Characterization of the response of in vitro cultured Myrtus communis L. plants to high concentrations of NaCl.

Effect of salt stress was examined in in vitro shoot cultures of Myrtus communis L. a species of the Mediterranean maquis. To determine the effects of high salt concentrations on myrtle plantlets and contribute toward understanding the mechanisms adopted from this species to counteract soil salinity, in vitro rooted shoots were transferred to a liquid culture medium containing 0, 125 or 250 mM NaCl for 30 days. After 15 and 30 days of in vitro culture, shoot and root growth, chlorosis and necrosis extension, chlorophylls, carotenoids, proline, arginine, cysteine and total sugars content, as well as guaiacol peroxidase (G-POD, EC 1.11.1.7) and ascorbate peroxidase (APX, EC 1.11.1.11) activities were determined. In treated plants shoot and root growth, as well as chlorophyll content, significantly decreased, while carotenoids content was not affected by the NaCl treatment. Among osmolytes, proline did not significantly increase, arginine and cysteine decreased, while total sugars were found to be higher in the treated plants than in the control. Enhancement of G-POD and APX activities was positively related to increasing salt concentrations in the culture media, regardless of the exposure time. Salt-treated plants did not show significant changes in lipid peroxidation or DNA fragmentation after 30 days salt treatment, regardless of the NaCl concentrations applied. The results represent a contribution towards understanding the mechanisms adopted by this species to high salinity.

Molecular cloning and characterization of a linalool synthase from lemon myrtle.

Using a homology-based PCR strategy, we identified a cDNA with sequence similarity to linalool synthase from lemon myrtle. Functional expression of the cDNA (designated BcLS) gene in Escherichia coli yielded an active enzyme capable of catalyzing the conversion of geranyl diphosphate to (-)-linalool, i.e., an acyclic monoterpene alcohol, and to lesser amounts of cyclic monoterpenes. The kinetic parameters of BcLS were similar to those of synthases producing cyclic monoterpenes. PCR analysis revealed that the BcLS gene transcript was ubiquitously expressed in lemon myrtle and was upregulated in response to jasmonic acid treatment. Although the physiological role of neryl diphosphate (NPP) dependency of BcLS remains unclear, the cyclization activity of BcLS was enhanced when NPP was used as substrate, resulting in predominant production of cyclic monoterpenes. These findings indicate that BcLS has novel specificity and kinetic parameters, but its physiological responses to stresses such as insect damage appear to be similar to known linalool synthases.