Biochemical and molecular tolerance of Carpobrotus acinaciformis L. halophyte plants exposed to high level of NaCl stress

Year 2019, Volume: 23 Issue: 1, 99 - 107, 25.03.2019

Sema Karakaş Dikilitaş , Murat Dikilitaş , Rukiye Tıpırdamaz

https://doi.org/10.29050/harranziraat.464133

Cited By: 2

Abstract

Carpobrotus acinaciformis L. plant is a kind of halophyte that is able
to survive in high salt conditions. It is important to determine its
physiological, biochemical and molecular limit of NaCl stress if one aims to
use it for phytoremediation purpose. In this study, the alkaline protocol of
the modified plant comet assay were used for rapid detection of DNA damage in
C. acinaciformis L. plants exposed to a series of  NaCl stress concentrations (0-, 50-, 100-,
200-, 300-, 400 and 500 mmol L^-1) in hydroponic conditions for 2
weeks. DNA damage was measured as the values of percentage of DNA in tails and
tail length. The halophyte C. acinaciformis L. did not show any dose response
up to 400 mmol L^-1 NaCl level in terms of DNA damages. DNA integrity
measured via comet assay showed that DNA preserved its original shape up to 400
mmol L^-1 NaCl level. However, the very high concentrations of NaCl
(400 and 500 mmol L^-1) caused DNA damages.  When physiological and biochemical parameters
such as proline, chlorophyll a and b, peroxidase (POX), catalase (CAT), H₂O₂,
malondialdehyde (MDA) contents were examined, oxidant molecules such as H₂O₂
(0.912-3.72 µmol g^-1 Fwt) and MDA (7.1-34 nmol g^-1 Fwt)
gradually increased along with the increase of NaCl concentrations, p<0.05.
On the other hand, antioxidant enzyme POX and an osmolyte molecule proline
slightly increased up to 400 mmol L^-1 NaCl level then slightly
decreased after that. Similar issues were obtained from that of protease enzyme
which indicates the power of protein hydrolysis in which a slight decrease
(182-95 Unit mg^-1 protein) over a dose of NaCl was evident.
Chlorophyll contents and CAT activity were not affected upon increase of NaCl
concentrations. This study showed that the halophyte C. acinaciformis L. can be
easily used to remove salt up to 400 mmol L^-1 NaCl concentrations
from a saline-affected soil. Measuring DNA damage is concluded as a very useful
parameter to find out what level of NaCl could be tolerated if a halophyte
plant is aimed to remediate the saline soils.

Keywords

Halophyte, NaCl stress, SCGE, Comet assay, DNA damage

NaCl stresine maruz bırakılan Carpobrotus acinaciformis L. halofit bitkisinin biyokimyasal ve moleküler tepkileri

Abstract

Carpobrotus acinaciformis L. bitkisi yüksek tuz koşullarında yaşayabilen bir
çeşit halofit bitkidir. Bu bitki fitoremediasyon
çalışmaları için kullanılmak üzere planlandığında, bu bitkinin tuz stresine
karşı fizyolojik, biyokimyasal ve moleküler sınırlarını belirlemek önem arz
etmektedir. C.
acinaciformis L.
bitkisinde DNA hasar seviyesini belirlemek için hidroponik koşullarda 2 hafta
süre ile tuz stresine (0-, 50-, 100-,
200-, 300-, 400 and 500 mmol L^-1)
maruz bırakılan bitkilerde modifiye edilmiş alkali bitki comet assay metodu
kullanılmıştır. DNA hasarı kuyruk uzunluğu ve kuyrukta DNA yüzdesi olarak ölçülmüştür.
Halofit C.
acinaciformis L 400 mmol L^-1 NaCl
seviyesine kadar DNA hasarı ile ilgili olarak doz tepkisi göstermemiştir. Comet
assay ile ölçülen yönteme göre halofit bitkilerin DNA bütünlüğünü 400 mmol
L^-1 NaCl  seviyesine kadar korunduğu gözlenmiştir. Fakat,
çok daha yüksek NaCl konsantrasyonları (400 ve 500 mmol L^-1)
DNA hasarına yol açmıştır. Prolin, klorofil a
ve b, peroksidaz (POX), katalaz
(CAT), H₂O₂, malondialdehid (MDA) içerikleri gibi
fizyolojik ve biyokimyasal parametreler incelendiğinde, oksidant moleküllerden
H₂O₂ (0.912-3.72 µmol g^-1 taze ağırlık)and
MDA (7.1-34 nmol g^-1 taze ağırlık) artan tuz konsantrasyonu ile
paralel olarak sıralı artış göstermiştir, p<0.05. Diğer yandan, antioksidant enzimlerden POX ve bir osmolit olan prolin 400
mmol L^-1 NaCl’ e kadar
hafifçe artış göstermiş daha sonra tekrar düşmüştür. Benzer durumlar protein
hidrolizini belirlemede kullanılan proteaz enzim (182-95 Unit mg^-1
protein) seviyesinde de görülmüş, artan
NaCl dozuna bağlı olarak enzim miktarı kademeli olarak azalmıştır. Klorofil
miktarı ve CAT enzim seviyesi NaCI konsantrasyon artışına bağlı olarak değişim
göstermemiştir. Bu çalışma, C. acinaciformis L. bitkisinin tuzdan
etkilenmiş topraklarda 400 mmol L^-1 NaCl’ e kadar olan tuz konsantrasyonunu
uzaklaştırmada rahatlıkla kullanılabileceğini ortaya koymuştur. DNA hasarını
ölçmek, tuzlu alanları ıslah etmede kullanılacak halofit bitkinin hangi
seviyede NaCl stresine dayanabileceğini belirlemede çok kullanışlı bir
parametre olarak kaydedilmiştir.

Keywords

NaCl stresi, Comet assay, SCGE, DNA hasarı

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