Topraktaki Kadmiyum (Cd) Birikimine Maruz Kalan Ekmeklik Buğdayda Kadmiyumun İndüklediği Bazı Genlerin DNA Metilasyonu ve İfadesindeki Değişikliklerin Karşılaştırılması

Year 2024, Volume: 29 Issue: 3, 830 - 841, 31.12.2024

Ilknur Colak , Gökçe Karadayı

https://doi.org/10.53433/yyufbed.1497910

Abstract

Doğal fenomenler ve antropojenik olaylar sonucunda toprakta kadmiyum (Cd) birikimi son yıllarda giderek artmakta ve bitkiler, yaşamları için gerekli olmayan bu ağır metale doğrudan maruz kalmaktadır. Çalışmamızda Cd stresine maruz bırakılan Triticum aestivum L. bitkisinde UGT-3 (UDP-Glikosiltransferaz-3), LTP-4 (Lipit Transfer Proteini-4), Plazma Membran PIP-1 (Plazma Membran Protein-1) genleri çalışılmıştır. Bu genlerde meydana gelen ekspresyon seviyelerindeki değişimlerin ve Real-Time PCR temelli kantitatif DNA metilasyon analizi (qAMP) kullanılarak ilk kez metilasyon yüzdelerinin belirlenmesi amaçlanmıştır. Bu kapsamda kontrol, 100, 250 ve 500 µM CdCl2 dozlarına maruz bırakılarak geliştirilen buğdayların kök ve gövdelerinden DNA ile RNA izolasyonu yapılmıştır. Daha sonra RNA örneklerinden elde edilen cDNA'lar ile gen ekspresyon analizi yapılarak gen ekspresyon düzeyleri belirlenmiştir. DNA metilasyon yüzdeleri qAMP tekniği uygulanarak saptanmıştır. Sonuç olarak UGT-3, LTP-4 ve PIP-1 genlerinde en yüksek metilasyon yüzdesinin hem gövdede hem de kökte 250 µM konsantrasyonda olduğu gözlenmiştir. UGT-3 geninde ekspresyon seviyesi gövdede 250 µM konsantrasyonda en yüksek oranı verirken, LTP-4 geninde kökte 250 µM konsantrasyonda aşırı ekspresyon gözlenmiştir. UGT-3, LTP-4 ve PIP-1 genlerinin metilasyon oranlarındaki değişimler bitkilerde kullanımı yeni bir teknik olan qAMP ile ilk kez araştırılmıştır. Genlerin ekspresyon seviyeleri ile metilasyon durumları arasında anlamlı bir ilişki olduğu bulunmuştur.

Keywords

Abiyotik stres, Epigenetik, Kadmiyum, qAMP, Triticum aestivum

Project Number

FYL-2020-8329

Comparison of DNA Methylation and Changes in the Expression of Certain Cadmium-induced Genes in Bread Wheat Exposed to Cadmium (Cd) Accumulation in Soil

Abstract

As a result of natural phenomena and anthropogenic activities, cadmium (Cd) accumulation in the soil has been increasing in recent years, and plants are directly exposed to this heavy metal, which is not essential for their life. In our study, UGT-3 (UDP-Glycosyltransferase-3), LTP-4 (Lipid Transfer Protein-4), Plasma Membrane PIP-1 (Plasma Membrane Protein-1) genes were investigated in Triticum aestivum L. plants exposed to Cd stress. It was aimed to determine the changes in expression levels in these genes and their methylation percentages for the first time using Real-Time PCR-based quantitative DNA methylation analysis (qAMP). In this context, DNA and RNA were isolated from the roots and stems of wheat grown by exposure to control, 100, 250 and 500 µM CdCl2 doses. Then, gene expression levels were determined by gene expression analysis with cDNAs obtained from RNA samples. DNA methylation percentages were determined by applying the qAMP technique. As a result, it was observed that the highest methylation percentage in the UGT-3, LTP-4 and PIP-1 genes was at 250 µM concentration in both the stem and the root. While the expression level of the UGT-3 gene was highest at a concentration of 250 µM in the stem, overexpression of the LTP-4 gene was observed at a concentration of 250 µM in the root changes in the methylation rates of UGT-3, LTP-4 and PIP-1 genes were investigated for the first time with qAMP, a new technique used in plants. A significant relationship was found out between the expression levels and methylation status of genes

Keywords

Abiotic stress, Cadmium, Epigenetics, qAMP, Triticum aestivum

Project Number

FYL-2020-8329

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