Species of penicillium are among the most widely ...

The 3rd Kurdistan Conference on Biological Sciences J. Duhok Univ.Vol.13,No.1, Pp 144-163, 2010 University of Duhok 4-6 May, 2010

TAXONOMIC STUDY ON PENICILLI FROM SOIL IN KURDISTAN REGION OF IRAQ. WAZEERA R. ABDULLAH and SAMIR K. ABDULLAH Dept. of Biology, College of Education, University OF Duhok, Kurdistan Region-Iraq (Accepted for publication: May 24, 2010)

ABSTRACT A taxonomic study on the genus Penicillium of the family Trichocomaceae (Eurotiales):Ascomycota inhabiting soils in Kurdistan region of Iraq was carried out using phenotypical and cultural characteristics. Twelve taxa have been identified . The identified species were P.aurantiogriseum, P.brevicompactum, P.camemberti, P.citrinum, P.corylophilum, P.digitatum, P.echinulatum, P.funiculosum, P.glabrum, P.roqueforti, P.rugulosum and P.variabile. A total of five species have been reported for the first time from Iraqi soil. The new records include P.aurantiogriseum, P.camemberti, P.corylophilum, P.echinulatum, and P.variabile. A brief description based on cultural and morphological characteristics is provided for each species. A dichotomous key in provided to facillate the identification of the reported species. KEY WORDS: Soil fungi, penicillium, Iraq

INTRODUCTION

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pecies of Penicillium are among the most widely distributed of all fungi.Virtually any environmental sample such as soil, water, air and food will yield viable spores of many species assigned to the genus Penicillium (Webster and Weber, 2007 ). Some species of Penicillium have a positive impact on our life in producing several industrial enzymes, organic acids, antibiotics, antifungal and antitumor agents (Fleming, 1944; Braim, 1960; Rolinson,1998; Jakobsen et al. 2002). Negative aspect of Penicillium included production of toxic secondary metabolites(mycotoxins), spoilage and deterioration of food and feeds and causing human and animal diseases (Pitt and Hocking, 1997; Vanittanakom et al. 2006; Kwon-Chung and Bennett, 1992). Taxonomy of Penicillium is not easy for unexperienced. The species are mostly similar in color and general colony appearance unlike Aspergillus. The reproductive structure characteristic of Penicillium is a conidiophore with a relatively delicate, distinct stipe terminating in a penicillus. The form of the penicillus determines, the primary taxonomic division of the genus into four subgenera (Pitt and Hocking, 1997). 1- Penicillium subgenus Aspergilloides: There is one branch point, conidia are borne from phialides which in turn are borne directly from the stipe (conidiophore ) as a single whorl or

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verticil. Such penicilli are termed monoverticillate. 2- Biverticillate penicilli have two branch points. In this category, there are two distinct subgenera: Penicillium subgenus Furcatum and Penicillium subgenus Biverticillum. In the first subgenus, the penicilli are irregular while in the second subgenus, penicilli are symmetrical (Pitt and Hocking, 1997). 3- Terverticillate in which metulae may in turn arise from a further verticil of branches, the rami i.e three branches . Terverticillate are assigned to the subgenus Penicillium. Moreover, biverticillate and quaterverticillate penicilli are also produced by some species (Frisvad and Samson, 2004). Most of the penicillium species are Known for their anamorphic states. Those that have teleomorphs in the form of cleistothecia assigned to the genera Eupenicillium , and Talaromyces in the family Trichocomaceae, order Eurotiales: phylum Ascomycota. Many species of Trichocomaceae have been isolated during several mycofloristic surveys on soil in the middle and south of Iraq (Tolba et al, 1957, ALDoory et al, 1959, Ismail and Abdullah, 1977; El-Dohlob and AL-Helfi ,1982; Abdullah and Zora, 1993; Abdullah and Abbas, 2008; Abdullah et al .1986, 2007a). The aim of the present study was to identify and describe member of the genus Penicillium inhabiting soils in Kurdistan region of Iraq and to provide a key for the reported species.


MATERIALS AND METHODS Collection of samples: Samples of soil were collected from different sites in three governorates(Duhok, Erbil,and Sulumania) in Kurdistan region of Iraq during the year 2006-2007. Approximately 500g of soil was removed with a sterile trowel from depth of 2-8cm from site after first scraping away the upper 0-2cm of surface soil. Soil samples were stored in polyethylene bags in a refrigerator at 5C˚. The samples were processed within 1 to 2 weeks after collection. Isolation of fungi : Four isolation methods were applied, standared dilution plate (Johnson et al, 1959), direct plating (Warcup, 1950), treatment with 70% ethanol( Warcup and Baker, 1963) and soil treatment with 5% acetic acid (Furuya and Naito, 1979). Two types of growth media were used for isolation of fungi viz. Malt extract agar (MEA) and Czapecks dox agar (CZ). Each medium was supplemented with 50mg/ml chlorophenicol to suppress bacterial growth. Plates for all methods and media (three replicates each ) were incubated at 25C˚ for 7 days in the dark. Single colonies were picked from the plates under a dissecting microscope and transferred to appropriate media to allow fungus development . Identification of penicillium isolates :Pure colonies were grown on five media according to Samson et al (2000). The media are as follows: Czapeck Yeast Extract Agar incubated for seven days at 25 C˚ (CYA25), Czapeck Yeast Extract Agar incubated for seven days at 37 C˚ (CYA37), Czapeck Yeast Extract Agar with 20% sucrose incubated for seven days at 25 C˚ (CY20S), Malt Extract Agar incubated for seven days at 25 C˚ (MEA) and Czapeck Dox solution Agar incubated for seven days at 25 C˚ (CZ). Ingredients and preparation of the above five media were mentioned in Pitt and Hocking (1977) and Samson et al .(2000) . For each culture five plates were used, tow of CYA and one each of CY20S, MEA and CZ. Each plate is inoculated at the centre and incubated in the dark for seven days. One CYA was incubated at 37 C˚.The rest are incubated at 250C . All measurements are taken after seven days incubation. Species identifications were according to the keys and descriptions provided by Pitt and Hocking (1997), Samson et al (2000) and Frisvad and Samson (2004).

RESULTS AND DISCUSSION 1-Penicillium aurantiogresum Dierckx, Ann.Soc. Scient. Brux. 25: 88, 1901. (Plate1. af). Colony diameters at 7 days, in mm : CYA25: 22mm, CYA37 no growth, CY20S 30mm, CZ 19mm, MEA 24-34mm,G25N 19mm. Colony Colors and Textures :Colonies on CYA25 grow restrictedly producing grey to dark blue green conidia with a granular to fasciculate colony surface, often with exudates droplets, mycelium white, reverse is orange brown. On MEA, conidia are blue green, plane or rarely radially sulcate low and relatively sparse, surface texture finely granular, mycelium bright yellow brown to reddish brown; reverse pale orange. Colonies on G25N radially sulcate, deep, dense, reverse pale yellow. Microscopic Characteristics: Stipes 200-350 x 3-4µm long, smooth-walled to finely roughened, bearing terminal terverticillate or less commonly biverticillate penicilli, phialides, ampulliform, mostly 8-10×2-3µm, metulae 7-8 x 2.5-3µm. Conidia globose to subglobose, 2.5-4µm in diameter, smooth- walled. Specimen examined: Dried and living culture (W4) soil at Prunus domestica, Vitis vinifera, Zawita-Badi, 30/4/2007. The two- stage branched (terverticillate) conidiophore, rough- walled stipe and smooth globose to subglobose conidia are the main characteristics of the species. The production grey to dark blue green conidia and restrictly growing on CYA and CZ at 25C˚, it is also acharacteristic feature of the species. The species has been reported to cause spoilage of a variety of stored fruits and vegetables (Pitt and Hocking, 1997). This is the first record for the species from Iraqi soil. 2-Penicillium brevicompactum Dierckx, Ann.Soc. Scient. Brux. 25: 88, 1901. (Plate 2. af). Colony diameters at 7 days, in mm : CYA25: 20mm, CYA37 no growth, CY20S 30mm, CZ 20-39mm, MEA 20mm,G25N 14-20mm. Colony Colors and Textures :Colonies on CZapeck agar, dull green, mycelium white, reverse yellowish to reddish brown, exudates usually present in minute droplets, often deeply embedded, but somtimes copious, pale to deep reddishbrown; also soluble green to reddish brown, mycelium white, reverse brown, plane or less-commonly radially sulcate, usually velutinous. Colonies on G25N are yellow

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reverse, other characters are similar to those on CYA. Microscopic Characteristics: Stipes 300-400x 4-6µm wide, smooth-walled, stipes terminating in acompact penicillus, terverticillate to quaterverticillat. Branches and metulae usually inflated, metulae 9-11x4-7µm. phialides flask shaped, 7-10x3-3.5µm. Conidia 2.5-4µm long; smooth to very finely roughened, globose to subglobose . Specimen examined: Dried and living culture (W4) soil at Prunus domestica, Vitis vinifera, Zawita-Badi, 30/4/2007, (W2) Pinus brutia, Zawita-Dohuk, 30/11/2007. The species is characterized by its restricted growth and large compact penicilli in a shape suggesting Aspergillus. The species has been previously reported in Iraq from soil at date palm plantations (Abdullah and Zora, 1993), from grains of Zea mays (Abdullah and AlMousawi,2006), from Baiza white cheese (Abdullah et al. 2007b) and from sediments of Shatt Al-Arab river at Basrah (Abdullah and Abbas, 2008). 3-Penicillium camemberti Thom, Bull. Bur. Anim. Ind. USDA, 82: 33, 1906. (Plate3. a-f). Colony diameters at 7 days, in mm : CYA25: 39mm, CYA37 no growth, CY20S 42mm, CZ 30mm, MEA 33mm,G25N 22mm. Colony Colors and Textures :Colonies on CZapeck agar at 25C° are grey green, mycelium white to yellow, reverse reddish brown; plane or lightly radially sulcate, convex, floccose, exudates somtimes present. Colonies on MEA are similar to those on CYA but without exudates.Colonies on G25N are similar to those on MEA. Microscopic Characteristics: Conidiophores borne from aerial hyphae, stipes 75-200 x 4µm long,rough-walled, rarely smooth-walled; typically bearing terminal bi-ter or quatertverticillate penicilli, sometimes irregular; metulae 9-14 x 2.5-3µm, giving rise to 3 to 6 phialides. Phialides ampulliform with short necks, 10-13 x 3µm. Conidia 4-5 x 3-4.5µm, globose to subglobose, smooth walled to finely roughened, white to slightly greenish. Specimen examined: Dried and living culture (W19) soil at Triticum aestivum, Berdiya – Zumar, 22/4/2007, (W15) Papaver rhoeas,Pistacia atlantica, Prunus amygdalus, Punica granatum, Qesrok-Dohuk, 30/5/2007.

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The species is distinguished by its white to slightly greenish, large globose to subglobose broadly ellipsoidal (4- 5 x 3- 4.5 μm) and smooth walled conidia. The unique habitat for the species is soft cheese and its surrounding environment. It has been also occasionally found on other sources such as meat (Pitt and Hocking, 1997). This may represent the first record for the species from Iraqi soil. 4-Penicillium citrinum Thom, Bull. Bur. Anim. Ind. USDA, 118: 61, 1910. (Plate 4. a-f). Colony diameters at 7 days, in mm : CYA25: 25mm, CYA37 no growth, CY20S 25mm, CZ 16mm, MEA 20mm,G25N 13mm. Colony Colors and Textures :Colonies on CYA25 blue green, mycelium white to greyish oranage, reverse yellow to orange, exudates clear,pale yellow, soluble pigment bright yellow or absent. Colonies on MEA are plane or radially sulcate, dull green, mycelium white to greyish orange, reverse yellow yellow to orange..Colonies on G25N are dull green, mycelium white; reverse pale, dull brown. Microscopic Characteristics: Conidiophores borne from surface hyphae, stipes100-200 x 23µm, smooth-walled with 3-5 divergent metulae in a whorl, metulae 11-18 x 2.5-3µm, bearing 610 phialides. Phialides flask-shaped,8-9 x 22.5µm. Conidia 2-3µm, produced in columns, globose to subglobose, smooth walled to finely roughened. Specimen examined: Dried and living culture (W16) soil at Pinus brutia, Bablo-Dohuk, 30/5/2007, (W114) Prunus domestica, Vitis vinifera, Solaf (Silav)-Dohuk, 5/4/2007. The species is readily distinguished by its penicilli which consists of 3- 5 divergent metulae bearing long columns of conidia. The species is a cosmopolitan and has been found on a variety of substrates (Pitt and Hocking, 1997). The species has been previously reported from Iraq from white cheese (Abdullah et al. 2007b). 5-Penicillium corylophilum Dierckx, In Raper and Thom. A manal of the penicillia, 341, 1949. (Plate 5. a-f). Colony diameters at 7 days, in mm : CYA25: 20-25mm, CYA37 no growth, CY20S 20mm, CZ 17-20mm, MEA 30mm,G25N 12-13mm. Colony Colors and Textures :Colonies on CYA25 are plane to deeply radially sulcate, low, moderately dense blue green to grey green, mycelium white, reverse dark grey.Colonies on MEA are similar to those on CYA25 but less


dense and growing some whate faster.Colonies on G25N are similar to those on CYA25; mycelium white, reverse pale. Microscopic Characteristics: Conidiophores borne from subsurface hyphae, stipes100-200 x 3-4µm in long, smooth-walled, often of unequall lenght, but soon terminating in whorl of 2-4 metulae, metulae 10-15 x 3-4µm. Phialides flask shaped,6-8 x 2.5-3µm. Conidia 2.5-3µm, subglobose to ellipsoidal, smooth walled . Specimen examined: Dried and living culture (W14) soil at Quercus sp, Dohuk – Kuzo, 21/3/2007. The distinctive features of the species are the one- stage branched conidiophore with metulae variable in length and the dark green reverse on MEA. This may represent the first report for the species from Iraqi soil. 6- Penicillium digitatum (Pers: Fr) Sacc., Fung. Ital., 894, 1881. (Plate 6. a-f). Colony diameters at 7 days, in mm : CYA25: 35mm, CYA37 no growth, CY20S 30mm, CZ 30mm, MEA 55mm,G25N 6mm. Colony Colors and Textures :Colonies on CZapeck agar are greyish green to olive, mycelium white, reverse brownish. Colonies on MEA are yellow to brown-green, reverse brown. Colonies on G25N mycelium are white, reverse olive. Microscopic Characteristics: Conidiophores borne from surface hyphae, stipes80-150 x 34µm in long, with thin, smooth-walled, few metulae and branches terminating in whorls of 36 phialides. Phialides are often solitary,cylindrical with a short neck, variable in size, 18-25 x 3-4µm. Conidia ellipsoidal to cylindrical, smooth walls,6.5-8 x 4-5µm, olivegreen in mass. Specimen examined: Dried and living culture (W106) soil at Prunus domestica, Punica granatum, Prunus amygdalus, Amadiya-Dohuk, 5/4/2007. The species is commonly found on rotted fruits of citrus species. The species can be recognized by its large (3.5- 8 x 3- 4 μm), ellipsoidal to cylindrical smooth olive green conidia. The species has been previously reported from Iraq (Mathur, 1968; Abdullah and Abbas, 2008). 7- Penicillium echinulatum Fassatiova, Acta Univ. Carol. Biol. 12: 326, 1977. (Plate7. a-f).

Colony diameters at 7 days, in mm : CYA25: 39mm, CYA37 no growth, CY20S 25-35mm, CZ 13-30mm, MEA 33mm,G25N 20mm. Colony Colors and Textures :Colonies on CZ agar are grow radially, grey green to dark green, mycelium white, reverse uncolored yellow, exudates absent or as color less droplets. Colonies on MEA are similar to those on CYA.Colonies on G25N are deep but dense; reverse pale, yellow brown, other characters are similar to CYA. Microscopic Characteristics: Conidiophores mononematous but loosely synuematous in marginal areas, hyaline, stipes200-400 x 33.5µm, rough-walled, terverticillate. metulae 813 x 3-4µm, cylindrical, smooth-walled or nearly so, bearing 4 to 8 phialides. Phialides 710 x 2-2.6µm.Conidia globose to subglobose 3.5-4.5µm, , rough-walled to echinulate. Specimen examined: Dried and living culture (W38) soil at Vitis vinifera, Prunus Sulaimaniya - Dabasha 23/5/2007. The species is characterized by its terverticillate conidiophore and globose to subglobose, greenish, rough- walled to echinulate conidia. The species mostly isolated from cheese and other foods, rich in lipids (Pitt and Hocking, 1997). The isolation of the species during our study apparently represents a new record for the Iraqi soil fungi. 8- Penicillium funiculosum Thom, Bull. Bur. Anim. Ind. USDA, 118: 69, 1910. (Plate 8. a-f). Colony diameters at 7 days, in mm : CYA25: 68mm, CYA37 80mm, CY20S 65mm, CZ 3040mm, MEA 75mm,G25N 4-8mm. Colony Colors and Textures :Colonies on CZ agar are grey-yellow grey, mycelium yellow vegetative, reverse orange brown. Clear exudates and pink soluble pigment produced by some isolates. Colonies on MEA are similar to those on CYA, but with richer sporulation. Colonies on G25N reverse pale to olive, other characters as on CZapeck agar. Microscopic Characteristics: Conidiophores borne from aerial hyphae, stipes30-50 x 23.5µm, walls smooth to finely roughened, bearing terminal biverticillate penicillin; metulae 10-12 x 2-2.5µm, terminating in a whorl of 3-6 phialides. Phialides lanceolate, 9-11 x 3µm.Conidia 3-3.5 x 2-2.5µm, subglobose to ellipsoidal, smooth to finely roughened. Specimen examined: Dried and living culture (W5) soil at Punica granatum, Malus sylvestris, Dosky-Dohuk, 30/11/2006, (W30) soil at

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Platanus sp, Shalal Gali Ali Beg-Erbil, 25/5/2007. The fungus is characterized by its rapid growth at 37C˚ and the formation of yellow well defined ropes of hyphae intermixed with grey to yellow grey conidiophores, with lanceolate to finely roughened conidia (2.5- 3.5 x 2- 2.5 μm in size). The fungus is very common in different agricultural commodities and in soil (Domsch et al. 1980). 9-Penicillium glabrum (Wehmer) Westling= Penicillium frequentans Westling. Ark. Bot. ll(1): 131, 1911. (Plate 9. a-f). Colony diameters at 7 days, in mm : CYA25: 35-38mm, CYA37 no growth, CY20S 36-40mm, CZ 34mm, MEA 40-43mm,G25N 19mm. Colony Colors and Textures :Colonies on CZapeck are dull green to dark green, mycelium white, reverse yellow to yellow orange, exudates produced centrally by some isolates, clear; yellow, bright yellow soluble pigment some times produced.Colonies on MEA are similar to those on CYA. Colonies on G25N mycelium are white, velutinous. Other characters are similar to CZapeck agar. Microscopic Characteristics: Conidiophores monoverticillate consisting of an unbranched stipes, 67-175 x 4-5µm long, walls smooth to finely roughened, terminating in a whorl of 1012 phialides, 9-12 x 3-3.5µm, flask shaped.Conidia produced in typical long columns, globose to subglobose, smooth to finely roughened, 3-3.5 µm in diameter . Specimen examined: Dried and living culture (W16) soil at Pinus brutia, Bablo-Dohuk, 30/5/2007. The species can be distinguished by its monoverticillate conidiophores with unbranched stipe, terminating in a whorl of 10- 12 phialides. The species isolated from different agricultural commodities, from soil and from indoor environment. (Domsch et al. 1980, Pitt and Hocking, 1997). Abdullah et al. (2007.a,b)isolated the species from soil and mud from the tidal zone of khawr Al- Zubair, canal southern Iraq and from white cheese respectively. 10-Penicillium roqueforti Thom, Bull. Bur. Anim. Ind. US Dept.Agric. 82: 35, 1906. (Plate10. a-f). Colony diameters at 7 days, in mm : CYA25: 27mm, CYA37 no growth, CY20S 40mm, CZ 21mm, MEA 61mm,G25N 20mm.

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Colony Colors and Textures :Colonies on CZ agar and MEA grow rapidly, blue green, mycelium inconspicuous white, reverse greenish, often changing to darker shades of green to black, plane or lightly radially sulcate, low, strictly velutinous.Colonies on G25N are similar to those on CYA. Microscopic Characteristics: Conidiophores ter-to quaterverticillate, stipes, 100-200 x 4-5µm long, the stipes typically ornamented with conspicuous warts, some trains less roughened at the apex of conidiophores stipes, but usually ornamented at the base. metulae 10-15 x 4µm, rough-walled, giving rise to clusters of 4-7 phialides 8-10 x 2.5-3µm, flask-shaped with a short neck. Conidia 4-5.5µm, globose to subglobose, smooth walled, loose columns. Specimen examined: from samples one of cheese, Dohuk-center, 1/3/2007. The species can be distinguished by its ter- to quaterverticillate conidiophore with stipes typically ornamented with conspicuous warts. Conidia are spherical, smooth- walled, dark green mostly 4- 6μm, occasionally up to 8μm in diameter. The species is a widely distributed spoilage fungus and can grow rapidly at low temperatures causing spoilage to stored foods in refrigerators. It is also a common cause of cheese spoilage, although it is best known for its role in the manufacture of Roquefort and related cheese types (Pitt and Hocking, 1997). The species has been reported recently from Iraq on white cheese (Baiza) by Abdullah et al. (2007b) and from sediments of Shatt Al-Arab river at Basrah (Abdullah and Abbas, 2008). 11- Penicillium rugulosum Thom, Bull. Bur. Anim. Ind. USDA 118, 60, 1910. (Plate11. a-f). Colony diameters at 7 days, in mm : CYA25: 15-22mm, CYA37 18-19, CY20S 20mm, CZ 17mm, MEA 30mm,G25N 5mm. Colony Colors and Textures :Colonies on CZapeck agar and MEA are yellowish-green to dark green, mycelium inconspicuous, mainly white but yellow or red encrusted hyphae often visible under magnification; exudates and soluble pigment usually absent; reverse dull olive or brown. Colonies on G25N are similar to the colors on CYA. Microscopic Characteristics: Conidiophores borne from surface, stipes, 70-100 x 3-4µm, smooth walls, often biverticillate. In whorls of 5 to 7, metulae 9-10 x 2-2.5µm. Phialides cylindrical with a conspicuous neck or


sometimes lanceolate, 10-12 x 2-2.5µm.Conidia ellipsoidal, conspicuously roughened, 3-3.5µm. Specimen examined: Dried and living culture (W115) soil at Punica granatum, Prunus persica,Ficus sp, Arbil – Kore, 23/6/2007, (W6) Punica granatum, Malus silvestris, DoskyDohuk, 30/11/2006. The species is characterized by its very low growth and production of biverticillate conidiophores with metulae in whorls of 5 to 7, terminating in cylindrical or sometimes lanceolate phialide. Conidia ellipsoidal 3- 3.5 x 2.5- 3μm, conspicuously roughened. Because of its very slow growth, so it is easily overlooked in isolation procedure. It has been reported on a wide range of agricultural commodities (Pitt and Hocking, 1997). The species has been recently reported in Iraq from surface sediments of Shatt Al-Arab river at Basrah (Abdullah and Abbas, 2008). 12- Penicillium variabile Sopp. Skr. VidenskSelsk. Christ iiana, Math-NaturVidensk. KI. 11: 169, 1912. (Plate12. a-f). Colony diameters at 7 days, in mm : CYA25: 35mm, CYA37 70mm, CY20S 75mm, CZ 50mm, MEA 70mm,G25N 9mm. Colony Colors and Textures :Colonies on CZapeck agar are deep orange to brownish yellow, mycelium yellow usually conspicuous,

reverse yellow to orange –brown, clear exudates occasionally produced.Colonies on MEA are similar to CYA but on MEA growing some what faster and with less yellow mycelium. Colonies on G25N colors similar to MEA. Microscopic Characteristics: Conidiophores often arise from aerial hyphae, biverticillate, stipes 60-200 x 3-4µm, smooth-walled, terminal in a whorl of 5-7 metulae. metulae 7-10 x 2.53µm. Phialides lanceolate, 9-12 x 23µm.Conidia ellipsoidal to uniform, smooth to irregular roughened, often with striations, variable in size 3- 4(6) µm, but mostly 3-3.5 x 22.5µm . Specimen examined: Dried and living culture (W116) soil at Punica granatum, Prunus persica,Ficus sp, Arbil – Kore, 23/6/2007. The species is characterized by its biverticillate conidiophore with smooth- walled stipe terminal in a whorl of 5- 7 metulae and lanceolate phialides with ellipsoidal to fusiform smooth to irregular roughened, often with striations variabile in size conidia. Pitt (1973) reported the maximum temperature for the species is about 37C˚. Our isolate showed a good growth at 37C˚. The species is known from a wide range of agricultural commodities (Pitt and Hocking, 1997). This is apparently the first report of the species from Iraqi soil.

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Plate (1): Penicillium aurantiogriseum a-colony on MEA, b- colony on CY20S, c- colony onG25N, d- colony on CYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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plate (2): Penicillium brevicompactum a-colony on MEA, b- colony on CY20S, c- colony onG25N, d- colony on CYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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Plate(3) : Penicillium camemberti a-colony on MEA, b- colony on CY20S, c- colony onG25N, d- colony onCYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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Plate (4): Penicillium citrinum a-colony on MEA, b- colony on CY20S, c- colony onG25N, dcolonyonCYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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plate (5): Penicillium corylophilium a-colony on MEA, b- colony on CY20S, c- colony onG25N, d- colony on CYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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plate (6): Penicillium digitatum a-colony on MEA, b- colony on CY20S, c- colony onG25N, d- colony on CYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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plate (7): Penicillium echinulatum a-colony on MEA, b- colony on CY20S, c- colony onG25N, d- colony on CYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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plate (8): Penicillium funiculosum a-colony on MEA, b- colony on CY20S, c- colony onG25N, d- colony on CYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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plate (9): Penicillium glabrum a-colony on MEA, b- colony on CY20S, c- colony onG25N, d- colony on CYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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plate (10): Penicillium roquforti a-colony on MEA, b- colony on CY20S, c- colony onG25N, d- colony on CYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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plate (11): Penicillium rugulosum a-colony on MEA, b- colony onCY20S, c- colony onG25N, d- colony on CYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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plate (12): Penicillium variabile a-colony on MEA, b- colony onCY20S, c- colony onG25N, d- colony on CYA25, e-conidiophores, f-conidia scale bar e&f =10um.

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Key to the identified species of Penicillium 1- conidiophore with smooth stipe……….……2 1- conidiophore with rough stipe…………...….9 2- conidiophores monoverticillate (simple unbranched) ………P.glabrum 2- conidiophores biverticillate or more…..…….3 3-conidiophores predominantly biverticillate or irregularly monoverticillate and biverticillate.......4 3- conidiophores predominantly terverticillate or quaterverticillate…………………………...…..5 4- Colonies on G25N less than 10mm diameter……………….………………….…....6 4- Colonies on G25N more than 10mm diameter………………………………………..8 5- conidiophore irregularly branched consisting of short stipes with few metulae. Conidia ellipsoidal to cylindrical, smooth olive green 3.5- 8 x 3- 4 μm………..……………P.digitatum 5- conidiophore compact consisting of a wide stipe 4- 6 μm wide, metulae usually inflated, conidia globose to subglobose slightly roughened 34.5 μm in diameter…………………......P.brevicompactum 6- Colonies often produce well defined ropes of hyphae (funicles) bearing short conidiophores………………...…..P.funiculosum 6- Funicles not produced………..………..…...7 7- Metulae in whorls of 5 to 7. Conidia ellipsoidal, conspicuously roughened, 3- 3.5 x 2.5- 3 μm…………..................…....P.rugulosum 7- Metulae in whorls of 5-7.Conidia ellipsoidal to fusiform, smooth to irregular roughened, variable in size, but mostly 3- 4(6) x 2- 2.5 μm……………………………….......P.variabile 8- Divergent metulae in a whorl 2- 4, often of unequal length. Colonies on MEA more than 25mm diameter………………………………..P. corylophilum 8- Metulae of equal length, in a whorl of 3- 5. Colonies on MEA less than 25mm diameter………………….……….….P.citrinum 9Conidia in mass white on CYA……………………….....…..P.camemberti 9- Conidia on CYA with shade of green……..10 10-Conidia echinulate……………..………….P.echinulatum 10- Conidia smooth to finely rough…………..11 11-Colonies on MEA growing rapidly, more than 40mm in diameter…..……...…P.roqueforti 11-Colonies on MEA growing slowly, less than 40mm in diameter…….……P.aurantiogriseum

162

EFERENCES - Abdullah, S. K. and Zora, S. E. (1993). Soil microfungi from date palm plantations in Iraq. Basrah J. sci. B, 11: 45-58. - Abdullah, S. K. and Al- Mousawi, K. A. Y. (2006). Diversity of fungal species associated with maize (Zea mays L.) cultivars grown in Iraq. Proc. 12th. Cong. Mediter. Phytopathol. Union, 11- 15 June, Rhodes Island, Greece PP. 62- 72. - Abdullah, S.K and Abbas, B.A (2008). Fungi inhabiting sediments of Shatt Al-Arab river and its creeks at Basrah, Iraq. Basrah J. Sci. B, 26: 68-81. - Abdullah, S. K. and Al- Khesraji, T. O. & Al- Edany, T. Y. (1986). Soil mycoflora of the southern desert of Iraq Sydowia, 39: 8- 16. - Abdullah, S. K., Al- Saadon, A. H. and Al- Salihy, M. H. (2007a). Fungi from the tidal zone of Khawr Alzubair Canal Southern Iraq. Marsh Bulletin 2: 1831. - Abdullah, S. K., Muhammed, A. A. H., Mustafa, K. M. and Issa, N.K. (2007b). Fungal contamination of Baiza (JaJi) white cheese during consumption stage in Duhok. J. Duhok Univ. 10: 39- 44. - Al- Doory, Y., Tolba, M. K. and Al- Ani, H. ( 1959) On the fungal flora of Iraqi soils II. Central Iraq. Mycologia 51: 429- 439. - Brian, P. W.(1960) Griseofulvin. Trans. Brit. Mycol. Soc. 43: 1-13. - Domsch, K. H., Gams, W. and Anderson, T. H. (1980). Compendium of soil fungi. Academic Press, London. - El- Dohlob, S. M. & Al- Helfi, M. A. (1982). Soil fungi of the south Iraq. Bas. Nat. Hist. Mus. Bull., 5: 23- 37. - Fleming, A. (1944). The discovery of penicillium. Brit. Med.Bull. 2, 4- 5. - Frisvad, J. C. & Samson, R. A. (2004). Polyphasic taxonomy of Penicillium subgenus Penicillium. A guide to identification of food and air-borne terverticillate Penicillia and their mycotoxins. Studies in Mycology, 49, 1- 173. - Furuya, K. and Naito, A. (1979). An effective method for isolation of Boothiella tetraspora from soil. Trans. Mycol. Soc. Japan 20: 309- 311. - Ismail, A. L. S. & Abdullah, S. K. (1977). Studies on the soil fungi of Iraq. Proc. Indian Acad. Sci. 86: 151154. - Jakobsen, M., Cantor, M. D. & Jespersen, L. (2002). Production of bread, Cheese and meat. In The Mycota X: Industrial Applications, ed. H. D. Osiewacz. Berlin: Springer- Verlag, PP. 3- 22. - Johnson, L. E., Curl, E. A., Bond, J. H. and Fribourgh, H. A. (1959). Methods for studying soil microfloraplant disease relationships. Burgess Pub. Co. Minneapolis, Mn. USA. - Kwon- Chung, K. J. & Bennett, J. E. (1992). Medical Mycology. Malvern, USA: Lea and Febiger. - Mathur, R. S. (1968) The Fungi and plant diseases of Iraq. Ministry of Agriculture, Iraq 90 pp.


- Pitt,J.I.(1973). An apprasial of identification methods for Penicillium species: novel taxonomic criteria based on temperature and water relations. Mycologia 65:1135-1157. - Pitt, J. I. & Hocking, A. D.(1997). Fungal and Food Spoilage 2nd edition. London, U.K. : Blackie Academic & Professional. - Rolinson, G. N. (1998). Forty years of β-lactam research. Journal of Antimicrobial Chemotherapy, 41, 589134. - Samson, R.A., Hoekstra, E.S., Frisvad, J.C. and Filtenborg,O.(2000). Introduction to food and airborne fungi. 6th edition. Centraalburea Voor Schimmelculture. Baarn. The Netherlands.

- Tolba, M. K., Al- Doory, Y. and Al- Wahhab, M.A. (1957). On the fungal flora of Iraqi soils I. Baghdad area; Proc. Third Arab Sci. Congr. 198- 214. - Vanittanakom, N., Cooper, C. R., Fisher, M.C. & Sirisanthana, T. (2006). Penicillium marneffei infection and recent advances in the epidemiology and molecular biology aspects. Clin. Microb. Rev, 19, 95- 110. - Warcup, J.H.(1950) soil plate method for isolation of fungi from soil. Nature. London. 66: 117-118. - Warcup, J. H. and Baker, K. F. (1963). Occurrence of dormant acospores in soil Nature (London) 197: 1317- 1318. - Webster, J and Weber, R. W. S. (2007). Introduction to Fungi. 3rd edition. Cambridge University Press. U.K.

‫الخالصة‬ ‫ من‬Eurotiales ‫ التابعة لرتبة‬Trichocomaceae ‫ من الفصيلة‬Penicillium ‫اجريت دراسة تصنيفية ألنواع الجنس‬ ‫ نوعا اعتمادا على الصفلت المظهرية والزرعية‬21 ‫ تم تشخيص‬.‫الفطريات الكيسية والمستوطنة لتربة اقليم كردستان العراق‬ : ‫ االنواع المشخصة هي‬. P.aurantiogriseum, P.brevicompactum, P.camemberti, P.citrinum, P.corylophilum, P.digitatum, P.echinulatum, P.funiculosum, P.glabrum, P.roqueforti, P.rugulosum and P.variable

‫منها خمسة انواع تسجل ألول مرة في العراق شملت‬ P.aurantiogriseum, P.camemberti, P.corylophilum, P.echinulatum and P.variable.

‫تم وصف االنواع اعتمادا على الصفات الزرعية والمظهرية فضال عن وضع مفتاح لتسهيل تشخيص االنواع المسجلة‬ . ‫في الدراسة‬

‫ثوختة‬ Eurotiales

‫لديف ثلة‬

Trichocomaceae

‫لة بنةماال‬

penicillium

‫ظةكولينا ظارظارتنى لةسةر جورةكانى تومخى‬

‫ جور هاتنا دياركرن لديف‬21 . ‫لة كةرووةكانى توورةكة ئةوا ئاكنجى دناظ ئاخيَن هةرميا كوردستانا عرياقىدا‬ :‫وجورييَن هاتينة دةستنيشانكرن ئةفةبون‬. ‫سالوخةتيَن مورفولوجى وجاندن‬ P.aurantiogriseum, P.brevicompactum, P.camemberti, P.citrinum, P.corylophilum, P.digitatum, P.echinulatum, P.funiculosum, P.glabrum, P.roqueforti, P.rugulosum and P.variable

‫ثيَنج جور بو ئيَكةمني جار هاتنة توماركرن لناظ ئاخيَن عرياقى كو ئةظيَن خوارىبون‬ P.aurantiogriseum, P.camemberti, P.corylophilum, P.echinulatum and P.variable.

‫جورةكانى هاتيَنة دةستنيشانكرن لديف سالوخةتيَن مورفولوجى وجاندن لةطةل دانانى كليلك بو ئاسانى‬ .‫دةستنيَشانكرن بو جورةكانى توماركرى لة ظةكولينى‬

163

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