Effect of of boron toxicity on miR393 expression at moderate and severe levels in Arabidopsis thaliana

Abstract

miR393, one of the microRNA (miRNA) families preserved in plants, plays a role in many critical developmental processes. In this study, expression levels of miR393 were determined in Arabidopsis thaliana exposed to 1 mM (1B) and 3 mM (3B) boric acid by Stem-Loop (SL) quantitative reverse transcription polymerase chain reaction method. In addition, genes targeted by miR393 in Arabidopsis thaliana and Gene Ontology Enrichment analysis of these genes were performed. After application of toxic levels of B in Arabidopsis thaliana plant, a decrease in the growth of Arabidopsis seedlings and chlorosis on the leaf tips of the seedlings were observed. While 1B application caused a 2.9 fold increase in miR393 expression, 3B application caused a 2.7 fold increase in this expression. According to the mature sequences (5p and 3p) at the 5 'and 3' ends of miR393, the main target genes are; Auxin signaling F-box, S-adenosyl-L-methionine-dependent methyltransferases superfamily protein, kinesin-like calmodulin-binding protein, Leucine-rich receptor-like protein kinase family protein, 1-deoxy-D-xylulose 5-phosphate synthase 3, ARM repeat superfamily protein, myb-like HTH transcriptional regulator family protein and bHLH, and WRKY33 transcription factors. On the other hand, according to the Gene Ontology (GO) Enrichment analysis, the main Biological Processes of the genes targeted by miR393 are as follows: The auxin-activated signaling pathway, the cellular response to auxin, various developmental processes, and different cellular responses. Molecular functions can be categorized as auxin binding, inositol hexakisphosphate binding and hormone and alcohol binding. As a result, growth retardation detected under B toxicity may be related to miR393 targeted auxin regulation and associated transcription factors such as bHLH.

Keywords

• Arabidopsis thaliana
• Boron
• Expression
• MiR393
• Toxicity

Orta ve şiddetli seviyelerde bor toksisitesinin Arabidopsis thaliana’da miR393 ekspresyonu üzerine etkisi

Abstract

Bitkilerde korunmuş olan mikroRNA (miRNA) ailelerinden biri olan miR393, birçok kritik gelişimsel süreçlerde rol oynamaktadır. Bu çalışmada, Stem-Loop (SL) kantitatif ters transkripsiyon polimeraz zincir reaksiyonu yöntemi ile 1 mM (1B) ve 3 mM (3B) borik aside maruz kalan Arabidopsis thaliana'da miR393’ün ekspresyon seviyeleri belirlenmiştir. Ayrıca, miR393’ün Arabidopsis thaliana’da hedeflediği genler ve bu genlerin Gen Ontoloji Zenginleştirme analizi yapılmıştır. Arabidopsis thaliana bitkisinde toksik seviyelerde B uygulaması sonrası, Arabidopsis fideciklerinin büyümesinde düşüş ve fideciklerin yaprak uçlarında klorosis gözlemlenmiştir. Arabidopsis thaliana’da 1B uygulaması miR393 ekspresyonu 2,9 kat artırırken, 3B uygulaması 2.7’lik bir artışa sebep olmuştur. miR393’ün 5’ve 3’ ucundaki olgun dizilere göre (5p ve 3p) başlıca hedef genleri; auxin sinyal F-box, S-adenosil-L-metiyonin- bağımlı metiltransferaz süper familya proteini, Kinezin benzeri kalmodulin-bağlanma proteini, Lösin-zengin reseptör benzeri protein kinaz familya proteini, 1- deoksi-D-ksilüloz 5-fosfat sentaz, ARM tekrar süper familya proteini, myb-benzeri HTH transkripsiyonel regülatör familya proteini ve bHLH, ve WRKY33 transkripsiyon faktörleridir. Öte yandan, miR393’ün hedeflediği genlerin Gen Ontoloji (GO) Zenginleştirme analizine göre başlıca Biyolojik Süreçleri şu şekildedir: Oksin ile aktifleşmiş sinyal yolağı, oksin uyarısına hücresel cevap, çeşitli gelişim süreçleri ve farklı hücresel cevaplardır. Moleküler Fonksiyonlar ise, oksin bağlayıcı, inositol heksakisfosfat bağlayıcı ve hormon ve alkol bağlayıcı şeklinde kategorize edilebilir. Sonuç olarak, B toksisitesi altında tespit edilen büyüme geriliği miR393 hedefli oksin regülasyonu ve bHLH gibi ilişkili transkripsiyon faktörleri ile ilgili olabilir.

Keywords

• Arabidopsis thaliana
• Bor
• Ekspresyon
• MiR399
• Toksisite

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