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					                               Summary

     A survey study was undertaken for the isolation and identification of
soil-borne fungi inhabiting different areas in Erbil, Iraq. A total of 15 soil
samples collected seasonally during August 2008-July 2009.
   Soil sampling situations were: Glkand, Agricultural Research Center in
Ankawa, field on Salahaddin summer resort Road, Agricultural College
Research in Grdarash, Mosul Road, field behind Technical Institute, Erbil
Citadel (Qalat), Sami-Abdulrhman (Erbil) park, garden in Rizgari Hospital,
garden in Erbil Teaching Hospital, Tayrawa (Mostofi) neighbourhood,
Mnara park, Salahaddin neighbourhood, Hawari hawri school, and College
of Science/Univeristy of Salahaddin.
   Fungi were isolated from soil during this study's period, by soil dilution
plate method on four types of selective media: Potato Dextrose Agar (PDA),
Malt Extract Tannic acid Agar (META), Dubo′s Cellulose Agar (DCA) and
Sabouraud Dextrose Agar (SDA).
   On PDA medium, eighty different genera of fungi were isolated during
the four seasons; (29, 27, 57, and 37) genera isolated in (summer, autumn,
winter and spring) respectively, the most frequently isolated fungi during
four seasons were Aspergillus spp. (539×103) colony forming units /g.soil,
followed   by    Penicillium   spp.    (215×103)cfu/g.soil,   Rhizopus   spp.
(115×103)cfu/g.soil, Emericella spp. (109×103)cfu/g.soil, Fusarium spp.
(47×103)cfu/g.soil, and Ulocladium botrytis (47×103)cfu/g. soil, while the
least frequently isolated fungal genera were Blakeslea tuningtam,
Clasterosporium cyperi, Idriella sp., Naranus cryptomeriae and Torula
alternata, (1×103)cfu/g.soil, for each one. On META medium, forty eight
different genera of fungi were isolated during the four seasons; (17, 17, 32
and 20) genera were isolated in (summer, autumn, winter and spring)



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respectively, the most frequent fungal species recorded during four seasons
were       Aspergillus      spp.      (695×103)cfu/g.soil,    Penicillium    spp.
(239×103)cfu/g.soil,      Emericella     spp.   (121×103)cfu/g.soil,   Ulocladium
botrytis (48×103)cfu/g.soil, and Yeast (36×103)cfu/g.soil. The least
frequently recorded genera were Harpographium sp., Helminthosporium sp.,
Monosporascus cannobolus, Peyronellaea sp., Pyrenochaeta terrestis,
Saksenae and Stigmella, (1×103) cfu/g.soil, for each one. On DCA medium,
sixty different genera of fungi were isolated during the four seasons, (26, 22,
42, and 40) genera isolated in (summer, autumn, winter and spring)
respectively, the highest incidence of the isolates found during the four
seasons were those of Aspergillus spp.(333×103)cfu/g.soil, followed by
Penicillium pp. (120×103)cfu/g.soil, Emericella spp. (81×103)cfu/g.soil,
Ulocladium botrytis (50×103)cfu/g.soil, Yeast (39×103)cfu/g.soil and
Stachybotrys spp. (37×103)cfu/g.soil.The least frequently recorded genera
were Ambelopsis vinacea, Ampulliferina persimplex, Annelophorella sp., and
Pythium catenulatum, (1×103)cfu/g. soil, for each one, and on SDA medium
with cyclohexamide, sixty two different genera were isolated during the four
seasons, (14, 20, 42 and 29) genera isolated in (summer, autumn, winter and
spring) respectively, the highest isolates found during the four seasons
belong to Trichophyton spp. (139×103)cfu/g.soil, followed by Aspergillus
spp. (124×103)cfu/g.soil and Yeast (68×103)cfu/g.soil, while least frequently
recorded     genera      were:     Cladosporium    sphaerosporium,     Corynascus
sepedonium, Cunninghamella elegans, Epicoccum andropogonis, Eroepsis
sp., Gonatobotrys simplex, Harpographium sp., Phialomyces macrosporus,
Staphylotrichum sp., Stilbum          penicilloides, Syncephalastrum racemosum
(1×103)cfu/g.soil, for each one.
    Fungi isolated from soil by hair baiting method, revealed that fifty three
species, belonging to twenty nine genera, among them: Ascotricha


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guaminsis, Aspergillus spp., Circenella musca, Cunninghamella spp.,
Emericella spp., Epidermophyton sp. and Fusarium spp.
    Fungi isolated from soil by washing method counted for ninety three
species belonging to fifty six genera, among them: Aspergillus spp.,
Circinella spp., Cunninghamella spp., Mucor spp., Mycellia sterilia,
Rhizopus spp.
   Aspergillus and Penicillium which isolated during the four seasons on
PDA, META, DCA and SDA media, were identified by using, Czapeks
yeast extract agar, Malt extract agar and Glycerol nitrate agar (G25%N)
media, incubated at different temperature were; 5,25 and 37 oC. Isolated
species of yeasts were identified by using API 20C AUX system.
    The study of antagonistic phenomenon show that all the four
antagonistic genera (Chaetomium gracile, Emericella nidulans, Gliocladium
virens and Trichoderma harzianum) were significantly inhibited the radial
growth of the soil borne plant pathogenic fungi tested, (Alternaria alternata,
Fusarium oxysporum, Rhizoctonia solani and Verticillium dahliae) with
different ratios.
    During the survey of soil borne fungi conducted by researches that
published in Kurdistan, the following genera were recorded for first time:
Ascotricha guamensis, Aspergillus clavati, A. penicilloides, Beltrania
rhombica, Blakeslea sp., Circinella muscae, Corynascus sepedonium,
Doratomyces stemonitis, Eurotium chevalieri, Fennellia nivea, Gibberella
sp., Gliocladium spp., Helicosporium sp., Isaria sp., Metarhizium
anisopliae, Myxotrichum chartarum, Penicillium steckii, P. vinaceum.,
Tritirachium sp. and Ulocladium botrytis.




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    List of Contents

Number Title                                            pages
           CHAPTER ONE
1          INTRODUCTION                                   1-3
           CHAPTER TWO
2          REVIEW OF LITERATURES                         4-21
2.1        Soil                                           4
2.1.1      Component of Soil                              4

2.2        Fungi                                          5
2.2.1      General characteristics of Fungi               5
2.2.2      Growth requirement of fungi                    8
2.2.3      Components of Fungal cell                      10
2.2.4      Fungal Pathogenesis                            12
2.2.5      Some common genera of Fungi                    13
2.2.5.1    Aspergillus spp.                               13
2.2.5.2    Penicillium spp.                               14
2.2.5.3    Mucor, Absidia and Rhizopus (Mucormycoses)     14
2.2.5.4    Fusarium spp.                                  15
2.2.5.5    Chaetomium spp.                                16
2.2.5.6    Stachybotrys spp.                              16
2.2.5.7    Ulocladium spp.                                17
2.2.6      Dermatophytes                                  17
2.2.6.1    Epidermophyton spp.                            18
2.2.6.2    Trichophyton spp.                              18

2.2.6.3    Microsporum spp.                               19

2.2.7      Candida spp.                                   19
2.3        Biocontrol Agents                              20




                                       IV
          CHAPTER THREE

3         MATERIALS AND METHODS                                  22-37
3.1       Media used for isolation and identification of fungi    22
3.1.1     Potato Dextrose Agar (PDA)                              22
3.1.2     Sabouraud Dextrose Agar (SDA)                           22
3.1.3.    Malt Extract Tannic acid Agar (META)                    23
3.1.4.    Dubo,s Cellulose Agar (DCA)                             23
3.1.5     Czapek (Dox) Agar                                       24
3.1.6     Yeast extract-Malt Extract Agar (YMA)                   24
3.1.7     API 20C AUX Kit (Biomerieux)                            25
3.1.8     Czapek concentrate                                      25
3.1.9     Czapek Yeast Extract Agar (CYE)                         25
3.1.10    Malt Extract Agar (MEA)                                 26
3.1.11    Glycerol Nitrate Agar (G25% N)                          26
3.1.12.   Stain (Lactophenol Cotton Blue)                         27
3.2       Soil Samples Collection                                 28
3.2.1     Physical analysis                                       30
3.2.1.1   Soil pH                                                 30
3.2.1.2   Electrical Conductivity (E.C.)                          30
3.2.1.3   Soil moisture                                           30
3.2.2     Chemical analysis (detection of soil minerals)          30
3.2.2.1   Organic Carbon (O.C.)                                   30
3.2.2.2   Total Nitrogen (T.N.)                                   30
3.2.2.3   Phosphorus (PO4)                                        31
3.2.2.4   Iron (Fe)                                               31
3.2.3     Isolation techniques                                    31
3.2.3.1   Standard dilution plate                                 31
3.2.3.2   Soil washing technique                                  31



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3.2.3.3   Hair baiting technique                                        32
3.3       Diagnosis                                                     33
3.3.1     Identification of the fungal genera                           33
3.3.1.1   Slide culture technique                                       33
3.3.2     Keys used for identification of the fungal genera             33

3.3.3     Identification of Aspergillus and Penicillium species         34

3.3.4     Identification of Dermatophytes & Non Dermatophytes fungi     34
3.3.5     Identification of yeast                                       35
3.3.5.1   Identification of Candida species                             35
3.3.5.2   API 20C AUX yeast system                                      35
3.3.6     Preservation of isolates                                      36
3.4       Antagonism                                                    37
          CHAPTER FOUR
4         RESULTS AND DISCUSSION                                      38-102
4.1       Fungi isolated from soil by dilution plate method             38
4.1.1     The identity of fungi on PDA medium                           38
4.1.2     The identity of fungi on META medium                          49
4.1.3     The identity of fungi on DCA medium                           52
4.1.4     The identity of fungi on SDA medium                           58
4.2       Fungi isolated from soil by hair baiting method               65
4.3       Fungi isolated from soil by washing method                    72
4.4       The identity of species of Aspergillus and Penicillium        78
4.5       The identity of yeast                                         89
4.6       Antagonisms                                                   97
          CONCLUSIONS AND RECOMMENDATIONS                             103-104
          REFERENCES                                                  105-133




                                      VI
       List of Tables


Tables Titles                                                                        P

4.1      Identification of soil Fungi isolated from fifteen location by dilution     46
         plate method on PDA medium

4.2      Identification of soil Fungi isolated from fifteen location by dilution     51
         plate method on META medium

4.3      Identification of soil Fungi isolated from fifteen location by dilution     56
         plate method on DCA medium

4.4      Identification of soil Fungi isolated from fifteen location by dilution     63
         plate method on SDA medium with cyclohexamide

4.5      Identification of soil Fungi isolated from fifteen location by using hair   70
         baiting technique

4.6      Identification of soil Fungi isolated from fifteen location by using        75
         washing method

4.7      Identification the species of Aspergillus spp.                              83


4.8      Identification the species of Penicillium spp.                              86

4.9      Identification the type of yeast by using API20C AUX System                 91


4.10     Antagonistic reaction of some genera of fungi with plant pathogenic         101
         genera of fungi




                                            VII
List of Figures

Figures Titles                                                                  P
                                                                               29
3.1     Erbil map and soil sampling sites
4.1     Some genera of fungi isolated on PDA medium from soil                   48
           A- Absidia corymbifera               B- Stemphylium sp.
           C- Circinella muscae                 D-Cunninghamella elegans
           E- Rhizoctonia solani                F- Corynascus sepedonium
4.2     Some genera of fungi isolated on DCA medium from soil                   57
           A- Chaetomium gracile                B- Stachybotrys elegans
           C- Alternaria alternata              D- Chaetomium succineum
           E- Gibberella acuminata               F- Tritirachium sp.
4.3     Some pathogenic fungi isolated on SDA medium with cyclohexamide         64
        from soil
           A- Histoplasma capsulatum            B- Trichophyton tonsurans
           C- Microsporum gypseum               D- Racquet hyphae
           E- Trichophyton rubrum               F- Microsporum canis
4.4     Some genera of fungi isolated from soil by using hair baiting technique 71
           A- Myxotrichum chartarum             B- Metarhizhium anisopliae
           C- Beltrania rhombica                D- Ascotricha guamensis
           E- Syncephalastrum racemosum          F- Emericellopsis minima
4.5     Some fungi isolated from soil by using washing method technique         77
           A- Doratomyces steminitis             B- Ulocladium botrytis
           C- Papulaspora irregularis            D- Helicosporium sp.
           E- Rhizopus stolonifer                F- Rhizomucor sp.
4.6     Aspergillus spp. identified according to (Pitt and Hocking, 1997).      85
        CYA, MEA, G25%N. A- A. terreus B- A. ustus
4.7     Penicillium spp. identified according to (Pitt and Hocking, 1997).      87
        CYA, MEA, G25%N.
        A- Penicillium funiculosum              B- Penicillium sclerotiginum
4.8     Species of Aspergillus and Penicillium                                  88
        A- Aspergillus clavati        B- A. ochraceus           C- A. fumigatus
        D- Penicillium expansum        E- P. variable           F- P. citrinum
4.9     Yeasts identified using API 20C AUX (Biomeriux) system                  92
        A- Rhodotorula glutinis B- Cryptococcus albidus C- Candida ciferrii
4.10    Antagonistic phenomenon against soil borne plant pathogenic fungi       102
        A- Trichoderma harzianum with Fusarium oxysporum.
        B- Trichoderma harzianum with Alternaria alternata.




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