Publication: Buğday (Triticum aestivum L.) ve arpa (Hordeum vulgare L.) bitkilerinde kadmiyum stresine bağlı genotoksik etkilerin belirlenmesi
Abstract
BUĞDAY (Triticum aestivum L.) VE ARPA (Hordeum vulgare L.) BİTKİLERİNDE KADMİYUM STRESİNE BAĞLI GENOTOKSİK ETKİLERİN BELİRLENMESİ Ağır metal kirliliği günümüzün küresel ölçekli sorunlarındandır. Son dönemde yapılan çalışmalar Kırgızistan’da da ağır metal kirliliği yaşandığını ve kirliliğin özellikle kadmiyum (Cd) kirliliği şeklinde öne çıktığını göstermiştir. Bu sebeple mevcut çalışmada Kırgızistan’da yaygın olarak ekilen buğday (Triticum aestivum L. var. intensivnaya) ve arpa (Hordeum vulgare L. var. alta) varyetelerine ait tohumlar temin edilip in vitro koşullarda çimlendirilerek Cd’un bitkiler üzerindeki genotoksik etkileri araştırılmıştır. Çalışmada amaca uygun olarak, çimlendirilen buğday ve arpa bitkileri kontrol grubu ve deney grubu bitkiler (50, 100, 200 ve 400 μM Cd uygulanan) olmak üzere ayrılmıştır. Uygulanan Cd stres şartları, belirtilen konsantrasyonlarda CdCl2 içeren Hoagland çözeltilerinin, bitkilere 1 ay boyunca günaşırı sıklıkta 20’şer ml hacimde verilmesiyle oluşturulmuştur. Hoagland içerisinde verilen Cd’un bitki yapraklarındaki akümülasyon değerleri ICP-OES cihazı kullanarak ölçülmüştür. Akümüle edilen Cd’un, araştırılan fizyolojik parametreler (yaprak uzunluğu, yaprak eni, yaprak yaş ve kuru ağırlıkları, yaprakta toplam klorofil miktarı) üzerindeki etkileri incelenmiştir. Ayrıca bitki yapraklarındaki genotoksik etkilerinin belirlenmesi amacıyla, modifiye edilmiş CTAB yöntemi ile DNA izolasyonu yapılmış ve sonrasında farklı ISSR (Inter Simple Sequence Repeat-Basit Tekrarlı Diziler Arası Polimorfizm) primerlerinin hedeflediği bölgeler PZR ile çoğaltılmıştır. PZR ile elde edilen amplikonlar sonrasında agaroz jel elektroforezi ile ortaya çıkan DNA bant profilleri kontrol grubu ile deney grubu karşılaştırılarak analiz edilmiştir. Elde edilen bant profillerinden yola çıkarak Cd stresine maruz kalmış bitkilerin genomik kararlılık değeri (GKY-Genomic Templete Stability (GTS)) hesaplanmış ve söz konusu veriye dayalı olarak bitkilerin hangi stres seviyesine dayanıklı/ duyarlı oldukları belirlenmiştir. Çalışma sonucunda ICP-OES ve DNA bant profil verileri birbirleriyle ilişkilendirilerek kadmiyumda doza bağlı genotoksik etkiler ortaya konmuştur.
DETERMINATION OF GENOTOXIC EFFECTS OF CADMIUM STRESS IN WHEAT (Triticum aestivum L.) AND BARLEY (Hordeum vulgare L.) Heavy metal pollution is one of the global scale problems of today. Recent studies have shown that heavy metal pollution is also experienced in Kyrgyzstan, and the pollution stands out especially cadmium (Cd) pollution. For this reason, in the present study, the genotoxic effects of Cd on plants were investigated by obtaining and in vitro germinating conditions wheat (Triticum aestivum L. var. intensivnaya) and barley (Hordeum vulgare L. var. alta) varieties widely planted in Kyrgyzstan. In accordance with the purpose of the study, germinated wheat and barley plants were divided into control group and experimental group plants (50, 100, 200 and 400 μM Cd applied).The applied Cd stress conditions were created by giving Hoagland solutions containing CdCl2 at the specified concentrations to the plants, in 20 ml every other day for one month. The accumulation of Cd in plant leaves were measured using ICP-OES. The effects of accumulated Cd on the investigated physiological parameters (leaf length, leaf width, leaf fresh and dry weights, total chlorophyll content in the leaf) were investigated. Additionally, the genotoxic effects of accumulated cadmium on plant leaves, and DNA were isolated with the modified CTAB method. Then the regions targeted by different ISSR (Inter Simple Sequence Repeat) primers were amplified by PCR. The DNA band profiles obtained by agarose gel electrophoresis after amplicons obtained by PCR were analyzed by comparing the control group with the experimental group. Based on the band profiles obtained, the genomic stability (Genomic Template Stability-GTS) of plants exposed to Cd stress was calculated and based on the data in question, it was determined to which stress level the plants were resistant/ sensitive. As a result of the study, dose-related genotoxic effects of cadmium were revealed by correlating ICP-OES and DNA band profile data.
DETERMINATION OF GENOTOXIC EFFECTS OF CADMIUM STRESS IN WHEAT (Triticum aestivum L.) AND BARLEY (Hordeum vulgare L.) Heavy metal pollution is one of the global scale problems of today. Recent studies have shown that heavy metal pollution is also experienced in Kyrgyzstan, and the pollution stands out especially cadmium (Cd) pollution. For this reason, in the present study, the genotoxic effects of Cd on plants were investigated by obtaining and in vitro germinating conditions wheat (Triticum aestivum L. var. intensivnaya) and barley (Hordeum vulgare L. var. alta) varieties widely planted in Kyrgyzstan. In accordance with the purpose of the study, germinated wheat and barley plants were divided into control group and experimental group plants (50, 100, 200 and 400 μM Cd applied).The applied Cd stress conditions were created by giving Hoagland solutions containing CdCl2 at the specified concentrations to the plants, in 20 ml every other day for one month. The accumulation of Cd in plant leaves were measured using ICP-OES. The effects of accumulated Cd on the investigated physiological parameters (leaf length, leaf width, leaf fresh and dry weights, total chlorophyll content in the leaf) were investigated. Additionally, the genotoxic effects of accumulated cadmium on plant leaves, and DNA were isolated with the modified CTAB method. Then the regions targeted by different ISSR (Inter Simple Sequence Repeat) primers were amplified by PCR. The DNA band profiles obtained by agarose gel electrophoresis after amplicons obtained by PCR were analyzed by comparing the control group with the experimental group. Based on the band profiles obtained, the genomic stability (Genomic Template Stability-GTS) of plants exposed to Cd stress was calculated and based on the data in question, it was determined to which stress level the plants were resistant/ sensitive. As a result of the study, dose-related genotoxic effects of cadmium were revealed by correlating ICP-OES and DNA band profile data.