Sarımsakta depolama süresince allinaz geni ifadesindeki değişikliğin belirlenmesi
Loading...
Files
Date
2020-02-19
Authors
Erşahin, Rukiye
Journal Title
Journal ISSN
Volume Title
Publisher
Bursa Uludağ Üniversitesi
Abstract
Sarımsak kendine özgü keskin ve karakteristik bir kokuya sahiptir. Sarımsağa kokusunu kükürtlü bileşiklerden allisin vermektedir. Allinaz geninin kodladığı enzim sarımsakta allin öncül kükürtlü bileşiği allisine dönüştürür. Bu yüksek lisans tez çalışmasının amacı, sarımsakta depolama süresince allinaz geni ifadesindeki değişimin Real-Time PCR analizi ile belirlenmesi ve kükürtlü bileşiklerden allin ve allisin içeriği ile karşılaştırılmasıdır. Tez çalışmasında, Kastamonu ve PI515971 sarımsak genotipleri 4°C ve 21°C’de 12 hafta depolanmıştır. Depolama sonunda alınan örneklerden, toplam RNA izolasyonu ve cDNA sentezi yapıldıktan sonra allinaz geni ifadesindeki değişimler aktin geni referans alınarak Real-Time PCR analizleri ile belirlenmiştir. Real-Time PCR analiz sonuçlarına göre, Kastamonu ve PI515971 sarımsak genotiplerinde 4°C ve 21°C’de depolanan örneklerde başlangıca göre göreceli allinaz gen ifade düzeyinde önemli bir artış görülmüştür. Kastamonu genotipinde, depolama öncesi örneklerinde 12,85 olan göreceli gen ifade düzeyi 4°C’de yaklaşık 2,5 kat artarak 32,37 düzeyine, 21°C’de yaklaşık 3 kat artarak 39,15 düzeyine çıkmıştır. 4°C ile 21°C uygulamaları arasında fark istatiksel olarak anlamlı bulunmuştur. PI515971 sarımsak genotipinde ise depolama öncesi örneklerinde göreceli allinaz ifadesi 18,08 iken 4°C’de yaklaşık 5 kat artarak 87,9 düzeyine, 21°C’de 5 kat artarak 88,9 düzeyine çıkmıştır. PI515971 sarımsak genotipinde, 4°C ile 21°C uygulamaları arasında istatistiksel olarak fark bulunmamıştır. Ayrıca allinaz gen ifadesindeki artış ile allin ve allisin miktarındaki artış arasında yüksek pozitif korelasyonlar saptanmıştır. Sonuç olarak allinaz gen ifadesine etki yönünden depolama sıcaklıkları arasında çok fazla fark olmadığı ve 4°C ile 21°C’ de depolamada göreceli allinaz gen ifade düzeyindeki artışın allisin ve allin miktarlarındaki artışla paralel olduğu görülmüştür.
Garlic has a distinctive pungent and characteristic odor. Allicin which is one of major sulfur compounds in garlic gives garlic odor. The enzyme coded by allinase gene coverts alliin precursor to the allicin. The aims of this master thesis were to determine the allinase gene expression during storage in garlic by Real-Time PCR analysis and to compare with alliin and allicin contents. In this thesis, Kastamonu and PI515971 garlic genotypes were stored at 4ºC and 21ºC for 12 weeks. After total RNA isolation and cDNA synthesis from the samples taken at the end of the storage, changes in relative allinase gene expression were determined by Real-Time PCR analysis by using actin reference gene. According to the results of Real-Time PCR analysis, significant increases in the relative allinase gene expression were observed in Kastamonu and PI515971 samples stored at 4ºC ve 21ºC compared to pre storage samples. The relative gene expression level of Kastamonu genotype was 12,85 in pre-storage samples and increased about 2,5 times to 32,37 at 4ºC, and increased approximately 3 times to 39.15 at 21ºC. A significant difference was found between 4ºC and 21ºC treatments. In the PI515971 garlic genotype, the expression was 18,08 in pre-storage samples and it increased about 5 times to 87,9 at 4ºC and increased about 5 times to 88,9 at 21ºC. There is no significant difference between 4ºC and 21ºC treatments in PI515971 garlic genotype. In addition, significant correlations were detected between increase of allinase expression level and increase of alliin and allicin contents. In conclusion, there is no significant difference between storage temperatures in terms of effect to allinase gene expression level and the increase in relative allinase gene expression during storage at 4ºC and 21ºC occurred in parallel with increase in contents of alliin and allicin.
Garlic has a distinctive pungent and characteristic odor. Allicin which is one of major sulfur compounds in garlic gives garlic odor. The enzyme coded by allinase gene coverts alliin precursor to the allicin. The aims of this master thesis were to determine the allinase gene expression during storage in garlic by Real-Time PCR analysis and to compare with alliin and allicin contents. In this thesis, Kastamonu and PI515971 garlic genotypes were stored at 4ºC and 21ºC for 12 weeks. After total RNA isolation and cDNA synthesis from the samples taken at the end of the storage, changes in relative allinase gene expression were determined by Real-Time PCR analysis by using actin reference gene. According to the results of Real-Time PCR analysis, significant increases in the relative allinase gene expression were observed in Kastamonu and PI515971 samples stored at 4ºC ve 21ºC compared to pre storage samples. The relative gene expression level of Kastamonu genotype was 12,85 in pre-storage samples and increased about 2,5 times to 32,37 at 4ºC, and increased approximately 3 times to 39.15 at 21ºC. A significant difference was found between 4ºC and 21ºC treatments. In the PI515971 garlic genotype, the expression was 18,08 in pre-storage samples and it increased about 5 times to 87,9 at 4ºC and increased about 5 times to 88,9 at 21ºC. There is no significant difference between 4ºC and 21ºC treatments in PI515971 garlic genotype. In addition, significant correlations were detected between increase of allinase expression level and increase of alliin and allicin contents. In conclusion, there is no significant difference between storage temperatures in terms of effect to allinase gene expression level and the increase in relative allinase gene expression during storage at 4ºC and 21ºC occurred in parallel with increase in contents of alliin and allicin.
Description
Keywords
Allium sativum L., Allicin, Allisin, Allinaz geni, Gen ifadesi, Real-Time PCR, Alliin, Allinase gene, Gen expression, Allin
Citation
Erşahin, R. (2020). Sarımsakta depolama süresince allinaz geni ifadesindeki değişikliğin belirlenmesi. Yayınlanmamış yüksek lisans tezi. Bursa Uludağ Üniversitesi Fen Bilimleri Enstitüsü.