Diterpenes biochemical profile and transcriptional analysis of cytochrome P450s genes in leaves, roots, flowers, and during Coffea arabica L. fruit development

Abstract

Lipids are among the major chemical compounds present in coffee beans, and they affect the flavor and aroma of the coffee beverage. Coffee oil is rich in kaurene diterpene compounds, mainly cafestol (CAF) and kahweol (KAH), which are related to plant defense mechanisms and to nutraceutical and sensorial beverage characteristics. Despite their importance, the final steps of coffee diterpenes biosynthesis remain unknown. To understand the molecular basis of coffee diterpenes biosynthesis, we report the content dynamics of CAF and KAH in several Coffea arabica tissues and the transcriptional analysis of cytochrome P450 genes (P450). We measured CAF and KAH concentrations in leaves, roots, flower buds, flowers and fruit tissues at seven developmental stages (30-240 days after flowering - DAF) using HPLC. Higher CAF levels were detected in flower buds and flowers when compared to fruits. In contrast, KAH concentration increased along fruit development, peaking at 120 DAF. We did not detect CAF or KAH in leaves, and higher amounts of KAH than CAF were detected in roots. Using P450 candidate genes from a coffee EST database, we performed RT-qPCR transcriptional analysis of leaves, flowers and fruits at three developmental stages (90, 120 and 150 DAF). Three P450 genes (CaCYP76C4, CaCYP82C2 and CaCYP74A1) had transcriptional patterns similar to CAF concentration and two P450 genes (CaCYP71A25 and CaCYP701A3) have transcript accumulation similar to KAH concentration. These data warrant further investigation of these P450s as potential candidate genes involved in the final stages of the CAF and KAH biosynthetic pathways.

Technical Report on the notification of infusion from coffee leaves (Coffea arabica L. and/or Coffea canephora Pierre ex A. Froehner) as a traditional food from a third country pursuant to Article 14 of Regulation (EU) 2015/2283

“…The presence of the diterpene cafestol in C. arabica leaves (45 mg/100g fresh weight, via HPLC analysis) was reported by Dias et al (2010), whereas the diterpene kahweol was not found in leaves of C. arabica (Dias et al, 2010;Ivamoto et al, 2017). Cafestol and kahweol were not detected in C. robusta leaves (Dias et al, 2010).…”

Metabolomics fingerprint of coffee species determined by untargeted-profiling study using LC-HRMS

“…As mentioned before, the literature is rich in coffee beans (green or roasted) phytochemical analyses but the composition of the coffee leaves is less described. Beverage quality of coffee bean extracts is of course highly related to the chemical compounds (Ivamoto et al, 2017). In addition to caffeine, other components of coffee beans like primary and secondary metabolites are important (Ivamoto et al, 2017).…”

“…Beverage quality of coffee bean extracts is of course highly related to the chemical compounds (Ivamoto et al, 2017). In addition to caffeine, other components of coffee beans like primary and secondary metabolites are important (Ivamoto et al, 2017). A complex combination of these chemicals determines beverage sensory characteristics.…”

“…Beverage quality is highly related to the chemical compounds in coffee beans. A complex combination of these chemicals determines all beverages or foods sensory characteristics (Ivamoto et al, 2017). Monitoring food quality is crucial and has to be a major concern in order to maintain and improve the standard of life.…”

An integrated analysis of mRNA and sRNA transcriptional profiles in Coffea arabica L. roots: insights on nitrogen starvation responses

“…With the development of functional genomics, large-scale transcriptome analysis based on expressed sequence tag (EST) sequencing, microarray hybridization, and RT-qPCR analysis of key genes have been extensively used to investigate the molecular basis of several features in Arabica coffee during fruit development stages (Joët et al 2009(Joët et al , 2014Ivamoto et al 2017a;Ivamoto et al 2017b), responses to abiotic (Marraccini et al 2012), and biotic (Cardoso et al 2014) stresses. More recently, next-generation sequencing technologies for quantitative determination of RNA levels (RNA-seq) have been incorporated into studies on the coffee transcriptome.…”