Oryza sativa Osmyb4 geni aktarılmış transgenik patateste Osmyb4 gen ifadesinin tuzluluk toleransına etkisi

Yıl 2020, Cilt: 35 Sayı: 1, 115 - 123, 14.02.2020

Gülsüm Aydın

https://doi.org/10.7161/omuanajas.624749

Cited By: 1

Öz

Bu çalışmada Osmyb4
geni ile transforme edilmiş patateste, MYB4’ün tuz toleransına olan potansiyel
etkileri araştırılmıştır. Daha önce yapmış olduğumuz bir çalışmada Osmyb4
geni aktarılmış transgenik patates bitkilerinin yüksek konsantrasyonda tuz
içeren ortamda, gen aktarılmamış bitkilerden daha iyi fizyolojik gelişim
gösterdiği tespit edilmiştir. Bu noktadan hareketle MYB4’ün patateste tuz toleransına
etkisini araştırmak amacıyla, transgenik bitkiler (TR) ve gen aktarılmamış
bitkiler (WT) 300 mM tuz stresine tabi tutularak elektrolit salınımı,
malondialdehit (MDA) miktarı, nispi su içeriği (RWC), hidrojen peroksit (H₂O₂)
konsantrasyonu, klorofil içeriği ve prolin miktarı belirlenmiştir. Bunların
yanı sıra literatürde tuz stresi ile ilişkili olduğu belirtilen NAC072,
NAC024, CDPK4 ve P5CS genlerinin ekspresyonu gerçek zamanlı
kantitatif PCR (qRT PCR) aracılığı ile incelenmiştir. Biyokimyasal analizler
sonucunda elde edilen veriler elektrolit salınımı, prolin miktarı ve H₂O₂
miktarı bakımından WT bitkiler ile TR bitkiler arasında anlamlı bir farklılık
olmadığını ortaya koymuştur. RWC transgenik S2 hattında transgenik olmayan
bitkilere kıyasla anlamlı oranda (P < 0.05) yüksek bulunmuştur. MDA miktarı
her iki transgenik hatta (S2 ve M48) WT bitkilere kıyasla anlamlı oranda düşük
bulunmuştur. qPCR analizi sonucunda elde edilen relatif ekspresyon değerleri,
S2’de NAC072 geninin ekspresyon seviyesinin WT ile karşılaştırıldığında
anlamlı olarak (P < 0.05) yüksek olduğunu göstermiştir. NAC024 ve P5CS
genlerinin ekspresyon seviyeleri de transgenik bitkilerde transgenik
olmayanlara oranla yüksek bulunmuştur. CDPK4 geninin ekspresyon seviyesi
ise S2 bitkileri ile WT bitkilerde birbirine yakın seviyede tespit edilmiştir.
Elde edilen veriler bir bütün olarak değerlendirildiğinde MYB4 transkripsiyon
faktörünün RWC, MDA miktarı ve stresle ilişkili çeşitli genlerin ekspresyon
seviyesini etkilemek suretiyle tuz stresi tolerans mekanizmasında rol
oynayabileceğine işaret etmiştir. 

Anahtar Kelimeler

Abiyotik stres, MYB4, patates, transgenik, tuzluluk

Destekleyen Kurum

Selçuk Üniversitesi

Proje Numarası

16401046

Teşekkür

Bu çalışma Selçuk Üniversitesi Bilimsel Araştırmalar Koordinatörlüğü tarafından 16401046 nolu proje ile desteklenmiştir.

Effect of Osmyb4 gene expression on salinity tolerance of potato transformed with Oryza sativa Osmyb4 gene

Öz

In this study, potential involvement of MYB4 in
salt tolerance of potato trasformed with Osmyb4 was investigated. In our
previous studies transgenic potato plants heterologously expressing Osmyb4
gene displayed better growth compared to non-transgenic plants upon exposure to
high salt concentrations. These results have led to a detailed analysis of
potential involvement of MYB4 in salt tolerance of potato. For this purpose
transgenic (TR) and non-transgenic (WT) plants were subjected to 300 mM salt
concentrations and electrolyte leakage, malonedialdehyde level (MDA), relative
water content (RWC), the amount of hydrogen peroxide (H₂O₂),
chlorophyll content and the level of proline was identified. The expression of
certain genes reported to be involved in salinity tolerance, namely, NAC072,
NAC024, CDPK4 and P5CS was also investigated by
quantitative real time PCR (qRT-PCR). The data obtained through biochemical
tests showed that there were no significant difference between WT and TR plants
with respect to electrolyte leakage, proline content and H₂O₂
content. RWC was significantly higher (P <0.05) in the transgenic line S2
compared to TR plants. MDA content in both transgenic lines (S2 and M48) were
found to be significantly lower compared to WT plants. Relative expression data
obtained by qPCR analysis revealed that expression of NAC072 was
significantly higher in S2 compared to WT (P < 0.05). Expression level of NAC024
and P5CS genes were also higher in transgenic plants compared to
non-transgenic WT. On the other hand expression level of CDPK4 was
similar in WT and transgenic lines. The results have indicated that MYB4
transcription factor may regulate salt stress tolerance mechanism in potato by
affecting RWC, MDA content and expression of certain stress related genes.

Anahtar Kelimeler

Abiotic stress, MYB4, Potato, Salinity, Transgenic

Proje Numarası

16401046

Kaynakça

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