039 MK352159 MK352334 Cuba 2006 Pérez–Ortega, S. s.n. hb. Pérez–Ortega
6761 MK352159 MK352413 Sri Lanka 2017 Weerakoon, G. Kn136 PDA
P. malcolmii Vezda & Kalb 1303 MK352170 MK352344 New Zealand 1994 Malcolm, W. in: Vezda, Lich. Rar. Exs. 200 GZU
P. martinii Swinscow & Krog 489 MK352129 MK352309 Tanzania 1989 Krog, H. 3T13/007 O
6740 MK352242 MK352409 Kenya 2014 Kirika, P. & Lumbsch, H.T. 4087 O
P. mauritiana (Taylor) Swinscow & Krog 487 MK352128 MK352307 Tanzania 1988 Krog, H. 2T12/037 O
488 MK352308 Mauritius 1991 Krog, H. & Timdal, E. MAU09/43 O
SE386 MG925900 MG925997 Mauritius 1991 Krog, H. & Timdal, E. MAU09/44 O
P. mediocris Swinscow & Krog 527 MK352144 MK352320 Tanzania 1988 Krog, H. 2T06/023 O
6346 MK352229 MK352399 Mauritius 2016 Diederich, P. 18571 hb. Diederich
6347 MK352230 MK352400 Mauritius 2016 Diederich, P. 18573 hb. Diederich
P. melanoglauca Zahlbr. 1038 MK352158 MK352333 Cuba 2006 Pérez–Ortega, S. s.n. hb. Pérez–Ortega
4042 MK352203 MK352374 Guatemala 2004 v.d. Boom, P. 33408 hb. v.d. Boom
4740 MK352213 MK352384 Venezuela 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-232 VEN
4743 MK352216 MK352387 Venezuela 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-247 VEN
6450 MK352235 MK352403 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-408 O
P. nemoralis Timdal & Krog 522 MK352142 La Réunion 1996 Krog, H. & Timdal, E. RE25/32 O
1434 MK352174 MK352347 South Africa 1996 Nordin, A. 4622 UPS:L:92604
P. neofoliata Elix 6745 MK352244 MK352411 Kenya 2015 Kirika, P. & Lumbsch, H.T. 4728 O
7245 MK352263 MK352429 Australia 1992 Elix, J.A. 32714 O
7249 MK352266 MK352432 Australia 1989 Elix, J.A. CANB
P. neotinica Kistenich & Timdal 505 MK352137 MK352316 Trinidad And Tobago 2008 Rui, S. & Timdal, E. 10774 O
1023 MK352149 MK352324 Cuba 2007 Tønsberg, T. 37923 BG
1438 MK352176 MK352349 Trinidad And Tobago 2008 Rui, S. & Timdal, E. 10763 O
P. neotinica 4742 MK352215 MK352386 Venezuela 2015 M.S. Dahl, J.E. Hernández M., S. Kistenich, E. Timdal & A.K. Toreskaas SK1-246 O
4769 MK352222 MK352393 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-402 O
P. ochroxantha (Nyl.) Zahlbr. 473 MK352118 MK352297 Peru 2006 Timdal, E. 10338 O
474 MK352119 MK352298 Peru 2006 Timdal, E. 10389 O
475 MK352120 MK352299 Trinidad And Tobago 2008 Rui, S. & Timdal, E. 10849 O
4049 MK352206 MK352377 Brazil 2015 Kistenich, S. & Timdal, E. SK1-47 O
4747 MK352218 MK352389 Brazil 2015 Dahl, M.S., Kistenich, S., Timdal, E. & Toreskaas, A.K. SK1-289 O
P. parvifolia (Pers.) Müll. Arg. 479 MK352124 MK352303 Tanzania 2008 Timdal, E. 10935 O
480 MK352125 MK352304 Trinidad And Tobago 2008 Rui, S. & Timdal, E. 10867 O
2049 MK352181 MK352353 Bolivia 2010 Flakus, A. & Quisbert, J. 20016 O
3561 MK352187 MK352359 South Africa 2014 Burrows, J. & Timdal, E. 14244 O
6365 MK352234 MK352402 Portugal 2015 v.d. Boom, P. 53877 hb. v.d. Boom
P. parvifoliella (Nyl.) Müll. Arg. 481 MK352126 MK352305 Peru 2006 Timdal, E. 10302 O
482 MG925902 MG925999 Indonesia 2000 Wolseley, P.A. s.n. BM:1104069
483 MK352127 MK352306 Thailand 1993 James, P.W. & Wolseley, P.A. 2491 BM
1004 MK412422 Thailand 1993 James, P.W. & Wolseley, P.A. 1847 BM
P. phaeobyssina (Vain.) Timdal 478 MK352123 MK352302 Trinidad And Tobago 2008 Rui, S. & Timdal, E. 10872 O
P. porphyromelaena (Vain.) Zahlbr. ch1 490 MK412416 MK412482 Thailand 1994 Wolseley, P. & Kanajriavanit, S. s.n. BM:1104012
498 MG925904 MG926001 La Réunion 1996 Krog, H. & Timdal, E. RE07/17 O
502 MK352135 MK352314 Japan 1995 Thor, G. 12941 UPS
1050 MK352164 MK352339 Kenya 2002 Killmann, D. & Fischer, E. s.n. hb. Killmann
1429 MK412444 Sri Lanka 2007 Jayalal, U. B9-4-3-3 PDA
7207 MK412464 MK412507 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-634 PDA
P. porphyromelaena ch2 491 MK412417 MK412483 Thailand 1993 Aguirre–Hudson, B. & Wolseley, P.A. 1663 BM
496 MK352133 Tanzania 1989 Krog, H. 4T16/019 O
503 MK352136 MK352315 Japan 2006 Thor, G. 21238 UPS
6436 MK412454 MK412497 Malaysia 2014 Paukov, A. 2233 B
7208 MK352255 MK352421 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-631 PDA
P. porphyromelaena ch2 7479 MK412475 MK412519 Japan 2017 Haugan, R. & Timdal, E. 16753 O:L:209897
P. porphyromelaena ch3 492 MK352130 MK352310 Thailand 1993 Aguirre, B., James, P.W. & Wolseley, P. 2857 BM
494 MK352132 MK352312 Thailand 1993 Aguirre, B., James, P.W. & Wolseley, P. 2481 BM
P. pseudocorallina Kistenich & Timdal 1034 MK412430 MK412491 Cambodia 2005 Kashiwadani, H. 47806 TNS
1418 MK412437 Malaysia 2012 Thüs, H., Wolseley, P. & Vairappan, C. M001a BORH
1419 MK412438 Malaysia 2012 Thüs, H., Wolseley, P. & Vairappan, C. M005 BORH
6356 MK412449 MK412495 Papua New Guinea 1992 Diederich, P. 11386 hb. Diederich
P. pyxinoides (Nyl.) Kistenich et al. 3574 MK352191 Brazil 2014 Cáceres, M., Haugan, R. & Timdal, E. 21024 O
7358 MK352274 USA 1991 Ryan, B. 27530 O
P. rappiana (Brako) Elix 6737 MK352240 MK352407 Australia 2005 Elix, J. 36867 O
7175 MK352250 MK352417 Panama 2010 v.d. Boom, P. 43820 hb. v.d. Boom
P. rosei Coppins & P. James 1299 MK352168 UK 1992 Coppins, B., James, P.W. & Poelt, J. Sc92/446 GZU
6339 MK352228 MK352398 France 2000 Diederich, P. 14602 hb. Diederich
7356 MK352272 MK352436 France 1990 Diederich, P. 9247 hb. Diederich
7357 MK352273 UK 1992 Coppins, B., James, P.W. & Poelt, J. Sc92/193 GZU
P. sabahana Kistenich & Timdal 1265 MK412434 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. S.B.oQ.3 BORH
1423 MK412441 Malaysia 2012 Thüs, H., Wolseley, P. & Vairappan, C. M089 BORH
1425 MK412442 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. D.8.02.4 BORH
6435 MK412453 MK412496 Malaysia 2014 Paukov, A. 2230 B
6457 MK412455 MK412498 Malaysia 2014 Paukov, A. 2229 B
P. santensis (Tuck.) Swinscow & Krog 2043 MK352179 MK352351 Bolivia 2009 Flakus, A. & Rodriguez, P. 15581 O
4038 MK352200 MK352371 Panama 2010 v.d. Boom, P. 44704 hb. v.d. Boom
4051 MK352207 MK352378 Brazil 2015 Kistenich, S. & Timdal, E. SK1-79 O
P. siamensis Kistenich & Timdal 448 MK412410 MK412477 Thailand 1993 Wolseley, P.A. & Boonpragob, K. 3245 BM
996 MK412418 MK412484 Thailand 1992 Wolseley, P.A. & Onsar 5590 BM
997 MK412419 MK412485 Thailand 1993 Aguirre–Hudson, B. & Wolseley, P.A. 1643 BM
1010 MK412423 MK412487 Thailand 1992 Wolseley, P.A. & Aguirre–Hudson, B. 5580 BM
P. sp. 1 7230 MK352259 MK352425 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-545 PDA
P. sp. 2 1017 MK352148 Malaysia 1997 Wolseley, P. s.n. BM:1104019
P. sp. 3 7227 MK352258 MK352424 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-555 PDA
P. sp. 4 7231 MK412471 MK412514 Sri Lanka 2017 Kistenich S. & Weerakoon, G. SK1-570 PDA
P. subhispidula (Nyl.) Kalb & Elix 501 MK352134 MK352313 Tanzania 1989 Krog, H. 4T15/007 O
6738 MK352241 MK352408 La Réunion 1996 Krog, H. & Timdal, E. RE36/15 O
6771 MK352249 MK352416 Sri Lanka 2017 Weerakoon, G. Hg29A PDA
P. swinscowii Timdal & Krog 476 MK352121 MK352300 Peru 2006 Timdal, E. 10190 O
525 MK352143 Mauritius 1991 Krog, H. & Timdal, E. MAU09/50 O
1025 MK352151 MK352326 Cuba 2007 Tønsberg, T. 37817 BG
1049 MK352163 MK352338 Kenya 2002 Killmann, D. & Fischer, E. s.n. hb. Killmann
4048 MK352205 MK352376 Brazil 2015 Kistenich, S. & Timdal, E. SK1-115 O
P. teretiuscula Timdal 1026 MK352152 MK352327 Cuba 2007 Tønsberg, T. 37814 BG
1306 MK352171 MK352345 Costa Rica 2003 Hafellner & Emmerer 1490 GZU
7474 MK352278 MK352438 Puerto Rico 1992 Harris, R.C. 27320 O
P. thaleriza (Stirt.) Brako 1048 MK352162 MK352337 Kenya 2003 Killmann, D. & Fischer, E. s.n. hb. Killmann
5465 MG925880 MG925982 South Africa 2014 Burrows, J. & Timdal, E. 14191 O
5466 MG925881 MG925983 South Africa 2015 Rui, S. & Timdal, E. 13877 O
5467 MK352226 MK352397 South Africa 2015 Rui, S. & Timdal, E. 13873 O

Molecular data and phylogenetic analysis

We obtained sequences for 140 phyllopsoroid specimens with 132 mtSSU and 106 ITS sequences (Tables 2, 3). Based on the initial RAxML analyses (not shown), 93 specimens were found to belong in Phyllopsora s. str. (Table 2) and were used in the subsequent analyses. The remaining 47 specimens did not belong in Phyllopsora s. str. (Table 3) and are referred to at the family level only due to many problems of generic affiliation.

The concatenated alignment had a length of 1,825 bp with 264 accessions including one specimen of Biatora beckhausii (Körb.) Tuck. and one of B. vacciniicola (Tønsberg) Printzen for rooting of the phylogenetic trees. The alignment contained ca. 20% missing data and is available from TreeBase (study no. 23881).

The software IQ-TREE suggested the following substitution models for four subsets: GTR+I+Γ for mtSSU and SYM+I+Γ for ITS1, 5.8S and ITS2. Bayesian phylogenetic analysis halted automatically after 40×106 generations, when the ASDSF in the last 50% of each run had fallen below 0.01. Following a burnin of 50%, we used 80,004 trees for the final Bayesian majority-rule consensus tree. The phylogenetic results generated by IQ-TREE vs. MrBayes showed no incongruences. The extended majority-rule consensus tree (Fig. 1; see Suppl. material 1: Fig. S1 for the uncollapsed version of the tree), based on the Bayesian topology with all compatible groups (BS ≥ 50 and/or PP ≥ 0.7), shows an overall good resolution of Phyllopsora species. Seventeen unidentified specimens did not associate with any known species in the phylogenetic tree (Fig. 1). Four unidentified specimens (1017, 7227, 7230, and 7231) were resolved on long branches, while the remaining 13 specimens grouped into three distinct, strongly supported clades (Fig. 1A–C). Clade A is resolved as sister to a clade consisting of P. cinchonarum and P. concinna, clade B is found in a clade with P. castaneocincta, P. foliata and P. neofoliata among others, and clade C is resolved as sister to P. neotinica.

Figure 1. 

Extended majority-rule consensus tree resulting from the MrBayes analysis of the mtSSU and ITS alignment with Bayesian PP ≥ 0.7 and/or IQ-TREE maximum likelihood BS ≥ 50 and branch lengths. Strongly supported branches (PP ≥ 0.95 and BS ≥ 75) are marked in bold; branches only supported with PP ≥ 0.7 or BS ≥ 50 are marked with a dot above the branch. Two species of Biatora were used for rooting. Accessions belonging to the same species are collapsed for convenience. Three clades are distinguished to facilitate the discussion of new species (A, B, C). ch = chemotype.

Newly generated sequences for specimens not belonging to Phyllopsora with voucher information and GenBank accession numbers. – indicates missing data.

Family Extract # mtSSU ITS Country Year Voucher Herb.
Malmideaceae 1268 MK400188 MK400239 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. D.1.10.3 BORH
Ramalinaceae 417 MK400189 MK400240 Thailand 1993 Wolseley, P.A. & David, F. 3347 BM:749829
423 MK400190 MK400241 Indonesia 2000 Wolseley, P. T9 LQ BM:1104053
427 MK400191 MK400242 Indonesia 2000 Wolseley, P. T13 LQ BM:1104062
432 MK400192 MK400243 Malaysia 1997 Wolseley, P. pkt. 8 BM:1104016
433 MK400193 MK400244 Thailand 1991 Wolseley, P.A. & Aguirre–Hudson, B. 5548 BM:749824
435 MK400194 MK400245 Indonesia 2000 Wolseley, P. T20 LMQ BM:1104013
1008 MK400195 Thailand 1993 Aguirre, James & Wolseley 2854 BM
1013 MK400196 Thailand 1993 James, P.W. & Wolseley, P.A. 1700b BM
1014 MK400197 MK400246 Thailand 1993 Aguirre, James & Wolseley 2478a BM:749861
1015 MK400198 MK400247 Thailand 1993 Aguirre, James & Wolseley 2715 BM:749853
1020 MK400199 Indonesia 2000 Wolseley, P. T6 LQ BM:1104066
1021 MK400200 Indonesia 2000 Wolseley, P. T1 BM:1104063
1266 MK400248 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. D.4.04.2 BORH
1270 MK400201 MK400249 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. M.1.12.oQ BORH
1275 MK400202 MK400250 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. D+40 BORH
1282 MK400203 MK400251 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. S.B.10.2 BORH
1284 MK400204 MK400252 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. D.7.09.1 BORH
1285 MK400205 MK400253 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. M.3.08.oQ.2 BORH
1287 MK400206 MK400254 Malaysia 2012 Wolseley, P., Thüs, H. & Vairappan, C. M.3.03.1 BORH
1426 MK400255 Malaysia 2013 Vairappan, C. L261 BM
1428 MK400207 MK400256 Thailand 1993 Aguirre, James & Wolseley 2477e BM:1031544
6056 MK400208 Malaysia 2014 Paukov, A. 2236 B
6057 MK400209 Malaysia 2014 Paukov, A. 2235 B
6762 MK400210 MK400257 Sri Lanka 2017 Weerakoon, G. Ne141 PDA
6768 MK400211 MK400258 Sri Lanka 2017 Weerakoon, G. WL60 PDA
6769 MK400212 MK400259 Sri Lanka 2017 Weerakoon, G. WL15/2 PDA
7186 MK400213 MK400260 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-651 PDA
7187 MK400214 MK400261 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-650 PDA
7188 MK400215 MK400262 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-564 PDA
7189 MK400216 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-566 PDA
7190 MK400217 MK400263 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-604 PDA
7191 MK400218 MK400264 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-602 PDA
7192 MK400219 MK400265 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-611 PDA
7193 MK400220 MK400266 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-558 PDA
7195 MK400221 MK400267 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-560 PDA
7196 MK400222 MK400268 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-673 PDA
Ramalinaceae 7198 MK400223 MK400269 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-659 PDA
7199 MK400224 MK400270 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-587 PDA
7202 MK400271 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-573 PDA
7204 MK400272 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-524 PDA
7206 MK400225 MK400273 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-666 PDA
7211 MK400226 MK400274 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-561 PDA
7215 MK400275 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-592 PDA
7222 MK400227 MK400276 Sri Lanka 2017 Weerakoon, G. 641 loc.31 PDA
7226 MK400277 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-628 PDA
7228 MK400228 Sri Lanka 2017 Kistenich, S. & Weerakoon, G. SK1-667 PDA

Discussion

In this study, we present the first revision of the genus Phyllopsora for Asia and Melanesia based on the integrative study of morphology, chemistry and DNA sequence data. We investigated 625 specimens of Phyllopsora collected from 18 countries and found the material to comprise at least 28 species of Phyllopsora s. str. (Figs 210) including three supported clades that we describe as species new to science. With this study, the genus Phyllopsora comprises 57 species.

Several species seem to be rather widespread throughout Asia and Melanesia, for instance, P. castaneocincta, P. confusa, P. isidiosa, and P. porphyromelaena (Table 1, Suppl. material 2: Table S1). In contrast, specimens of, for example, P. cuyabensis, P. mediocris and P. neofoliata, are rarely collected and reported from few countries (Table 1, Suppl. material 2: Table S1). Thus, their distribution range requires further studies.

Among the 28 species of Phyllopsora, eight are reported as new for Asia and Melanesia (Table 1). One of these new species is P. africana (Fig. 2A). This species has recently been found to be morphologically and chemically heterogeneous, comprising three chemotypes (Kistenich et al. in press). In addition to the known isidiate morph, a lacinulate morph was detected among P. africana material by Kistenich et al. (in press). Moreover, they described two new chemotypes. The lacinulate morph occurred in specimens of chemotype 1 and 3, but has so far never been found in those of chemotype 2. Specimens of chemotype 2, however, were shown to be morphologically cryptic to the sister species P. swinscowii (Kistenich et al. in press). In this study, we added twelve specimens of P. africana to our phylogeny (mainly lacinulate specimens of chemotype 3), but are not able to disentangle the difficult nature of this species complex. While we found most specimens of P. africana to roughly group according to chemotype in the phylogenetic tree (Suppl. material 1: Fig. S1), one specimen of P. africana chemotype 1 (7224) was more closely related to P. swinscowii (Suppl. material 1: Fig. S1). This raises the question of whether the two species should be synonymized based on their morphological and chemical similarity in combination with the short branches in the phylogenetic tree (Suppl. material 1: Fig. S1). We refrain from synonymizing them here, awaiting more data.

Figure 2. 

Species of Phyllopsora occurring in Asia and Melanesia. A Phyllopsora africana (Kistenich & Weerakoon SK1-517) B P. breviuscula (Kistenich & Weerakoon SK1-601) C P. buettneri (Thor 13183). Scale bars: 2 mm.

The two species P. cuyabensis (Fig. 4B) and P. longiuscula (Fig. 7B) are reported as new for the Asian continent. Specimens of both species are morphologically congruent with their Neotropical representatives. In the phylogenetic tree (Suppl. material 1: Fig. S1), however, the respective Asian accessions sit on rather long branches, clearly distinct from the Neotropical specimens. In these cases, there seem to exist genetically different populations for Neotropical and Asian specimens and more specimens should be collected to investigate the extent of genetic variation.

The genus Phyllopsora was recently shown to be polyphyletic by Kistenich et al. (2018b). The typical growth form, which characterizes this genus, has evolved multiple times independently in the family Ramalinaceae. These findings corroborate the morphological co-evolution in tropical lichens already indicated by Lakatos et al. (2006). Hence, molecular methods are often the only means of reliably assigning specimens to Phyllopsora or rather to its morphologically similar relatives (e.g., Bacidia De Not., Bacidina Vězda, Eschatogonia, Parallopsora). It is thus not surprising that several of our sequenced specimens (Table 3) were extraneous to Phyllopsora s. str. We did not assign those specimens to genus level, but all but one belong in the Ramalinaceae. The non-Ramalinaceae specimen appears to belong in the Malmideaceae. This indicates that correct taxonomic assignment even at family level using morphology may prove challenging in certain cases. Furthermore, about a quarter of the total material investigated could not be identified, partly because many of those unidentified specimens were sterile and deficient in lichen substances. Unfortunately, we were not able to generate sequences of all unidentified specimens in the course of this study.

New species

The three new species, P. pseudocorallina, P. sabahana and P. siamensis, fall into distinct and well-supported clades in the phylogeny (Fig. 1A–C). They were originally assumed to comprise Asian populations of the species P. porphyromelaena, P. corallina and P. imshaugii, respectively, based on morphology and/or chemistry. Their sequence data, however, revealed them as separate species clearly distinct from their look-alikes (Fig. 1). Phyllopsora pseudocorallina (Fig. 9B) is distinguished from its namesake, i.e. P. corallina, by forming a partly more rosulate thallus. Poorly developed specimens, however, might be difficult to assign to the correct species. Specimens of P. sabahana (Fig. 9C) are challenging to identify based on morphology only. The species is morphologically and chemically almost identical to P. porphyromelaena chemotype 1. It differs only in forming slightly smaller ascospores. Thus, sterile specimens cannot be identified without DNA sequence data. Phyllopsora siamensis (Fig. 10B) is described from material collected in Thailand and we have not been able to detect this species in collections from other countries. The specimens resemble P. imshaugii in morphology and chemistry, but may be readily distinguished by forming larger ascospores. See also the remarks in the Taxonomy section.

In addition, we found sequences of the four unidentified specimens with extraction numbers 1017, 7227, 7230, and 7231 to be resolved on rather long branches (Fig. 1, Suppl. material 1: Fig. S1). Hence, we could not assign them to any other Phyllopsora species, for which DNA sequences of the mtSSU or ITS region were available, based on molecular data, either. It is possible that these specimens represent several new species. In this study, however, we refrain from describing them as new species pending the collection of more material. Even though specimens 1017 and 7230 are clustered together in a clade with short branches (Suppl. material 1: Fig. S1), they are morphologically quite distinct and more specimens are needed to support the hypothesis that they belong to the same species.

Unconfirmed species records

Despite investigating about 600 phyllopsoroid specimens, we were not able to find in our material any specimens belonging to seven species (i.e., P. chlorophaea, P. corallina, P. isidiotyla, P. mauritiana, P. nemoralis, P. pyxinoides, and P. swinscowii) previously reported from India, South Korea, Sri Lanka, Taiwan, Thailand, and Vietnam (Table 1), respectively. We have only investigated a few collections from especially India, South Korea, Taiwan, and Vietnam (Suppl. material 2: Table S1), though. Also for the other countries, collections are limited to certain areas and we cannot exclude the species’ occurrence in other parts of the respective countries. About 23% of the investigated material could not be identified to species level and it is possible that some of these unidentified specimens represent a poorly developed individual of any of these seven species. Regarding P. corallina, for instance, we found two candidate specimens from Papua New Guinea, but DNA sequence data is necessary to resolve their species status unambiguously. Alternatively, some of these species records might be based on misidentifications. In the case of P. swinscowii, we have shown the species to be morphologically identical to the isidiate morph of P. africana, a very widespread species. It is therefore possible that the records of P. swinscowii indeed represent P. africana. In general, we have repeatedly experienced difficulties in correctly identifying species of Phyllopsora based on morphology only. For many of the species records, it remains unclear whether anatomical studies and/or chemical investigations were performed as part of the identification process or not. Especially P. chlorophaea, P. corallina and P. isidiotyla may be difficult to identify without TLC or even sequence data.

Taxonomy

This taxonomy section is a result of the integrative species delimitation process primarily based on the conclusions from the statistically inferred species delimitation analyses combined with morphological and chemical evaluations as performed in the global Phyllopsora study by Kistenich et al. (in press). The additional material of the present study complements the global dataset for the phylogenetic analysis (Fig. 1, Suppl. material 1: Fig. S1) and revealed three new species, which were mainly delimited by forming separate clades on long branches compared to their neighboring clades.

Distribution references for Asia and Melanesia are cited in Table 1; for all other distributions, references are cited below.

Phyllopsora africana Timdal & Krog

Description

Timdal and Krog (2001), Elix (2009).

Distribution

Africa (Timdal and Krog 2001), Asia, Australia (Elix 2009).

Remarks

See discussion above and Kistenich et al. (in press) for taxonomic discussion. The species (Fig. 2A) is one of the most common in our material, represented by 59 collections (Suppl. material 2: Table S1). We found both isidate and lacinulate morphs as well as representatives of all three chemotypes (i.e., chemotype 1 contains chlorophyllopsorin and argopsin; chemotype 2 contains methyl 2,7-dichloropsoromate and methyl 2,7-dichloronorpsoromate; chemotype 3 contains chlorophyllopsorin, methyl 2,7-dichloropsoromate, methyl 2,7-dichloronorpsoromate, and argopsin) among the material. It is the phylogenetic sister to P. swinscowii (Fig. 1). The species is new to Asia and Melanesia, i.e. to Indonesia, Japan, Malaysia, Papua New Guinea, The Philippines, The Solomon Islands, Sri Lanka, Thailand, and Vanuatu.

Phyllopsora breviuscula (Nyl.) Müll. Arg.

Description

Timdal and Krog (2001), Elix (2009).

Distribution

Pantropical (Brako 1991, as P. parvifolia var. breviuscula; Timdal and Krog 2001; Elix 2009).

Remarks

Paleotropical material of this species (Fig. 2B) tends to be more narrow-lobed and ascending than Neotropical material. Species delimitation analyses by Kistenich et al. (in press) show that the taxon could be split into three or four entities, but as all sequenced specimens fell into one well-supported clade, and morphologically intermediate specimens exist, we still treat the taxon as one variable species. It is the phylogenetic sister to P. mauritiana (Fig. 1). The species is new to New Caledonia, The Philippines, and Vietnam.

Phyllopsora buettneri (Müll. Arg.) Zahlbr.

Description

Swinscow and Krog (1981), Timdal and Krog (2001), Timdal (2008b, as P. buettneri chemotypes 1 and 2), Elix (2009).

Distribution

Pantropical (Brako 1991, as P. buettneri var. buettneri; Timdal and Krog 2001; Elix 2009).

Remarks

We recognize five chemotypes of this species, three occurring in Asia and Melanesia (Fig. 2C). Chemotype 1 (pannarin and zeorin) was found in material from Japan, Sri Lanka, and Thailand; chemotype 3 (dechloropannarin, zeorin) in material from China, Japan, Sri Lanka, and Thailand; and chemotype 5 (pannarin and an unknown compound, no zeorin) in material from Papua New Guinea and Sri Lanka (Suppl. material 2: Table S1). The specimen of chemotype 1 from Japan, and one of the two from Sri Lanka, differed from typical specimens of the chemotype in lacking zeorin. Chemotype 5 is described here for the first time, and contains an unknown compound with Rf values similar to argopsin in solvent system B’ but with a distinct blue UV366 fluorescence (not quenching) on the chromatograms after development. Chemotype 2 (pannarin, phyllopsorin, zeorin) is Neotropical and chemotype 4 (argopsin, norargopsin, zeorin) is known from the Norfolk Islands. We were unable to sequence specimens of chemotype 4 and 5, but specimens of chemotype 1–3 are resolved in a clade with chemotype 1 and 2 of P. porphyromelaena (Fig. 1, Suppl. material 1: Fig. S1). The species is new to China, Japan, and Sri Lanka.

Phyllopsora castaneocincta (Hue) Kistenich & Timdal

Description

Timdal and Krog (2001), Elix (2009), both as P. kiiensis.

Distribution

Africa (Timdal and Krog 2001), Asia, Australia (Elix 2009).

Remarks

This is one of the most common species in our material (Suppl. material 2: Table S1). It is usually easily recognized by the well-developed squamulose thallus on a reddish brown prothallus and by containing furfuraceic acid (Fig. 3A), but care is needed as about 10% of the examined specimens were actually deficient in lichen substances. It is new to Cambodia, Malaysia, Nepal, New Caledonia, Papua New Guinea, The Solomon Islands, Taiwan, and Thailand.

Figure 3. 

Species of Phyllopsora occurring in Asia and Melanesia. A Phyllopsora castaneocincta (Kirika, Mugambi, & Lumbsch 3011) B P. chodatinica (Paukov 2232) C P. cinchonarum (Thor 21521). Scale bars: 2 mm.

Phyllopsora chodatinica Elix

Description

Elix (2006, 2009).

Distribution

Australasia (Elix 2009) and Oceania.

Remarks

This species resembles P. porphyromelaena and P. sabahana, to which it is closely related in the phylogenetic tree (Fig. 1), but differs in the presence of xanthones and the absence of argopsin and norargopsin. Kistenich et al. (in press) showed that probably all Neotropical records of this species (e.g., by Timdal 2008b, 2011) belong in another species, P. neotinica. Phyllopsora chodatinica (Fig. 3B) is new to Malaysia, New Caledonia, and Vanuatu.

Phyllopsora cinchonarum (Fée) Timdal

Description

Brako (1989, as Squamacidia janeirensis), Timdal (2008b), Elix (2009, as Triclinum cinchonarum).

Distribution

Central and South America (Brako 1989; Timdal 2008b), Asia, Australia (Elix 2009).

Remarks

The species is recognized by the squamulose thallus on a white prothallus, long isidia, and the presence of lobaric acid (Fig. 3C). Several additional compounds are reported, for example atranorin, fumarprotocetraric acid, and a scarlet pigment. In our Asian material, we have encountered only lobaric acid (always major), atranorin (minor to absent), and some unknown compounds (minor to absent). It is the phylogenetic sister to the Neotropical P. concinna (Fig. 1).

Phyllopsora confusa Swinscow & Krog

Description

Swinscow and Krog (1981), Timdal and Krog (2001), Elix (2009).

Distribution

Pantropical (Brako 1991; Timdal and Krog 2001; Elix 2009).

Remarks

This species is characterized by the small, lacinulate squamules lacking lichen substances (Fig. 4A), but may be difficult to separate from, for example, P. foliata and P. mediocris. It is also possible that some of the specimens we have left undetermined belong in this species. We have sequenced nine specimens from Asia and Melanesia (Table 2), in addition to the holotype from Kenya, and those specimens make up the core in our concept of this species. The accessions of P. confusa form a strongly supported clade with P. loekoesii in the phylogenetic tree (Fig. 1). The two specimens from Papua New Guinea (6360 and 6361) fall in between the P. confusa and P. loekoesii clade (Suppl. material 1: Fig. S1) and show an intermediate morphology. Further specimens are needed to investigate the possible synonymy of these two species. Phyllopsora confusa is new to Indonesia, Japan, Malaysia, Taiwan, and Thailand.

Figure 4. 

Species of Phyllopsora occurring in Asia and Melanesia. A Phyllopsora confusa (Kistenich & Weerakoon SK1-532) B P. cuyabensis (Aguirre, James & Wolseley 2467a) C P. dolichospora (Weerakoon Si113B). Scale bars: 2 mm.

Phyllopsora cuyabensis (Malme) Zahlbr.

Description

Timdal (2008b).

Distribution

Central and South America (Brako 1991; Timdal 2008b), Asia.

Remarks

The species is represented by a single specimen from Thailand in our material (Fig. 4B). The Asian accession (450) falls into a strongly supported clade with sequences from four Neotropical specimens, although resolved on a long branch as sister to all Neotropical accessions (Suppl. material 1: Fig. S1). Being morphologically identical to the Neotropical specimens, it is unclear whether this specimen (450) represents a new species or merely genetic variation within P. cuyabensis. Additional sequences of Asian specimens are necessary to evaluate this possibility further. The species is sister to a clade comprising P. byssiseda, P. fendleri and P. kalbii (Fig. 1). The species is new to Asia.

Phyllopsora dolichospora Timdal & Krog

Description

Timdal and Krog (2001).

Distribution

Africa (Timdal and Krog 2001), Asia.

Remarks

This species (Fig. 4C) is morphologically and chemically (furfuraceic acid) similar to P. furfuracea, to which it is closely related (Fig. 1), but differs in forming longer ascospores and in containing additional substances (methyl furfuraceiate and methyl homofurfuraceiate). Judging from the number of examined specimens (Suppl. material 2: Table S1), P. dolichospora seems to be more common than P. furfuracea in Asia, although the number of reports (Table 1) suggests the opposite. This, however, might be a result of morphological misidentifications when TLC has not been run. The species is new to Japan and Papua New Guinea.

Phyllopsora foliata (Stirt.) Zahlbr.

Description

Elix (2009).

Distribution

Asia, Australia (Elix 2009).

Remarks

This rarely reported Australian species (Fig. 5A) is here confirmed from Japan and Sri Lanka mainly based on our DNA sequences (both mtSSU and ITS), which were compared with sequences obtained from Australian material (Table 2). It is new to Japan.

Figure 5. 

Species of Phyllopsora occurring in Asia and Melanesia. A Phyllopsora foliata (Kistenich & Weerakoon SK1-627) B P. furfuracea (Wolseley & Aguirre-Hudson 4025) C P. gossypina (Kistenich & Weerakoon SK1-524). Scale bars: 2 mm.

Phyllopsora furfuracea (Pers.) Zahlbr.

Description

Timdal and Krog (2001), Timdal (2008b), Elix (2009).

Distribution

Pantropical (Brako 1991; Timdal and Krog 2001; Elix 2009).

Remarks

Despite widespread reports in the literature, we were able to confirm the presence of this species (Fig. 5B) in Papua New Guinea, Sri Lanka, and Thailand, only. In the phylogenetic tree, P. furfuracea forms a clade with P. dolichospora and P. foliatella (Fig. 1).

Phyllopsora gossypina (Sw.) Kistenich, Timdal, Bendiksby & S. Ekman

Description

Hue (1909).

Distribution

Apparently pantropical.

Remarks

The species (Fig. 5C) was included in the genus Crocynia until recently (Kistenich et al. 2018b), and not originally a part of our taxon sampling; hence the few specimens examined. The accession from Sri Lanka (7201) clusters together with specimens of C. molliuscula (Suppl. material 1: Fig. S1), from which it is morphologically and chemically different. Further specimens need to be investigated to inform about its relationship to C. molliuscula. The species is the phylogenetic sister to P. imshaugii (Fig. 1).

Phyllopsora halei (Tuck.) Zahlbr.

Description

Swinscow and Krog (1981, as P. pannosa), Timdal and Krog (2001).

Distribution

North America (Brako 1991), Africa (Timdal and Krog 2001), Asia.

Remarks

This species (Fig. 6A) was previously known from the type collection from North America (Louisiana), East Africa (Ethiopia, Kenya, Tanzania), and a few reports from Asia (Table 1). We here confirm its presence in Asia, based on DNA sequences from material from Sri Lanka compared with sequences from Kenya and Tanzania (Suppl. material 1: Fig. S1). Three chemotypes of this species are known (Timdal and Krog 2001), differing in terpenoid patterns and presence of an unknown compound. Our two specimens from Sri Lanka belong in chemotype 3 of Timdal and Krog (2001). The species is the phylogenetic sister to P. amazonica (Fig. 1). It is new to Sri Lanka.

Figure 6. 

Species of Phyllopsora occurring in Asia and Melanesia. A Phyllopsora halei (Weerakoon 1008) B P. isidiosa (Diederich 13210) C P. kalbii (Aguirre, James & Wolseley 2695). Scale bars: 2 mm.

Phyllopsora himalayensis G.K. Mishra, Upreti & Nayaka

Description

Mishra et al. (2011).

Distribution

India (Mishra et al. 2011).

Remarks

The species was not studied by us due to lack of response from LWG to our repeated loan requests.

Phyllopsora isidiosa Kistenich & Timdal

Description

Kistenich et al. (in press).

Distribution

Pantropical, also occurring in temperate Asia and North America (Kistenich et al. in press).

Remarks

The species (Fig. 6B) is treated in detail by Kistenich et al. (in press). It is closely related to, for instance, P. dolichospora, P. foliatella, and P. furfuracea (Fig. 1).

Phyllopsora kalbii Brako

Description

Brako (1991), Timdal and Krog (2001).

Distribution

North, Central, and South America (Brako 1991), Africa (Timdal and Krog 2001), Asia.

Remarks

The species (Fig. 6C) was reported from India by Mishra et al (2011), and we confirm its presence in Asia by DNA sequences (mtSSU and ITS; 456) from material from Thailand. The species is sister to a clade comprising P. byssiseda and P. fendleri (Fig. 1). It is new to Thailand.

Phyllopsora loekoesii S.Y. Kondr., E. Farkas, S.–O. Oh & Hur

Description

Kondratyuk et al. (2016).

Distribution

Asia.

Remarks

The species (Fig. 7A) was recently described from South Korea by Kondratyuk et al. (2016), and we report it as new to Japan and Nepal. Our sequences were compared to unpublished sequences of the holo- and isotype kindly provided to us by Sergey Kondratyuk. Our accessions form a strongly supported clade together with accessions of P. confusa (Fig. 1), from which it is difficult to distinguish. See also remarks for P. confusa.

Figure 7. 

Species of Phyllopsora occurring in Asia and Melanesia. A Phyllopsora loekoesii (Sharma, Olley & Cross AC5) B P. longiuscula (Weerakoon Kn136) C P. mediocris (holotype, Moberg 1481a-1, Tanzania). Scale bars: 2 mm.

Phyllopsora longiuscula (Nyl.) Zahlbr.

Description

Brako (1991).

Distribution

Central and South America (Brako 1991), Asia, Australia (Kistenich et al. in press).

Remarks

In the concept of Brako (1991) and Timdal (2011), this species is lacinulate. Kistenich et al. (in press), however, extend the concept to include the isidiate species P. intermediella, which they synonymize. The Asian material we have examined is lacinulate. The species (Fig. 7B) is the phylogenetic sister to P. thaleriza (Fig. 1). It is new to Asia (Sri Lanka, Thailand, and Vietnam).

Phyllopsora mediocris Swinscow & Krog

Description

Swinscow and Krog (1981), Timdal and Krog (2001).

Distribution

Africa (Timdal and Krog 2001), Asia.

Remarks

The species (Fig. 7C) was previously known from East Africa and the Mascarenes (Timdal and Krog 2001). Although not sequenced, we here report it as new to Asia based on a specimen (Moberg 2750, UPS) from Sri Lanka (Suppl. material 2: Table S1). The species is the phylogenetic sister to P. parvifolia (Fig. 1).

Phyllopsora neofoliata Elix

Description

Elix (2006, 2009).

Distribution

Africa (Kistenich et al. in press), Asia, Australia (Elix 2009).

Remarks

This originally Australian species is reported as new to Africa (Kenya) by Kistenich et al. (in press) and here as new to Asia (Sri Lanka; Fig. 8A). Both the African and Sri Lankan specimens were sequenced (mtSSU and ITS) and found to conform with sequences of an isotype (O L-1319). The species is generally identified by the squamulose, lacinulate thallus containing furfuraceic acid.

Figure 8. 

Species of Phyllopsora occurring in Asia and Melanesia. A Phyllopsora neofoliata (Weerakoon WL21) B P. parvifolia (Kistenich & Weerakoon SK1-661) C P. parvifoliella (James & Wolseley 2491). Scale bars: 2 mm.

Phyllopsora parvifolia (Pers.) Müll. Arg.

Description

Elix (2009).

Distribution

Pantropical, but mainly Neotropical, extending into the temperate zones in North and South America and in Europe (Brako 1991, as P. parvifolia var. parvifolia; Kistenich et al. in press).

Remarks

Despite several reports from Asia and Melanesia (Table 1), we have seen only a single specimen of this species (Sri Lanka, Kistenich & Weerakoon SK1-661, PDA, not sequenced; Fig. 8B) from the area. The species is the phylogenetic sister to P. mediocris (Fig. 1). It is new to Sri Lanka.

Phyllopsora parvifoliella (Nyl.) Müll. Arg.

Description

Timdal (2008b).

Distribution

Central and South America (Brako 1991; Timdal 2008b), Asia.

Remarks

This squamulose, isidiate species contains atranorin and parvifoliellin (Fig. 8C); characters it shares with P. concinna and P. rappiana. The molecular phylogeny (Fig. 1) shows that the three species are not closely related, though; rather P. parvifoliella belongs in a clade together with P. africana, P. ochroxantha, and P. swinscowii. We have sequenced material from Indonesia and Thailand, and here report the species as new to Asia and Melanesia, i.e. from Indonesia, Papua New Guinea, The Philippines, and Thailand.

Phyllopsora porphyromelaena (Vain.) Zahlbr.

Description

Timdal and Krog (2001), Elix (2009), both as P. albicans.

Distribution

Pantropical (Brako 1991, as P. buettneri var. glauca chemical stains I and III; Timdal and Krog 2001; Elix 2009).

Remarks

This is the most common species in our material from Asia and Melanesia (Fig. 9A), despite previous records only from India, The Philippines, and Taiwan (mostly as P. albicans). Two chemotypes are previously recognized, and a third is recognized here. Chemotype 1 (argopsin and norargopsin) and chemotype 2 (argopsin and pannarin) are both widely distributed in Asia and Melanesia, but chemotype 3 (zeorin and three unknown compounds) is restricted to Thailand (Suppl. material 2: Table S1). The unknown compounds move in Rf-classes A:3–4, B’:4–5, C:5 (major compound); A:6, B’:6, C:5–6 (minor compound); and A:3, B’:3, C:5 (minor compound).

In the phylogenetic tree (Fig. 1, Suppl. material 1: Fig. S1), accessions of chemotypes 1 and 2 group into a weakly supported clade with P. buettneri, while accessions of chemotype 3 form a clade with P. chodatinica and the P. buettneri/P. porphyromelaena clade. Additional specimens of chemotype 3 should be sequenced to find out whether it indeed represents a chemical strain of P. porphyromelaena or rather a distinct species.

The species is morphologically very similar to P. sabahana; see that species for discussion. It is possible that some specimens listed as P. porphyromelaena chemotype 1 in Suppl. material 2: Table S1, especially those from Malaysia, represent P. sabahana. It is new to Fiji, Indonesia, Japan, Malaysia, New Caledonia, Papua New Guinea, South Korea, Sri Lanka, and Thailand.

Figure 9. 

Species of Phyllopsora occurring in Asia and Melanesia. A Phyllopsora porphyromelaena (Kistenich & Weerakoon SK1-572) B P. pseudocorallina sp. nov. (holotype, Kashiwadani 47806) C P. sabahana sp. nov. (holotype, Wolseley, Thüs & Vairappan S.B.oQ.3). Scale bars: 2 mm.

Phyllopsora pseudocorallina Kistenich & Timdal, sp. nov

MycoBank No: 829572
Fig. 9B

Diagnosis

Differs from P. corallina in having a more rosulate thallus and in substitutions in the mtSSU and ITS sequences.

Type

CAMBODIA, Siem Reap: around Ta Nei temple, Angkor Wats complex, 13°27'N, 103°53'E, ca. 30 m alt., on rock (sand stone), 2005-12-20, H. Kashiwadani 47806 (TNS!—holotype) [TLC: no lichen substances; DNA: MK412430 (mtSSU), MK412491 (ITS)].

Description

Thallus effuse or forming irregular rosettes up to 1 cm diam., squamulose; squamules medium sized, up to 1 mm wide, adnate to ascending, elongate, contiguous or partly imbricate, crenulate to incised, plane to weakly convex, medium green, glabrous on the upper side, faintly pubescent along the margin; isidia common, attached marginally to the squamules, cylindrical, simple or slightly branched, up to 0.1 mm wide and 0.6 mm long; upper cortex formed by thick-walled hyphae with rounded lumina (type 2), 20–30 µm thick; cortex and medulla not containing crystals (PD–, K–); prothallus indistinct to partly well developed, white.

Apothecia common, up to 1.5 mm diam., rounded when young, later often becoming irregular, simple or sometimes somewhat conglomerate, plane to moderately convex, yellowish to medium brown, with an indistinct, usually slightly paler, glabrous to finely pubescent margin; excipulum pale brown to colourless, K–; hypothecium pale brown to colourless, K–; epithecium colourless; no crystals in apothecium; ascospores narrowly ellipsoid to shortly bacilliform, simple, 6–10 × 2.5–3 µm (n=30). Conidiomata not seen.

Chemistry

No lichen substances.

Distribution

Cambodia, Malaysia, Papua New Guinea, The Seychelles.

Etymology

The specific epithet refers to its morphological and chemical similarity to P. corallina.

Remarks

The species is morphologically, anatomically, and chemically very similar to P. corallina. There is, however, a tendency of P. pseudocorallina being more rosulate, i.e., composed of more radiating and elongated marginal lobes. The phylogenetic tree (Fig. 1), however, shows the two species not to be closely related: P. corallina is resolved in a clade with P. glaucella and P. rappiana as sister to P. martinii, while P. pseudocorallina appears in a clade with P. castaneocincta and P. neofoliata among others (Fig. 1). The new species is widely distributed in Asia and also found on The Seychelles. It is unclear whether P. corallina occurs in Asia at all or if previously reported specimens of P. corallina rather represent specimens of P. pseudocorallina.

Additional specimens examined

MALAYSIA, Sabah: Malaysian Borneo, Maliau, “Knowledge Trail”, pristine lowland dipterocarp forest; on stem (ca. 20 m high) of fallen tree on crushed bridge, 2012, H. Thüs, P. Wolseley & C. Vairappan M001a (BORH) [DNA: MK412437 (mtSSU)]; same locality data, H. Thüs, P. Wolseley & C. Vairappan M001b (BORH); same locality data, H. Thüs, P. Wolseley & C. Vairappan M005 (BORH) [DNA: MK412438 (mtSSU)]. PAPUA NEW GUINEA, Madang: Manam island, near Bogia, in gardens near Budua, 4°07'S, 145°00'E, 50 m alt., epiphytic, 1992-07-22, P. Diederich 11386 (hb. Diederich) [DNA: MK412449 (mtSSU), MK412495 (ITS)]. THE SEYCHELLES, Mahé: W of Anse Royale, Le Jardin du Roi, 4.74642S, 55.50297E, 150–200 m alt., parkland and neighbouring forest, on rock, 2015-07-26, P. Diederich 17809 (hb. Diederich); Port Glaud, near Sauzier Waterfall, 4.65847S, 55.41403E, 20–70 m alt., on tree, 2015-07-28, P. Diederich 17897 (hb. Diederich).

Phyllopsora sabahana Kistenich & Timdal, sp. nov.

MycoBank No: 829571
Fig. 9C

Diagnosis

Differs from P. porphyromelaena in having smaller ascospores and in substitutions in the mtSSU and ITS sequences.

Type

MALAYSIA, Sabah: Malaysian Borneo, SAFE-project Area, mostly Macaranga dominated secondary forest, 2012, P. Wolseley, H. Thüs & C. Vairappan S.B.oQ.3 (BORH!—holotype) [TLC: argopsin (major), norargopsin (minor); DNA: MK412434 (mtSSU)].

Description

Thallus effuse, squamulose; squamules medium sized, up to 0.8 mm wide, ascending, elongated, often imbricate, incised to deeply divided, plane to weakly convex; upper side pale green to medium green, glabrous, epruinose; margin concolorous with upper side, often finely pubescent; lacinules common, developing from lobe-tips; upper cortex formed by thick-walled hyphae with cylindrical lumina (type 1), 30–40 µm thick, containing crystals dissolving in K (PD+ orange, K–); medulla containing crystals partly dissolving in K (PD+ orange, K–); prothallus well developed, reddish brown.

Apothecia not common, up to 2 mm diam., rounded to irregular, simple or soon becoming conglomerate, weakly to moderately convex, yellowish brown, more or less immarginate even when young; excipulum pale brown to colourless, K–; hypothecium medium brown, K–; epithecium pale brown to colorless; no crystals in apothecia; ascospores narrowly ellipsoid, simple, 6–8 × 2–2.5 µm (n=20). Conidiomata not seen.

Chemistry

Argopsin (major), norargopsin (minor). Medulla and upper cortex PD+ orange, K–, C–, KC–.

Distribution

Malaysia (Borneo).

Etymology

The specific epithet refers to its occurrence in Sabah, Malaysia.

Remarks

The species is morphologically and chemically very similar to P. porphyromelaena chemotype 1, and is close to be regarded as a morphologically cryptic species. It may, however, be distinguished in forming smaller ascospores (6–8 × 2–2.5 vs. 8–13 × 2–4 µm). Apothecia are not common in neither species, however, and the measurements are based on only 20 spores from each species (the holotype of P. sabahana and two specimens of P. porphyromelaena from La Réunion). In the phylogenetic tree (Fig. 1), the five accessions of P. sabahana form a strongly supported clade as sister to the Neotropical species P. neotinica, from which it may readily be distinguished in its composition of lichen substances (P. neotinica contains xanthones). So far, P. sabahana is only known from Borneo.

Additional specimens examined

MALAYSIA, Sabah: Malaysian Borneo: Maliau Basin, surroundings of Agathis Camp, pristine lowland Dipterocarp forest, 2012, P. Wolseley, H. Thüs & C. Vairappan, C. M089 (BORH) [DNA: MK412441 (mtSSU)]; Danum valley, pristine lowland Dipterocarp forest, 2012, P. Wolseley, H. Thüs & C Vairappan D.8.02.4 (BORH) [DNA: MK412442 (mtSSU)]; Ranau district, Kinabalu park, Tambuyukon trail, Kera camp (loc. T089), 6°12.742'N, 116°43.609'E, 728 m alt., epiphytic, 2014-12-08, A. Paukov 2229 (B) [DNA: MK412455 (mtSSU), MK412498 (ITS)] & 2230 (B) [DNA: MK412453 (mtSSU), MK412496 (ITS)].

Phyllopsora santensis (Tuck.) Swinscow & Krog

Description

Timdal (2008b), Elix (2009).

Distribution

North, Central, and South America (Brako 1991, as P. corallina var. santensis; Timdal 2008b), Asia, Australia (Elix 2009).

Remarks

The species was previously reported from Japan, Papua New Guinea, and The Philippines (Table 1), and is here reported from four localities in Thailand (Fig. 10A). We were unable to produce DNA sequences from our material, and the identification is based on typical morphology and presence of argopsin (major) and noragopsin (minor). New to Thailand.

Figure 10. 

Species of Phyllopsora occurring in Asia and Melanesia. A Phyllopsora santensis (Aguirre, James & Wolseley 2485) B P. siamenses sp. nov. (holotype, Wolseley & Boonpragob 3245) C P. subhispidula (Weerakoon 1248). Scale bars: 2 mm.

Phyllopsora siamensis Kistenich & Timdal, sp. nov.

MycoBank No: 829573
Fig. 10B

Diagnosis

Differs from P. imshaugii in having more well developed squamules, larger ascospores, and in substitutions in the mtSSU and ITS sequences.

Type

THAILAND, Lampang: Doi Khun Tan National Park, loc. T118, 18°25'N, 99°14'E, 1000 m alt., hill evergreen forest, 1993-01-11, P.A. Wolseley & K. Boonpragob 3245 (BM 749856!—holotype) [TLC: norstictic acid; DNA MK412410 (mtSSU), MK412477 (ITS)].

Description

Thallus effuse, crustose to squamulose; squamules small, up to 0.4 mm wide, adnate, isodiametrical, more or less scattered when young, later contiguous or fusing, more or less crenulate, plane to weakly convex; upper side medium green, somewhat shiny, epruinose, glabrous; margin concolorous with upper side, often pubescent; isidia common, attached marginally to the squamules, cylindrical, simple or slightly branched, up to 0.15 mm wide and 1.5 mm long; upper cortex formed by thick-walled hyphae with rounded lumina (type 2), 15–30 µm thick, containing a few scattered crystals dissolving in K; medulla containing crystals dissolving in K and recrystallizing by forming acicular, red crystals, PD+ yellow, K+ red; prothallus well developed, thick, white.

Apothecia seen in the holotype only, up to 1.5 mm diam., more or less plane when young, soon becoming weakly to moderately convex, medium brown, rounded to irregular, simple, when young with a rather thick, paler, weakly pubescent margin, later becoming more or less immarginate; excipulum pale brown in the rim, darker brown in inner part; hypothecium dark brown, K–; crystals present in inner part of exciple and in hypothecium, dissolving in K and recrystallizing by forming acicular, red crystals; epithecium pale brown to colourless, K–; ascospores narrowly ellipsoid or fusiform to bacilliform, simple, 15–22 × 3.5–4.5 µm (n=20). Conidiomata not seen.

Chemistry

Norstictic acid (major), atranorin (minor to trace or absent). Medulla PD+ yellow, K+ red, C–, KC–.

Distribution

Thailand.

Etymology

The specific epithet refers to its occurrence in Thailand.

Remarks

The species is morphologically and chemically very similar to P. imshaugii. Phyllopsora siamensis, however, may be distinguished by forming slightly larger squamules and longer ascospores (15–22 × 3.5–4.5 vs 10.5–14.5 × 3–4 µm; the latter measurements are based on 40 spores in the type material from Jamaica) than P. imshaugii. So far, P. imshaugii is only known to occur in the Neotropics, while P. siamensis is solely known from Thailand. In the phylogenetic tree (Fig. 1), the four accessions of P. siamensis cluster in a strongly supported clade as sister to a clade comprising P. cinchonarum and P. concinna, from which the new species is readily distinguished by its chemistry. Phyllopsora imshaugii and P. siamensis are the only Phyllopsora species known to contain norstictic acid; the major compound of the two other species are lobaric acid and parvifoliellin, respectively.

Additional specimens examined

THAILAND, Chiang Mai: Doi Suthep National Park headquarters walk, loc. 62.4, 18°48'N, 98°54'E, 1050 m alt., tropical mixed deciduous forest, on Lauraceae, 1993-03-27, B. Aguirre–Hudson & P.A. Wolseley 1643 (BM 749866) [DNA: MK412419 (mtSSU), MK412485 (ITS)]; Uthai Thani: Khao Nang Rum, Cathouse site, 15°29'N, 99°18'E, 650 m alt., tropical mixed deciduous forest, 1992-01-07, P.A. Wolseley & B. Aguirre–Hudson 5580 p.p. (BM 1031552 p.p.) [DNA: MK412423 (mtSSU), MK412487 (ITS)]; Khao Nang Rum, Khao Kiew, 15°27'N, 99°20'E, 1250 m alt., oak/chestnut forest, 1992-01-23, P.A. Wolseley and Onsar 5590 (BM 749833) [DNA: MK412418 (mtSSU), MK412484 (ITS)].

Phyllopsora subhispidula (Nyl.) Kalb & Elix

Description

Timdal and Krog (2001).

Distribution

Africa (Timdal and Krog 2001), Asia.

Remarks

This species resembles closely the more common P. buettneri, but differs in forming isidia, not lacinules (Fig. 10C). It contains argopsin (major), norargopsin (minor), zeorin (major), and sometimes atranorin (trace), similar to chemotype 4 of P. buettneri. Phylogenetically (Fig. 1), the two species are not closely related, though. Phyllopsora subhispidula is sister to the clade comprising P. nemoralis and P. hispaniolae/P. rosei (Fig. 1). It is new to Asia (Sri Lanka).

Key to the phyllopsoroid genera in Asia and Melanesia

1 Apothecia zeorine, surrounded by a thalline sheath Physcidia p.p.
Apothecia biatorin 2
2(1) Tholus non-amyloid or with an indistinct conical amyloid structure; ascospores filiform, spirally arranged in ascus; thallus and apothecia with red or purple patches caused by non-crystalline, acetone-insoluble pigment Krogia
Tholus with a distinct amyloid conical structure (Bacidia type); ascospores ellipsoid to filiform, not spirally arranged in ascus; thallus and apothecia without red patches 3
3(2) Upper and lower cortices formed by a single layer of isodiametric cells, continuous over the edge of the areolae/squamule Eschatogonia
Upper cortex multicellular or poorly differentiated; lower cortex absent 4
4(2) Ascospores ellipsoid to fusiform, simple or rarely pseudoseptate 5
Ascospores bacilliform to filiform, septate or pseudoseptate 6
5(4) Apothecia and prothallus blackish brown to black; isidia lacking; thallus containing fumarprotocetraric acid Phyllopsora’ cfr. nigrocincta (Malmideaceae)
Apothecia brown; prothallus white to dark reddish brown; isidia present or absent; if fumarprotocetraric acid present, then isidia present Phyllopsora
6(4) Thallus sorediat 7
Thallus not sorediate 8
7(6) Squamules mostly adnate, bursting into convex soralia, containing atranorin and divaricatic acid Phyllopsorasorediata
Squamules ascending, with labriform soralia, containing methyl barbatate and often terpenoids Phyllopsoraglaucescens
8(6) Thallus large, subfoliose, isidiate Physcidiacylindrophora
Thallus crustose to squamulose, not isidiate 9
9(8) Thallus formed by ascending squamules, lacinulate, containing stictic acid Parallopsora sp.*
Thallus crustose or formed by adnate squamules, not lacinulate, not containing lichen substances 10
10(9) Thallus crustose Sporacestra
Thallus squamulose Aciculopsora

Key to the species of Phyllopsora in Asia and Melanesia

1 Thallus pruinose, rosulate, broad-lobed 2
Thallus not pruinose, effuse to rosulate, narrow to broad-lobed 3
2(1) Thallus lacinulate, containing pannarin, dechloropannarin or rarely argopsin P. buettneri
Thallus isidiate, containing argopsin P. subhispidula
3(1) Upper cortex absent or poorly developed 4
Upper cortex well developed 5
4(3) Species always apotheciate; isidia lacking; apothecia plain to concave, with a pale margin; lichen substances present P. gossypina
Species apotheciate or not; isidia often present; apothecia convex, more or less immarginate; lichen substances absent P. cuyabensis
5(3) Medulla K+ red (norstictic acid) P. siamensis
Medulla K– 6
6(5) Medulla PD+ orange to red 7
Medulla PD– 10
7(6) Prothallus white or absent; lacinules absent P. santensis
Prothallus brown; lacinules present or absent 8
8(7) Squamules isodiametrical or shortly elongate, more or less adnate, containing chlorophyllopsorin or methyl 2,7-dichloronorpsoromate; isidia or lacinules present P. africana
Squamules elongate, ascending, lacking chlorophyllopsorin and methyl 2,7-dichloronorpsoromate; lacinules present 9
9(8) Ascospores narrowly ellipsoid, 6–8 × 2–2.5 µm P. sabahana
Ascospores narrowly ellipsoid to fusiform or bacilliform, 8.0–12.5 × 2.5–3.5 µm P. porphyromelaena
10(6) Thallus isidiate 11
Thallus phyllidiate, lacinulate or apotheciate 20
11(10) Prothallus white or absent 12
Prothallus brown 15
12(11) Isidia often more than 1 mm long, mainly simple; ascospores bacilliform to acicular, 26–41 × 2–3 µm; containing lobaric acid P. cinchonarum
Isidia shorter than 1 mm, globular to coralloid; ascospores ellipsoid to shortly bacilliform, less than 12 µm long; containing only atranorin or no lichen substance 13
13(12) Isidia globular; thallus containing atranorin; crystals present in medulla and hypothecium P. himalayensis
Isidia cylindrical to coralloid; thallus and apothecia lacking lichen substances and crystals 14
14(13) Isidia becoming coralloid; species crustose, effuse; areoles up to 0.1 mm diameter P. isidiosa
Isidia cylindrical or weakly branched; species squamulose, effuse or rosulate; squamules up to 1 mm diameter P. pseudocorallina
15(11) Thallus crustose, consisting of more or less scattered areoles or sometimes isidia only 16
Thallus squamulose 17
16(15) Ascospores narrowly ellipsoid, 7–13 × 2–3 µm; containing furfuraceic acid only P. furfuracea
Ascospores bacilliform, 16–25 × 2–3 µm; containing furfuraceic acid and 2–3 related compounds P. dolichospora
17(15) Prothallus thick, forming a cushion with colonizing areoles along the periphery 18
Prothallus thin, not forming a cushion 19
18(17) Thallus containing atranorin and terpenoids P. halei
Thallus containing furfuraceic acid or rarely no compounds P. castaneocincta
19(17) Isidia globular or shortly cylindrical; thallus pale green, containing atranorin or no lichen substances P. kalbii
Isidia cylindrical; thallus dark green to brown, containing atranorin and parvifoliellin P. parvifoliella
20(10) Thallus containing xanthones P. chodatinica
Thallus not containing xanthones 21
21(20) Thallus containing furfuraceic acid P. neofoliata
Thallus not containing lichen substances 22
22(21) Prothallus brown, well developed 23
Prothallus white or absent 27
23(22) Thallus rosulate or composed of elongated squamules 24
Thallus effuse, composed of more or less isodiametrical squamules 26
24(23) Thallus phyllidiate; phyllidia mainly occurring in central part of thallus P. parvifolia
Thallus lacinulate 25
25(24) Squamules long, linear, deeply incised to branched P. breviuscula
Squamules short, crenulate to narrowly incised P. mediocris
26(23) Thallus crustose, consisting of closely adnate areoles and ascending lacinules P. longiuscula
Thallus squamulose, consisting of ascending squamules, breaking into lacinules P. confusa
27(22) Thallus phyllidiate; phyllidia mainly occurring in central part of thallus P. parvifolia
Thallus lacinulate 28
28(27) Squamules closely adnate, elongated, linear, somewhat branched P. loekoesii
Squamules ascending, short, not branched 29
29(28) Ascospores narrowly ellipsoid to fusiform, 11–20 × 2–3 µm P. foliata
Ascospores narrowly ellipsoid to shortly bacilliform, 9–11 × 2–2.5 µm P. confusa

Acknowledgements

We are grateful to the following herbaria for providing loans of material for our study: B, E, H, TNS, and UPS, in addition to the private herbarium of P. Diederich. We would like to thank S. Kondratyuk for providing sequences of the type material of P. loekoesii. ET, MB and SK thank Siri Rui for curatorial assistance at the Natural History Museum, University of Oslo (Norway). Special thanks are due to everyone who accompanied the authors in the field in Indonesia, Malaysia, Sri Lanka and Thailand. We acknowledge the Sabah Biodiversity Council, the administration units at Danum Valley and Maliau Conservation Areas for granting permits to access and work in the studied sites in the framework of the NHM “Danum-Maliau Quantitative Inventory” (JKM/MBS.1000-2/2 (77), MBMC 2012/15) and for the provision of logistic support. GW is thankful for the Research funding received from National Geographic Society (USA) and Dilmah Conservation (Sri Lanka) to conduct fieldwork in Sri Lanka in 2017. SW and GW are grateful for the collecting and export permits from Forest and Wildlife departments of Sri Lanka. The Science library at the University of Oslo (Norway) is acknowledged for its financial support to SK for open access publishing.

References

  • Altekar G, Dwarkadas S, Huelsenbeck JP, Ronquist F (2004) Parallel metropolis coupled Markov chain Monte Carlo for Bayesian phylogenetic inference. Bioinformatics 20: 407–415. https://doi.org/10.1093/bioinformatics/btg427
  • Aptroot A (1997) Lichen biodiversity in Papua New Guinea, with report of 173 species on one tree. Bibliotheca Lichenologica 68: 203–213.
  • Aptroot A, Diederich P, Sérusiaux E, Sipman HJM (1997) Lichens and lichenicolous fungi from New Guinea. Bibliotheca Lichenologica 64: 1–220.
  • Aptroot A, Saipunkaew W, Sipman H, Sparrius L, Wolseley P (2007) New lichens from Thailand, mainly microlichens from Chiang Mai. Fungal diversity 24: 75–134.
  • Aptroot A, Sparrius LB (2003) New microlichens from Taiwan. Fungal diversity 14: 1–50.
  • Brako L (1989) Reevaluation of the genus Phyllopsora with taxonomic notes and introduction of Squamacidia, gen. nov. Mycotaxon 35: 1–19.
  • Brako L (1991) Phyllopsora (Bacidiaceae). Flora Neotropica: 1–66.
  • Buaruang K, Boonpragob K, Mongkolsuk P, Sangvichien E, Vongshewarat K, Polyiam W, Rangsiruji A, Saipunkaew W, Naksuwankul K, Kalb J (2017) A new checklist of lichenized fungi occurring in Thailand. MycoKeys 23: 1–91. https://doi.org/10.3897/mycokeys.23.12666
  • Chernomor O, von Haeseler A, Minh BQ (2016) Terrace aware data structure for phylogenomic inference from supermatrices. Systematic Biology 65: 997–1008. https://doi.org/10.1093/sysbio/syw037
  • Culberson CF, Johnson A (1982) Substitution of methyl tert.-butyl ether for diethyl ether in the standardized thin-layer chromatographic method for lichen products. Journal of Chromatography 238: 483–487. https://doi.org/10.1016/S0021-9673(00)81336-9
  • Culberson CF (1972) Improved conditions and new data for identification of lichen products by standardized thin-layer chromatographic method. Journal of Chromatography 72: 113–125. https://doi.org/10.1016/0021-9673(72)80013-X
  • Elix JA (2006) Additional lichen records from Australia 58. New records from Norfolk Island. Australasian Lichenology 59: 23–29.
  • Elix JA (2009) Phyllopsoraceae. In: McCarthy PM (Ed.) Flora of Australia. ABRS and CSIRO Publishing, Canberra and Melbourne, 41–59.
  • Elix JA, McCarthy PM (1998) Catalogue of the lichens of the smaller Pacific Islands. Bibliotheca Lichenologica 70: 1–361.
  • Hoang DT, Chernomor O, von Haeseler A, Minh BQ, Vinh LS (2018) UFBoot2: Improving the Ultrafast Bootstrap Approximation. Molecular Biology and Evolution 35: 518–522. https://doi.org/10.1093/molbev/msx281
  • Hue A (1909) Lichenum generis Crocyniae Mass. plerasque species juxta archetypa specimina morphologice et anatomice descripsit. Memoires de la Société Nationale des Sciences Naturelles et Mathématiques de Cherbourg 37: 223–254.
  • Joshi Y, Nguyen TT, Wang XY, Lőkös L, Koh YJ, Hur J–S (2011) Contribution to the lichen mycota of South Korea. Mycotaxon 116: 61–74. https://doi.org/10.5248/116.61
  • Kalb K, Elix J (1995) The lichen genus Physcidia. Bibliotheca Lichenologica 57: 265–296.
  • Kalyaanamoorthy S, Minh BQ, Wong TKF, von Haeseler A, Jermiin LS (2017) ModelFinder: fast model selection for accurate phylogenetic estimates. Nature Methods 14: 587. https://doi.org/10.1038/nmeth.4285
  • Katoh K, Standley DM (2013) MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability. Mol Biol Evol 30: 772–780. https://doi.org/10.1093/molbev/mst010
  • Kauff F, Lutzoni F (2002) Phylogeny of the Gyalectales and Ostropales (Ascomycota, Fungi): among and within order relationships based on nuclear ribosomal RNA small and large subunits. Mol Phylogenet Evol 25: 138–156. https://doi.org/10.1016/S1055-7903(02)00214-2
  • Kearse M, Moir R, Wilson A, Stones–Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C (2012) Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28: 1647–1649. https://doi.org/10.1093/bioinformatics/bts199
  • Kistenich S, Bendiksby M, Ekman S, Cáceres MES, Hernández JE, Timdal E (in press) Towards an integrative taxonomy of Phyllopsora (Ramalinaceae). The Lichenologist.
  • Kistenich S, Rikkinen J, Thüs H, Vairappan C, Wolseley PA, Timdal E (2018a) Three new species of Krogia (Ramalinaceae, lichenized Ascomycota) from the Paleotropics. MycoKeys: 69–88. https://doi.org/10.3897/mycokeys.40.26025
  • Kistenich S, Timdal E, Bendiksby M, Ekman S (2018b) Molecular systematics and character evolution in the lichen family Ramalinaceae (Ascomycota: Lecanorales). Taxon 67: 871–904. https://doi.org/10.12705/675.1
  • Kondratyuk SY, Lőkös L, Halda JP, Upreti DK, Mishra GK, Haji Moniri M, Farkas E, Park JS, Lee BG, Liu D (2016) New and noteworthy lichen-forming and lichenicolous fungi 5. Acta Botanica Hungarica 58: 319–396. https://doi.org/10.1556/ABot.58.2016.3-4.7
  • Logesh AR, Chinlampianga M, Shukla AC, Upreti DK (2017) Studies on Lichens of Mizoram, Northeast India. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences 87: 445–457. https://doi.org/10.1007/s40011-015-0592-z
  • Menlove JE (1974) Thin-layer chromatography for the identification of lichen substances. British Lichen Society Bulletin 34: 3–5.
  • Mishra GK, Upreti DK, Nayaka S, Haridas B (2011) New taxa and new reports of Phyllopsora (lichenized Ascomycotina) from India. Mycotaxon 115: 29–44. https://doi.org/10.5248/115.29
  • Moon KH (2013) Lichen-forming and lichenicolous fungi of Korea. Korean Lichen Research Institute (KoLRI), 139 pp.
  • Müller J (1894) Conspectus systematicus specierum lichenum Novae Zelandiae. Bulletin de l‘Herbier Boissier 2 (Appendix 1): 1–114.
  • Nguyen L–T, Schmidt HA, von Haeseler A, Minh BQ (2015) IQ-TREE: A Fast and Effective Stochastic Algorithm for Estimating Maximum-Likelihood Phylogenies. Molecular Biology and Evolution 32: 268–274. https://doi.org/10.1093/molbev/msu300
  • Ohmura Y, Kashiwadani H (2018) Checklist of lichens and allied fungi of Japan. National Museum of Nature and Science Monographs 49: 1–110.
  • Streimann H (1986) Catalogue of the lichens of Papua New Guinea and Irian Jaya. Bibliotheca Lichenologica 22: 1–145.
  • Streimann H, Sipman H (1994) New lichen records from the island of New Britain in Papua New Guinea. Fragmenta Floristica et Geobotanica 39: 369–382.
  • Timdal E (2011) The lichen genus Phyllopsora (Ramalinaceae) in the West Indies. Bibliotheca Lichenologica 106: 319–351.
  • Timdal E, Krog H (2001) Further studies on African species of the lichen genus Phyllopsora (Lecanorales). Mycotaxon 77: 57–90.
  • Upreti DK, Divakar PK, Nayaka S (2002) Notes on species of the lichen genus Phyllopsora in India. Bibliotheca Lichenologica 86: 185–191.
  • Wei JC (1991) An Enumeration of Lichens in China. International Academic Publishers, Beijing, 278 pp.

1 unpublished data