Fungal Diversity Occurrence of zoosporic and terrestrial fungi in accumulated rainfall water in the Riyadh region (Saudi Arabia)

1* 2 M.A. EI-Nagdy and L.A. Nasser

IBotany Department, Faculty of Science, Assiut University, Assiut, Egypt 2Girls College of Education, Riyadh, Saudi Arabia

El-Nagdy, M.A. and Nasser, L.A. (2000). Occurrence of zoosporic and terrestrial fungi in accumulated rainfall water in the Riyadh region (Saudu Arabia). In: Aquatic Mycology across the Millennium (eds K.D. Hyde, W.H. Ho and S.B. Pointing). Fungal Diversity 5: 175-183.

Twenty-one belonging to ten genera of zoosporic fungi were recovered from 23 surface water samples collected from ponds of accumulated rainfall in the Riyadh region (Saudi Arabia) using seeds as baits at 20 ± 2 C. The most common genera were Allomyces (3 species), Aqualinderella (I species), Dictyuchus (2 species) and Pythium (3 species). The remaining six genera were less frequent. Four species (Allomyces macrogynus, Aqualinderella fermentans, Pythium rostratum and Pythium ultimum) were recorded for the first time in Saudi Arabia. Fifty-one species and one variety of terrestrial fungi belonging to 21 genera were also identified on glucose and cellulose Czapek's Dox agar incubated at 28 C. The most common species were Aspergillus flavus, A. fumigatus. A. niger, A. flavus var. columnaris, Acremonium stictum, Cladosporium herbarum, Fusarium oxysporum and Penicillium corylophilum. Some fungi such as Chaetomium globosum and Memnoniella echinata were isolated only on cellulose agar medium.

Key words: rainfall, Riyadh, Saudi Arabia, terrestrial fungi, zoosporic fungi.

Introduction The occurrence of zoosporic fungi in different water habitats from various geographical regions of the world have been intensively studied (e.g. Dick, 1972; Johnson, 1974; Hasija and Batra, 1978; Rattan et al., 1980; EI-Hissy and Khallil, 1989; EI-Nagdy and Abdel-Hafez, 1990; EI-Hissy, 1994; EI-Hissy et al., 1996). In Saudi Arabia, there have only been one study on the occurrence and distribution of this group of fungi in various habitats (EI-Nagdy et al., 1992). The occurrence and distribution of lower freshwater fungi and terrestrial fungi in accumulated rainfall in the Riyadh region have not yet been studied, however, in the Riyadh region have not yet been studied. This investigation focuses on studying freshwater fungi from accumulated rainfall in this region.

Materials and methods Twenty-three surface water samples were collected during January 1998

175 from some ponds of the accumulated rainfall in the Riyadh region (Saudi Arabia). This region is mainly a mountainous area, and has a cold winter with an annual rainfall of between 5-12 mL. Temperature and pH value of the collected water samples were recorded at the time of sampling. Total soluble salts and organic contents were determined according to the method described by Jackson (1958).

Zoosporic fungi Aliquots of 20 mL of each rainfall sample were introduced into each of five sterile Petri dishes. Six sterilized sesame seed were placed in each Petri dish as baits and plates were left at room temperature (~20 C) for 24 h. Colonized seeds were then transferred to Petri dishes containing sterile water and incubated at 20 ± 2 C for 5 weeks during which the colonized seeds with zoosporic fungi were transferred several times to sterile Petri dishes containing sterile water. The zoosporic fungi that appeared on the baits were examined, identified, counted and the numbers were calculated per 30 seeds in every sample.

Terrestrialfungi Two mL of each water sample were transferred aseptically into each of ten sterile Petri dishes. Modified CZAPEK's Dox agar medium in which glucose (10 g/L) or powder cellulose (20 g/L) was used for isolation of glucophilic and cellulose-decomposing fungi respectively. Rose Bengal (1/15000) was used as a bacteriostatic agent. Five plates were used for each medium and the plates were incubated at 28 C or 7-15 days during which the developing fungi were counted, examined and identified. The numbers were calculated per 1 mL water in every sample.

Results and discussion Chemical analysis show that the organic contents of the water samples were poor and ranged between 0.18-0.86%, the total soluble salts fluctuated between 0.15-0.97%. The results of pH values reveal no appreciable differences. All were alkaline with a pH from 7.1-8.05. Water temperatures ranged between 20 and 22 C. EI-Nagdy et al. (1992) found that the accumulated rainfall and mud collected from the Taif region (Saudi Arabia) were generally poor in organic matter and varied between 0.03-0.42%, 0.86-2.9% and their contents of total soluble salts ranged between 0.02-0.88%; 0.14-0.74% respectively. Also, Abdel-Hafez (1982a) found that the organic contents and total soluble salts in Saudi Arabia desert soils were generally low and fluctuated between 0.02-0.32% and 0.05-0.46% respectively.

176

n Fungal Diversity Zoosporic fungi recovered on sesame seeds baits Twenty-one species of aquatic zoosporic fungi belonging to ten genera were collected from rainfall water using sesame seeds as baits (Table 1). Most of these species have previously been recorded from accumulated rainfall water and mud in Taif, Saudi Arabia (El-Nagdy et al., 1992) Allomyces macrogynus, Aqualinderella fermentans, Pythium rostratum and Pythium ultimum were recorded for the first time in Saudi Arabia. Allomyces was the most prevalent and was recovered from 95.7% of the samples yielding 22.8% of total fungi. A. arbuscula and A. moniliformis were the most common with A. macrogynus was less frequent. This is in accordance with the results previously obtained from accumulated rainfall water and mud in Taif region which is situated in the western highland region of Saudi Arabia (El-Nagdy et al., 1992). These species have also been encountered, with varying frequencies, from the River Nile system (El-Hissy and El-Nagdy, 1983 and El-Hissy et al., 1996) ground water of the Kharga Oases in the Western Desert of Egypt (El-Nagdy and Abdel-Hafez, 1990). Reports indicate that A. abuscula and A. moniliformis have are wide spread in tropical and warm regions (Bhargava and Singh, 1965). Allomyces was also found in water samples from India (Sirvastava, 1967). Aqualinderella (1 species), Dictyuchus (2 species) and Pythium (3 species) were also isolated with a high frequency of occurrence. They were recovered from 91%, 87%, 56.5% of the water samples, contributing to 14%, 17%, 7% of total fungal population respectively. All the identified species in these genera have been isolated previously, but with different frequencies from various water habitats in Egypt, Saudi Arabia and other countries (Alabi, 1973; Lui and Volz, 1976; Sati and Khulbe, 1980; Chowdhery and Rai, 1980; Prabhuji, 1984; Klich and Tiffany, 1985; El-Nagdy and Abdel-Hafez, 1990; El-Nagdy et al., 1992; El• Hissy, 1994; El-Hissy et al., 1996). had a moderate occurrence and was represented by S. anisospora, S. ferax and S. hypogyna. also had a moderate frequency and contributed the broadest spectrum of species (6 taxa) of which Achlya prolifera, A. dubia and A. debaryana were the most common. Similar results were reported by El-Nagdy et al. (1992) from the accumulated rainfall water and mud in Taif, Saudi Arabia. The recoverable Saprolegnia and Achlya species were encountered, but with various incidences, from various water habitats in Egypt (EI-Nagdy and Abdel-Hafez, 1990; Khallil et al., 1993; El-Hissy et al., 1996) and many parts of the world (Alabi, 1973; Khulbe, 1980; Misera, 1982; Klich and Tiffany, 1985; El-Hissy, 1994). The aquatic taxa identified during this investigation have previously been isolated from the Nile system. The aquatic mycota of freshwater in Saudi

177 Table 1. Total counts (calculated per 30 seeds in every sample), and number of isolations of zoosporic fungal genera and species from 23 water samples using sesame seeds as baits at 20 ± 2e.

Fungi Total count Number of isolations Achlya 61 11 M A. debaryana Humphrey 7 4 L A. dubia Coker 8 5 L A. prolifera e.G.Nees 16 10 M A. proliferoides Coker 5 2 R A. racemosa Hildebrand 7 4 L Achlya sp. (non. sexual) 18 7 M Allomyces 61 22 H A. arbuscula Butler 37 18 H A. macrogynus Emerson and Wilson II 6 M A. moniliformis Coker and Braxton 13 12 H Aphanomyces laevis De Bary 3 2 R Aqualinderellafermentans Emerson and 38 21 H Weston Brevilegnia bispora Couch 3 3 L Calyptralegnia ripariensis Hohnk 5 4 L Dictyuchus 46 20 H D. carpophorus Zopf 8 3 L D. sterilis Coker 38 19 H Pythium 21 13 H P. rostratum Butler 12 8 M P. thalassium Atkins 7 6 M P. ultimum Trow 2 2 R Saprolegnia 27 10 M S. anisospora De Bary 3 2 R S.ferax (Gruith.) Thuret 8 5 L S. hypogyna (Pringsheim) De Bary 7 4 L Saprolegnia sp. (non-sexual) 9 7 M Thraustotheca clavata (De Bary) 2 2 R Humphery Total count 267

I Occurrence remarks (OR): H = high occurrence (> II samples; out of 23); M = moderate occurrence (6-11 samples); L = low occurrence (3-5 samples); R = rare occurrence «3 samples).

Arabia is similar to the Nile system in Egypt.

Terrestrial fungi Fifty-one species and one variety of terrestrial fungi belonging to 21 genera were also identified (Table 2).

178 0.43.52.1- 136.328.832.120.442.810.40.80.40.22.62.8NCI3.97.80.13.88.21.4M5.4R1.61.27.23.223NCIRLMORlHOR110187HTC1213311104165220541216118102311623818247201253121131511 Cellulose 52.145.636.510.213.24.10.27.10.15.35.12.90.50.41.51.61.21.1- 169.80.22.16.60.50.44.2221.71.5- Glucose Fungal Diversity ChaetomiumAlternariaG.A.F. SmithalternatawentiiacuminatumcandidusterreChurch.usWehmerThom.Link:(Fr.)EllisKreisslerFr.andA.Everh.clavatus Desmazieres CephalosporiumAspergillusBotyotrichumCladosporiumCurvulariaHumicolaI AcremoniumOccurrenceA.A.flavusF.C.A. solanisydowitenuissimamoniliformeveriscoloroxysporiumglobosumrugulosusnidulansustusspiralenigerherbarumcladosporioidesgrisealunata(Bain.)Vanvar.Link:(Mart.)(Bain.Zopfremarksatrogriseumstrictum(Eidam)ThomKunze:Tieghem(Vuill.)columnaris(Pers.:Fr.)curtipes(Kunze:Fr.)TraaenSchlechtendahlFr.Sheldon(Wakker)Thom.andSacc.(OR):(Fresenius)andDorotomycesMemnoniellaWint.W.Fr.TiraboschiSart.)Sacc.andA.fumigatusF.RaperLinkVanGamsRaperBoedijnWiltshireHequisetiThom.Church.Beyma= dehighandmicrosporusandVriesechinata(Corda)FennelloccurrenceFreseniusSacc.(Riv.)(Sacc.)(>Galloway11 samples;MortonTC andout of 23); M = moderateA.Fusariumtamariioccurrencenumbersamples).Kitaof isolations(6-11 samples);(NCI) ofLgenera= lowandoccurrencespecies from(3-523 samples);water samples.R = rare occurrence «3 Table 2. Total counts (TC, calculated per mL water in every sample) of terrestrial fungi, and Fungi

179 0.10.81.21.1- Table 2.53.320214.23.20.20.40.8NCI8.4R1.1M8.6L1.1NCIRMHOR!OR'H10TC45132102012111813741615213(Continued).36151 246.]Cellulose 32.112.315.411.216.90.10.40.2- 76.73112.78.40.91.20.50.21.4-- ] Glucose PhomaRhizopusMyceliaTrichodermaTotalTrichurusMucorP.P. countvariabileglomeratajenseniioxalicumwaksmaniichrysogenumduclauxicorylophilumcitrinumHochafelsteriliastoloniferspiralisvirideZaleskiThorn.DelacroixSoppCurrie(white(Corda)ZaleskiHasselbringThorn.DierckxPers.:Ehrenb.andScopulariopsisStachybotrysPenicilliumandPaecilomycesWollenWeberFr.Thorn.M.P.dark)funiculosummartensiiracemosuscircinelloideschartarumvariotibrevicaulisandBiourgeFeseniusThorn.VanBainier(Ehrenb.Tiegh.(Sacc.) Fr.)BainierTCHughes P. humicola Gilrn(j.nand Abbott Fungi

Glucophilic fungi (recovered on glucose-Czapek's agar) Forty-five species and one variety belonging to 17 genera were isolated on glucose-Czapek's agar (Table 2). The most frequent genera were Aspergillus (13 species + 1 variety), Cladosporium (2 species), Fusarium (5 species), Mucor (2 species) and Penicillium (10 species). From the above genera the most prevalent species were Aspergillus fumigatus, A. flavus, A. flavus var. columnaris, A. niger, A. terreus, Cladosporium herbarum, Fusarium oxysporum, F. solani. Mucor racemousus, Penicillium chrysogenum, P. corylophilum and P. funiculosum. These genera and species were previously isolated, but with various numbers and frequencies, from different substrata in Saudi Arabia such as rainfall water and mud (El-Nagdy et al., 1992), soil (Abdel-Hafez, 1982a) and ferns (Abdel-Hafez, 1984). The remaining species were less frequent (Table 2). Almost all of the terrestrial taxa recovered in this study had been found in various habitats in Egypt (Abdel-Hafez and Bagy,

180 Fungal Diversity 1985; EI-Hissy et al., 1990; Moharrum et al., 1990; EI-Nagdy and Abdel• Hafez, 1990) and other countries (Barlocher and Kendrick, 1974; Bettucci et al., 1993; Bettucci and Roquebert, 1995). Terrestrial fungi in aquatic habitats are likely to originate from air (Sparrow, 1968), as well as from living or dead animal and plant, soil and litter being in contact with water (Park, 1972).

Cellulose-decomposingfungi (recovered on cellulose-Czapek's agar) Forty species belonging to 15 genera were isolated from water samples on cellulose-Czapek's agar (Table 2). The results were similar to those on glucose agar and the most frequent genera were Acremonium, Aspergillus, Chaetomium, Fusarium and Penicillium. The most prevalent species were Acremonium strictum, Aspergillus flavus, A. fumigatus, A. niger, A. terre us, Chaetomium globosum, Fusarium oxysporium, F. solani, Penicillium chrysogenum, P. corylophilum and P. citrinum. These species have also been isolated from soils, water and other substrata in Saudi Arabia, Egypt and other countries (Abdel-Hafez et al., 1978; Abdel-Hafez, 1982b; Abdel-Kader et al., 1983; Abdel-Hafez and Bagy, 1985). The remaining genera and species were less frequent (Table 2). Most species recovered on cellulose Czapek's agar were reported to be cellulose decomposing (Tribe, 1961, 1966; Stewart and Walsh, 1972; EI-Nagdy and Abdel-Hafez,1990).

Acknowledgements The authors are indebted to F.T. El-Hissy for valuble discussions.

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(Received 16 July 1999, accepted 27 June 2000)

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