Using The Allium Test of 4-Methyl Umbelliferone: Physiological, Cytogenetic, Biochemical and Anatomical Evaluation

Abstract

In this study, the dose dependent effects of exogenous 4-methyl umbelliferone (4-MU) on various physiological, cytogenetic, anatomical and biochemical parameters using Allium cepa L. as test material were researched. The physiological parameters examinated were germination percentage, root length, rootlet number and fresh weight; the cytogenetic parameters were micronucleus (MN) frequency, chromosome aberration (CA) and mitotic index (MI); the biochemical parameters were free proline content, malondialdehyde (MDA) level, catalase (CAT) and superoxide dismutase (SOD) activities. And the structural changes in root tip cells were investigated with anatomical sections. Structural changes in onion root tip cells were examined by taking anatomical sections. For these purposes, bulbs were divided into four groups as one control and three applications. The bulbs in the control group were treated with distilled water; the bulbs in the application groups were treated with 125 µM, 250 µM and 500 µM doses of 4-MU for 7 days. 4-MU application caused a decrease in the physiological parameters compared to the control group. This treatment created an increase in the frequency of MN and CA, and a reduce in the MI. In addition, 4-MU application induced an increase as dose-dependent in CAT and SOD activities and MDA and free proline contents compared to the control group. Moreover, after all 4-MU application, anatomical changes such as MN formation in epidermis cells, epidermis and cortex cell deformations, accumulation of some chemical compounds in cortex, unclear transmission tissue and necrose were identified and these root anatomical changes were found to reach maximum levels at 500 µM 4-MU. As a result, 4-MU had a negative effect on all the investigated parameters and it was determined that the Allium test material could be a useful bioindicator for monitoring these effects.

Keywords

• Allium cepa L.
• Antioxidant enzymes
• Germination
• Lipid peroxidation
• 4-methyl umbelliferone
• Cytogenetic

4-Metil Umbelliferon’un Allium Testi Kullanılarak; Fizyolojik, Sitogenetik, Biyokimyasal ve Anatomik Değerlendirilmesi

Abstract

Bu çalışmada, test materyali olarak Allium cepa L. kullanılarak çeşitli fizyolojik, sitogenetik, anatomik ve biyokimyasal parametreler üzerine dışsal 4-metil umbelliferonun (4-MU) doza bağlı etkileri araştırıldı. İncelenen fizyolojik parametreler: çimlenme yüzdesi, kök uzunluğu, kökçük sayısı ve taze ağırlık; sitogenetik parametreler: mikronükleus (MN) sıklığı, kromozom anormalliği (KA) ve mitotik indeks (Mİ); biyokimyasal parametreler: serbest prolin içeriği, malondialdehit (MDA) düzeyi, katalaz (KAT) ve süperoksit dismutaz (SOD) aktiviteleridir. Soğan kök ucu hücrelerinde meydana gelen yapısal değişimler anatomik kesitler alınarak incelendi. Bu amaçlar için, soğanlar bir kontrol ve üç uygulama olmak üzere dört gruba ayrıldı. 7 gün süresince kontrol grubundaki soğanlar distile su ile muamele edilirken; uygulama gruplarındaki soğanlar 4-MU’nun 125 µM, 250 µM ve 500 µM dozları ile muamele edildi. 4-MU uygulaması kontrol grubuyla kıyaslandığında fizyolojik parametrelerde bir azalmaya neden oldu. Bu uygulama, KA ve MN sıklığında bir artış ile Mİ'de bir azalma meydana getirdi. Ayrıca, 4-MU muamelesi kontrol grubuyla karşılaştırıldığında KAT ve SOD aktiviteleri ile MDA ve serbest prolin içeriklerinde de doza bağlı olarak bir artışa neden oldu. Ayrıca, tüm 4-MU uygulamaları sonrasında, epidermis hücrelerinde MN oluşumu, epidermis ile korteks hücre deformasyonları, kortekste bazı kimyasal bileşiklerin birikmesi, belirgin olmayan iletim dokusu ve nekroz gibi kök anatomik değişimleri tespit edildi ve bu anatomik değişimlerin 500 µM 4-MU'da maksimum seviyelere ulaştığı bulundu. Sonuç olarak, 4-MU incelenen tüm parametreler üzerinde olumsuz bir etki gösterdi ve bu etkilerin izlenmesi için Allium test materyalinin yararlı bir biyoindikatör olabileceği tespit edildi.

Keywords

• Allium cepa L.
• Antioksidan enzimler
• Çimlenme
• Lipid peroksidasyonu
• 4-metil umbelliferon
• Sitogenetik

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