Songklanakarin Journal of Science and Technology SJST-2013-0136.R1 MOHD RAZIKIN First report of pteridocolous discomycetes, Lachnum lanariceps and L. oncospermatum, on decayed tree fern in Bukit Bendera (the Penang Hill), Pulau Pinang, Malaysia For Review Only Journal: Songklanakarin Journal of Science and Technology Manuscript ID: SJST-2013-0136.R1 Manuscript Type: Original Article Date Submitted by the Author: 14-Mar-2014 Complete List of Authors: MOHD RAZIKIN, MUHAMMAD ZULFA; UNIVERSITI SAINS MALAYSIA, Nagao, Hideyuki; UNIVERSITI SAINS MALAYSIA, Zakaria, Rahmad; UNIVERSITI SAINS MALAYSIA, Keyword: Agricultural and Biological Sciences For Proof Read only Page 1 of 12 Songklanakarin Journal of Science and Technology SJST-2013-0136.R1 MOHD RAZIKIN 1 2 3 1 Abstract 4 5 6 2 Bukit Bendera is 833m above sea level and situated in the Northern part of Penang Island, 7 8 3 Malaysia. Generally an average temperature is between 20 to 27 °C, which is about 5°C 9 10 4 cooler than at the sea level. The hill dipterocarp forest dominates Bukit Bendera and tree fern 11 12 13 5 scatteredly grows at higher altitude. Two Lachnum spp. were observed as pteridocolous cup 14 15 6 fungi on decayed rachides of several tree fern species, Cyathea contaminans, C. latebrosa, 16 17 7 and C. hymenodes . Lachnum oncospermatum is characterized by a wrinkled apothecium and 18 For Review Only 19 8 branched stipe. The hairs contain brown coloured resinous materials and are finely granulated. 20 21 9 Lachnum lanariceps is characterized by a central and cylindrical stipe and hairs containing 22 23 24 10 pale yellow pigment with red or garnet resinous matter. These 2 Lachnum species are new to 25 26 11 Malaysia. 27 28 29 12 30 31 32 13 Key words Cyathea · Lachnum · pteridocolous discomycetes · tree fern · Malaysia 33 34 35 14 36 37 38 15 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 For Proof Read only Songklanakarin Journal of Science and Technology SJST-2013-0136.R1 MOHD RAZIKIN Page 2 of 12 1 2 3 1 4 5 2 Introduction 6 7 8 3 Lachnum is a genus of fungi belonging to Hyaloscyphaceae, Helotiales, 9 10 4 Leotiomycetidae, Leotiomycetes, Pezizomycotina and Ascomycota. Twenty two genera have 11 12 5 been treated in synonym with Lachnum Retz., which are, Arenaea Penz. & Sacc., Belonidium 13 14 15 6 Mont. & Durieu, Belonidium sect. Lasiobelonium Sacc., Capitotricha (Raitv.) Baral, 16 17 7 Chaetoscypha Syd., Dasypezis Clem., Dasyscyphus Nees ex Gray, Dasyscyphus subgen. 18 For Review Only 19 8 Capitotricha Raitv., Dyslachnum Clem., Erinella Quél, Erinella Sacc., Erinellina Seaver, 20 21 9 Erioscypha Kirschst., Erioscyphella Kirschst., Helolachnum Torrend, Hyphoscypha Bres., 22 23 24 10 Lachnaster Höhn., Lachnella Boud., Lachnobelonium Höhn., Lasiobelonium (Sacc.) Sacc. & 25 26 11 P. Syd., Pezizellaster Höhn. and Trichopezizella Dennis & Raitv. (Spooner, 1987). 27 28 29 12 Lachnum is a very large genus with cosmopolitan distribution not only in the 30 31 13 temperate zone such as UK, Europe (Dennis, 1949), USA (Seaver, 1951) and Japan (Nagao, 32 33 14 1996; 2008; Nagao and Doi, 1996), but also in the tropical zone such as Central and South 34 35 15 America (Haines, 1980; 1992), Taiwan (Wu et al ., 1998; Wu and Wang, 2000) and in the 36 37 38 16 Southeast Asia and Australasia (Dennis, 1958; Spooner, 1987), some new species were earlier 39 40 17 recorded in Java (Penzig and Saccardo, 1904). Lachnum species live and grow on plants. The 41 42 18 forms of substrate are dead and decaying leaves, stems or woody substrate, and herbaceous 43 44 19 stems or leaves. The categories of plants are pteridophytes, angiopserms and gymnosperms 45 46 47 20 (Spooner, 1987). 48 49 50 21 Lachnum is characterized by the finely granulate hairs on apothecia, asci with a conical apex 51 52 22 stained blue in Melzer’s reagent, and an ectal excipulum composed of prismatic cells 53 54 23 (Spooner, 1987). Among the species, longer ascospores are recognized on palm specific 55 56 57 58 59 60 For Proof Read only Page 3 of 12 Songklanakarin Journal of Science and Technology SJST-2013-0136.R1 MOHD RAZIKIN 1 2 3 1 species, L. pritzelianum and L. palmae ranging 53-60 µm and 62-77 µm, respectively 4 5 2 (Spooner, 1987). 6 7 8 3 Tree ferns are recognized as the fern with a tall trunk-like rhizome and they are distributed in 9 10 4 the tropical lowland to submontane environments (Piggott, 1988; Large and Braggins, 2004). 11 12 13 5 Decaying ferns are known as good substrates for large number of microfungi, not only 14 15 6 parasitic but also for saprophytic species (Bøhler 1974; Holm and Holm 1978; Haines, 1980). 16 17 7 Several tropical ferns; Alsophila , Blechnum , Cyathea , Dicksonia , Gleichenia , and Papuapteris 18 For Review Only 19 8 have been recognized as the host plant of several species of pteridocolous Hyaloscyphaceae 20 21 9 (Dennis, 1958; Haines, 1980; 1992; Spooner, 1987; Nagao, 1996; 2008; Nagao and Doi, 22 23 24 10 1996; Wu et al ., 1998; Wu and Wang, 2000). In Southeast Asia, decaying rachides of tree 25 26 11 ferns were specifically colonized by these pteridocolous species recorded in Java (Penzig and 27 28 12 Saccardo, 1904) and the Philippine Islands (Dennis, 1958; Spooner, 1987) 29 30 31 13 In Bukit Bendera, tree fern species, Cyathea contaminans was found at the edge of the forest 32 33 14 besides roads and Cyathea latebrosa was widely distributed at the hill areas and beside 34 35 36 15 streams with greater atmospheric condition. However, up until now there has not been 37 38 16 recognition of pteridocolous fungus from Bukit Bendera. Two species of pteridocolous 39 40 17 Lachnum were observed on decayed rachides of Cyathea spp. in Bukit Bendera. The objective 41 42 18 of this research is to study the biodiversity of saprophytic microbes in Bukit Bendera. 43 44 45 19 46 47 48 20 Materials and Method 49 50 51 21 Collection Sites 52 53 54 55 56 57 58 59 60 For Proof Read only Songklanakarin Journal of Science and Technology SJST-2013-0136.R1 MOHD RAZIKIN Page 4 of 12 1 2 3 1 Decayed rachides of tree fern were collected along the jeep road and beside a stream in Bukit 4 5 2 Bendera between 600m and 800m elevation. Seven specimens on Cyathea spp. were obtained 6 7 3 on 15 December 2011 and 11 January 2012. 8 9 10 4 Observations of Micro-morphology 11 12 13 5 Fruiting bodies on the samples were immediately observed to determine the type of apothecia. 14 15 16 6 Dried specimens were dehydrated and sectioned freehand using a razor-blade. Sections were 17 18 7 mounted in distilledFor water or Shear’s Review solution for light microscopicOnly observation. 3% KOH-1% 19 20 8 phloxine and Melzer’s reagent were properly added to detect septa formation and positive 21 22 9 iodine reaction in the ascus pore respectively. A squashed section was used to observe asci, 23 24 ascospores, paraphyses and the texture of apothecium and stipe in detail. 25 10 26 27 28 11 29 30 31 12 Results and Discussion 32 33 34 13 35 36 37 14 1. Lachnum oncospermatum (Berk. & Broome) M. L. Wu, J. H. Haines and Y. Z. Wang, 38 39 15 Mycotaxon 67:346, 1998. 40 41 16 42 43 17 ≡ Peziza oncospermatis Berk. & Broom, J. Linn. Soc. Bot. 14:105, 1875. 44 45 ≡ Dasyscyphus oncospermatis 46 18 Berk. & Broom, Syll. Fung. 8:465, 1889. 47 48 19 ≡ Atractobolus oncospermatis (Berk. & Broom) O. Kuntze, Revis. Gen. Pl. 3(2): 446, 1898. 49 50 20 ≡ Aranea oncospermatis (Berk. & Broom) Petch, Ann. R. Bot. Gard. Peradiniya 6: 164, 1917. 51 52 21 53 54 22 Apothecia lobed, branched stipes, centric, scattered on the decayed rachides of Cyathea sp. 55 56 57 23 Apothecia ranged in size between 0.4-0.7 mm diameter. The apothecia are covered with hair 58 59 60 For Proof Read only Page 5 of 12 Songklanakarin Journal of Science and Technology SJST-2013-0136.R1 MOHD RAZIKIN 1 2 3 1 containing pale yellow pigment. The hairs bearing brown coloured resinous materials and are 4 5 2 finely granulated. The sizes of asci ranged from 40.1-49.1 × 3.1-3.8 µm and cylindric clavate 6 7 3 with 8 ascospores, the apex stained blue in Melzer’s reagent. Ascospores dimensions ranged 8 9 10 4 from 11.4-17.6 × 1.4-2.0 µm, narrowly fusoid, acute at the ends, straight or slightly curved 11 12 5 and non-septate. Paraphyses were range between 1.2-1.7 µm at the widest point, narrowly 13 14 6 lanceolate, hyaline, straight, sometime branched and the length of paraphyses was longer than that 15 16 7 of asci. 17 18 For Review Only 19 8 Morphology of the ascospores of pteridocolous discomycetes is known to be acute and 20 21 9 aseptate. The length of ascospores of L. oncospermatum is shorter than L. lanariceps . Colour 22 23 of resinous matter on hairs of L. oncospermatum is also different from L. lanariceps . Colour 24 10 25 26 11 of resinous matter of L. oncospermatum is brown in colour while L. lanariceps is red or red- 27 28 12 brown (Spooner, 1987; Wu et al ., 1998 ). Apothecia of L. oncospermatum are branched 29 30 13 whereas those of L. lanariceps single. The morphological differences in these two species are 31 32 14 confirmed between this examination.