RT-PCR amplification procedures of mRNA for the detection of gene expression in compost

The deliberately or accidental release of volatile organic compounds (VOCs) leads to serious environmental damage. One particular concern is the air contamination by the emissions into the atmosphere of solvents used during the varnishing of unfinished items of the wood-working industry. Microorganisms have evolved different pathways to utilise as carbon source each compounds present on the complex mixtures. The objective of the present study was to show a method to predict the expression of catabolic gene of degradative microorganisms in organic matter of a pre-adapted compost. In particular, the aim of this study was to optimize a qualitative procedure of co-extraction of purified DNA/RNA from complex organic matter and to demonstrate its applications to detect xylene monooxygenase gene expression by RT-PCR. A DNA/RNA extraction protocol based on combination of sample lyophilization pre-treatment and cetyl-trimethylammonium bromide (CTAB) - phenol/chloroform extraction procedure was optimized for the recovery of purified nucleic acids. PCR and RT-PCR protocols were established to detect xylene monooxygenase gene expression starting from differentially induced organic matrices obtained by VOC(s) biofiltration technology. This work standardized a methodology to follow microbial degradation activities into heterogeneous organic solid media and offers a quick method to follow specific biological activities during solid and/or semisolid organic substrates biotransformations.