Ecology and Systematics of Filamentous Fungi on Nypa Fruticans

Indian Journal of Marine Sciences Vol. 35(4), December 2006, pp. 297-307

Biodiversity and ecological observations on filamentous fungi of mangrove palm Nypa fruticans Wurumb (Liliopsida – Arecales) along the Tutong River, Brunei

K.D. Hyde & V.V. Sarma1*# Centre for Research in Fungal Diversity, Department of Ecology & Biodiversity, The University of Hong Kong, Hong Kong SAR, PR China *[Email: [email protected]] Received 10 July 2006; revised 21 August 2006

Biodiversity and ecology of higher filamentous fungi on Nypa fruticans in Brunei were examined during 1999. Forty-six taxa were recorded including 33 ascomycetes and 13 anamorphic taxa in 25 genera. Linocarpon was the most speciose genus (6 species) followed by Aniptodera and Astrosphaeriella (4 each). The mycota mostly comprised typical marine species (e.g. Aniptodera, Lulworthia, Savoryella). Linocarpon appendiculatum (13%) and L. bipolaris (13%) were most frequently recorded, while Oxydothis nypae (9%), Astrosphaeriella striatispora (6.4%), Trichocladium nypae (6.4%) and Linocarpon nypae (5.4%) were also common. Most of the fungi (50%) were rare in their occurrence. More diversity was found on fronds than on leaves. Linocarpon appendiculatum, L. bipolaris, Neolinocarpon globosicarpum and Oxydothis nypae were more frequently recorded on fronds than other fungi, while Linocarpon bipolaris (13.5%), Astrosphaeriella striatispora (12.2%), Trichocladium nypae (8.1%) and Linocarpon appendiculatum (8.1%) were more frequently recorded on leaves. Examination of vertical distribution of fungi showed that the submerged parts have a rich fungal diversity followed by intertidal parts and finally the aerial parts. [Key words: Frequency of occurrence, horizontal distribution, fungi, mangrove fungi, vertical distribution, Tutong river, Brunei, biodiversity]

Introduction Materials and Methods Saprobic marine fungi decay various substrates Study area such as grasses of salt marshes and trees of The Tutong River in Brunei feeds into the South 1-3 mangroves in the sea and intertidal substrates . Nypa China Sea on the northwest side of Borneo. Samples fruticans Wurumbm, belonging to class Liliopsida were collected from four sites along the Tutong River. and order Arecales, is a palm common in the Data on the physico-chemical conditions of the sites mangroves of Asia and forms extensive stands. The (lat. 04°47´N; long. 114°41´E) were published in fungi isolated on this palm have been shown to Fryer et al.17. There was generally a gradient of produce enzymes capable of cellulose and lignin salinity from 0 to 32 ‰, but at times of high river 4 degradation . There have been several studies on the flow, all sites had salinity of 0 ‰. biodiversity of fungi on this intertidal palm and Marine site (Site I) - The Tutong River at this site 5-14 several new genera and species have been described . is approximately 50 m wide and 8.5 m deep at the More than 40 species are known only from this host mid-point. The riparian vegetation at this point was and many of these fungi have been thought to be host- dominated by Avicennia alba, Nypa fruticans and 2,15,16 specific . We therefore studied the saprobic fungi Sonneratia caseolaris. The tidal range is on N. fruticans along a river in Brunei to establish approximately 2 m. 1) if different fungi occur at different salinities and Brackish site A (Site II) - The canal at this point is 2) if fungi are vertically distributed. approximately 20 m wide and 2 m deep in the deepest part. The water is also tidal. A riparian strip ______approximately 50 m in width, borders the river. The 1Present address: Shri A.M.M. Murugappa Chettiar Research Centre, Taramani, Chennai – 600 113, India; strip is inundated in times of high river flow (when Ph: 91-44-2243 0937; Fax: 91-44-2243 0369 the water was effectively fresh) and during high tides. #Corresponding author The vegetation was a mixture of Barringtonia cf. 298 INDIAN J. MAR. SCI., VOL. 35, NO. 4, DECEMBER 2006

racemosa, Bruguiera gymnorrhiza, Cerbera odallam, species richness. Calculations were carried out Ficus cf. microcarpa, Gluta velutina, Heritiera according to Magurran18 and Ludwig & Reynolds19. globosa, Hibiscus tiliaceus, Nypa fruticans and Margalef Index on diversity (Da) was calculated as Sonneratia caseolaris. follows: Brackish site B (Site III) - This site was 100 m upstream of Brackish site A, and has very similar Margalef index Da = (S-1)/InN vegetation. However, the topography of the site is where Da = Margalef Index, S = the number of more that of a floodplain. The channel at this point is species; N = the total number of fungal occurrences, less distinct and not as deep as at Brackish site A (approximately 1 m maximum). Plant growth is not n(n-1) restricted to the riverbanks, and some plants grow in Simpson's index D = Σ ――― the middle of the stream. N(N-1) Freshwater site A & B (Site IV) - This area is a where n = Number of occurrences of ith species, floodplain with no defined channel. Maximum depth N = Number of occurrences of all the species is approximately 1 m. The riverine vegetation is dominated by species associated with disturbance ni ni such as Dillenia suffruticosa, Ischaemum barbatum, Shannon index H' = ─Σ — In — Scleria sp. and Vitex pubescens together with some N N regenerating peat swamp species. H Collection of samples Evenness = ―— Totally 120 samples were collected and all had a InN similar size of 20 cm. We randomly collected and examined 120 samples (60 fronds and 60 leaves) of where H = Shannon index, N = Number of occur- Nypa fruticans and examined them for saprobic rences of all the species filamentous fungi following incubation. Samples were collected from submerged, intertidal and terrestrial Results and Discussion zones (at each level 5 leaves and 5 fronds) at four Fungal diversity different sites (Sites I-IV) with differing salinities Five samples of leaves and fronds were collected along the Tutong River. The samples were collected each from three different vertical levels and at four during July 1999. different sites along a horizontal range totaling 120 Abiotic measurements, pH, salinity, temperature samples examined. Ninety-two samples supported and dissolved oxygen (DO2), of the water at each site sporulating fungi while 28 did not. Forty-six fungal 17 are provided in Fryar et al . Samples were briefly taxa were identified of which 33 were ascomycetes washed in the river to remove excess mud and algae and 13 anamorphic taxa. Linocarpon was the most and placed individually into zip-lock bags and speciose genus with 6 species, followed by incubated at 27ºC. After two weeks, the samples were Aniptodera, Annulatascus and Astrosphaeriella observed with a dissecting microscope for fungal (4 each) and Anthostomella (3) (Table 1). Most taxa fruiting bodies on the surface of the wood. Samples were aquatic fungi, i.e., marine or freshwater fungi were observed again after 2 months of incubation. (e.g. Aniptodera, Lulworthia, Savoryella), while a few Abundance was measured as presence/absence on a were probably terrestrial fungi (e.g. Ansthostomella sample. That is, if a fruiting body of one species was eructans, Sporidesmium crassisporum). Many of the seen more than once on a sample, it was counted only species identified usually occur on palms20. once. Accordingly the mycota on Nypa fruticans is unique 2,11,15,16,21 Statistical analysis with a number of fungi specific to this host . Based on the percentage occurrence of different In addition to several genera often recorded on palms species they were grouped as very frequent (>10%), (e.g Anthostomella, Astrosphaeriella, Oxydothis, frequent (> 5%-10%), infrequent >1% < 5%) and rare Linocarpon), the genus Annulatascus has been (<1%). recorded for the first time on Nypa fruticans. The Shannon-Weiner diversity and evenness indices genus Annulatascus has been recently reported from were calculated for each site along with Margalef mangroves of Brunei by Fryar et al.17. Nypa fruticans HYDE & SARMA: FILAMENTOUS FUNGI 299

Table 1 — Frequency of occurrence of filamentous fungi on leaves and fronds of Nypa fruticans at Brunei S.No. Name of the species Leaves Fronds Total % occurrence 1 Aniptodera chesapeakensis Shearer & M.A. Mill. 1 1 2 1.0 2 A. intermedia K.D. Hyde & Alias - 1 1 0.5 3 A. mangrovei K.D. Hyde 2 1 3 1.0 4 A. nypae K.D. Hyde - 4 4 2.1 5 Annulatascus velatispora K.D. Hyde 1 - 1 0.5 6 Annulatascus palmietensis K.D. Hyde, Goh & T.D. Steinke 2 4 6 3.1 7 Annulatascus sp.1 - 2 2 0.5 8 Annulatascus sp.2 - 1 1 0.5 9 Anthostomella nypensis K.D. Hyde, Alias & B.S. Lu 1 - 1 0.5 10 Anthostomella eructans - 1 1 0.5 11 Anthostomella sp. 1 - 1 0.5 12 Astrosphaeriella aquatica K.D. Hyde - 1 1 0.5 13 A. nypae K.D. Hyde 4 2 6 3.1 14 A. striatispora (K.D. Hyde) K.D. Hyde 9 3 12 6.2 15 Astrosphaeriella sp. 1 - 1 0.5 16 Carinispora nypae K.D. Hyde - 3 3 1.6 17 Frondicola tunitricuspis - 5 5 2.6 18 Herpotrichea nypicola K.D. Hyde & Alias - 1 1 0.5 19 Helicascus nypae K.D. Hyde 1 1 2 1.0 20 Lignincola laevis Höhnk 1 - 1 0.5 21 Linocarpon angustatum K.D. Hyde & Alias - 3 3 1.6 22 L. appendiculatum K.D. Hyde 6 18 24 12.5 23 L. bipolaris K.D. Hyde 10 15 25 13.0 24 L. livistonae (Henn.) K.D. Hyde 2 - 2 1.1 25 L. longisporum K.D. Hyde 2 - 2 1.1 26 L. nypae (Henn.) K.D. Hyde 4 8 12 6.2 27 Lulworthia grandispora Meyers - 2 2 1.0 28 Lulworthia sp. - 1 1 0.5 29 Marinosphaera mangrovei K.D. Hyde 1 - 1 0.5 30 Neolinocarpon globosicarpum K.D. Hyde - 7 7 3.6 31 Oxydothis nypae K.D. Hyde & Nakagiri 2 18 20 10.4 32 Phomatospora sp.-like - 1 1 0.5 33 Savoryella aquatica K.D. Hyde 3 - 3 1.6 34 S. lignicola E.B.G. Jones & R.A. Eaton - 1 1 0.5 35 Cancellidium applanatum Tubaki 2 - 2 1.0 36 Dictyochaeta sp. - 1 1 0.5 37 Endophragmia sp. 1 - 1 0.5 38 Helicorhoidon nypicola K.D. Hyde & Goh 5 - 5 2.6 39 Papulospora sp. - 1 1 0.5 40 Phialogeniculata sp. - 1 1 0.5 41 Phoma sp. - 1 1 0.5

…Contd

300 INDIAN J. MAR. SCI., VOL. 35, NO. 4, DECEMBER 2006

Table 1 — Frequency of occurrence of filamentous fungi on leaves and fronds of Nypa fruticans at Brunei — Contd. S.No. Name of the species Leaves Fronds Total % occurrence 42 Sporidesmium crassisporum M.B. Ellis 5 1 6 3.1 43 Trichocladium nypae K.D. Hyde & Goh 6 6 12 6.2 44 Trichocladium sp. 1 - 1 0.5 45 Unidentified hyphomycete 1 - 1 1 0.5 46 Unidentified hyphomycete 2 - 1 1 0.5 No. of fungal occurrences 74 118 192 No. of samples examined 60 60 120 No. of samples supporting sporulating fungi 39 53 92 No. of samples without sporulating fungi 21 7 28 No. of species 25 32 46 No. of species found only on any one substrate & common to both 14 21 11 Average no. of fungi per sample 1.23 1.96 1.6 Simpson's Index D 0.05627 0.07373 0.0586 Simpson's Index of Diversity 1-D 0.94373 0.92627 0.9414 Simpson's Reciprocal index 1/D 17.77 13.56 17.06 Shannon index 3.8528 3.8663 4.1054 Evenness 0.8954 0.8104 0.7711 Margalef index 5.6118 6.4979 8.559 Table 2 — Very frequent and frequently recorded fungi on two different substrata . Very frequent Frequent Fronds Linocarpon appendiculatum (15.2%) Linocarpon nypae (6.8%) Oxydothi nypae (15.2%) Neolinocarpon globosicarpum (5.9%) L. bipolaris (12.7%) Trichocladium nypae (5.1%) Leaves L. bipolaris (13.5%) Linocarpon appendiculatum (8.1%) Astrosphaeriella striatispora (12.2%) Trichocladium nypae (8.1%) Sporidesmium crassisporum (6.7%) Helicorhoidion nypicola (6.7%) All the samples put together Linocarpon bipolaris (13%) Trichocladium nypae (6.2%) L. appendiculatum (12.5%) Astrosphaeriella striatispoa (6.2%) Oxydothis nypae (10.4%) Linocarpon nypae (6.2%) may be considered as one of the mangrove hosts sporulating fungi of which 25 species and 74 fungal supporting richest fungal diversity in mangroves as occurrences were recorded (Table 1). Linocarpon could be seen from the present and previous studies11. bipolaris (13.5%) and Astrosphaeriella striatispora The diversity may increase as new mangrove (12.2%) were very frequent while Trichocladium formations in other countries are investigated. nypae (8.1%), L. appendiculatum (8.1%), Helicorhoidion nypicola (6.7%) and Sporidesmium Substratum recurrence crassisporum (6.7%) were frequent (Table 2). Others Sixty frond samples examined supported 118 were either infrequent or rare in their occurrence. fungal occurrences comprising 32 fungal species Of the 46 taxa recorded in the present study, 32 (Table 2). Oxydothis nypae (15.2%), were recorded on fronds of which 21 were found only L. appendiculatum (15.2%) and L. bipolaris (12.7%) on fronds (Table 1). Twenty-five species were were frequently occurring fungi on fronds (Table 3). recorded from leaves of which 14 were found only on Thirty-nine leaf samples (out of 60) supported leaves. There were 11 overlapping species. HYDE & SARMA: FILAMENTOUS FUNGI 301

Table 3 — Horizontal distribution of filamentous fungi on Nypa fruticans, at Brunei S. No. Name of the species Site 1 Site 2 Site 3 Site 4 No. of sites of occurrence of each fungus 1 Aniptodera chesapeakensis Shearer & M.A. Mill - 1 - 1 2 2 A. intermedia K.D. Hyde & Alias - - - 1 1 3 A. mangrovei K.D. Hyde 1 1 - 1 3 4 A. nypae K.D. Hyde 2 2 - - 2 5 Annulatascus velatispora K.D. Hyde 1 - - - 1 6 Annulatascus palmietensis K.D. Hyde, Goh & T.D. Steinke 3 1 1 1 4 7 Annulatascus sp.1 - 2 - - 1 8 Annulatascus sp.2 - - 1 - 1 9 Anthostomella nypensis K.D. Hyde, Alias & B.S. Lu - - - 1 1 10 Anthostomella eructans - - - 1 1 11 Anthostomella sp. 1 - - - 1 12 Astrosphaeriella aquatica K.D. Hyde 1 - - - 1 13 A. nypae K.D. Hyde 1 - 1 4 3 14 A. striatispora (K.D. Hyde) K.D. Hyde 2 9 - 1 3 15 Astrosphaeriella sp. - - 1 - 1 16 Carinispora nypae K.D. Hyde - 1 2 - 2 17 Frondicola tunitricuspis - 1 3 1 3 18 Herpotrichea nypicola K.D. Hyde & Alias - - 1 - 1 19 Helicascus nypae K.D. Hyde - 1 - 1 2 20 Lignincola laevis Höhnk 1 - - - 1 21 Linocarpon angustatum K.D. Hyde & Alias 1 - 2 - 2 22 L. appendiculatum K.D. Hyde 6 5 5 8 4 23 L. bipolaris K.D. Hyde 6 1 8 10 4 24 L. livistonae (Henn.) K.D. Hyde - 2 - - 1 25 L. longisporum K.D. Hyde 1 1 - - 2 26 L. nypae (Henn.) K.D. Hyde 2 4 2 4 4 27 Lulworthia grandispora Meyers - 2 - - 1 28 Lulworthia sp. - 1 - - 1 29 Marinosphaera mangrovei K.D. Hyde - - 1 - 1 30 Neolinocarpon globosicarpum K.D. Hyde 5 - 2 - 2 31 Oxydothis nypae K.D. Hyde & Nakagiri 8 2 5 5 4 32 Phomatospora sp.-like - - - 1 1 33 Savoryella aquatica K.D. Hyde 1 1 1 - 3 34 S. lignicola E.B.G. Jones & R.A. Eaton 1 - - - 1 35 Cancellidium aplanatum Tubaki - - 1 1 2 36 Dictyochaete sp. - - - 1 1 37 Endophragmia sp. - - 1 - 2 38 Helicorhoidon nypicola K.D. Hyde & Goh 2 1 2 - 3 39 Papulospora sp. - - 1 - 1 40 Phialogeniculata sp. - 1 - - 1 41 Phoma sp. - - - 1 1 …Contd.

302 INDIAN J. MAR. SCI., VOL. 35, NO. 4, DECEMBER 2006

Table 3 — Horizontal distribution of filamentous fungi on Nypa fruticans, at Brunei — Contd. S. No. Name of the species Site 1 Site 2 Site 3 Site 4 No. of sites of occurrence of each fungus 42 Sporidesmium crassisporum M.B. Ellis 2 2 2 3 43 Trichocladium nypae K.D. Hyde & Goh 5 5 - 2 3 44 Trichocladium sp. 1 - - - 1 45 Unidentified hyphomycete 1 - - - 1 1 46 Unidentified hyphomycete 2 - - 1 - 1 No. of species 21 22 21 21 No. of fungal occurrences 52 47 44 49 No. of samples examined from each site and the total 30 30 30 30 Number of samples supporting sporulating fungi 23 26 20 23 Number of samples without sporulating fungi 7 4 10 7 Average number of fungi per sample 1.73 1.56 1.46 1.63 Simpson's Index D 0.064 0.063 0.060 0.08 Simpson's Index of Diversity 1-D 0.936 0.937 0.938 0.917 Simpson's Reciprocal index 1/D 15.6 15.9 16.6 12.1 Shannon index 3.736 3.733 3.811 3.682 Evenness 0.957 0.985 1.007 0.945 Margalef index 5.062 5.454 5.285 5.139

Linocarpon appendiculatum and Oxydothis nypae compared to other hosts excepting hosts such as were recurrent on fronds, while Astrosphaeriella Phragmites australis25,26, thus providing more width striatispora and A. nypae were recurrent on leaves. and depth for fruit body formation. Even though the Linocarpon bipolaris and Trichocladium nypae were leaf litter in the tropical forests gets moist it does not recorded with more or less the same percentage get inundated (albeit for longer periods). Whereas in occurrence on both the substrata. Nypa fruticans may the case of Nypa fruticans and Phragmites australis be considered as one of the mangrove hosts the leaves and leaf sheaths often get inundated. This supporting richest fungal diversity in mangroves. condition (availability of enough moisture for longer Yanna et al.20 illustrated similar tissue-recurrences of periods) coupled with relatively thick leaves of fungi on terrestrial palm fronds. N. fruticans or leaf sheaths in the case of Phragmites The average number of fungi and diversity per australis pose a conducive environment for the sample was greater on fronds than on leaves as formation of microscopic fruit bodies of ascomycetes. exemplified by the diversity indices (Table 1). It is Frequency of occurrence understandable that fronds with a greater width and Frequency of occurrence was calculated by thickness may offer a better substratum for fungal including the fungal occurrences on all 120 samples colonization than leaves, which are thinner. More than and is tabulated in Table 2. It was found that 70% of the species belonged to Ascomycota. This is Linocarpon bipolaris (13%), L. appendiculatum akin to other mangrove hosts as far as fronds are (12.5%), were the very frequent fungi, while concerned. However, the interesting observation in Oxydothis nypae (10.4%), Astrosphaeriella the present study is that more than 70% of recorded striatispora (6.2%), Trichocladium nypae (6.2%), species on leaves also belonged to Ascomycota. Linocarpon nypae (6.2%) were the "frequent" fungi. Anamorphic fungi are usually recorded in large The common species are thought to have better numbers on leaves of mangrove or terrestrial competitive capabilities thus accounting for their plants22-24. The reason for recording a greater number dominance27. Twenty-three fungi were recorded only of ascomycetes on leaves of Nypa fruticans may be once and are considered as rare in their occurrence on attributed to the thickness of the leaves when this host (Tables 1 and 2). It is common to find so HYDE & SARMA: FILAMENTOUS FUNGI 303

many rare species in a study of this type25,26,28, but taxa were recorded (Table 3). Oxydothis nypae their ecological importance is unknown16. Linocarpon (15.4%), Linocarpon appendiculatum (11.5%), L. and Oxydothis recorded more frequently on Nypa bipolaris (11.5%), Neolinocarpon globosicarpum fruticans have filiform and hyaline ascospores. (9.6%) and Trichocladium nypae (9.6%) were Among marine fungi that have filiform ascospores are recorded in greater numbers than others (Table 4). species belong to Lindra and Lulworthia29. It is Fungi recorded only at site I were Astrosphaeriella intriguing to note that Linocarpon and Oxydothis, aquatica, Lignincola laevis, Savoryella lignicola, which typically occur on terrestrial palms, have Trichocladium sp., Anthostomella sp. and adapted to marine conditions. Nypa fruticans might Annulatascus sp.1 (Table 3). have originally been a terrestrial palm that could have At Site II, 47 fungal occurrences belonging to 22 adapted itself to mangrove environments in the species were recorded. Astrosphaeriella striatispora geological timescale30. Species belonging to (19.6%), Trichocladium nypae (10.6%) and Linocarpon, Neolinocarpon and Oxydothis might L. appendiculatum (10.6%) were very frequent have reached marine environment along with the host (Table 4). Linocarpon livistonae, Lulworthia sp. and Nypa fruticans. Phialogeniculata sp., were recorded at Site II only Astrosphaeriella nypae, Linocarpon appendicula- (Table 3). tum, L. nypae and Oxydothis nypae were also reported At Site III, 44 fungal occurrences belonging to 21 to be very frequently occurring on Nypa fruticans in species were recorded. Linocarpon bipolaris (18.2%), the Philippines31, in Brunei7 and at Kuala Selangor in L. appendiculatum (11.4%) and Oxydothis nypae Malaysia11. However Lignincola laevis and (11.4%) were very frequent (Table 4). Annulatascus Astrocystis selangorensis G.J.D. Smith and K.D. sp.2, Astrosphaeriella sp., Herpotrichea nypicola, Hyde (reported as Rosellinia sp. in the above studies) Marinosphaera mangrovei, Endophragmia sp., that were very frequent in the above studies were Papulospora sp., Trichocladium sp., and unidentified either absent or recorded with very low percentage hyphomycete 1 were recorded only at Site III occurrence in the present study. (Table 3). Horizontal distribution At Site IV, 49 fungal occurrences belonging to 21 There have been two previous studies examining species were recorded. Linocarpon bipolaris (20.4%), the fungi on submerged substrates in estuaries along a L. appendiculatum (16.3%) and Oxydothis nypae freshwater to marine gradient17,32, however these (10.2%) were recorded in greater numbers (Table 4). studies investigated non-palm hosts. In this study 30 Aniptodera intermedia, Anthostomella nypensis, samples (5 each of leaves and fronds, each from Dictyochaeta sp., Phoma sp., Phomatospora sp.-like, submerged, intertidal and terrestrial zones) were Anthostomella eructans, Annulatascus velatisporus collected and examined from each of 4 sites17. and Unidentified Hyphomycete 2 were recorded only At the first site 52 fungal occurrences belonging to 21 at Site IV (Table 3).

Table 4 — Very frequent and frequently recorded fungi at different sites Very frequent Frequent Site I Oxydothis nypae (15.4%) Neolinocarpon globosicarpum (9.6%) Linocarpon appendiculatum (11.5%) Trichocladium nypae (9.6%) Linocarpon bipolaris ((11.5%) Astrosphaeriella striatispora (19.6%) Linocarpon nypae (8.5%) Site II L. appendiculatum (10.6%) Trichocladium nypae (10.6%) Linocarpon bipolaris (18.2%) Frondicola tunitricuspis (6.8%) Site III Oxydothis nypae (11.4%) Linocarpon appendiculatum (11.4%) Linocarpon bipolaris (20.4%) Astrosphaeriella nypae (8.2%) Site IV L. appendiculatum (16.3%) Linocarpon nypae (8.2%) Oxydothis nypae (10.2%) 304 INDIAN J. MAR. SCI., VOL. 35, NO. 4, DECEMBER 2006

The species composition at the four sites recorded at Site III (Table 4). Thus it can be dominated by different fungi is discernible except in mentioned that some marine fungi are tolerant to less the cases of Sites III and IV. In the first site Oxydothis saline conditions and that some freshwater species are nypae (15.4%) was dominant, while at Site II tolerant to more saline conditions17. The number of Astrosphaeriella striatispora (19.6%) was dominant. ascomycetes was greater at Sites I and II and less at Linocarpon bipolaris occurred abundantly (18.2% Sites III and IV. Correspondingly the numbers of and 20.4%, respectively) at Sites III and IV (Table 4). anamorphic taxa were less at Sites I and II and greater It is interesting to find that the species belonging to at Sites III and IV. A somewhat similar trend has been 17 Annulatascus are recorded on Nypa fruticans in the observed by Fryar et al. for unidentified decaying present study that were hitherto recorded only from wood in the same mangrove forest. submerged wood in freshwater and recently in Vertical distribution brackish water17. Salinity does not seem to have much Three levels were chosen: submerged, intertidal effect on the mycota as marine fungi occurred in all and terrestrial for vertical distribution study (at each the 4 sites, while species of Annulatascus also level 40 samples were examined). Thirty-two species occurred at all 4 sites. Differences in the species and 99 fungal occurrences were recorded from 40 diversity among the 4 sites were not significant samples collected from the submerged parts (Table 5). although the average number of fungi per sample was Linocarpon bipolaris (17.2%), L. appendiculatum more at Site I followed by Site IV and II with the least (11.1%) were very frequent at level I (submerged)

Table 5 —Vertical distribution of filamentous fungi on Nypa fruticans, at Brunei S. No. Name of the species Submerged Intertidal Terrestrial 1 Aniptodera chesapeakensis Shearer & M.A. Mill 1 1 - 2 A. intermedia K.D. Hyde & Alias 1 - - 3 A. mangrovei K.D. Hyde 2 1 - 4 A. nypae K.D. Hyde 2 2 - 5 Annulatascus velatispora K.D. Hyde - 1 - 6 Annulatascus palmietensis K.D. Hyde, Goh & T.D. Steinke 4 2 - 7 Annulatascus sp.1 - 2 - 8 Annulatascus sp.2 - 1 - 9 Anthostomella nypensis K.D. Hyde, Alias & B.S. Lu - 1 - 10 Anthostomella eructans - - 1 11 Anthostomella sp. - - 1 12 Astrosphaeriella aquatica K.D. Hyde - 1 - 13 A. nypae K.D. Hyde 3 3 - 14 A. striatispora (K.D. Hyde) K.D. Hyde 9 3 - 15 Astrosphaeriella sp. 1 - - 16 Carinispora nypae K.D. Hyde 1 2 - 17 Frondicola tunitricuspis 5 - - 18 Herpotrichea nypicola K.D. Hyde & Alias - 1 - 19 Helicascus nypae K.D. Hyde 2 - - 20 Lignincola laevis Höhnk 1 - - 21 Linocarpon angustatum K.D. Hyde & Alias - 1 2 22 L. appendiculatum K.D. Hyde 11 10 3 23 L. bipolaris K.D. Hyde 17 8 - 24 L. livistonae (Henn.) K.D. Hyde - 1 1 25 L. longisporum K.D. Hyde 2 - - …Contd. HYDE & SARMA: FILAMENTOUS FUNGI 305

Table 5 —Vertical distribution of filamentous fungi on Nypa fruticans, at Brunei S. No. Name of the species Submerged Intertidal Terrestrial 24 L. livistonae (Henn.) K.D. Hyde - 1 1 25 L. longisporum K.D. Hyde 2 - - 26 L. nypae (Henn.) K.D. Hyde 2 6 4 27 Lulworthia grandispora Meyers 2 - - 28 Lulworthia sp. 1 - - 29 Marinosphaera mangrovei K.D. Hyde 1 - - 30 Neolinocarpon globosicarpum K.D. Hyde 3 2 2 31 Oxydothis nypae K.D. Hyde & Nakagiri 6 5 9 32 Phomatospora sp.-like 1 - - 33 Savoryella aquatica K.D. Hyde 3 - - 34 S. lignicola E.B.G. Jones & R.A. Eaton 1 - - 35 Cancellidium aplanatum Tubaki 2 - - 36 Dictyochaete sp. 1 - - 37 Endophragmia sp. 1 - - 38 Helicorhoidon nypicola K.D. Hyde & Goh 4 1 - 39 Papulospora sp. 1 - - 40 Phialogeniculata sp. - 1 - 41 Phoma sp. 1 - - 42 Sporidesmium crassisporum M.B. Ellis - 2 4 43 Trichocladium nypae K.D. Hyde & Goh 6 4 2 44 Trichocladium sp. - - 1 45 Unidentified hyphomcete 1 - 1 - 46 Unidentified hyphomycete 2 1 - - No. of species 32 25 11 No. of fungal occurrences 99 63 30 No. of samples examined from each site and the total 40 40 40 No. of samples supporting sporulating fungi 38 30 24 No. of samples without sporulating fungi 2 10 16 Average number of fungi per sample 2.475 1.575 0.75 Simpson's Index D 0.0608 0.0593 0.1241 Simpson's Index of Diversity 1-D 0.9392 0.9907 0.8759 Simpson's Reciprocal index 1/D 16.44 16.86 8.058 Shannon index 4.0005 3.8507 3.4097 Evenness 0.8706 0.9295 1.0025 Margalef index 6.745 5.793 2.646 followed by Astrosphaeriella striatispora (9.1%), appendiculatum (15.9%), L. bipolaris (12.7%), Oxydothis nypae (6.1%) and Trichocladium nypae L. nypae (9.5%) and Oxydothis nypae (7.9%) were (6.1%), which were recorded frequently (Table 6). recorded in greater numbers than others. Eight species Eighteen species were recorded only at level I were recorded only at level II (Table 5). (Table 5). At level III (terrestrial zone) 11 species and 30 Twenty-five species and 63 fungal occurrences fungal occurrences were recorded. Only 24 out of 40 were recorded at intertidal level (level II). Linocarpon samples supported sporulating fungi and 16 did not 306 INDIAN J. MAR. SCI., VOL. 35, NO. 4, DECEMBER 2006

Table 6 —Very frequent and frequently recorded fungi at different vertical levels Very Frequent Frequent Submerged Linocarpon bipolaris (17.2%) Astrosphaeriella striatispora (9.1%) Linocarpon appendiculatum (11.1%) Oxydothis nypae (6.1%) Trichocladium nypae (6.1%) Intertidal Linocarpon appendiculatum (15.9%) Linocarpon nypae (9.5%) L. bipolaris (12.7%) Oxydothis nypae (7.9%) Helicorhoidion nypicola (6.3%) Terrestrial Oxydothis nypae (30%) Linocarpon nypae (13.3%) Sporidesmium crassisporum (13.3%) Linocarpon appendiculatum (10%) show any fruition. Oxydothis nypae is the very found that samples collected from lower tidal level frequent taxon (30%), followed by Linocarpon nypae harboured relatively more fungal taxa (19) than at the (13.3%) and Sporidesmium crassisporum (13.3%) that upper tidal level (16). Most of the fungi occurred at were recorded in more numbers than others (Table 6). both levels. Further the differences in their percentage Anthostomella eructans, Anthostomella sp., occurrence were insignificant. In the present study, Trichocladium sp. were recorded only from the however, 3 levels were chosen and the diversity was terrestrial parts in the present study (Table 5). richest at the submerged level, followed by the The statistical analysis shows significant intertidal level, while the highest tide level showed differences among the three vertical levels of which a the least diversity. rich diversity was found at the submerged level To conclude, this study shows that fronds have rich (level I) followed by the intertidal level (level II) and fungal diversity when compared to leaves; submerged finally the terrestrial zone (level III) (Table 5). parts have a higher fungal diversity when compared to The number of samples supporting sporulating intertidal or aerial parts; and all the sites along a fungi decreased through submerged parts (2.42 per salinity gradient showed equal fungal diversity sample), to intertidal parts (1.5) and terrestrial parts although very frequently occurring fungi were (0.75). Mostly members of the Halosphaeriales different in some sites. (marine fungi) were recorded on the submerged parts. A greater number of unitunicate ascomycetes Acknowledgement colonized the submerged parts (Table 5). While a First author is delighted to contribute this article in mixed mycota was found at the intertidal parts, the honor of Raghukumars who have done exemplary diversity was less at the terrestrial zone where typical work in the field of marine mycology. Sally Fryar and palm fungi and a few hyphomycetes were recorded. Gavin Smith are thanked for collecting the samples Only 4 fungi occurred in all three zones i.e. from Brunei. Helen Leung is thanked for technical throughout the tidal range viz., Linocarpon nypae, help. Neolinocarpon globosicarpum, Oxydothis nypae and References Trichocladium nypae. Of these Oxydothis nypae 1 Hyde, K.D., A comparison of intertidal mycota of five occurred more on terrestrial parts, while T. nypae mangrove tree species, Asian. Mar. Biol., 7 (1990) 93-107. occurred more on the submerged parts. A large 2 Hyde, K.D. & Lee, S.Y., Ecology of mangrove fungi and number of fungi (18) occurred in the submerged parts their role in nutrient recycling. What gaps occur in our only, followed by 10 fungal taxa only from intertidal knowledge?, Hydrobiologia, 295 (1995) 107-118. parts and 3 fungal taxa only from the terrestrial parts. 3 Poon, M.O.K. & Hyde, K.D., Biodiversity of intertidal Unlike Rhizophora33 a greater number of species and estuarine fungi on Phragmites at Mai Po marshes, Hong Kong, Bot. Mar., 41 (1998) 141-155. diversity was found on the submerged parts, than 4 Bucher, V.V.C., Hyde, K.D., Pointing, S.B. & Reddy, C.A., intertidal parts in the present study. Only one study on Production of wood decay enzymes, mass loss and lignin vertical distribution of fungi on Nypa fruticans is solubilization in wood by marine ascomycetes and their available for comparison31. In this study31 it was anamorphs, Fungal Divers, 15 (2004) 1-14. HYDE & SARMA: FILAMENTOUS FUNGI 307

5 Hyde, K.D., A new amphisphaeriaceous fungus from 20 Yanna, Ho, W.H. & Hyde, K.D., Fungal succession on intertidal fronds of Nypa fruticans, Trans. Mycol, Soc, Jpn., fronds of Phoenix hanceana in Hong Kong, Fungal Divers., 32 (1991) 265-271. 10 (2002) 185-211. 6 Hyde, K.D., Helicascus kanaloanus, Helicascus nypae sp. 21 Sarma, V.V. & Hyde, K.D., A review on frequently nov. and Salsuginea ramicola gen. et sp. nov. from intertidal occurring fungi in mangroves, Fungal Divers., 8 (2001) mangrove wood, Bot. Mar., 34 (1991) 311-318. 1-34. 7 Hyde, K.D., Fungi from decaying intertidal fronds of Nypa 22 Kirschner, R., Piepenbring, M & Chen, C.J., Some fruticans, including three new genera and four new species. cercosporoid hyphomycetes from Taiwan, including a new Bot. J. Linn. Soc., 110 (1992) 95-110. species of Stenella and new reports of Distocercospora 8 Hyde, K.D., Fungi from Nypa fruticans: Nipicola pachyderma and Phacellium paspali, Fungal Divers., carbospora gen. et sp. nov. (Ascomycotina), Cryptogam. 17 (2004) 57-68. Bot., 2 (1992) 330-332. 23 Lee, S., Mel'nik, V., Taylor, J.E. & Crous, P.W., Diversity 9 Hyde, K.D., Fungi from palms. V. Phomatospora nypae sp. of saprobic hyphomycetes on Proteaceae and Restionaceae nov. and notes on marine fungi from Nypa fruticans in from South Africa, Fungal Divers., 17 (2004) 91-114. Malaysia. Sydowia, 45 (1993) 199-203. 24 Schubert, K. & Braun, U., Taxonomic revision of the genus 10 Hyde, K.D. & Alias, S.A., Linocarpon angustatum sp.nov. Cladosporium s.l. 4. Species reallocated to Asperisporium, and Neolinocarpon nypicola sp. nov. from petioles of Nypa Dischloridium, Fusicladium, Passalora, Pseudo- fruticans, and a list of fungi from aerial parts of Nypa palm, asperisporium and Stenella, Fungal Divers., 20 (2005) Mycoscience 40 (1999) 145-149. 187-208. 11 Hyde, K.D. & Alias, S.A., Biodiversity and distribution of 25 Van Ryckegem, G. & Verbeken, A., Fungal ecology and fungi associated with decomposing Nypa fruticans. succession on Phragmites australis in a brackish tidal Biodiver. Conser., 9 (2000) 393-402. marsh. I. Leaf sheaths, Fungal Divers, 19 (2005) 157-187. 12 Hyde, K.D. & Nakagiri, A., A new species of Oxydothis 26 Van Ryckegem, G. & Verbeken, A., Fungal ecology and from the mangrove palm Nypa fruticans, Tran. Mycol. Soc. succession on Phragmites australis in a brackish tidal Jpn., 30 (1989) 69-75. marsh. II. Stems. Fungal Divers., 20 (2005) 209-233. 13 Hyde, K.D. & Sutton, B.C., Nypaella frondicola gen. et sp. nov., Plectophomella nypae sp. nov. and Pleurophomopsis 27 Fryar, S.C., Booth, W., Davies, J., Hodgkiss, I.J. & Hyde, nypae sp. nov. (Coelomycetes) from intertidal fronds of K.D., Evidence of in situ competition between fungi in Nypa fruticans, Mycol. Res., 96 (1992) 210-214. freshwater, Fungal Divers., 18 (2005) 59-71. 14 Chatmala, I., Sakayaroj1, J., Somrithipol, S. & 28 Luo, J., Yin, J.F., Cai, L., Zhang, K. & Hyde, K.D., Phongpaichit, S, Marine hyphomycetes of Thailand and Freshwater fungi in Lake Dianchi, a heavily polluted lake in Cumulospora varia sp. nov., Fungal Divers., 17 (2004) 1-9. Yunnan, China, Fungal Divers., 16 (2004) 93-112. 15 Hyde, K.D., Sarma, V.V. & Jones, E.B.G. Morphology and 29 Tsui, C.K.M. & Hyde, K.D., Freshwater mycology. Fungal taxonomy of higher marine fungi: In Marine mycology-A Diversity Research Series 10 (Fungal Diversity Press, practical approach, edited by K.D. Hyde and S.B. Pointing, University of Hong Kong, Hong Kong) (2003) pp 350. Fungal Divers. Res. Ser.1, 172-204, (Fungal Diversity Press, 30 Tomlinson, P. B., The botany of mangroves, (Cambridge University of Hong Kong, Hong Kong), 2000, pp 377 Univ. Press, Cambridge) 1986, pp 433. 16 Hyde, K.D., Bussaban, B., Paulus, B., Crous, P.W., Lee, S., 31 Besitulo, A.D., Sarma, V.V. & Hyde, K.D., Mangrove fungi Mckenzie, E H.C., Photita, W. & Lumyong, S.. Biodiversity from Siargao Island, Philippines, In: Fungi in marine of saprobic fungi. Biodiver. Conser. (in press) (2006) environments, edited by K.D. Hyde, Fungal Diversity 17 Fryar, S.C., Booth, W., Davies, J., Hodgkiss, I.J. and Hyde, Research Series 7, 267-283 (Fungal Diversity Press, K.D., Distribution of fungi on wood in the Tutong River, University of Hong Kong, Hong Kong), 2002, pp 397. Brunei, Fungal Divers, 17 (2004) 17-38. 32 Tsui, C.K.M. & Hyde, K.D., Biodiversity of fungi on 18 Magurran, A.E., Ecological diversity and its measurement, submerged wood in a stream and estuaries in the Tai Ho (Princeton University, New Jersey), 1988, pp 192 Bay, Hong Kong, Fungal Divers., 15 (2004) 171-186. 19 Ludwig, J.A. & Reynolds, J.F., Statistical ecology - A 33 Hyde, K.D., A study of the vertical zonation of intertidal primer on methods and computing, (John Wiley, New York) fungi on Rhizophora apiculata at Kampong Kapok 1988, pp 337 mangrove, Brunei, Aquat. Bot., 36 (1990) 255-262.