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

Ilknur Colak; Gökçe Karadayı

Makale detayları

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

Yazarlar:

Ilknur Colak | Gökçe Karadayı

Yayıncı:

Van Yuzuncu Yıl University

Ana Başlıklar:

Epigenetik

Dergi:

Yüzüncü Yıl Üniversitesi Fen Bilimleri Enstitüsü Dergisi Vol.29 Issue 3, pp.830-841

Belge Tipi:

Makale

Yayın Tarihi:

12/2024

Belge Dili:

İngilizce

Document ID (Org No.):

2001338118

pISSN:

1300-5413

Anahtar kelimeler:

Özet:

 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.

 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.

 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.

 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 cDNAlar 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.

Makale detayları

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

Yazarlar:

Ilknur Colak | Gökçe Karadayı

Yayıncı:

Van Yuzuncu Yıl University

Ana Başlıklar:

Epigenetik

Dergi:

Yüzüncü Yıl Üniversitesi Fen Bilimleri Enstitüsü Dergisi Vol.29 Issue 3, pp.830-841

Belge Tipi:

Makale

Yayın Tarihi:

12/2024

Belge Dili:

İngilizce

Document ID (Org No.):

2001338118

pISSN:

1300-5413

Abiotic stress

Cadmium

Epigenetics

qAMP

Triticum aestivumAbiyotik stres

Epigenetik

Kadmiyum

qAMP

Triticum aestivum

Sayı

Makale detayları

Yazarlar: Ilknur Colak | Gökçe Karadayı

Yayıncı: Van Yuzuncu Yıl University

Ana Başlıklar: Epigenetik

Dergi: Yüzüncü Yıl Üniversitesi Fen Bilimleri Enstitüsü Dergisi Vol.29 Issue 3, pp.830-841

Belge Tipi: Makale

Yayın Tarihi: 12/2024

Belge Dili: İngilizce

Document ID (Org No.): 2001338118

pISSN: 1300-5413

Sayı

Yazarlar:

Ilknur Colak | Gökçe Karadayı

Yayıncı:

Van Yuzuncu Yıl University

Ana Başlıklar:

Epigenetik

Dergi:

Yüzüncü Yıl Üniversitesi Fen Bilimleri Enstitüsü Dergisi Vol.29 Issue 3, pp.830-841

Belge Tipi:

Makale

Yayın Tarihi:

12/2024

Belge Dili:

İngilizce

Document ID (Org No.):

2001338118

pISSN:

1300-5413