7 61 G. A. Shaker Bull. Iraq nat. Hist. Mus. (2010) 11 (2): 61-68 IDENTIFICATION OF PATHOGENIC FUNGI ASSOCIATED WITH WATER HYACINTH IN SELECTED REGIONS IN THE MIDDLE AND SOUTH OF IRAQ Goner A. Shaker Iraq NaturalHistory Museum,University of Baghdad, Baghdad, Iraq ABSTRACT To identify the fungi associated with water hyacinth (Eichhornia crassipes [Mart.] Solms), an aquatic weed, which presents in Tigris river from Baghdad south ward. Five regions from middle and south of Iraq (Al-Noumanya, Saeid Bin-Jubier, Al-Azizia, Al-Reyfay and Al-Hay) were selected for this study. Twelve fungal species were isolated. Alternaria alternata, Acremonium sp and Cladsporium herbarum, were the most frequently species (91.66 % ,50 % and 25 %) respectively. The fungi Alternaria alternata, Acremonium sp and Phoma eupyrena were more aggressive to water hyacinth as (91.66%,83,33%, and 75%) in pathogenicity test. INTRODUCTION Water hyacinth is one of tropical and semitropical plants, its first apperance in Iraq in the mid of eighties of the 20th century, and entered as ornamental plant in the some nurseries in Baghdad province, and had been taken from nurseries on the riverbank of (Kanat Al-Gaish) east of Baghdad, which drain in Diala river near of its firth in Tigris river south of Baghdad and then gradually transferred to Diala river and at last to the colum of Tigris river. The plant is known as aquatic weed belonged to the monocotyledonous family Pontederiaceae (Alison, 2000). The Amazon Basin, indicated by many authors as the center of origin of this plant (Barreto,1991).It infested approximately 62,000 ha of water resources in Mexico, the chemical and mechanical control have been used in Mexico since 1958 to manage water hyacinth (Martnies et al. 1998). Several fungal pathogens have been reported to attack water hyacinth in various parts of the world (Alison,2000 Butt et al, 2001 El-Morsy, 2004 Martnies and Charuattan,1998 Naseema et al,2008 and Praveena&Naseema, 2004). The aim of this study is to isolate and identify the fungi associated with water hyacinth (Eichhornia crassipes [Mart.] Solms) in the middle and south of Iraq and evaluate their pathogenicity. MATERIALS AND METHODS Isolation: The isolation of fungal species from infected parts of the plant such as leaves, swollen leaf bases were collected from (Al-Noumanya, Saeid Bin-Jubier, Al-Azizia, Al- Reyfay and Al-Hay) in middle and south of Iraq, the plant samples stored in plastic tanks in the laboratory and then infected samples were subjected for isolation technique ,Briefly; small pieces (2-3)mm2 surface sterilized in 1.5% sodium hypochlorite solution for (1-2) minutes, rinsed with sterile water and then cultured on Potato Dextrose Agar (4 pieces/plate) and incubated at 27°C.Fungal growth checked for purity, the fungi were subculture to serve as inoculum source. 62 Identification of Pathogenic Fungi Associated Pathogenicity test: Based on Koch postulate healthy leaves of water hyacinth plants, cleaned spotless of similar size leaves were collected, washed with tap water and placed on surface of wet-cotton in perti-dish, a (5 mm diam.)inoculum's plugs of each fungi culture placed reversely on the bottom of the leaf, each fungus isolate was replicated three times and the control treatment used (PDA) plug only, and then left in room temperature until the symptoms were appeared, the data had been recorded, and the treated specimens were ranked on the basis of the disease severity and assessed as follows: 0= leaves healthy ,1= a few spots or slight necrosis as (1-25%) from leaf size, 2= the spotting take over (26-50%), 3= the spotting take over (51-75%), 4= blighting take over (76-100%) (El-Morsy, 2004). Disease index (DI) = . × ….. . × . × . × 100 (Praveena and Naseema, 2004). RESULTS AND DISCUSSION Identification: Twelve fungi were isolated and indentified depending on the keys for each fungus, and depending on the morphology on conidia and conidiophores (Fig. 2) which formed on the pure fungal growth in Petri-dishes (Barnett et al., 1972, Booth, 1971, and Ellis,1971). The most common known species were Alternaria alternata, Acremonium sp, Cladosporium herbarum, and Fusarium oxysporum.Of these fungi, Alternaria alternata (91.66% of colonies) in Al- Noumanya, (66.66%) in Al-aziza as represented in (Table 1.). This is the First survey for fungi associated with water hyacinth in Iraq. Table 1.The fungi species isolated from leave blades and swollen leaf bases of water hyacinth from selected regions of middle and south of Iraq. 5 4 3 2 1 Localities L S L S L S L S L S (x) Fungal pathogen 41.66 - 16.66 50 16.66 100 33.33 - 50.00 - Acremonium sp 66.66 100 33.33 50 50 - 16.66 100 91.66100 Alternaria alternata - - - - 16.66 - - - 25 - Cladosprium herbarum - - 0.33 - - - - - 25- Chaetomium sp - - - - 0.33 - - - - - - Phoma eupyrena 16.66 - - - 0.33 - - - - - Thielaviopsis sp - - - - - - - - 0.33 - Geotrichum sp - - - - - - - - 0.33 - Nigospora sp - - 16.66 - - - - - - - Rhizoctonia solani - - - - 16.66 - - - 0.33 - Stemphyllium botryosum - - 41.66 - - - - - 0.33 25 Fusarium oxysporum - - - - 33.33 - - - - - Rhizopus sp 1=Al-Noamanya, 2=Saeid Bin Jubeier, 3=Al-Hay, 4-Al-Reyphay5-Al-Azizia. L =Leaf Blades, S=Swollen leaf blades. 63 G. A. Shaker Pathogenicity test: The fungi Alternaria alternata, Acremonium sp and Phoma eupyrena, infected the inoculated leaves and showed the symptoms (Fig.3) and they are efficient agent as result of (DI), Alternaria alternata (91.66%), Phoma eupyrena (83.33%) and Acremonium sp (75%), in comparing with the rest Fungi (Table.2), and the yellowing and blighting of the leaves are more than others. According to (Butt et al., 1971), fungi have damaged effect to water hyacinth and are considered to be as potential bioherbicide agent, and more extensive and wide researches needed to determine the best and more effective fungal pathogens economically and ecologically. (x) = isolation frequency calculated according the following formula: The no. pieces colonized by the fungus X 100Total no. cultured pieces Table 2.Disease index of fungi on the leave blades invitro condition. (DI) = . × ….. . × . × . × 100 ACKNOWLEDGEMENT I appreciate and thank Dr. Mohammad Kadhum Mohammad from the museum for his efforts and helping and to provide the samples to carry out the research. LITERATURE CITED Alison, W. 2000. An Evaluation of fungal Isolates for the Biological Control of water hyacinth, EichhorniaCrassipes.UF Journal of Undergraduate Research,1(8):, Issue 8- may 2000. Barnett, H.L. and Hunter, B. B., 1972.Illustrated genera of imperfect fungi. Burgess Pub. Co., Minneapolis, Minnesota, USA. Booth, C. 1971. The genus Fusarium –Commonwealth mycological Institute, Kew, Surrey, England.pp-237. Butt, T.M., Jackson, C., and Magon, N. 2001. Introduction – Fungal Biological Control Agents: Problems and Potential. CAB International. Ellis, M. B., 1971. Dematiaceoushyphomycetes.Commonwealth mycological institute, Kew, survey, England.pp- 608. El-morsy, E. M. 2004.Evaluation of micro fungi for the biological control of water hyacinth in Egypt.Fungal Diversity, Damietta Province, Egypt. Fungal Diversity 16:35-51. Species (DI) % Alternaria alternata 91.66 Phoma eupyrena 83.33 Acremonium sp 75 Chaetomium sp 50 Stemphyllium botryosum 50 64 Identification of Pathogenic Fungi Associated Martines, M.Jimenez and Charuattan, R. 1998. Survey and Evaluation of Mexican Native fungi for Potential Biocontrol of water hyacinth – Journal Aquat. Plant manage. 36: 145 – 148. Nassema, A., Dhanya, B., Anjanadevi, I.P., Sheena, K.G., and Swapna Alex. 2008. Isolation and regeneration of protoplasts from the mycelium of FusariumPallidoroseum – a potential biocontrol agent of water hyacinth [Eichhorniacrassipes (Mart.) Solms]. Journal of Tropical Agriculture 46(1 – 2): 55 – 57 Praveena, R. and Naseema, A. 2004.Fungi occurring on water hyacinth [EichhorniaCrassipes(Mart.) Solms] in Kerala.Jornal of Tropical Agriculture 42(1-2): 21-23. 65 G. A. Shaker 66 Identification of Pathogenic Fungi Associated 67 G. A. Shaker 68 Identification of Pathogenic Fungi Associated Bull. Iraq nat. Hist. Mus. (2010) 11 (2): 61-68 المناطق من وسط بعض شب النیل فيالتي تصیب عتشخیص الفطریات وجنوب العراق كونر عبدالوھاب شاكر جامعة بغداد -مركز بحوث و متحف التاریخ الطبیعي الخالصة Waterعشب النیل اجري ھذا البحث لتشخیص الفطریات التي تصیب نبات hyacinth (Eichhornia crassipes [Mart.] Solms) ، ھو دغل مائي یتواجد في اختیرت لھذه الدراسة خمسة مواقع من وسط . نھر دجلة باتجاه جنوب بغداد تم ). العزیزیة، الرفاعي، الحي، سعید بن جبیر، النعمانیة(وجنوب العراق Alternaria alternata, Acremonium andوكانت الفطریات ، فطر) ١٢(عزل Cladsporium flvum, 25 و% 50 و % 91.66( ھي االكثر تكرارا % ( . على التوالي Acremonium sp., Phoma eupyrena Alternaria ,اظھرت الفطریات alternata, امراضیة اشد في نبات عشب النیل وبالنسب .على التوالي%) ٧٥، ٨٣.٣٣، ٩١.٦٦(