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Effects of the Nematophagous Fungi Arthrobotrys Oligospora Fresen and Trichoderma Harzianum Rifai on Nematodes Infecting Banana, Tomato and Lime Plants

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Effects of the Nematophagous Fungi Arthrobotrys Oligospora Fresen and Trichoderma Harzianum Rifai on Nematodes Infecting Banana, Tomato and Lime Plants Effects of the Nematophagous Fungi Arthrobotrys oligospora Fresen and Trichoderma harzianum Rifai on Nematodes Infecting Banana, Tomato and Lime Plants Suad Abdel Gamiel Mohamed Ahmed B.Sc. in Crop Protection Faculty of Agricultural Sciences University of Cairo, Egypt (1981) M.Sc. in Crop Protection (Plant Pathology) Faculty of Agricultural Sciences University of Gezira, (1997) A Thesis Submitted to the University of Gezira in Fulfilment of the Requirements for the Award of the Degree of Doctor of Philosophy in Crop Protection (Plant Pathology) Plant Protection Department Faculty of Agricultural Sciences University of Gezira September, 2013 Effects of the Nematophagous Fungi Arthrobotrys oligospora (Fresen) and Trichoderma harzianum (Rifai) on Nematodes Infecting Banana, Tomato and Lime Plants By Suad Abdel Gamiel Mohamed Ahmed Supervision committee: Name Position Signature Prof. El Nour El Amin Abdel Rahman Main Supervisor ………… Prof. Ahmed El Bashir Mohamed El Hassan Co-Supervisor ………… Date: September, 2013 Effects of the Nematophagous Fungi Arthrobotrys oligospora (Fresen) and Trichoderma harzianum (Rifai) on Nematodes Infecting Banana, Tomato and Lime Plants By Suad Abdel Gamiel Mohamed Ahmed Examination Committee: Name Position signature Prof. El Nour El Amin Abdel Rahman Chairman ……………… Prof. Abel Mageed Yassin Abel Mageed External Examinar……………… Dr. Mohamed Hamza Zain Alabdeen Internal Examiner……………… Date of Examination: 29, September, 2013 DEDICATION To my beloved and kind family, my late dear father, my late dear mother, Brothers, sisters, my late dear sister Sharifa and late dear brothers Mohamed and Ahmed. To my kind husband El Badri Yassin ACKNOWLEDGEMENT First of all my thanks to the Almighty ALLAh who gave me strength ,power and patience to complete this study and bring it to its end. My sincere gratitude , appreciation and a lot of thanks to my supervisor Prof. El Nour El Amin for his keen guidance, interest . kind close supervision and encouragement. I am also grateful to my co-supervisor Dr. Ahmed Elbashier Mohamed Elhassan for hidiscussions during the progress of the study. Special gratitude and appreciation are due to the staff of plant pathology section and plant pathology lab. for they had always assisted me during my lab. work. Special thanks also due to pesticides residue lab. , specially Mr. Saeed Hamid for the very valuable help and assistance. Deep sincere gratitude to staff of Nematology lab. specially Mr. Sir El Khatim Momhamed, Iman and Samia Yousif for their valuable assistance during preparing, washing and counting nematodes under study. I am deeply indebted to Pof. Dafalla Ahmed Dawoud for his good ideas and advises. I am extremely grateful to my family for their continual encouragement and strong support. My sincere thanks and deep appreciation are extended to my faithful friends and colleagues for their valuable encouragement and assistance offered during the course of this study. My greatest appreciations to any unnamed person , who contributed in one way or another at time of difficulties. I am indebted to the Agricultural Research Corporation for the unlimited support and for giving me the chance to do this work. My appreciation and gratitude are extended to the government of Sudan for funding and financially supporting this study. Last but not least I wish to express my thanks and appreciative acknowledgement to my husband El Badri for his sacrifice and moral support. Effects of the Nematophagous Fungi Arthrobotrys oligospora Fresen and Trichoderma harzianum Rifai on Nematodes infecting Banana, Tomato and Lime Plants Doctor of Philosophy in Plant Protection(Plant Pathology) (September, 2013) Department of Crop Protection Faculty of Agriculture Sciences, University of Gezira ABSTRACT Nematophagous fungi are the fungi which attack nematodes, many of them are plant pathogens. These fungi worldwide in distribution and have been reported from many countries This study has been carried out to search for nematophagous fungi in Gezira including Sudan. soil and their capability to attack nematodes. Random samples collected from Gezira soil grown with tomato, banana and lemon crops were put on Corn Meal Ajar media (CMA) for the growth of the fungi and nematodes. Using Digital Microscope many types of trapping nematodes had been seen, Such as those with adhesive nets, adhesive knob and presence of fungal spores The samples labeled, and kept in laboratory for further inside the dead body of the nematode. The fungi has been identified as Arthrobotrys oligospora and .study in the glass house Trichoderma harzianum. The study investigated the capability of those fungi to attack and destroy nematodes. The nematophagous fungi A. oligospora was found to attack the nematode similis It was also noticed that, the nematode had been captured by adhesive knops Radophilus and after that the nematode struggled until death. The fungus A. oligospora penetrates the nematode cuticle and consumed all the body content of the nematode. Also the same fungus had been seen capturing the nematode Xiphenema sp. by adhesive net and had been held at two points and sometimes at several points. The fungus T. harzianum has been tested for its potential as a biological control agent and was found to suppress nematode population densities when it was added in different concentrations (105, 104 and103 spores/ml) to 50 Meloidojyne javanica nematodes in soil planted with tomato. The fungus reduced the numbers of nematode galls on tomato roots compared with the untreated control and resulted in excellent plant vegetative growth. Growing of lemon plants on soil artificially infested with Xiphenema sp. nematodes and treated with different concentrations of the fungus A. oligospora inocula significantly increased the number of lemon plant leaves, stem length, root length and also increased the fresh and dry root weight compared to the same soil without fungal inoculation. Also the different concentrations of the same fungus resulted in excellent vegetative growth of banana plants when grown in soil artificially infested with the nematode R. similis. Effect of different rates of the fungus T. harzianum metabolites on the nematode Xiphenema sp. had been studied. The different rates of the fungal metabolites (10-1, 10-2 and 10-3) after 24 hours gave comparable percentage of mortality, 16.8%, 14.19% and 11.65% respectively, but significantly better than the untreated control that resulted in 3.78% mortality. After 72 hours, the highest concentrations of the fungal metabolites resulted in significantly high nematodes mortality (66.24%) compared with the lowest one (42 93%) and the untreated control (8.87%). The study recommended that, Nematophagous fungi, if given more attention may be useful as biological control which can decrease cost of nematicides and conserve the ecosystem. على النيماتودا التى تصيب .harzianum Rifai T وA. oligospora Fresenتأثير الفطريات القاتله للنيماتودا نباتات الموز، الطماطم و الليمون دكتوراة الفلسفة في وقاية المحاصيل )امراض النبات( قسم وقاية المحاصيل، كلية العلوم الزراعية، جامعة الجزيرة ملخص الدراسة الفطريات القاتله للنيماتودا هي تلك التي تهاجم النيماتودا والتي كثيرا منها مسببا ﻻمراض النبات. وهذه الفطريات كثيرة اﻻنتشار وسجلت قي كثير من البلدان بما فيها السودان. أجريت هذه الدراسه للبحث عن بعض الفطريات التي تقوم بمهاجمة انواع مختلفة من النيماتودا. جمعت عينات عشوائية من مناطق مختلفة بالجزيرة زرعت بمحاصيل الطماطم,الموز والليمون. وضعت العينات في بيئة مناسبة لنمو الفطر والنيماتودا معا. باستخدام المجهر الرقمي ,شوهدت عديد من طرق هجوم الفطر علي النيماتودا, منها صنع الفطر لشبكه من الميسيليوم واصطياد النيماتودا بها, أيضا هناك بعض الفطريات تقوم بعمل حلقات منقبضة و غير منقبضة وتعمل علي خنق النيماتودا بها حتي تموت. فطريات أخري تخترق جدار النيماتودا وتقوم باستهﻻك اﻷحشاء الداخلية للنيماتودا وتكوين جراثيمها داخل جسم النيماتودا. تم التعرف علي هذه الفطريات بعد عزلها بالمعمل ,كما تم التعرف علي النيماتودا أيضا وتم حفظ العينات بالمعمل وذلك ﻹجراء الدراسة الﻻزمة بالمشتل. الفطريات التي تم التعرف عليها هي , Arthrobotrys oligosporaو Trichoderma harzianum. خﻻل هذه الدراسة تمت معرفة مدي مقدرة تلك الفطريات علي مكافحة الديدان ألثعبانيه. لغرض هذه الدراسة تم جمع عينات التربة من مناطق مختلفة هي مزرعة هيئة البحوث الزراعيه, منطقة باشكار )قسم المسلميه(, وادي شعير, أم سنط و حنتوب. اوضحت هذه الدراسه النشاط العالي للفطرA. oligospora علي مهاجمة بعض الديدان الثعبانيه خاصة Radophillus similis المعروفه بمقدرتها علي مهاجمة نبات الموز وتؤدي الي نقصان اﻻ نتاجيه. اوضحت الدراسه ان الديدان الثعبانيه تقبض بالتصاقها بالحلقات المنقبضه وتستمر الديدان الثعبانيه بالمقاومه حتي تموت. الفطر A. oligospora يخترق جدار الديدان الثعبانيه ويستهلك كل محتوياتها. كذلك يصطاد هذا الفطر الديدان الثعبانيه من نوع .Xiphenema sp ويلتصق بها بواسطة شبكه في اكثر من نقطه. وهذا النوع من الديدان الثعبانيه معروف بمهاجمته ﻻشجار الليمون. الفطر .T harzianum تم اختياره في مجال المكافحه الحيويه كعامل مؤثر علي تقليل كثافة الديدان الثعبانيه Meloidojyne javanica عندما اضيف بتركيزات مختلفه and 103) spores/ml 104 ,105( مع عدد 50 من هذه الديدان في تربه زرعت بنباتات طماطم. اوضحت النتائج مقدرة هذا الفطر علي تقليل اﻻنتفاخات في جذور الطماطم مقارنة بالشاهد كما اوضحت النتائج مقدرة هذا الفطر في تحسين النمو الخضري للنبات مما يوضح مقدرته فى مكافحة الديدان الثعبانيه . أدت اضافة الفطر A. oligospora بتركيزات مختلقه ) (and 10-3 2-10 ,1-10) مع عدد 50 من الديدان الثعبانيه Xiphenema sp الي التربه الي زيادة أوراق نبات الليمون ,طول السيقان, طول الجذور و الوزن الجاف والرطب للجذور لموسمين مقارنة مع الشاهد. كذلك أدي نفس الفطر عندما أضيف بتركيزات مختلفة and 10- 10-2 ,10-1) (3 مع عدد 50 من الديدان الثعبانيه R. similis. الي تربه زرعت بنبات الموز لتحسين النمو الخضرى للنبات من زياده لعدد اﻻوراق,طول السيقان وزيادة الوزن الجاف والرطب للجذور.أدي وضع تركيزات مختلفه من metabolites الفطر T. harzianum (3-10 2-10 ,1-10) مع أعداد معينه من الديدان الثعبانيه .Xiphenema sp في أطباق بتري الى نسبه عاليه من قتل الديدان الثعبانيه تراوحت بين 11.65%- 16.8% بعد 24 ساعه مقارنة مع 3.78% للشاهد وتم الحصول علي نفس النتائج بعد 48 و 72 ساعه. ادي اعلي تركيز إلي نسبة موت بلغت 66.24% بينما أدي التركيز اﻷقل والشاهد الي 42.93% و 8.87% علي التوالي. توصي الدراسه بانه اذا ما وجدت الفطريات القاتله للنيماتودا مزيدا من اﻻهتمام يمكن ان تفيد في المكافحه الحيويه وتقلل من تكلفة المبيدات النيماتوديه وتحمي المنظومه البيئيه.
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