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

Yıl 2017, Cilt: 18 Sayı: 2, 175 - 183, 01.12.2017

Nihal Gören Sağlam

https://doi.org/10.23902/trkjnat.319185

Öz

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. 

Anahtar Kelimeler

Arabidopsis thaliana, GUS, GFP, senesens

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

Öz

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.

Anahtar Kelimeler

Arabidopsis thaliana, GUS, GFP, senescence

Kaynakça

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