Introduction
Graphidaceae is the largest family of tropical lichen fungi, with well over 2000 species known and more than 3000 predicted (Lücking et al. Reference Lücking, Johnston, Aptroot, Kraichak, Lendemer, Boonpragob, Cáceres, Ertz, Ferraro and Jia2014, Reference Lücking, Hodkinson and Leavitt2017). The family was traditionally divided into eight genera based on ascoma and ascospore types but recent revisions have substantially changed genus concepts in Graphidaceae, now recognizing c. 80 genera (Staiger Reference Staiger2002; Frisch et al. Reference Frisch, Kalb and Grube2006; Rivas Plata et al. Reference Rivas Plata, Lumbsch and Lücking2012; Lumbsch et al. Reference Lumbsch, Kraichak, Parnmen, Rivas Plata, Aptroot, Cáceres, Ertz, Feuerstein, Mercado-Díaz and Staiger2014; Lücking et al. Reference Lücking, Hodkinson and Leavitt2017). In its revised definition, the type genus, Graphis, was circumscribed as having lirellate, carbonized ascomata and distoseptate, hyaline, I+ amyloid ascospores (Staiger Reference Staiger2002; Lücking Reference Lücking2009; Lücking et al. Reference Lücking, Archer and Aptroot2009). However, it was subsequently shown that Graphis sensu Staiger included two distantly related lineages (Rivas Plata et al. Reference Rivas Plata, Hernández, Lücking, Staiger, Kalb and Cáceres2011), a finding already anticipated by Staiger (Reference Staiger2002). This separation was recently formalized by reinstating the genus Allographa for the second lineage (Lücking & Kalb Reference Lücking and Kalb2018). The latter differs from Graphis in the usually prominent, robust, mostly massively carbonized lirellae, generally larger ascospores, often lack of secondary substances, and type B hymenial inspersion sensu Lücking (Reference Lücking2009). With one exception (Graphis ferruginea Vain.), Allographa now contains all species of Graphis sensu Staiger with pigmented lirellae, including its type species, A. lutea Chevall.
On 7 November 2015, the first author posted a picture of an orange Graphis sensu Staiger from Java (Fig. 1) on the Facebook page Lichens Connecting People (https://www.facebook.com/groups/150880938305901). This group page was founded in 2011 by Anna Voytsekhovich and currently has over 3700 members from 79 countries worldwide. Around 90% of the group members are amateurs, with c. 10% being professional lichenologists from 46 countries. Since 2011, more than 6000 lichen-related posts have been published on this social media page, often depicting unknown and potentially undescribed taxa from remote places. The orange Graphis sensu Staiger was immediately recognized by the senior author as unusual, since this group of species was not known from the eastern Palaeotropics at the time (Fig. 1). Subsequent study revealed that it differed from other species with pigmented lirellae in the very large, transversely septate ascospores and therefore represented an undescribed taxon. At around the same time, another Graphis sensu Staiger with pigmented lirellae was collected in the backyard of the second author's home in Sri Lanka and was initially considered to be conspecific with the material from Java; however, a closer look revealed that both differed in important features, including ascospore size and septation and pigment chemistry, and therefore the material from Sri Lanka also represented an undescribed taxon. These discoveries underline the importance of social media in facilitating communication between collectors and experts to quickly identify potentially undescribed species from almost anywhere in the world, as well as the importance of sometimes just looking in one's own backyard to reveal new taxa.
Original post of the new species from Java in the Facebook group Lichens Connecting People. In colour online.
Herein we describe both species formally in the recently segregated genus Allographa and provide a key to all species of the genus with pigmented lirellae.
Material and Methods
The material from Indonesia (Java) was studied at the Department of Biology in Universitas Padjadjaran, using an Olympus SZ61 dissecting microscope. Sections of thallus and lirellae were examined with an Olympus CX31 compound microscope. The material from Sri Lanka was studied at the Field Museum, Chicago, and at the Botanischer Garten und Botanisches Museum, Freie Universität Berlin, using Leica MS5, Motic K400, Olympus SZX12 and Leica Zoom 2000 dissecting microscopes. Sections of thallus and lirellae mounted in tap water were examined under Olympus BH-2, VistaVision VWR V036 and Zeiss Axioscop compound microscopes. Secondary chemistry was assessed using KOH spot tests and verified with standardized thin-layer chromatography (TLC) using solvent system C (Orange et al. Reference Orange, James and White2010). Lugol's solution (Fluka 62650) was used for iodine reactions.
Results and Discussion
Allographa kamojangensis Jatnika, Noer & Lücking sp. nov.
MycoBank No.: MB 829357
Differing from Allographa firferi in the much larger ascospores and the orange, K+ immediately purple-violet pigment, and from A. lutea in the completely carbonized excipulum and the larger ascospores.
Type: Indonesia, West Java, Kamojang, 07°07′30″S, 107°48′00″E, on bark and twig of Arthocarpus heterophyllus, 5 November 2015, M. Feisal Jatnika L.BIOUNPAD.G.0023 (UNPAD—holotype!).
(Fig. 2A–C)
Thallus corticolous, epiperidermal, up to 6 cm diam., continuous, white-grey, uneven; prothallus not observed. Thallus in section 100–200 µm thick, with prosoplectenchymatous cortex, 20–30 µm thick, and photobiont layer, 50–70 µm thick, nubilous with numerous small crystals.
A–C, Allographa kamojangensis (holotype); A, thallus in situ; B, lirellae, inset showing section through lirellae with completely carbonized excipulum; C, ascospore. D–F, A. jayatilakana (holotype); D & E, thallus with lirellae; F, ascospore. Scales: B, D & E = 1 mm; B inset = 0·5 mm; C & F = 10 µm. In colour online.
Ascomata lirellate, irregularly branched, prominent, 2–4 mm long, 0·3–0·4 mm wide, 0·1–0·2 mm high; disc concealed; labia mostly entire but becoming striate in older lirellae, apically exposed, black, laterally covered by a yellow-orange pruina. Excipulum completely and massively carbonized, laterally 50–100 µm wide, basally up to 250 µm high; periphysoids absent; hypothecium 20–30 µm high, hyaline to greyish. Hymenium 250–300 µm high, clear; paraphyses unbranched, apically smooth. Asci 200–250 × 40–60 µm, oblong. Ascospores 8 per ascus, 13–15-septate, 100–140 × 20–25 µm, oblong, 4–6 times as long as wide, with thickened septa and lens-shaped to rectangular lumina, hyaline, I+ violet-blue.
Secondary chemistry
No substances detected by TLC in the thallus; lirellae covered with orange anthraquinone, K+ immediately purple-violet in microscopic section.
Etymology
The epithet refers to the type locality, a geothermic field and popular tourist location in West Java.
Distribution and ecology
The original vegetation cover of the type locality in the Kamojang geothermal area corresponds to broadleaf evergreen forest but most of the area has been deforested (Budi et al. Reference Budi, Harijadi, Suryo, Beni and Yudi2009), and the material was collected on semi-exposed trees in a crop plantation. The species is likely to be more common in the upper strata of forest remnants in the area.
Remarks
Allographa kamojangensis is generally very similar to the neotropical A. chrysocarpa and the neotropical-African palaeotropical A. ochracea (Staiger Reference Staiger2002; Lücking et al. Reference Lücking, Chaves, Sipman, Umaña and Aptroot2008; Lücking & Kalb Reference Lücking and Kalb2018), sharing the completely carbonized lirellae with orange, K+ immediately purple-violet pruina. It differs from both in the transversely septate ascospores. In A. chrysocarpa the ascospores are richly muriform and comparatively longer and narrower (120–190 × 17–23 µm, 6–8 times as long as wide), whereas in A. ochracea they are terminally muriform (appearing transversely septate when not carefully checked) and distinctly narrower (70–120 × 10–15 µm, 7–8 times as long as wide). The only other species with transversely septate ascospores in this group are the neotropical A. firferi and A. lutea (Lücking et al. Reference Lücking, Chaves, Sipman, Umaña and Aptroot2008; Lücking & Kalb Reference Lücking and Kalb2018); A. firferi differs in the yellow, K+ yellow then slowly purple-violet pigment, whereas A. lutea has an apically carbonized excipulum only. Both also have distinctly smaller and narrower ascospores (70–100 × 11–15 µm, 6–7 times as long as wide).
Allographa jayatilakana Weerakoon, Arachchige & Lücking sp. nov.
MycoBank No.: MB 829360
Differing from Allographa flavominiata in the much shorter ascospores, from A. firferi in the terminally muriform ascospores, and from A. ochracea in the yellow-orange, K+ yellow then slowly purple-violet pruina.
Type: Sri Lanka, Western Province, Colombo City, Hokandara, garden of Weerakoon residence, 06°52′49″N, 79°58′00″E, 75 m, on bark of ornamental Ficus benjamina, 28 April 2015, G. Weerakoon GW 100 (PDA—holotype!; B, BM—isotypes!).
(Fig. 2D–F)
Thallus corticolous, epiperidermal, up to 10 cm diam., continuous, white-grey, uneven; prothallus not observed. Thallus in section 100–150 µm thick, with prosoplectenchymatous cortex, 20–30 µm thick, and photobiont layer, 50–70 µm thick, nubilous with numerous small crystals.
Ascomata lirellate, unbranched to sparsely branched, prominent, 1–5 mm long, 0·3–0·4 mm broad, 0·3–0·4 mm high; disc concealed; labia mostly entire but becoming striate in older lirellae, apically exposed, black, laterally covered by a yellow to yellow-orange pruina. Excipulum completely and massively carbonized, laterally 50–100 µm wide, basally up to 250 µm high; periphysoids absent; hypothecium 20–30 µm high, hyaline to greyish. Hymenium 140–170 µm high, clear; paraphyses unbranched, apically smooth. Asci 140–160 × 20–30 µm, oblong. Ascospores 8 per ascus, terminally muriform, with 15–19 transverse septa and usually one or two of the (sub)terminal cells on each side with a longitudinal septum, 80–120 × 9–12 µm, oblong with rounded proximal and acute terminal end, 8–10 times as long as wide, with thickened septa and lens-shaped lumina and with gelatinous caps at both ends, rounded at the proximal end, fin-like at the distal end, hyaline, I+ violet-blue.
Secondary chemistry
No substances detected by TLC in the thallus; lirellae covered with unknown yellow anthraquinone, K+ yellow slowly turning violet, in microscopic section with vanishing K+ yellow efflux and pigment granules then turning purple-violet.
Etymology
This new species honours the memory of the late Jayatilaka Bandara Weerakoon, father of the second author and owner of the property where the species was collected.
Distribution and ecology
The original vegetation of the type locality would have been tropical wet evergreen forest, but today most of the region has undergone land use change through urbanization and agriculture (Myers et al. Reference Myers, Mittermeier, da Fonseca and Kent2000; Gunawardene et al. Reference Gunawardene, Daniels, Gunatilleke, Gunatilleke, Karunakaran, Nayak, Prasad, Puyravaud, Ramesh and Subramanian2007). Before being further converted into a private property garden, the location was a monoculture plantation of Cocos nucifera. As with the preceding taxon, this species is likely to be more common in the upper strata of forest remnants in the area.
Remarks
The yellow anthraquinone, K+ yellow slowly turning purple-violet pruina is shared by the new species and Allographa firferi, A. flavoaltamirensis and A. flavominiata, all described from Costa Rica (Lücking et al. Reference Lücking, Chaves, Sipman, Umaña and Aptroot2008). Allographa flavoaltamirensis differs in the inspersed hymenium and muriform ascospores, whereas A. flavominiata possesses terminally muriform ascospores but these are much larger (100–170 × 12–15 µm). Allographa firferi has ascospores of about the same size as the new species, but slightly broader, and they lack longitudinal septa. The neotropical-African palaeotropical A. ochracea (Lücking et al. Reference Lücking, Chaves, Sipman, Umaña and Aptroot2008; Lücking & Kalb Reference Lücking and Kalb2018) agrees with the new species in lirella anatomy and ascospore size and septation, but has a different, orange, K+ immediately purple-violet pigment.
Key to the species of Allographa with pigmented lirellae
1 Excipulum apically carbonized only; pigment orange, K+ immediately purple-violet; hymenium clear; ascospores transversely septate, 90–100 × 11–13 µm, 7–8 times as long as wide; Neotropics ……… Allographa lutea Chevall. Excipulum completely carbonized; pigment, hymenium and ascospores ……… variable ……… 2
2(1) Hymenium strongly inspersed; labia usually striate; pigment yellow, K+ yellow slowly turning purple-violet; ascospores regularly muriform, 80–100 × 10–17 µm, 6–8 times as long as wide; Neotropics……… Allographa flavoaltamirensis (Sipman & Lücking) Lücking & Kalb Hymenium clear; labia, pigment and ascospores variable ……… 3
3(2) Ascospores regularly muriform, 120–190 × 17–23 µm, 6–8 times as long as wide; pigment orange, K+ immediately purple-violet; Neotropics ……… Allographa chrysocarpa (Raddi) Lücking & Kalb Ascospores transversely septate or terminally muriform only, their size variable; pigment variable ……… 4
4(3) Ascospores transversely septate (check carefully) ……… 5 Ascospores terminally muriform (check carefully) ………6
5(4) Ascospores 100–140 × 20–25 µm, 4–6 times as long as wide; pigment orange, K+ immediately purple-violet; eastern Palaeotropics ……… Allographa kamojangensis Jatnika et al. Ascospores 70–100 × 11–15 µm, 6–7 times as long as wide; pigment yellow, K+ yellow slowly turning purple-violet; Neotropics ……… Allographa firferi (Lücking) Lücking & Kalb
6(4) Lirellae soon striate, with complete thalline cover that may rupture to expose the pigmented pruina; pigment orange, K+ immediately purple-violet; ascospores 80–100 × 10–15 µm, 6–8 times as long as wide; Neotropics ……… Allographa miniata (Redinger) Lücking & Kalb Lirellae usually remaining entire, with the pigmented pruina exposed; pigment and ascospores variable ……… 7
7(6) Ascospores 100–170 × 12–15 µm, 8–12 times as long as wide; pigment yellow, K+ yellow slowly turning purple-violet; Neotropics ……… Allographa flavominiata (Moncada & Lücking) Lücking & Kalb Ascospores 70–120 × 9–15 µm, 7–8 times as long as wide; pigment variable ……… 8
8(7) Pigment yellow, K+ yellow slowly turning purple-violet; eastern Palaeotropics ……… Allographa jayatilakana Weerakoon et al. Pigment orange, K+ immediately purple-violet; Neotropics and African Palaeotropics…. Allographa ochracea (C. W. Dodge) Lücking & Kalb
[≡ Graphis subchrysocarpa Lücking]
Author ORCIDs
Gothamie Weerakoon: 0000-0001-8290-2910. Robert Lücking: 0000-0002-3431-4636.
Data obtained for this study were gathered as part of several projects funded by the National Science Foundation: “TICOLICHEN” (DEB-0206125 to The Field Museum; PI Robert Lücking); “Neotropical Epiphytic Microlichens – An Innovative Inventory of a Highly Diverse yet Little Known Group of Symbiotic Organisms” (DEB-715660 to The Field Museum; PI R. Lücking) and “ATM – Assembling a Taxonomic Monograph: The Lichen Family Graphidaceae” (DEB-1025861 to The Field Museum; PI T. Lumbsch, CoPI R. Lücking). GW is grateful for research funding from Dilmah Conservation and the National Geographic Society (CS-R001-17) and the opportunity to use the research facilities at the Field Museum, Chicago. She also extends her gratitude to Prof. Siril Wijesundara and Pat Wolseley. MFJ thanks Ruchyat Partasasmita, chief of the Department of Biology, as well as Joko Kusmoro and Ririn Eka Permatasari. Two anonymous reviewers are thanked for comments that helped to improve the manuscript.
