Bakır Stresi altında Nohut Bitkisinde (Cicer arietinum L.) Stresle İlişkili Gen Ekspresyon Seviyelerinde Meydana Gelen Değişimlerin Belirlenmesi ve Hücresel H2O2 ile İlişkisi

Bu çalışmanın amacı, nohut (Cicer arietinum) bitkisinde farklı zaman ve konsantrasyonlarda bakıra maruz kalmış bitki yapraklarındaki stres genlerinin ekspresyon seviyelerindeki değişimlerin belirlenmesidir. Ayrıca; gen ekspresyonlarındaki değişim ile hücresel H2O2 arasındaki ilişki tespit edilmeye çalışılmıştır. Bu bağlamda hücrede oksidatif stresin göstergesi olan melondialdehit (MDA) miktarı ve reaktif oksijen türlerinden (ROS) H2O2 miktarları tespit edilmiştir. Ayrıca antioksidan savunma elemanlarından Metallothionein (MT), Catalase (CAT) ve superoksit dismutaz (Cu/Zn-SOD) enzimlerinin gen ekspresyonlarındaki değişim house-keeping gen olarak seçilmiş aktinin ekspresyon düzeyi baz alınarak tespit edilmiştir. Çalışma sonucunda MDA içeriğinin süre ve konsantrasyona bağlı olarak belirgin bir şekilde arttığı, stres alakalı gen ekpresyonlarının bütün konsantrasyonlarda kontrolden yüksek düzeyde eksprese edildiği ancak en yüksek ekspresyon gerçekleştikten sonra süre ve konsantrasyondan arttıkca ekpsreyon düzeylerinde bir azalma olduğu belirlenmiştir. Bu durum stres anındaki kararlı bir artış gösteren hücresel H2O2 ile ilişkilendirilmiştir. Çalışma sonucunda Cu maruziyetinin oskidatif strese neden olarak stres alakalı genlerin ekpresyonlarını indüklediği tespit edilmiştir. Ayırca hücresel H2O2’nin belirli konsantrasyona kadar gen ekpresyonunu up-regule ederken belirli konsantrasyondan sonra down regule etmiş olduğu düşünülmektedir. bu calısmanın sonucları sayesinde; tarım alanlarındaki bitkiler belirli konsantrasyonda H2O2 ye maruz bırakılarak biyotik ve ya abiyotik streslere karsı oksidadatif stres cevabını daha erken evrede vermesi sağlanabilir. Bu sayede, daha dayanıklı ürünler elde edilmesine bağlı olarak kimyasal zirai ilac kullanımı asgari düzeye indirilebilir.

Anahtar Kelimeler:

cicer arietinum, Cu, H2O2, Oksidatif Sinyalizasyon, stres alakalı genler

The Evulation of Stress Related Gene Expression Level and Relationship to Cellular H2O2 in Chickpea (Cicer arietinum L.) Under Copper Stress

The aim of this study is to determine the changes in expression levels of stress genes in chickpea (Cicer arietinum) plant leaves exposed to copper (Cu) at different times and concentrations. Also; the relationship between the changes in gene expression and cellular H2O2 was investigated. In this context, the amount of malondialdehyde (MDA) and reactive oxygen species (ROS) hydrogen peroxide (H2O2) levels were determined. Furthermore, the changes in gene expressions of Metallothionein (MT), Catalase (CAT) and superoxide dismutase (Cu / Zn-SOD) enzymes were determined based on the actin expression level that selected as a housekeeping gene. It was determined that MDA content increased significantly due to time and concentration, In all duration and concentrations, the expression of stress-related genes significantly differed from the control group. Hleowever, a decrease has been determined by all gene expressions after the highest expression. This phenomenon is associated with cellular H2O2, which shows a steady increase in stress. At the end of the study, it was concluded that the elevating duration and concentration of Cu induced oxidative stress and caused the expression of stress-related genes. Furthermore, cellular H2O2 might be acting as a signal molecule that, up-regulate gene expressions until a certain concentration and down-regulate until a certain concentration. Thanks to the results of this study; Plants in agricultural areas can be exposed to a certain concentration of H2O2 to provide an earlier response to oxidative stress against biotic or abiotic stresses. In this way, the use of chemical pesticides can be minimized due to obtaining more durable products.

Keywords:

Cicer arietinum, Cu, H2O2, Oxidative signaling, Stress-related genes,

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