Separation and Identification of Volatile Compounds from Liquid Cultures of Trichoderma harzianum by GC-MS using Three Different Capillary Columns

Abstract

A simple, fast, repeatable and less laborious sample preparation protocol was developed and applied for the analysis of biocontrol fungus Trichoderma harzianum strain FA1132 by using gas chromatography-mass spectrometry. The match factors for sample spectra with respect to the mass spectra library of fungal volatile compounds were determined and used to study the complex hydrocarbons and other volatile compounds, which were separated by using different capillary columns with nonpolar, medium polar and high polar stationary phases. To date, more than 278 volatile compounds (with spectral match factor at least 90%) such as normal saturated hydrocarbons (C7-C30), cyclohexane, cyclopentane, fatty acids, alcohols, esters, sulfur-containing compounds, simple pyrane and benzene derivatives have been identified. Most of these compounds have not previously been reported. The method described in this paper is a more convenient research tool for the detection of volatile compounds from the cultures of T. harzianum.

Formation of 6-n-pentyl-2H-pyran-2-one (6-PAP) and other volatiles by different Trichoderma species

“…The finding that T. harzianum was incapable of 6-PAP formation is not consistent with several earlier studies (Claydon et al 1987;Serrano-Carreon et al 1992;Scarselletti and Faull 1994;Bonnarme et al 1997;Whitaker et al 1998;El-Hasan et al 2007;Vinale et al 2008, Souza Ramos et al 2008Siddiquee et al 2012). However, in the opinion of Samuels (2006), Dodd et al (2003), and Polizzi et al (2011), T. harzianum strains used in mentioned works were misclassified strains of T. atroviride and T. viride.…”

“…harzianum was analyzed here on PDA medium, whereas in the other studies, T. harzianum for 6-PAP production was cultivated in potato dextrose broth (PDB) or malt extract agar (MEA) medium (Scarselletti and Faull 1994;El-Hasan et al 2007;Siddiquee et al 2009Siddiquee et al , 2012. Therefore, the inability of T. harzianum to produce 6-PAP could also be due to the difference in substrate composition used in present study.…”

“…6-PAP was detected in the headspace of all T. atroviride and T. viridescens isolates, as well as in individual cultures of T. citrinoviride (4 isolates), T. hamatum (8 isolates), and T. viride (3 isolates). Previously, these compounds had been described as being produced by T. viride (Collins and Halim 1972;Bonnarme et al 1997), T. harzianum (Claydon et al 1987;Serrano-Carreon et al 1992;Scarselletti and Faull 1994;Bonnarme et al 1997;Whitaker et al 1998;El-Hasan et al 2007, Souza Ramos et al 2008Vinale et al 2008;Siddiquee et al 2012), T. koningii (Cutler et al 1986;Simon et al 1988), and T. atroviride (Reithner et al 2005(Reithner et al , 2007Stoppacher et al 2010;Polizzi et al 2011).…”

“…These compounds, with the exception of 2-heptanone, were also found in several cultures of T. atroviride , T. citrinoviride , T. hamatum , T. harzianum , T. koningii , T. viride , T. viridescens , and T. virens (Table 3). Previously, 2-heptanone was reported for T. viride (isolate T60), T. atroviride , and T. harzianum (Wheatley et al 1997;Fiedler et al 2001;Stoppacher et al 2010;Polizzi et al 2011;Siddiquee et al 2012). 2-nonanone has been described to be produced by T. aureoviride , T. atroviride , and T. harzianum (Bruce et al 2000;Stoppacher et al 2010;Polizzi et al 2011;Siddiquee et al 2012).…”

“…Previously, 2-heptanone was reported for T. viride (isolate T60), T. atroviride , and T. harzianum (Wheatley et al 1997;Fiedler et al 2001;Stoppacher et al 2010;Polizzi et al 2011;Siddiquee et al 2012). 2-nonanone has been described to be produced by T. aureoviride , T. atroviride , and T. harzianum (Bruce et al 2000;Stoppacher et al 2010;Polizzi et al 2011;Siddiquee et al 2012). 1-octen-3-ol and 3-octanone have been found before in culture samples of T. harzianum and T. atroviride (Fiedler et al 2001;Nemčovič et al 2008;Stoppacher et al 2010;Polizzi et al 2011;Siddiquee et al 2012).…”