In vitro co-culture of Solanum tuberosum hairy roots with Meloidogyne chitwoodi: structure, growth and production of volatiles

Abstract

Meloidogyne spp., commonly known as root- knot nematodes (RKNs), are economically important plant sedentary endoparasites that cause galls on susceptible hosts. The Columbia root-knot nematode (CRKN), M. chitwoodi, is a quarantine A2 type pest by the European and Mediterranean Plant Protection Organization since 1998. This nematode has been found associated with economi- cally important crops such as potato and tomato, causing severe damage and making the agricultural products unac- ceptable for the fresh market and food processing. In vitro co-culture of host and parasite offers an advantageous experimental system for studying plant-RKN interactions. The structure, growth and production of volatiles of Sola- num tuberosum hairy roots (HR) and of S. tuberosum HR/ CRKN co-cultures were compared. HR were induced by inoculation of aseptic potato tuber segments with Rhizo- bium rhizogenes. Co-cultures were initiated by inoculating HR with sterilized CRKN eggs. Infection with CRKN induced the RKN symptomatology in the HR and several nematode life stages were observed by light and scanning electron microscopy. Potato HR and HR/CRKN co-culturesexhibited similar growth patterns, evaluated by measuring fresh and dry weight and by the dissimilation method. Volatiles, isolated by distillation–extraction and analyzed by gas chromatography (GC) and gas chromatography coupled to mass spectrometry, revealed that palmitic acid (37–52 %), n–pentadecanal (10–16 %) and linoleic acid (2–16 %) were the main constitutive components of S. tu- berosum HR, and of the HR/CRKN co-cultures (24–44, 8–22 and 4–18 %, respectively). S. tuberosum HR/CRKN co-cultures can be considered a suitable biotechnological tool to study RKN infection mechanism by mimicking what occurs under field conditions.