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Fundaatila2015.Pdf (11.97Mb) EGE ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ (DOKTORA TEZİ) FARKLI Hericium İZOLATLARININ EN UYGUN YETİŞTİRME KOŞULLARI VE ORTAMLARININ BELİRLENMESİ VE MOLEKÜLER KARAKTERİZASYONU Funda ATİLA Tez Danışmanı : Prof. Dr. Yüksel TÜZEL Bahçe Bitkileri Anabilim Dalı Bilim Dalı Kodu : 501.01.00 Sunuş Tarihi : 02.06.2015 Bornova-İZMİR 2015 EGE ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ ETİK KURALLARA UYGUNLUK BEYANI E.Ü. Lisansüstü Eğitim ve Öğretim Yönetmeliğinin ilgili hükümleri uyarınca Yüksek Lisans Tezi / Doktora Tezi olarak sunduğum “Farklı Hericium izolatları için en uygun yetiştirme koşulları ve ortamlarının belirlenmesi üzerine araştırmalar” başlıklı bu tezin kendi çalışmam olduğunu, sunduğum tüm sonuç, doküman, bilgi ve belgeleri bizzat ve bu tez çalışması kapsamında elde ettiğimi, bu tez çalışmasıyla elde edilmeyen bütün bilgi ve yorumlara atıf yaptığımı ve bunları kaynaklar listesinde usulüne uygun olarak verdiğimi, tez çalışması ve yazımı sırasında patent ve telif haklarını ihlal edici bir davranışımın olmadığını, bu tezin herhangi bir bölümünü bu üniversite veya diğer bir üniversitede başka bir tez çalışması içinde sunmadığımı, bu tezin planlanmasından yazımına kadar bütün safhalarda bilimsel etik kurallarına uygun olarak davrandığımı ve aksinin ortaya çıkması durumunda her türlü yasal sonucu kabul edeceğimi beyan ederim. 02 / 06 / 2015 İmzası Funda ATİLA vii ÖZET FARKLI Hericium İZOLATLARININ EN UYGUN YETİŞTİRME KOŞULLAR VE ORTAMLARININ BELİRLENMESİ VE MOLEKÜLER KARAKTERİZASYONU ATİLA, Funda Doktora Tezi, Bahçe Bitkileri Anabilim Dalı Tez Danışmanı: Prof. Dr. Yüksel TÜZEL Haziran 2015, 253 sayfa Bu çalışmada, 8 Hericium izolatının (HE-Ankara, HE-Denizli, HE, HE- Trabzon, HE-İzmit, HA, HC, HE-Amerika) misel ve şapka gelişimi açısından, ortam, sıcaklık, pH gibi faktörlerin etkilerinin belirlenmesi bölgemizde en kolay ve ucuz olarak bulunabilecek lignoselülozik açıdan zengin farklı tarım ve odun artıkları ile hazırlanan ortamların Hericium türlerinin üretiminde kullanım olanaklarının araştırılması ve bu ortamların verim ve mantar kalitesine etkisinin belirlenmesi, Hericium üretimi yapılan yetiştirme ortamında, üretim dönemi süresince meydana gelen kimyasal değişimlerin gözlenmesi ve 8 Hericium izolatı arasındaki genetik ilişkilerin tespit edilmesi amaçlanmıştır. Hericium izolatlarının misel gelişimleri 7 farklı sıcaklık (5, 10, 15, 20, 25, 30 ve 35oC) ve 5 farklı pH (5, 6, 7, 8 ve 9) gözlenmiştir. Şapka oluşumu ile ilgili denemelerde, sıcaklık denemeleri 15, 20 ve 25oC olmak üzere 3 farklı sıcaklıkta, ortam denemeleri ise, meşe talaşı (MT) ile buğday kepeği (BK), pamuk küspesi (PK) ve zeytin pirinasının (ZP) %10, 20 ve 30 oranlarında karıştırılması ile hazırlan 7 farklı yetiştirme ortamında (80MT:20BK, 90MT:10PK, 80MT:20PK, 70MT:30PK, 90MT:10ZP, 80MT:20ZP, 70MT:30ZP) yürütülmüştür. Farklı sıcaklık ve yetiştirme ortamlarının Hericium izolatlarının verim, BE, ortalama mantar ağırlığı, şapka boyutları, şapka rengi, besin elementi içeriği ve antioksidan aktivitesi ve fenol içeriği üzerine etkileri araştırılmıştır. 8 Hericium arasındaki genetik farklılıklar ISSR ve SRAP markör sistemlerinin tek başlarına ve kombinasyonları kullanılarak değerlendirilmiştir. Hericium izolatlarının optimum misel gelişimi için uygun sıcaklık 25oC ve optimum pH 5 belirlenmiştir. Taslak ve şapka oluşumu için ise optimum sıcaklık 20oC olarak belirlenmiştir. Ortam denemelerinde en iyi verim %20 ve 30 oranında PK eklenen ortamlardan alınır iken, 80MT:20BK ortamında ise en düşük verim ve en yüksek protein ve besin elementi içeriğine sahip şapkalar elde edilmiştir. Antioksidan içeriği 15oC’de gelişen şapkalarda en yüksek iken, fenol içeriği üzerinde sıcaklığın etkisi bulunmamıştır. Yetiştirme ortamlarının ise antioksidan ve fenol içeriği üzerinde etkili olmadığı tespit edilmiştir. Hericium üretimi yapılan yetiştirme ortamlarının içeriğinde üretim sezonu boyunca değişiklik meydana geldiği ve üretim sezonu sonunda bu kompostun protein ve mineral madde içeriğinin başlangıçtakine göre arttığı belirlenmiştir. Ayrıca yetiştirme ortamının besin içeriğinin mantar besin içeriğine etkisinin olmadığı bulunmuştur. İzolatlar 2 büyük gruba ayrılmış, HA izolatı bu gruplardan birinde tek başına bulunurken, diğer 7 izolat aynı grupta yer almıştır. Anahtar sözcükler: Hericium spp, SRAP, ISSR, pamuk küspesi, zeytin pirinası, buğday kepeği ix ABSTRACT DETERMINATION OF OPTİMAL GROWİNG CONDITIONS AND GROWING SUBSTRATE FOR DIFFERENT HERICIUM ISOLATES AND MOLEKULER CHARACTERIZATION ATİLA, Funda PhD in Horticulture Department. Supervisor: Prof. Dr. Yüksel TÜZEL June 2015, 253 pages In the present study, it was aimed to determinete the effects of temperature, pH and substrate on mycelial growth and pinhead and fructification of Hericium isolates (HE-Ankara, HE-Denizli, HE, HE-Trabzon, HE-İzmit, HA, HC, HE-Amerika), to investigate the possibilities of using different agricultural and timbered wastes rich in lignocelluloses and easily available in a large amount in our region, in the cultivation of H. erinaceus, to determinate effects of temperatures and substrates on the yield and quality of mushrooms, to observe chemical changes in substrates during cultivation period and to determinate genetic relation among 8 Hericium isolates. Myceliel growth of isolates of Hericium was observed in 7 different temperatures (5,10,15,20,25,30,35 oC and 5 different pH (5,6,7,8,9). Experiments on pinhead and fructification temperatures were carried out at 15, 20 and 25oC degrees, while for substrate experiments were conducted with 7 different substrates prepared with oak sawdust, wheat bran, cotton and olive waste materials (80OS:20BK, 90 OS:10 CSH, 80 OS:20 CSH, 70 OS:30CSH, 90 OS:10 OPC, 80 OS:20 OPC, 70 OS:30OPC). The influences of temperatures and growing substrates on yield, BE’s, avarage mushroom weight, fruitbody size and color, nutritional composition, antioxidant activity and phenol content of Hericium isolates were investigated. Genetic diversity among 8 Hericium isolates was evaluated individually and combinated using ISSR and SRAP marker systems. The temperature suitable for optimal mycelial growth was determined to be 25oC, the most favorable growthn observed at pH 6. The temperature suitable for optimal pinhead and fruitbody formation was determined to be 20oC. Oak sawdust medium supplemented with 20 and 30 % cotton seed hulls was the best for yield and BE’s of the mushroom , while 80MT+20WB showed significantly lowest yield with maximum crude protein and mineral element content. The maximum antioxidant activity was obtained when it was cultivated at 15oC while the temperature didnt be effective in phenol content of mushrooms. Antioxidant activity and fenol content werent influenced by substrates. Content of substrate changed considerably during Hericium isolates growing period. At the end of the cultivation, protein and mineral contents in spend substrate were incresed as compared with the original substrate. The nutritional content of mushroom wasnt influenced by the nutritional content of substrate. Two major clades were revealed within this isolates. Clade 1 consisted only of HA isolate, while Clade II included all other 7 isolates. Key words: Hericium spp., ISSR, SRAP, cotton hulls, olive pres-cake, wheat bran xi TEŞEKKÜR Doktora çalışmamda desteğini hiç bir zaman esirgemeyen ve her türlü imkanı sağlayan değerli Hocam Prof. Dr. Yüksel TÜZEL’e, doktora tezimin şekillenmesi ve yürütülmesi sırasında, değerli fikir ve görüşlerini benimle paylaşan Prof. Dr. Aysun PEKŞEN’e, tez izleme jürisindeki değerli katkılarından dolayı Prof. Dr. Hatice ÖZAKTAN’a ve beni mantarcılık konusunda doktora yapmaya yönlendiren hocam Prof. Dr. Kaya BOZTOK’a teşekkürlerimi borç bilirim. Tohumluk misel temininde yardımcı olan Agro Mantarcılık Şirketinin sahibi Mehmet ATMACA’ya ve Erkel Misel LTD’ye, çalışmamın moleküler analizlerinin yürütülmesi konusunda yardımlarını esirgemeyen Yrd. Doç Dr. Birsen ÇAKIR’a, Dr. Deniz EROĞUL ve ZYM Tansel Aşçıoğlu’na, antioksidan ve fenol analizinde, yardımlarını esirgemeyen Doç. Dr. Fatih ŞEN’e, bölüm olanaklarından faydalanmamı sağlayan bölüm başkanımız Prof. Dr. Ercan ÖZZAMBAK’a, makro ve mikro element analizlerinde Cartagena Politeknik Üniversitesinin imkanlarından yararlanmamı sağlayan Prof. Dr. Juan Fernandez Hernandez ve Prof. Dr. Angel Faz’a, araştırmamı destekleyen Ege Üniversitesi Bilimsel Araştırmalar Proje Komisyonu’na çok teşekkür ederim Doktora çalışmamda bana güç veren ve yardımcı olan eşim Cenker ATİLA’ya, göstermiş oldukları sabır, anlayış ve özveriden dolayı sevgili annem Makbule ÇAVUŞLAR ve babam Halim ÇAVUŞLAR ve kızım Ela Deniz ve oğlum Alper’e yürekten teşekkürlerimi sunarım. xiii İÇİNDEKİLER Sayfa ÖZET ............................................................................................................... …vii ABSTRACT ..........................................................................................................ix TEŞEKKÜR .........................................................................................................xi ŞEKİLLER DİZİNİ ........................................................................................... xvii ÇİZELGELER DİZİNİ ....................................................................................... xxv SİMGELER VE KISALTMALAR DİZİNİ ......................................................xxix 1.GİRİŞ ................................................................................................................... 1 2.ÖNCEKİ ÇALIŞMALAR ..................................................................................
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