Chromatographic Methods for Metabolite Profiling of Virus- and Phytoplasma-Infected Plants of Echinacea purpurea

Abstract

This study was focused on the effects of virus and phytoplasma infections on the production of Echinacea purpurea (L.) Moench secondary metabolites, such as caffeic acid derivatives, alkamides, and essential oil. The identification of caffeic acid derivatives and alkamides was carried out by means of high-performance liquid chromatography-diode array detection (HPLC-DAD), HPLC-electrospray ionization-mass spectrometry (ESI-MS), and MS(2). Quantitative analysis of these compounds was carried out using HPLC-DAD. The results indicated that the presence of the two pathogens significantly decreases (P < 0.05) the content of cichoric acid, the main caffeic acid derivative. Regarding the main alkamide, dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamide, a significant decrease (P < 0.05) in the content of this secondary metabolite was observed in virus-infected plants in comparison with healthy plants, while in the phytoplasma-infected sample the variation of this secondary metabolite was not appreciable. The % relative area of the E/Z isomers of this alkamide was also found to change in infected samples. The gas chromatography (GC) and GC-MS analysis of E. purpurea essential oil enabled the identification of 30 compounds. The main significant differences (P < 0.05) in the semiquantitative composition were observed for three components: limonene, cis-verbenol, and verbenone. The results indicate that the presence of virus and phytoplasma has an appreciable influence on the content of E. purpurea secondary metabolites, which is an important issue in defining the commercial quality, market value, and therapeutic efficacy of this herbal drug.

Purple coneflower viruses: species diversity and harmfulness

“…The petals showed contractions, deformations and mottling [10,11]. In 2009, CMV-infected plants showed stunting; leaves with yellow mosaics, ring, line patterns and malformations; small fl owers with pale stripes on red petals [12]. Mo reover, Italian scientists have studied genetic modifi cations of the CMV isolates, taken from aromatic, offi cinal and ornamental plants in the Northern Italy.…”

Chicoric acid: chemistry, distribution, and production

“…A HPLC method (INA-Institute for Nutraceutical Advancement method 106.000) from NSF (National Science Foundation) International (Ann Arbor, MI, USA) is accessible as well. Clear chicoric acid mass spectra examples can be found in Shiga et al (2009) and Mulinacci et al (2001); mother and fragmented masses have been reported in numerous papers with various MS settings, so one of these references can be used for comparison with a comparable analysis (Mulinacci et al, 2001; Baur et al, 2004; Lee and Scagel, 2009; Shiga et al, 2009; Pellati et al, 2011; Ribas-Agusti et al, 2011; Carazzone et al, 2013). …”

“…Pathogens can have a negative impact on plant growth but may increase phenolic accumulation. For example pathogen attack (cucumber mosaic virus and phytoplasma-prokaryote) decreased chicoric acid levels in E. purpurea roots (Pellati et al, 2011). Foliar application of carboymethyl chitin glucan (a fungal elicitor produced by Penicillium ) increased production of chicoric acid in roots of E. purpurea (Hudec et al, 2007).…”

Chemical composition of acid lime leaves infected with &lt;i&gt;Candidatus&lt;/i&gt; Phytoplasma aurantifolia

“…Other compounds that increased in infected leaves were β-ocimene (I = 3.463%, H = 2.304%), trans-caryophyllene (I = 1.828%, H = 0.853%), cis-geraniol (Nerol) (I = 0.887%, H= 0.330%), and β-myrcene (I = 0.670%, H = 0.434%). Phytoplasma infection is known to change the cell metabolism leading to an increase of certain compounds and the reduction of others [4,[25][26][27].…”