M. Ercan ÖZZAMBAK, Emrah ZEYBEKOĞLU, İzzet GÜN, Tuğba KILIÇ

Spathiphyllum’un in vitro mikro çoğaltımı üzerine şeker konsantrasyonlarının etkileri

Fotoototrofik mikro çoğaltım, doku kültüründe gereksinim duyulan karbonun karbondioksitten karşılanmasıdır. Bu yöntemin temeli besin ortamına organik karbon kaynağı olarak şekerin ilave edilmemesi veya konsantrasyonunun azaltılması koşuluna dayanmaktadır. Fotoototrofik mikro çoğaltım, doku kültürünün en önemli sakıncaları olan vitrifikasyon, dış koşullara adaptasyon güçlüğü, yoğun işgücü kullanımı, yüksek maliyet, kontaminasyon riski gibi sorunları azaltabilmekte ya da ortadan kaldırabilmektedir. Bu çalışma ile Spathiphyllum wallisii’nin farklı şeker konsantrasyonları içeren besin ortamlarındaki sürgün çoğaltma-köklendirme kabiliyetinin in vitro fotoototrofik çoğaltıma uygunluk durumu belirlenmeye çalışılmıştır. Sürgün çoğaltma denemesinde 8-10 mm uzunluğunda iki yapraklı in vitro sürgünler MS + 0.5 mg/l BAP + 0.1 mg/l NAA ortamında; köklendirme denemesinde ise 25 mm boyundaki in vitro sürgünler MS + 0.1 mg/l NAA ortamında kültüre alınmış olup; her iki ortam da 0, 10, 20, 30 g/l konsantrasyonlarında şeker içermektedir. Sürgün çoğaltma aşamasında, çoğalma oranı, kardeş sürgün adedi ve uzunluğu, sürgün yaş ağırlığı, yaprak renk değerleri ve gelişme puanı bakımından şekerli (10, 20, 30 g/l) ve şekersiz (0 g/l) ortamlar arasında istatistiksel olarak önemli farklılıklar bulunmuştur. 10, 20, 30 g/l şeker konsantrasyonlarında çoğalma sırasıyla 4.4, 4.6, 5.4 sürgün/eksp., 0 g/l şeker konsantrasyonunda 0.7 sürgün/eksp. olarak gerçekleşmiştir. Köklendirme aşamasında da ortamda şeker bulunması incelenen özellikler üzerine pozitif yönde etki göstermiştir. 20, 30 g/l şeker konsantrasyonlarında köklenme oranı %100 olup; kök sayısı sırasıyla 16.1-8.8 adet/eksp., yaş kök ağırlığı ise 518.9-440.6 mg olarak belirlenmiştir. Bu çalışma koşullarında fotoototofik mikro çoğaltmanın tam olarak gerçekleşmediği; Spathipyllum’un in vitro sürgün çoğaltma ve köklendirilmesi için besin ortamında 20 ya da 30 g/l konsantrasyonlarında şeker bulunması gerektiği sonucuna varılmıştır.

Effects of sugar concentrations on in vitro micropropagation of spathiphyllum

Photoautotrophic micropropagation can be described as a tissue culture technique in which, all required carbon is supplied from carbon dioxide. Preparing the medium without or with low concentration of sugar, which is a source of organic carbon, is the main condition. Decreasing some problems as vitrification and contamination, lowering high input and labour costs and improving the acclimatization capacity of in vitro plantlets are the main advantages of photoautotrophic micropropagation. The present study was carried out to determine the compatibility of spathiphyllum for in vitro photoautotrophic micropropagation by proliferation and rooting ability of microshoots on different sugar concentrations. In shoot multiplication trial, micro shoots in 8-10 mm length with two leafs were planted in MS + 0.5 mg/l BAP + 0.1 mg/l NAA medium and in rooting experiment shoots in 25 mm length were transferred on MS + 0.1 mg/l NAA. Both media prepared with four different sugar concentrations (0, 10, 20, 30 g/l). In multiplication stage, significant differences were found between medium with sugar and without sugar at proliferation rate, new shoot number/exp. and length, shoot fresh weight, leaf colour values and growth score parameters. 10, 20, 30 g/l sugar concentrations gave the highest proliferation rate with means of 4.4, 4.6, 5.4 shoots per explant and 0 g/l doses gave 0.7 shoot/exp. respectively. A positive relationship was found between the increase in sugar concentration and rooting rate, root number, shoot fresh weight, root dry weight. 100% rooting percentage, 16.1-8.8 root number per explant 518.9-440.6 mg root fresh weight were verified for 20, 30 g/l sugar respectively. In this study conditions, photoautotrophic micropropagation was not be occurred exactly and the presence of 20 or 30 g/l sugar concentrations was necessary for efficient in vitro proliferation and rooting of Spathiphyllum.

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