Gene Cloning and Gene Expression Characteristics of Alcohol Dehydrogenase in Osmanthus fragrans var. semperflorens

Dangquan Zhang

doi:10.9755/ejfa.2018.v30.i10.1825

Abstract

Because of a lot of aroma components in the flowers, Osmanthus fragrans is widely used as ornamental tree species and its flowers is used as materials of spices and food additives. Alcohol dehydrogenase (ADH) plays an important regulation role in growth, development, stress resistance and aroma synthesis of plants. Using the flowers of O fragrans var. semperflorens as materials, the full-length cDNA of ADH gene (termed OfADH) was successfully cloned through rapid amplification of cDNA ends (RACE), which has the 1128 bp open reading frame (ORF) in length and codes 375 amino acids. The results of conserved domain analysis show that the OfADH belongs to the zinc-dependent medium-chain dehydrogenase /reductase (MDR) superfamily, contains 22 NAD(P) binding sites in the liver_alcohol_DH_like conserved domains, and also has the PLN02740 (Alcohol dehydrogenase-like) domain. The results of semi-quantitative RT-PCR indicate that the expression level of OfADH in O. fragrans flowers is consistent with the rule of aroma formation and release, while the expression of OfADH in leaves is positively correlated with high temperature stress, suggesting that OfADH has obvious functional diversity in regulation of aroma synthesis and response to high temperature stress.

Highlights

Alcohol dehydrogenase (ADH), a zinc metalloprotease, existing in all kinds of organs and cells of plants, animal and microorganisms widely (Jacquelyn LM et al, 2013; Kluver N et al, 2014), is one of the rates limiting enzyme of short chain alcohols metabolism in organism (Cheung et al, 2017; Naser and Lawrey, 2018)
In the presence of NAD+ and NADH, ADH is able to catalyze a reversible reaction between primary alcohols and aldehydes, between ethanol and aldehydes (Gottlieb et al, 1982)
In the fatty acid derivatives metabolic pathway of apple aroma (Robert, et al, 2007), hexanal transformed by linoleic acid hydroperoxide could generate 2-methylbutanol which can produce 2-isoamyl acetate by the alcohol acyltransferase in presence of ADH (Dudareva et al, 2008)

Summary

INTRODUCTION

Alcohol dehydrogenase (ADH), a zinc metalloprotease, existing in all kinds of organs and cells of plants, animal and microorganisms widely (Jacquelyn LM et al, 2013; Kluver N et al, 2014), is one of the rates limiting enzyme of short chain alcohols metabolism in organism (Cheung et al, 2017; Naser and Lawrey, 2018). ADH plays an important role in the growth and development process of humans, animals and plants. It exhibits obvious biological functional diversity (Alpat et al, 2010). Studies have indicated that alcohol dehydrogenase played an important role in the formation and diversity of plant aroma (Kmita et al, 2018; Liu and Liu, 2010; Shen et al, 2017). In the mature process of fruits, the expression level of ADH has important influence on the diversity of aroma components, especially the content of aldehydes and alcohols (Iyit, 2018; Jiang et al, 2018; Khan et al, 2018). This study will provide the basis for exploring the regulation mechanism and biological mechanism of ADH gene in aroma synthesis of O. fragrans var. semperflorens

MATERIALS AND METHODS

Methods

RESULTS AND ANALYSIS