Türk Ekmeklik Buğday (T. aestivum L.) Çeşidi Gerek 79'da Kuraklığa-Duyarlı Genlerin mRNA Farklılık Gösterim Analizi İle Tanımlanması

Öz

Buğday, dünyadaki en önemli tahıl ürünlerinden biridir. Dünyanın birçok bölgesi, gıda ve hayvan yemi kaynağı olarak buğdaya bağımlıdır. Kuraklık stresi buğdayın gelişimini olumsuz etkiler ve üretimini büyük ölçüde azaltır. Kuraklık tepkisi çok sayıda gen, transkripsiyon faktörleri, miRNA'lar, hormonlar, proteinler, kofaktörler, iyonlar ve metabolitleri içeren karmaşık bir genetik mekanizmadır. Kuraklığa dayanıklılık mekanizmalarının genetik temellerinin anlaşılması, buğdayda bu özelliğin genetik ıslahı için oldukça önemlidir. Buğday Türkiye için de önemli bir tahıl ürünüdür. Bu çalışmada, ekmeklik buğdayda (Triticum aestivum cv. Gerek 79) kuraklık stresini düzenleyen genlerin belirlenmesi amaçlanmış ve kontrol ve kuraklık stresi altındaki (%62,4 RWC) bitkilerin fide yapraklarında mRNA farklılık gösterimi (mRNA DD) kullanılarak gen ekspresyon profillemesi yapılmıştır. Karşılaştırmalı profilleme çalışması, farklı şekilde eksprese edilen toplam 20 cDNA bandı göstermiş ve bu bantlardan 10 tanesi klonlanmış ve dizilenmiştir. Düşük kaliteli okumalara sahip olanlar elenmiştir. G1 ve G2 olarak adlandırılan kalan iki cDNA'nın nükleotid dizileri benzerlik analizine tabi tutulmuştur. G1 ve G2 sırasıyla Triticum aestivum ve diğer bazı bitkilerin mor asit fosfataz mRNA dizisine ve glikosiltransferaz ailesi 92 protein benzeri dizisine yüksek derecede homoloji göstermiştir. Mor asit fosfatazların, abiyotik ve biyotik streslere karşı bitki tepkilerinde yer aldığı gösterilmiştir. Benzer şekilde, glikosiltransferazların bitkilerde hücresel homeostaz ve detoksifikasyon yollarındaki işlevlerinin yanı sıra pirinçte termotolerans üzerinde rolü olduğu bildirilmiştir. Bu raporlar ve bulgularımız, G1 ve G2 cDNA klonlarının daha fazla araştırılması için bir temel oluşturmuştur. Mor asit fosfataz ve glikosiltransferaz ailesi 92 protein benzeri diziye karşılık gelen bu gen parçalarının kuraklık koşulları altında RNA düzeyinde gen ekspresyonu değişikliklerinin araştırılması devam etmektedir. Bu genlerin ileri karakterizasyonu kuraklık tepkisindeki işlevlerinin anlaşılmasında önemli olabilir.

Anahtar Kelimeler

• Buğday
• Kuraklık
• mRNA farklılık gösterimi
• gen ekspresyonu

Identification of Drought-Responsive Genes in Turkish Bread Wheat (T. aestivum L.) Cultivar Gerek 79 by mRNA Differential Display Analysis

Öz

Wheat is one of the most important cereal crops in the world. Many parts of the world depend on wheat as a source of food and animal feed. Drought stress negatively affects its development and greatly reduce its production. Drought response is a complex genetic mechanism involving multiple genes, transcription factors, miRNAs, hormones, proteins, co-factors, ions, and metabolites. The understanding of genetic basis of the drought tolerance mechanisms is very important for genetic improvement of this trait in wheat. Wheat is also an important cereal crop in Turkey. In this study, it is aimed to identify drought stress-regulated genes in bread wheat (Triticum aestivum cv. Gerek 79) and gene expression profiling using mRNA differential display (mRNA DD) was performed for seedling leaves of control and drought-stressed plants (62.4% of RWC). The comparative profiling study showed a total of 20 differentially-expressed cDNA bands and 10 of them were cloned and sequenced. The inserts having poor quality reads were eliminated. The nucleotide sequences of the remaining two cDNAs named G1 and G2 were subjected to similarity analysis. G1 and G2 showed a high degree of homology to mRNA sequence of purple acid phosphatase and glycosyltransferase family 92 protein-like sequence of Triticum aestivum and some other plants, respectively. Purple acid phosphatases have been shown to be involved in plant responses to abiotic and biotic stresses. Similarly, the role of glycosyltransferases on thermotolerance has been reported in rice besides their functions in cellular homeostasis and detoxification pathways in plants. These reports and our findings have laid a foundation for further investigation of G1 and G2 cDNA clones. The investigation of differential expression of these gene fragments corresponding to purple acid phosphatase and glycosyltransferase family 92 protein-like sequence under drought conditions at the RNA level is ongoing. Further characterization of these genes could be important in understanding the functions of these gene/s in drought response.

Anahtar Kelimeler

• Wheat
• Drought
• mRNA differential display
• gene expression

Kaynakça

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