Isolation of ascorbate peroxidase (APX) gene in lentil (Lens culinaris Medik.) and expression analysis under drought stress conditions

Year 2022, Volume: 59 Issue: 3, 439 - 447, 30.09.2022

Melike Bakır Cebrail Yıldırım

https://doi.org/10.20289/zfdergi.1007041

Abstract

Objective: The objective of this study was to isolate partial cDNA that belongs to the ascorbate peroxidase (APX) gene of lentil (Lens culinaris Medik.) and to express LcAPX gene in lentil seedlings under drought stress conditions.
Material and Methods: To identify the relationships between drought stress and LcAPX gene expression, lentil seedlings grown for 2 weeks were subjected to drought stress through not irrigating for 6, 13, and 20 days. Effects of drought stress were determined by measuring the stem relative water content (RWC). Gene expression changes in lentil seedlings were determined with real-time RT-qPCR.
Results: The LcAPX gene expression levels of both drought-tolerant Firat-87 and drought-sensitive Ozbek cultivars varied with the severity of drought stress. The gene expression of LcAPX reached the highest level in Firat-87 cultivar on the 6th day, whereas a significant increase was observed only on the 20th day of the Ozbek cultivar, and this increase was relatively low as compared to the Fırat-87 cultivar.
Conclusion: From the study conducted, it was concluded that time-dependent changes of the expression of LcAPX gene indicates that LcAPX gene had a highly specific gene expression profile and complex regulation in lentil drought response.

Keywords

Ascorbate peroxidase, drought stress, gene expression, lentil, RT-qPCR

Supporting Institution

Erciyes Üniversitesi

Project Number

6684

Thanks

This study was supported by Erciyes University, Scientific Research Projects Unit (Project no: 6684). Thanks are extended to Betül-Ziya Eren Genome and Stem Cell Center of Erciyes University for their supports provided throughout the experiments.

Isolation of ascorbate peroxidase (APX) gene in lentil (Lens culinaris Medik.) and expression analysis under drought stress conditions

Abstract

Amaç: Bu çalışmada, mercimekte (Lens culinaris Medik.) askorbat peroksidaz (APX) geninin partial cDNA klonu izole edilmiş ve LcAPX geninin kuraklık stresi koşullarında mercimek fidelerinde değişen gen ifadesi seviyesi belirlenmiştir.
Materyal ve Yöntem: Kuraklık stresi ve LcAPX gen ifadesi arasındaki ilişkiyi anlamak için, 2 hafta süre ile yetiştirilen mercimek fidelerine 6, 13 ve 20 gün süre ile sulamama şeklinde kuraklık stresi uygulanmıştır. Kuraklık stresinin etkileri, sap nispi nem içeriği (RWC) ölçülerek belirlenmiştir. Mercimek fidelerinde meydana gelen gen ifadesi değişimleri eş zamanlı kantitatif PCR (Real-time qPCR) ile belirlenmiştir.
Araştırma Bulguları: Hem kuraklığa dayanıklı Fırat-87 hem de kuraklığa duyarlı Özbek çeşitlerinin LcAPX gen ekspresyon seviyeleri, kuraklık stresinin şiddetine göre değişiklik göstermiştir. LcAPX gen ekspresyonu Fırat-87 çeşidinde 6. günde en yüksek seviyeye ulaşırken, Özbek çeşidinin sadece 20. gününde önemli bir artış gözlenmiş ve bu artış Fırat-87 çeşidine göre nispeten düşük kalmıştır.
Sonuç: Sonuç olarak, LcAPX geninin ekspresyonunun gün bazında değişmesi, LcAPX geninin mercimeğin kuraklığa tepkisinde oldukça spesifik bir gen ekspresyon profiline ve karmaşık bir regülasyona sahip olduğunu göstermektedir.

Keywords

Askorbat peroksidaz, kuraklık stresi, gen ekspresyonu, mercimek, RT-qPCR

Project Number

6684

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