Arabidopsis thaliana (L.) Heynh. YAPRAKLARINDA GUS VE GFP RAPORTÖR GENLERİ KULLANILARAK SENESENS İLE İLİŞKİLİ GENLERİN ANALİZİ

Abstract

Yaprak senesensinin düzenlenme ve işleyiş yapısı birçok biyokimyasal ve düzenleyici yolağın aktivasyonunu içeren kompleks bir olaydır. Daha önce yapılan çalışmalar sonucunda, yaprak senesensinin moleküler mekanizmasını anlamak için senesens ile ilişkili bazı genler (SAGs=Senescence Associated Genes) belirlenmiştir. Bu çalışma ANAC018, ANAC019, MYB75 (PAP1) ve MYB2 genlerinin senesens ile ilişkili olabilme potansiyelini araştırmak için planlanmıştır. Bu amaçla GUS (β-glukuronidaz) ve GFP (Yeşil Fluoresans Protein) işaretleyicileri ile ANAC018, ANAC019, MYB75 (PAP1) ve MYB2 promotör genlerini içeren Arabidopsis thaliana (L.) Heynh. (ekotip Columbia) bitkisi kullanılmıştır. GUS ve GFP analizinde promotörün ne zaman ve hangi hücre ya da hücre tiplerinde aktif olduğuna bakılmıştır. Karanlığın teşvik ettiği senesens modeli oluşturulmuş ve GUS boyama ve konfokal mikroskopi ile genlerin aktif olduğu zaman ve hücre tipleri belirlenmiştir. Elde edilen veriler ışığında ANAC018, ANAC019, MYB75 (PAP1)ve MYB2 genlerinin senesens sırasında, epidermis, epidermal hücrelerin nükleusları, mezofil ve stoma hücrelerinde aktif oldukları belirlenerek bu genlerin yaprak senesensinde rol oynadıkları ortaya konulmuştur. 

Keywords

• Arabidopsis thaliana,GUS,GFP,senesens

Analysis of Senescence-Related Genes Expressions Using Gus and Gfp Reporter Genes in Arabidopsis thaliana (L.) Heynh. Leaves

Abstract

The regulation and functional steps of leaf senescence is a complex phenomenon involving the activation of many biochemical and regulatory pathways. As a result of previous studies, some genes related to senescence (SAGs = Senescence Associated Genes) have been identified to understand the molecular mechanism of leaf senescence. This study was designed to investigate the association potential of ANAC018, ANAC019, MYB75 (PAP1) and MYB2 genes with senescence. For this purpose, GUS (β-glucuronidase) and GFP (Green Fluorescent Protein) markers and Arabidopsis thaliana (L.) Heynh. (ecotype Columbia) plants containing ANAC018, ANAC019, MYB75 (PAP1), MYB2 promoter genes were used. GUS and GFP analyses were used to determine when the promoter is active in which cells or cell types. Dark-induced senescence model was designed and the time and the cell types where the genes were active were determined by using GUS staining and confocal microscopy. The results showed that ANAC018, ANAC019, MYB75 (PAP1) and MYB2 genes were active in epidermis, nucleus of epidermal cells, mesophyll and stomata cells during senescence and these genes were found to play a role in leaf senescence.

Keywords

• Arabidopsis thaliana,GUS,GFP,senescence

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