Bor Toksisitesine Yanıt Olarak Bor Toleranslı Triticum Urartu Genotipinin Transkriptomik Analizi
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Dosyalar
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bor toksisitesi özellikle kurak ve yarı kurak tarım bölgelerinde buğday ürününde
büyüme ve tane veriminde azalmaya yol açmaktadır. Bu nedenle, bor’a toleranslı
buğday çeşitlerinin geliştirilmesi için bor toksisitesine toleransı daha yüksek olan
buğday yabani akrabalarının (WWR'ler) incelenmesi ve kullanılması faydalı olacaktır.
Ayrıca, bor toksisitesi stres toleransını geliştirmek için WWR'leri verimli bir şekilde
kullanmak için, bor toksik büyüme koşulları altında bunlarda meydana gelen
transkriptomik değişiklikleri anlamak önemlidir. Bu yaklaşım doğrultusunda, bu
çalışmada, bor toksisitesine toleranslı bir Triticum urartu genotipinin transkriptomik
yanıtı RNA dizileme yoluyla araştırılmış ve RT-qPCR yoluyla B toksisitesi toleransını düzenlemede rol oynadığı düşünülen bazı genlerinin ekspresyonunu incelenmiştir.
Fizyolojik büyüme analizinde, gövde yaş ağırlığı hariç, T. urartu, PI662222 genotipinin
kök-gövde uzunluğu, kök yaş ağırlığı ve kök-gövde kuru ağırlığı dâhil olmak üzere
diğer tüm büyüme parametreleri, kontrol çeşidi Bolal 2973'e kıyasla yüksek bordan (10
mM) daha az etkilenmiştir. Ayrıca, bor toksisitesi, genotipin B tolerans seviyesinden
bağımsız olarak gövdelere kıyasla kökler üzerinde daha azaltıcı bir etkiye sahip
olmuştur. Moleküler analizde, çalışmada toplam 2.254 yeni gen ve 7402 yeni transkript
tanımlanmıştır. İki gövde kütüphanesinde toplam 32.777 genin diferansiyel olarak
düzenlenirken, bunlar arasında 654 gende önemli ölçüde diferansiyel ekspresyonu
gözlenmiştir. Önemli ölçüde düzenlenmiş bu genlerden 441'inin aşağı düzenlendiği,
213 genin ise yukarı düzenlendiği tespit edilmiştir. RT-qPCR analizi için transkriptom
dizileme sonuçlarına göre farklı mekanizmalarda yer alan ve önemli ölçüde diferansiyel
ekspresyon gösteren dört gen rastgele seçilmiştir. Kontrol ile karşılaştırıldığında Bor
toksik büyüme koşulu altında çalışılan dört gen arasında, α-DOX, GELP, FTSH 9,
genleri aşağı regülasyon gösterirken, UXS1 geni yukarı regülasyon göstermiştir.
Çalışılan tüm genlerin RT-qPCR yoluyla diferansiyel ekspresyonu, RNA dizileme
sonuçlarıyla uyumludur.
Bu tezde tanımlanan B toksisitesi stresine duyarlı aday genlerin
tanımlanmasıyla, buğday ve diğer tahıl ürünlerinde gelecekteki omik araştırmaları
kolaylaştırılacaktır. Araştırılan tez, T. urartu genotipinin bor toksisitesine toleransı ve
transkriptomundaki değişiklikler üzerine yapılan ilk araştırmadır. Elde edilen sonuçlar,
yüksek bor’a yanıt veren tanımlanmış genlerin bor toksisite toleransı sağlamadaki
işlevlerini doğrulamak üzere işlevsel olarak karakterize etmek için transgenik deneylere
ihtiyaç olduğunu göstermektedir. Sonuçlarımız, evcilleştirilmiş buğdayın genetik
çeşitliliğindeki dar boğazı genişletmek için T. urartu’dan farklı genlerin
kullanılabileceğini ortaya koyarken, moleküler ıslah veya genetik mühendisliği
aracılığıyla buğdayın yüksek bor konsantrasyonuna toleransını geliştirmeye olanak
sağlayabilir.
One of the main problems brought by the increasing population and climate change in today's world is the need for an increased food supply over the years. However, increasing the food supply is getting difficult due to the reduction in crop yields under a continuously changing environment. Thus, it is required to develop new crops with more adaptation toward environmental change. Wild genetic resources are known for their greater adaptation in harsh growth conditions and thus, should be used for the development of genotypes with greater tolerance levels. Wheat is an important agricultural crop whose production is largely affected by excess or deficiency of mineral elements in the soil. Boron is one of those elements whose toxicity leads to a reduction in growth and grain yield in wheat crop, especially in arid and semi-arid agricultural regions. Thus, it will be beneficial to study and utilize wheat wild relatives (WWRs) with greater boron toxicity tolerance to develop boron-tolerant wheat cultivars. However, to efficiently utilize WWRs for developing boron toxicity stress tolerance, it is important to understand transcriptomic changes occurring in them under boron-toxic growth conditions. In line with this approach, in this study, we explored the transcriptomic response of a boron toxicity-tolerant, Triticum urartu genotype, PI662222 via RNA sequencing, and studied the expression of some of its genes that seems to be involved in regulating its B toxicity tolerance via RT-qPCR. In physiological growth analysis, except shoot fresh weight, all the other growth parameters of T. urartu, PI662222 genotype including root-shoot length, root fresh weight, and root shoot dry weight were less affected by high boron (10 mM) as compared to check cultivar, Bolal 2973. Moreover, regardless of the genotype's level of boron tolerance, boron toxicity had a greater depleting effect on roots than on shoots. In molecular analysis, total 2,254 novel genes and 7402 novel transcripts were identified in the study. While a total of 32,777 genes were differentially regulated in the two shoot libraries, significant differential regulation was observed in 654 genes. Among these, 213 and 441 genes were up- and down-regulated respectively. Four significantly differentially expressed genes involved in different mechanisms were randomly selected based on transcriptome sequencing results for RT-qPCR analysis. Among the studied four genes, while α-DOX, GELP, and FTSH 9 genes showed downregulation, UXS1 gene showed upregulation under boron toxic growth conditions as compared to Control. The differential expression of all the studied genes via RT-qPCR was in line with the RNA sequencing results. The future omics research in wheat and other cereal crops will be facilitated by the identification of the putative genes for boron toxicity stress responsiveness found in this thesis. As per the available literature, this was the first study to determine the boron toxicity tolerance of T. urartu genotype and changes in its transcriptome. The obtained results suggested the need for more investigation including transgenic experiments to functionally characterize the identified high boron-responsive genes and to confirm their function in conferring B toxicity tolerance. The results directed towards the utilization of several T. urartu genes to increase the restricted genetic diversity of cultivated wheat. Molecular breeding or genetic engineering-based techniques can be employed to effectively utilize the obtained data to increase boron toxicity tolerance in modern wheat cultivars.
One of the main problems brought by the increasing population and climate change in today's world is the need for an increased food supply over the years. However, increasing the food supply is getting difficult due to the reduction in crop yields under a continuously changing environment. Thus, it is required to develop new crops with more adaptation toward environmental change. Wild genetic resources are known for their greater adaptation in harsh growth conditions and thus, should be used for the development of genotypes with greater tolerance levels. Wheat is an important agricultural crop whose production is largely affected by excess or deficiency of mineral elements in the soil. Boron is one of those elements whose toxicity leads to a reduction in growth and grain yield in wheat crop, especially in arid and semi-arid agricultural regions. Thus, it will be beneficial to study and utilize wheat wild relatives (WWRs) with greater boron toxicity tolerance to develop boron-tolerant wheat cultivars. However, to efficiently utilize WWRs for developing boron toxicity stress tolerance, it is important to understand transcriptomic changes occurring in them under boron-toxic growth conditions. In line with this approach, in this study, we explored the transcriptomic response of a boron toxicity-tolerant, Triticum urartu genotype, PI662222 via RNA sequencing, and studied the expression of some of its genes that seems to be involved in regulating its B toxicity tolerance via RT-qPCR. In physiological growth analysis, except shoot fresh weight, all the other growth parameters of T. urartu, PI662222 genotype including root-shoot length, root fresh weight, and root shoot dry weight were less affected by high boron (10 mM) as compared to check cultivar, Bolal 2973. Moreover, regardless of the genotype's level of boron tolerance, boron toxicity had a greater depleting effect on roots than on shoots. In molecular analysis, total 2,254 novel genes and 7402 novel transcripts were identified in the study. While a total of 32,777 genes were differentially regulated in the two shoot libraries, significant differential regulation was observed in 654 genes. Among these, 213 and 441 genes were up- and down-regulated respectively. Four significantly differentially expressed genes involved in different mechanisms were randomly selected based on transcriptome sequencing results for RT-qPCR analysis. Among the studied four genes, while α-DOX, GELP, and FTSH 9 genes showed downregulation, UXS1 gene showed upregulation under boron toxic growth conditions as compared to Control. The differential expression of all the studied genes via RT-qPCR was in line with the RNA sequencing results. The future omics research in wheat and other cereal crops will be facilitated by the identification of the putative genes for boron toxicity stress responsiveness found in this thesis. As per the available literature, this was the first study to determine the boron toxicity tolerance of T. urartu genotype and changes in its transcriptome. The obtained results suggested the need for more investigation including transgenic experiments to functionally characterize the identified high boron-responsive genes and to confirm their function in conferring B toxicity tolerance. The results directed towards the utilization of several T. urartu genes to increase the restricted genetic diversity of cultivated wheat. Molecular breeding or genetic engineering-based techniques can be employed to effectively utilize the obtained data to increase boron toxicity tolerance in modern wheat cultivars.
Açıklama
Anahtar Kelimeler
Abiotic Stress, Boron Toxicity, Gene Expression, Triticum Urartu, Whea, Abiyotik Stres, Bor Toksisitesi, Buğday, Gen Ekspresyonu, Transkriptomik Analiz, RT-qPCR, Transcriptomic Analysis
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Uyar, G. S., (2023). Bor Toksisitesine Yanıt Olarak Bor Toleranslı Triticum Urartu Genotipinin Transkriptomik Analizi. (Yüksek Lisans Tezi). Selçuk Üniversitesi, Fen Bilimleri Enstitüsü, Konya.
