### APPENDIX III: DRAFT RFP FOR POPULATION GENETICS

GLOBEC CALL FOR RESEARCH PROPOSALS CONCERNING GENETIC IDENTIFICATION AND CHARACTERIZATION OF ZOOPLANKTON POPULATIONS

Context and Need for Proposals

The GLOBEC initiative seeks fundamental information about basic mechanisms that determine the abundance and distribution of zooplankton populations (including holoplankton, meroplankton and ichthyoplankton) and, most importantly, the variability of these populations about their average values. Obtaining this fundamental information depends critically on the ability to characterize zooplankton field samples according to taxonomic and population genetic criteria. Ideally such characterization would be rapid and accurate enough to allow real-time automated sorting of live zooplankton field samples. In reality, current methods of molecular genetics cannot simply be coupled to plankton pumps and flow cytometers to achieve this ideal. New molecular technology must be developed for this task. Existing molecular techniques can, however, be applied immediately to studies of zooplankton systematics and population genetics in order to provide a foundation for the development of new technology and data on the coupling of oceanographic and population processes. This is a call for proposals that, in the context of the GLOBEC program, address the development and application of technology for improved resolution of zooplankton taxa and populations through all life stages from egg to adult.

Research Areas of Interest

Three research areas of interest have been identified:

1. application of existing methods of molecular biology and genetics to the identification and characterization of zooplankton species and populations;

2. development of new technology, amenable to shipboard practice, for the rapid and accurate identification of preserved, intact or live zooplankton;

3. investigations into the genetic bases of phenotypic variation that might allow adaptation of zooplankton populations to global climate change.

A range of existing molecular techniques -- allozyme electrophoresis, RFLP analysis, sequence-specific oligonucleotide probes, DNA fingerprinting, and immunofluorescent probes -- are available to discriminate morphologically similar species and to analyze population genetic structure. Proposals that apply these techniques to zooplankton species are solicited. Characterizing species-specific markers through studies of molecular systematics will provide a foundation for unambiguous identification and enumeration of zooplankton species in GLOBEC field studies. Assessing intraspecific variation in molecular markers will allow sampling of conspecific zooplankton populations on spatial/temporal scales appropriate to physical oceanographic features or processes that may influence population mixing or recruitment success. There is a basic need to assess which molecular markers are most suitable, in terms of informational content and sampling efficiency, to which conceptual problems.

Proposals to develop new technology that could be applied to the problem of identifying and sorting zooplankton field samples should address one or more of the following aspects of the problem: 1) reducing existing or new biotechnology to shipboard practice; 2) extracting molecular information from preserved zooplankton samples, allowing rapid preservation of field samples for genetic analysis as well as retrospective studies of historical zooplankton collections; 3) labeling of intact zooplankton at the species level; 4) labeling of live zooplankton at the species level; 5) automating zooplankton identification, sorting, and enumeration through molecular or physical methods such as optical image analysis or a combination of these.

Proposals that assess the genetic bases of variation in phenotypes likely to determine responses of zooplankton populations to global climate change are solicited. Such phenotypes include: 1) characters important in recruitment success (e.g., duration of the pelagic larval phase, competency for metamorphosis, response to environmental cues for settlement); or 2) physiological, behavioral or life-history traits likely to be influenced directly by global warming.

Selection of Target Species or Model Systems

Zooplankton species selected for study under this call for proposals should meet one of the following criteria: 1) they are important in marine ecosystems likely to be targets of GLOBEC field studies; 2) they are particularly suited to the development of new technology for zooplankton identification; 3) they are amenable to study of fundamental mechanisms governing abundance and distribution. These guidelines are intended to give investigators broad latitude in choosing appropriate species yet serve the overall goals of the GLOBEC program.

Training Component of Research Proposals

A major impediment to accomplishing the GLOBEC agenda is the need for cross-disciplinary training of marine biologists at all professional levels in molecular and population genetics. Proposals incorporating funding for collaborative research and for graduate students, post-doctoral scholars or sabbatical-leave faculty are encouraged.