Identification of asteroid genera with species capable of larval cloning.

Asexual reproduction in larvae, larval cloning, is a recently recognized component of the complex life histories of asteroids. We compare DNA sequences of mitochondrial tRNA genes (Ala, Leu, Asn, Pro, and Gln) from larvae in the process of cloning collected in the field with sequences from adults of known species in order to identify asteroid taxa capable of cloning. Neighbor-joining analysis identified four distinct groups of larvae, each having no, or very little, sequence divergence (p distances ranging from 0.00000 to 0.02589); thus, we conclude that each larval group most likely represents a single species. These field-collected larvae cannot be identified to species with certainty, but the close assemblage of known taxa with the four larval groups indicates generic or familial identity. We can assign two of the larval groups discerned here to the genera Luidia and Oreaster and another two to the family Ophidiasteridae. This study is the first to identify field-collected cloning asteroid larvae, and provides evidence that larval cloning is phylogenetically widespread within the Asteroidea. Additionally, we note that cloning occurs regularly and in multiple ways within species that are capable of cloning, emphasizing the need for further investigation of the role of larval cloning in the ecology and evolution of asteroids.

Amino Acid Uptake and Metabolism by Larvae of the Marine Worm Urechis caupo (Echiura), a New Species in Axenic Culture.

Axenic (bacteria-free) larval cultures of the marine echiuran worm, Urechis caupo, were reliably obtained by aseptically removing gametes directly from the gamete storage organs. Trochophore larvae only removed neutral amino acids from seawater as measured by high-performance liquid chromatography (HPLC). There was no detectable uptake, as measured by HPLC, of acidic or basic amino acids. Kinetic analysis showed that the transport system for alanine in 4-day-old larvae had a Kt of 4-6 µM and a Jmax of 9-10 pmol larva-1 h-1. Following a 50-min exposure, the majority of the radioactivity (95%) from 14C-alanine was found in the trichloroacetic acid-soluble fraction. Very little label appeared as acid-insoluble material, and there was no detectable lipid biosynthesis from 14C-alanine. Approximately 12% of the total alanine transported was released in the form of 14CO2. Thin-layer chromatography of intracellular free amino acid pools demonstrated that aspartic acid and glutamic acid were radiolabeled from the alanine precursor. A comparison of the energy acquired from the transport of alanine, with the metabolic rate of 4-day-old larvae, revealed that 51% of the metabolic demand could be provided by the transport and complete catabolism of this single amino acid at a concentration of 595 nM in seawater.