Molecular detection of native and invasive marine invertebrate larvae present in ballast and open water environmental samples collected in Puget Sound

Non-native marine species have been and continue to be introduced into Puget Sound via several vectors including ship's ballast water. Some non-native species become invasive and negatively impact native species or near shore habitats. We present a new methodology for the development and testing of taxon specific PCR primers designed to assess environmental samples of ocean water for the presence of native and non-native bivalves, crustaceans and algae. The intergenic spacer regions (IGS; ITS1, ITS2 and 5.8S) of the ribosomal DNA were sequenced for adult samples of each taxon studied. We used these data along with those available in Genbank to design taxon and group specific primers and tested their stringency against artificial populations of plasmid constructs containing the entire IGS region for each of the 25 taxa in our study, respectively. Taxon and group specific primer sets were then used to detect the presence or absence of native and non-native planktonic life-history stages (propagules) from environmental samples of ballast water and plankton tow net samples collected in Puget Sound. This methodology provides an inexpensive and efficient way to test the discriminatory ability of taxon specific oligonucleotides (PCR primers) before creating molecular probes or beacons for use in molecular ecological applications such as probe hybridizations or microarray analyses. This work addresses the current need to develop molecular tools capable of diagnosing the presence of planktonic life-history stages from non-native marine species (potential invaders) in ballast water and other environmental samples.     

PHYTOCHEMICAL STUDIES IN THE FAMILY MALVACEAE. I. COMPARISON OF ESSENTIAL OILS OF SIX SPECIES BY GAS-LIQUID CHROMATOGRAPHY

When the chemical constitution of the essential oils of six species within three genera of the family Malvaceae were investigated, the essential oils of Gossypium hirsutum L. and Cienfuegosia rosei Fryxell were found to contain 67.0 % and 58.2 % hydrocarbons, respectively. Those of Hibiscus lasiocarpus (Cav.) and Callirhoe involucrata (Nutt.) Gray contained equal percentages of hydrocarbons (14 %), but that of Cienfuegosia drummondii (A. Gray) Lewt. contained only 2.9 % hydrocarbons. Beta-bisabolol was the major oxygenated component in G. hirsutum (13.7 %), Hibiscus militaris (Cav.) (55.0 %), and Hibiscus lasiocarpus (32.6 %), but the major constituents in the essential oil of C. involucrata were myrtenal (32.0 %) and trans-2-cis-6-nonadienal (12.0 %), and the major oxygenated component in C. rosei was an unknown sesquiterpene alcohol (17.4 %) with chemical and physical properties resembling those of beta-bisabolol. The concentration of total oil was 150.0 ppm in G. hirsutum; 50.0 ppm in H. militaris; 46.3 ppm in C. rosei; 41.0 ppm in H. lasiocarpus; 25. 4 ppm in C. drummondii; and 10.0 ppm in C. involucrata, all based on wet weight.