Scholarly Studies Program

OFFICE OF FELLOWSHIPS AND GRANTS

SCHOLARLY STUDIES PROGRAM

Proposal Cover Sheet

Name: S.I. Address & Phone:

Principal Investigator:

Robert Hershler Department of Invertebrate Zoology National Museum of Natural History NHB E-518 (202) 786-2077

Co-Principal Investigator(s).

Hsiu-Ping Liu Biology Department Montclair State University Upper Montclair, NJ 07043

Collaborator(s): None

Proposal Title: Phylogeny of Pyrgulopsis Springsnails, a Major Component of North American

Aquatic Biodiversity: Implications for Systematics. Biogeography, and

Conservation

Start Date and Duration of Project: September 1, 1998,. two years

Budget Request: Year 1 $28,970 Year 2 $33,536 Total $62,506

P.I.'s Unit Organization Code: 3380

Signatures: Date:

Principal Investigator 3/23/98

Co-Principal Investigator 3/23/98 NON-SPECIALIST SUMMARY

We request funding to continue and expand a multidisciplinary and collaborative research program on the evolution and biogeography of western American springsnails of the family

Hydrobiidae. Although the West provides an outstanding theater for such studies because of its complex and historically dynamic landscape, previous work focused on a single group, fishes.

Hydrobiids are an equally suitable study group and also offer distinct advantages over fishes as a consequence of the small size and limited dispersal abilities of these snails. Prior funding from the Scholarly Studies Program enabled us to construct a robust phylogenetic hypothesis for a component of this fauna (genus Tryonia, 21 species) based on mitochondrial DNA sequences

(mtCOI gene), thus removing the major "roadblock" that had prevented full utilization of hydrobiid snails in evolutionary and biogeographic studies. (The morphological characters provided by these snails have thus far proved insufficient in this respect.) Our study was also encouraging as patterns of snail distribution and evolution were partly congruent with those documented for pupfish (Cyprinodon) and provided evidence that an important regional drainage, the Amargosa River basin, is not only a biotic, but also a geologic composite, in congruence with the complex tectonic history of the Death Valley region.

Based on the successful and exciting results of the Tryonia project, we propose a similar phylogenetic analysis, but much more comprehensive biogeographic analysis of another western hydrobiid, Pyrgulopsis, the largest genus of nonmarine snails in North America (123 extant species). Our study will determine whether this anatomically heterogeneous and confusing group represents a clade (a group of species that includes an ancestral species and all of its descendants), delineate evolutionary structure within the group, provide needed taxonomic stability, and evaluate whether morphological characters are congruent with a molecular-based phylogeny of these snails. We anticipate that the few broadly ranging congeners will be revealed as large species complexes - we will document these discoveries by publishing formal taxonomic descriptions and share relevant information with regulatory agencies seeking to conserve and manage aquatic ecosystems in the West. Finally, we will use our phylogenetic hypothesis as the basis for an analysis of historical relationships of areas inhabited by these snails (vicariance biogeography), which will be evaluated for congruence with relevant geologic history and contrasted with previous such studies of other regional organisms.

We will conduct fieldwork to obtain fresh material for DNA analysis. We will sample most of the diversity in this group, including various undescribed congeners which can provide biogeographic resolution of otherwise unrepresented geographic areas. The fossil record suggests a minimally Miocene age for both Pyrgulopsis and Tryonia, and thus we assume that the mtCOI gene will be similarly informative for this study. We tested this by analyzing a small sub-set of Pyrgulopsis and related groups, the results of which confirmed the resolving power of this gene and indicated encouraging congruence with morphological-based hypotheses concerning snail relationships. As a large number of characters will be necessary to well resolve the phylogeny of this huge group, we will sequence this entire gene, as well as a second gene

(probably mtC0II).

This project benefits from the complementary skills of the participants. Hershler has been been conducting field surveys and studying the systematics and morphology of western hydrobiids for 20 years while Liu is expert in molecular analyses and is interested in applying these techniques to evolutionary processes of mollusks.

ii INTRODUCTION. This requests funding to continue and expand research on the evolution and biogeography of western American springsnails of the family Hydrobiidae. The

West is extremely fertile ground for such studies because aquatic elements are profoundly isolated by inhospitable deserts and mountain ranges, live in extremely restricted and/or harsh environments, and provide distributional and phylogenetic patterns which have been molded by the extremely complex and dynamic Cenozoic history of the region (Minckley et al. 1986).

Whereas early biogeographic treatments of fishes (e.g., Hubbs and Miller 1948; Hubbs et al.

1974) focused on dispersal opportunities provided by a highly integrated late Pleistocene

("pluvial") regional drainage, Minckley et al. (1986) accepted great antiquity (Oligocene-

Miocene) of this fauna and consequently emphasized the complex role of geological events in effecting vicariance. In this seminal work they provided a model for historical relationships of western areas (Minckley et al. 1986:figs. 15.4, 15.5) and set the stage for a new era in the study of regional biogeography, yet few pertinent phylogenetic hypotheses have since been generated and these have focused almost exclusively on fishes (e.g., Echelle and Dowling 1992; Smith

1993; but also see Hendrickson 1986).

Hydrobiid snails, the most diverse family of freshwater snails in North America (Turgeon et al. 1998), share many attractive features for evolutionary and biogeographic studies with fishes, as well as providing distinct advantages over them. The regional age of both groups is minimally Paleogene (Taylor 1985; Minckley et al. 1986), and thus these organisms are suitable for evaluation of old biogeographic relationships. Hydrobiids are obligate to water and disperse slowly, features which link them tightly with drainage history (Taylor and Bright 1987) and make them ideal tools for evaluating biotic response to vicariance. Hydrobiids exhibit a much greater degree of local endemism than fishes, and they sometimes comprise "species flocks,"

1 including several spectacular faunas in spring systems (Taylor 1966b). These tiny snails often persist in poorly watered basins devoid of ichthyofauna and do not appear to be subject to introgression, which commonly occurs in the more mobile fishes (Smith 1992), and thus they can provide a more comprehensive and a clearer biogeographic signal than their vertebrate counterparts.

Despite the unique and compelling features of western hydrobiids, an absence of rigorously proposed phylogenetic hypotheses has limited the use of these animals in evolutionary and biogeographic studies, although they have figured prominently in development of provocative, albeit non-phylogenetic scenarios for western drainage history (Taylor 1985, 1987;

Taylor and Bright 1987). The few, morphology-based phylogenetic studies of hydrobiids resulted in poorly resolved trees and indicated that most characters are homoplasic (e.g., Ponder et al. 1993; Hershler 1994), and although results may be improved by more finely discriminating characters and their states (Hershler and Ponder 1998), there is a need for infusion of additional data sets. As a first step toward removing this "roadblock," we obtained funding from the

Scholarly Studies Program in 1994 to examine monophyly and phylogenetic structure within one of the larger western genera, Tryonia, using allozymes and, ultimately, mitochondrial DNA sequences, and have produced extremely promising results (Hershler, Mulvey and Liu 1998;

Hershler, Liu and Mulvey 1998). Partial sequences from the mitochondrial cytochrome-c- oxidase subunit I (mtC0I) gene suggested the presence of cryptic species and permitted generation of well-resolved trees which indicated that the genus, as currently constituted, is polyphyletic. Subsequent anatomical studies of these taxa (which had never been monographed) revealed congruent heterogeneity in female genitalic groundplan (RH, unpublished), suggesting that molecular analyses can have an important predictive function in such cases. Phylogenetic

2 structure within the clade composed of "true" Tryonia (16 species) generally corresponded with timing of pertinent vicariant events beginning in the late Tertiary and exhibited some commonality with biogeographic pattern described for pupfish (Cyprinodon; Minckley et al.

1986; Echelle and Dowling 1992). Based on this analysis, the Amargosa River basin (whose lower segment is Death Valley) was identified as a biotic and geologic composite (fide Platnick and Nelson 1984), in congruence with the complex tectonic history of the area.

PROPOSAL. We propose to continue this research program in the form of a collaborative, multidisciplinary study of the evolution and biogeography of Pyrgulopsis, the most diverse genus of nonmarine mollusks in North America (131 Recent species, 123 of which are extant; Hershler 1994, 1995, 1998; Thompson 1995), which is broadly distributed in the western portion of the continent, from the Rio Grande basin west to the California coast, and from the Snake-Columbia River basin south to the Bolson de Mapimi in northern Mexico (Figure

1)1. This study has three main goals: (a) To evaluate monophyly and phylogenetic structure of this large, heterogeneous group using mtDNA sequences, and to study patterns of morphological evolution by mapping characters onto the resulting trees. (b) To revise the taxonomy of the genus using the results of this analysis and other available data. We anticipate that this study will show a general pattern of non-conspecificity across major drainage divides, and consequently reveal numerous previously unrecognized species. Information concerning these novelties (and associated changes in composition of previously described species) will be shared with a consortium of land managers seeking to conserve and manage aquatic ecosystems at risk

Although 12 eastern American species currently are allocated to the genus (Hershler 1994; Thompson 1995), recent morphological studies indicate that this fauna is a distinct clade that is not closely related to Pyrgulopsis (RH, unpublished).

3 in the West, within the spirit of a Memorandum of Understanding (MOU) regarding springsnail conservation that was recently developed with the National Museum of Natural History. (c) To use the resulting phyogenetic hypothesis as the basis for an analysis of vicariance biogeography of Pyrgulopsis.

STUDY GROUP AND RATIONALE. We chose Tryonia as the first subject of this research program because at that time, it was the only sizable western hydrobiid genus that had been well documented in the literature. During the past four years Hershler has worked hard to provide a similar baseline level of knowledge for Pyrgulopsis, a group of small snails (1-8 mm) having a smooth or weakly sculptured shell (Figure 2). He published a monographic synthesis of the group (Hershler 1994), filled in gaps with a descriptive paper (Hershler 1995), and completed a comprehensive treatment of the large Great Basin fauna, which included descriptions of 58 new species (Hershler 1998).

Current State of Systematics. Pyrgulopsis is loosely defined on the basis of a combination of morphological features (Hershler 1994) and monophyly within the North

American subfamily Nymphophilinae (Thompson 1979) has not been demonstrated as only a single outgroup was used in the sole published phylogenetic analysis of the genus (Hershler

1994:fig. 31). In contrast to the modest diversity in shell form (Figure 2), penial variation within the genus is marked (Figure 3), exceeding that among all other nymphophilines. Many congeners are strongly differentiated by autapomorphies, and placement of some of these within the genus is controversial (e.g., Taylor 1987; Hershler 1994). Species groupings in use (Gregg and Taylor 1965; Taylor 1987; Hershler 1994) are based on very few characters that often are non-congruent. The penis offers a potentially large suite of useful characters (e.g., number, size, position, and fusion of glandular units), but intra-specific variation in these is common and there

4 also are unresolved questions regarding their homology. Taxonomic revision of broadly distributed and/or morphologically variable species (notably P. gibba, P. kolobensis, P. micrococcus, P. wongi) will be necessary, as evidenced by a recent allozymic study of P. wongi from the western Great Basin, which revealed fixed-allele differences among populations from different drainages (Hamlin 1996), suggesting the presence of cryptic species.

Distribution and Biology. In contrast with Tryonia, whose inland occurrences are highly disjunct owing to its restriction to thermal waters, Pyrgulopsis is uniformly distributed in the region. The Great Basin contains the greatest diversity of species (80), followed by a far smaller fauna in the Colorado River drainage (20 species). Most congeners are endemic to single springs or spring systems and allopatry of congeners is typical, although two or even three species occasionally are sympatric. Pyrgulopsis is a locally abundant member of benthic communities throughout the West, and grazes on periphyton (Mladenka 1992). Females are oviparous, depositing egg capsule on hard substrates, and larval development is direct, with "crawl-away" young emerging from the egg capsules. These snails are not uniformly distributed within drainage systems, but instead are typically concentrated near spring sources (e.g., Noel 1954), a pattern which surely contributes to their tendency for local differentiation. A few species alternatively live in lakes, rivers, and large streams. While many species are restricted to basin

floor habitats, others also live along montane slopes, suggesting the possibility of upland

headwater transfers across drainage divides. Pyrgulopsis is well-represented in the fossil record

(Taylor 1966a) and dates minimally to the Miocene, based on P. truckeensis from the Truckee

Formation (Gregg and Taylor 1965).

PRELIMINARY RESULTS. Inasmuch as the fossil record suggests that both Tryonia

and Pyrgulopsis arose in the Miocene or earlier, we assume that the COI gene also will provide

5 an appropriate phylogenetic signal for Pyrgulopsis. We tested this by sequencing an approximately 600 base pair segment of this gene from seven congeners (representing most of the morphological variation within the group) and four eastern members of the Nymphophilinae

(placed in "Cincinnatia," Notogillia, and Spilochlamys). Two Australian taxa (Ascorhis,

Phrantela) considered among the more plesiomorphic hydrobiids (Ponder and Clark 1988;

Ponder et al. 1993) were used as outgroups.

Total aligned sequences consisted of 584 base pairs, of which 152 were parsimony- informative. All characters were weighed equally as there was no evidence of base pair composition bias and saturation among taxa. Using maximum parsimony, two trees of 502 steps were obtained, one of which is shown in Figure 4. Taxa were well-resolved and branches had high bootstrap support. These results also are encouraging because they are congruent with morphological evidence suggesting that eastern species allocated to the genus ("Pyrgulopsis" sp.) are not closely related to western snails. The results also suggest an answer to the question of direction of evolution of penial morphology within the genus. The sister relationship between

Pyrgulopsis and "Cincinnatia," and of these with Spilochlamys and Notogillia, indicates that ancestral Pyrgulopsis was endowed with a highly glandular penis and simple-pened congeners

(e.g., clade composed of P. owensensis, P. amargosae, P. stearnsiana, and Pyrgulopsis sp.) are reductive.

MATERIALS AND METHODS. Taxa. Sampling of all 123 extant species assigned to

Pyrgulopsis is logistically impractical if not impossible (some species are endemic to private lands whose owners have a record of overt hostility toward field biologists!). We instead propose to sample about 80 described congeners, which will be selected to span morphological variation within the genus, provide coverage from all drainages occupied by these snails, and to

6 test monophyly of previously proposed species groups and locally endemic assemblages. We also will sample 15-20 unreported species of Pyrgulopsis (identified from USNM holdings) which provide biogeographic coverage of areas lacking nominate forms (e.g., lower Columbia

River basin, interior drainages of northern Mexico) or represent divergent morphology.

Representatives of the other seven genera of Nymphophilinae (as well as eastern "Pyrgulopsis") will be analyzed to evaluate monophyly and phylogenetic position of Pyrgulopsis within the subfamily. Outgroups will consist of Australasian hydrobiids and other primitive members of the family.

Fieldwork. No additional collecting is necessary for the morphological component of this study as the large USNM collection (>1,400 lots) will be used for this purpose. However, this collection was fixed in formalin, as necessary for anatomical study, and is not suitable for DNA analysis. Small samples (ca. 10 individuals) of fresh material, placed directly into ethanol, must be collected for this purpose. Most species of Pyrgulopsis have a restricted range and for these one to three populations (including topotypes) will be sampled. For the few species having wide distributions that cross major drainage divides, 10-15 populations will be sampled. Hershler will do most of the collecting during a series of 10-day field trips based out of major western cities.

(Liu will participate in one of these trips to gain more familiarity with these animals and their habitats.) Fieldwork is expected to proceed rapidly owing to the familiarity that Hershler has with regional habitats (having visited most of them previously) and the need to collect but a few specimens from each site. Hershler is thoroughly familiar with appropriate state and federal permitting regulations and will obtain such approvals as necessary. Additional material of

Pyrgulopsis, other nymphophilines, and outgroups will be obtained through a network of colleagues that Hershler has established. We anticipate that 200 populations will be sampled for

7 analysis. Voucher material from each population sampled will be cataloged and curated into the

USNM collection.

DNA sequencing. Since we will attempt to resolve the phylogeny of a very large number of species, it is unlikely that the partial sequence of mtCOI gene, which provides about 150 informative characters, will suffice. Therefore, early laboratory study will focus on increasing the size of the molecular data set by designing primers to amplify the entire COI gene

(approximately 1500 base pairs). We will also pursue similar study of the mtC0II gene (ca.

1000 base pairs), which we assume will present a similarly informative phylogenetic signal as it is a sub-unit of the same mitochondrial gene as COI. If the mtC0II gene does not prove suitable upon preliminary analysis, we will pursue other genes such as COIII, 16S, or ND genes (NADH- dehydrogenase). Based on published mtDNA sequences on mollusks (Cypraea, Katharina,

Mytilus; GENBANK) and our previous work on hydrobiids, we anticipate that little effort or expense will be needed for primer development. DNA extraction, amplification, purification, and sequencing will follow protocols established in Hershler, Mulvey and Liu (1998). For several species, several specimens from single populations will be analyzed to evaluate variation at this level. Otherwise, one or two specimens will be used for each population.

Phylogenetic Analysis. Data will consist of approximately 1500 base pairs for the COI gene and 1000 base pairs for the COII gene (or other selected gene). A thorough analysis of base composition substitution patterns and dynamics will precede phylogenetic analysis (Ritland and

Eckenwalder 1992). Our preliminary mtCOI sequence data did not exhibit significant deviations in base composition bias and did not indicate saturation among taxa. The homologous data will be analyzed using PAUP* (Swofford 1998). If we generate multiple trees of shortest length, information will be synthesized in the form of strict consensus trees, and we will also use

8 maximum likelihood models (Huelsenbeck and Crandall 1997) to select preferred topologies.

Bootstrapping (Felsenstein 1985) will be used to estimate the reliability of branches on the shortest trees. The literature provides no general consensus as to whether separate data sets should be combined or partitioned in phylogenetic analysis (Bull et al. 1993; Chippendale and

Wiens 1994; Huelsenbeck et al. 1994; Wiens and Chippendale 1994). Since COI and COII both are mitochondrial genes and sub-units of cytochrome c oxidase, we will combine these data in our analysis. If a different second gene is used instead of COII, we will analyze these data sets together as well as separately.

Hershler already has assembled a large database of more than 50 morphological characters for each species of Pyrgulopsis, expanded and modified from that used in an earlier analysis (Hershler 1994). He also has been generating a similar database for other members of the Nymphophilinae (Hershler and Thompson 1996; RH unpublished). Upon obtaining a robust, phylogenetic hypothesis based on mtDNA data, we will map morphological traits onto the tree in order to study patterns of character evolution and congruence with molecular data using

MacClade (Maddison and Maddison 1992).

Biogeographic Analysis. Taxon-area relationships will be evaluated using the principles of vicariance biogeography (Platnick and Nelson 1978; Rosen 1978). Lack of concordant patterns of relationships within the genus will be interpreted as possible evidence of dispersal. A broadly distributed ancestral snail fauna is assumed, perhaps tied to the Eocene-Oligocene erosional surface of low relief that covered much of the West (Epis and Chapin 1978; Gresens

1981). The fossil record of Pyrgulopsis will be reviewed, based on original study of available institutional material and scrutiny of pertinent literature, in order to pinpoint minimal age of the group in given regions and help constrain timing of branching events. Regional geologic and

9 hydrographic history will be synthesized from available literature, much of which recently has been summarized (Minckley et al. 1986; Burchfiel et al. 1992; Hershler and Sada 1998).

Biogeographic patterns documented for other regional biota will be evaluated for possible congruence with that of Pyrgulopsis.

DELIVERABLES. The following publications will be prepared as deliverables: (a) An

assessment of monophyly and phylogenetic structure within Pyrgulopsis based on DNA

sequence data. This paper will also evaluate congruence between this phylogeny and

morphological variation within the group, and discuss implications of these results for

systematics of these and related snails specifically, and aquatic gastropods generally. (b) An

analysis of vicariance biogeography of Pyrgulopsis, based on our phylogenetic hypothesis. (c) A

taxonomic revision of those Pyrgulopsis species whose scope and content must be altered in the

wake of our analysis. This will include formal descriptions of new species revealed by our

studies. (d) An informal report summarizing species-level taxonomic changes resulting from this

project, together with associated species distribution information. This will be circulated to

relevant land management agencies as discussed above.

PERSONNEL. Hershler brings a wealth of experience with hydrobiids generally and

Pyrgulopsis in particular, having collected these snails and published extensively on their

morphology and systematics since 1985. Liu is expert in DNA laboratory methodologies and

analysis of sequence data, having done this work on two other hydrobiid projects in collaboration

with Hershler (e.g., "Tryonia project," and a broader study of hydrobiid phylogeny). The budget

includes a small amount of funds enabling us to work together in the field and laboratory, which

is an important component of the project as we both wish to learn from the other's areas of

expertise.

10 Both investigators will devote about 33% of their time over the next two years to the project. Hershler will perform most of the necessary fieldwork, refine and assemble his morphological database for Pyrgulopsis, and review fossil material, pertinent geological literature, and comparable biogeographic studies of other western biota. Liu will perform all laboratory analyses with the aid of an undergraduate assistant (for whom funding is requested).

Together they will perform the phylogenetic analyses and map morphological characters onto resulting trees.

TIMETABLE. This two-year project will begin September 1, 1998. Specimens will be collected and solicited during the first 15 months. During this time Hershler will also complete his survey of fossil material, review appropriate literature, and prepare a rough summary of pattern and timing of geological events that may have effected vicariance of these snails. Liu will devote the first three months to developing and refining protocols for analysis of the COI and COII (or other suitable) genes, and then sequence the COII gene for all taxa during the next nine months. She will then sequence the COI gene during the first six months of the second year. The next three months will be spent analyzing data and performing phylogenetic analyses, after which manuscript preparation will ensue.

11 ci < .

S' ,---,/, . C— / ,,/ / _./-/ I

0 100 200 300 400 Kilometers

FIGURE 1. Map of western North America showing distribution of Pyrgulopsis.

12 FIGURE 2. Shell form diversity among Pyrgulopsis species.

13 FIGURE 3. Variation in penial form and glandular ornament (hachured areas) among Pyrgulopsis species. Dense stipple defines pigmented areas.

14 "Cincinnatia" winkleyi 94

"Cincinnatia" sp. (Florida)

Pyrgulopsis kolobensis 54 54

Pyrgulopsis wongi

Pyrgulopsis owensensis 71 52

Pyrgulopsis sp. (Nevada) 57 100

78 Pyrgulopsis amargosae

Pyrgulopsis stearnsiana

Spilochlamys gravis 97

Notogillia wetherbyi

"Pyrgulopsis" sp. (Georgia)

Ascorhis victoriae

Phrantela marginata

FIGURE 4. One of two shortest trees (502 steps, C1=0.65) based on maximum parsimony analysis of mtC01 sequence data from species of Pyrgulopsis, other nymphophiline taxa, and two outgroups (Ascorhis, Phrantela). Numbers are bootstrap percentages for well-supported clades (1000 replicates).

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of Mines & Mineral Resources Bulletin 116:50 pages.

Taylor, D.W. and R.C. Bright. 1987. Drainage history of the Bonneville basin. Pp. 239-256 in

R.S. Koop and R.E. Cohenour, editors, Cenozoic geology of western Utah: sites for precious

metal and hydrocarbon accumulations. Utah Geological Association Publication 16.

Thompson, F.G. 1979. The systematic relationships of the hydrobioid snail genus

Nymphophilus Taylor 1966, and the status of the subfamily Nymphophilinae. Malacological

Review 12:41-49.

Thompson, F.G. 1995. A new freshwater snail from the Coosa River, Alabama (Gastropoda:

Prosobranchia: Hydrobiidae). Proceedings of the Biological Society of Washington 108:502-

507.

Turgeon, D.D., and 14 co-authors. 1998. Common and scientific names of aquatic invertebrates

from the United States and Canada: Mollusks. Second edition. American Fisheries Society

Special Publication. In press.

20 Wiens, J.J. and P.T. Chippendale. 1994. Combining and weighting characters and the prior

agreement approach revisited. Systematic Biology 43(4):564-566.

21 BUDGET

Year 1 Year 2

Travel: Hershler RT airfares $ 1420 $1050 Per diem, 40 days @ $80 ($50+30) $ 3200 $3200 Vehicle rental, 30 days @ $75 $ 2250 $2250 Liu RT airfares $ 264 0 Per diem, 10 days @ $80 ($50+30) $ 800 0

Contractual Services: Liu $ 5813 $5813 Undergraduate Assistant (to Liu) $ 5723 $5723

Supplies laboratory analyses: 250 samples @ $30 (COII gene) $7500 0 250 samples @ $54 (COI gene) 0 $13500

other expendable laboratory supplies (centrifuge tubes, gloves, tips, bottles, etc.) $ 2000 $2000

Equipment: None requested

Total Year 1 $28970 Total Year 2 $33536

TOTAL $62506.00

22 BUDGET EXPLANATION

Travel. Hershler will make three, 10-day trips each year to collect material. During the first year, he will fly to Salt Lake City (RT airfare, $264), Boise ($618), and San Francisco

($214); while he will work out of El Paso ($302), Phoenix ($208), and Las Vegas ($216) during the second year. He will require use of a four-wheel drive vehicle to confidently travel along poor-quality roads in remote areas. In addition, he will spend 10 days each year learning laboratory methodologies in Liu's laboratory and working with her on data analysis and manuscript preparation (RT airfare, Newark, NJ, $324). Liu will join Hershler on one field trip

(first year, Salt Lake City, RT airfare $264) in order to gain more familiarity with hydrobiids and their habitats. All per diem is at the general CONUS rate. Hershler will also use special trust and federal contract funds available to him to help fund colleagues who will provide additional material as needed.

Contractual Services. Liu is a junior-level faculty member on a nine-month salary schedule. One month of summer funding is requested to support her contribution to the project at a level equivalent to her current salary ($5333 + 9% fringe). Montclair State University

(MSU) will match 25% of Liu's salary during the semesters in the form of a reduced teaching load. In 1995 - 1996, Liu was funded half time through a NMNH Research Initiative award (to

Hershler) to work on "Deep Phylogeny of Hydrobiid Gastropods." During these two years, she manually sequenced roughly 144,000 base pairs (1800 base pairs x 80 specimens). In this study, we propose to sequence 625,000 base pairs (2500 base pairs x 250 specimens). Given this increased work load we request two years of support for an undergraduate assistant at a level equivalent to salary provided by MSU (30 weeks x 20 hours/week @ $5.25/hour + 9% fringe for

23 fall/spring semesters; 10 weeks x 40 hours/week @ 5.25 + 9% fringe during summers) to help

Liu process samples. With use of an automatic sequencer and the undergraduate assistant help, we are confident that we will complete the proposed project in the proposed time frame.

Supplies (and Laboratory Analyses). Based on sampling of 200 populations and the need to analyze multiple specimens from various populations, we assume that a total of 250 specimens will be analyzed. Costs for laboratory analyses as follows: DNA extraction, $1.00; PCR amplification, $3.00; agarose gel and photo, $1.00; purification of PCR product, $1.00; sequencing and polyacrylarnide gel, $12.00.

Each gene will be sequenced in both a forward and backward direction and four reactions will be required to confidently sequence the large COI gene. Thus, for COI, costs are $6

(extraction, amplification, etc.) + $12 x 4 (sequencing) = $54; while for COII costs are $6 + $12 x 2 = $30. The sequencing cost at MSU ($12) is much less than at many other institutions.

Equipment—The MSU Department of Biology is fully equipped for this study. Liu will have full access to major equipment that includes an ABI automatic sequencer, thermal cyclers, power supplies, horizontal gel rigs, variable intensity UV transilluminator, autoclave, Polaroid photo documentation apparatus, pH meter, Ultracold freezers (-80C), freezers (-20C), refrigerators, and computers with appropriate software for data analysis.

03/23/98 17:35

MONTCLAIR Department of Biology STATE UNIVERSITY

March 23, 1998

Roberta Rubinoff, Director Office of Fellowships and Grants Smithsonian Institution 955 L'Enfant Plaza, Suite 7000 Washington, D.C. 20560

Dear Ms. Rubinoff:

The Biology department of Montclair State University has hired Dr. Hsiu-Ping Liu as an Assistant Professor starting September 1, 1998.

The Biology Department and the College of Science and Mathematics are fully equipped, and Dr. I.iu has full access to all the equipment needed, for this research. This includes an ABI automatic sequencer, thermal cyders.. power supplies, horizontal gel rigs, variable intensity UV transillurninator, autoclave. Polaroid photo documentation apparatus, p1-I meters. Ultracold freezers, freezers, refrigerators, and computers with appropriate software. She will also be receiving start-up funds to purchase more equipment that she needs to do her research.

Dr. Liu will be receiving released time in her faculty teaching load through FSIP (Faculty Scholarship Incentive Program). The amount of released time constitutes 25% of her contract teaching load.

If you have any additional questions, please contact me at 973-655-5107 or thorugh e- mail at: lustigman satum.montelair.edu.

Sincerely,

Bonnie Lustigman, Ph.D. Chair of Biology

UPPER MON ICLAIF1, NEW JERSEY 07043 OPPORT UNIT YIAFFIRMA JIVE ACTION INSTITUTION CURRICULUM VITAE ROBERT HERSHLER

Current Position. Research Zoologist and Curator, NMNH, GS-14. November 1993 to present.

Previous Positions. Associate Curator, NMNH, GS-13. August 1990 to November 1993. Associate Curator, NMNH, GS-12. December 1987 to August 1990. Associate Curator, NMNH, GS-11. September 1985 to December 1987. Research Associate. Southwest Texas State University, May 1984 to February 1985 (funded by contract from U.S. Fish and Wildlife Service). Consulting Research Biologist. Australian Museum. July 1983 to March 1984. (funded by grant to W.F. Ponder)

Education. B.S. State University of New York at Stony Brook, 1975. Ph.D. Johns Hopkins University, 1983.

Professional Societies. American Malacological Union Desert Fishes Council Unitas Malacologica

Major professional interests. Malacology/ Gastropoda/ systematics/ morphology/ phylogenetic analysis/ biogeography/ aquatic ecosystems/ conservation/ New World

Awards and Honors Sustained Superior Performace Award, National Museum of Natural History, 1989.

Pre- and post-doctoral students.

Smithsonian Pre-doctoral Fellow Ellen Strong, 1996-1997

Smithsonian Post-doctoral Fellow Susan Bandoni, 1991-1992

26 Grants in the past 5 years.

California springsnails: taxonomic clarification and threatened/endangered status of selected taxa. 1993. California Department of Fish and Game, $10,000.

Evolution of freshwater snails of the genus Tryonia in the Death Valley region. 1993. Office of the Director, NMNH, $16,219.

Taxonomic validation for species of hydrobiid snails currently proposed for listing as endangered or threatened. 1994. National Biological Survey, $638.

Evolution of the North American springsnail genus Tryonia. 1994-1995. Scholarly Studies Program, Smithsonian Institution, $25,000.

Aquatic biodiversity in the West: Great Basin springsnails. 1994-1997. National Biological Survey, $78,676; Bureau of Land Management, $27,348.

Deep phylogeny of hydrobiid gastropods. 1995-6. NMNH Research Initiatives, $47,900.

Talks presented in past 2 years.

1996. Annual Meeting of the American Malacological Union (Chicago, IL). "Status of taxonomic research on non-marine gastropods of the United States," Hershler R. (presenter) and B. Roth.

1997. GBASH 97, Great Basin Aquatic System History (Salt Lake City, UT). "Springsnail biogeography," Hershler, R. (presenter) and D. W. Sada.

Publications

Peer reviewed research articles and books.

1980. Hershler, R. and G. M. Davis. The morphology of Hydrobia truncata: relevance to systematics of Hydrobia. Biological Bulletin 158: 195-219.

1984. Hershler, R. Hydrobiid snails of the Cuatro Cienegas basin, Mexico: systematic relationships and ecology of an unique fauna. In Biota of Cuatro Cienegas, Coahuila, Mexico, P. Marsh, editor, Journal of the Arizona-Nevada Academy of Sciences 19: 61- 76.

1985. Hershler, R. Systematic revision of the Hydrobiidae (Gastropoda: Rissoacea) of the Cuatro Cienegas basin, Coahuila, Mexico. Malacologia 26: 31-123.

27 1986. Hershler, R. and G. Longley. Phreatic hydrobiids (Gastropoda: Prosobranchia) from the Edwards (Balcones Fault Zone) Aquifer region, south-central Texas. Malacologia 27: 127-172.

1986. Hershler, R. and G. Longley. Hadoceras taylori, a new genus and species of phreatic Hydrobiidae (Gastropoda: Rissoacea) from south-central Texas. Proceedings of The Biological Society of Washington 99: 121-136.

1986. Hershler, R. and W. L. Minckley. Microgeographic variation in the Banded Spring Snail (Hydrobiidae: Mexipyrgus) from the Cuatro Cienegas basin, Coahuila, Mexico. Malacologia 27: 357-374.

1987. Hershler, R. Redescription of Assiminea infima Berry 1947 from Death Valley, California. Veliger 29: 274-288.

1987. Hershler, R. and F. G. Thompson. North American Hydrobiidae (Gastropoda: Rissoacea): redescription and systematic relationships of Tryonia Stimpson, 1865 and Pyrgulopsis Call and Pilsbry, 1886. Nautilus 101: 25-32.

1987. Hershler, R. and G. Longley. Phreatoceras, a new name for Hadoceras Hershler and Longley, 1986 (Gastropoda) non Strand, 1934 (Cephalopoda). Proceedings of the Biological Society of Washington 100: 402.

1987. Hershler, R. and G. Longley. Phreatodrobia coronae, a new species of cavesnail from southwestern Texas. Nautilus 101: 133-139.

1987. Hershler, R. and D. Sada. Springsnails (Gastropoda: Hydrobiidae) of Ash Meadows, Amargosa basin, California-Nevada. Proceedings of the Biological Society of Washington 100: 776-843.

1988. Hershler, R. and J. J. Landye. Arizona Hydrobiidae. Smithsonian Contributions to Zoology 459: 1-63.

1988. Hershler, R. and L. Hayek. Shell variation of springsnail populations in the Cuatro Cienegas basin, Mexico: preliminary analysis of limnocrene fauna. Nautilus 102: 56-64.

1988. Hershler, R. and L. Hubricht. Notes on Antroselates Hubricht, 1963 and Antrobia Hubricht, 1971 (Gastropoda: Hydrobiidae). Proceedings of the Biological Society of Washington 101: 730-740.

1988. Hershler, R. and F. G. Thompson. Notes on morphology of Amnicola limosa (Say, 1817)(Gastropoda: Hydrobiidae) with comments on status of the subfamily Amnicolinae. Malacological Review 22: 81-92.

28 1988. Hershler, R. Holsingeria unthanksensis, a new genus and species of aquatic cavesnail from eastern North America. Malacological Review 22: 93-100.

1989. Hershler, R. Springsnails (Gastropoda: Hydrobiidae) of Owens and Amargosa River (exclusive of Ash Meadows) drainages, Death Valley system, California-Nevada. Proceedings of the Biological Society of Washington 102: 176-248.

1989. W. F. Ponder, R. Hershler and B. J. Jenkins. An endemic radiation of hydrobiid snails from artesian springs in northern South Australia: their taxonomy, physiology, distribution and anatomy. Malacologia 31: 1-141.

1990. Hershler, R. and J. R. Holsinger. Zoogeography of North American hydrobiid cavesnails. Stygologia 5: 1-12.

1990. Hershler, R. and F. G. Thompson. Antrorbis breweri, a new genus and species of hydrobiid cavesnail from Coosa River basin, northeastern Alabama. Proceedings of the Biological Society of Washington 103: 197-204.

1990. Hershler, R. and W. L. Pratt. A new Pyrgulopsis (Gastropoda: Hydrobiidae) from southeastern California, with a model for historical development of the Death Valley hydrographic system. Proceedings of the Biological Society of Washington 103: 279- 299.

1990. Hershler, R., J. R. Holsinger and L. Hubricht. A revision of the North American freshwater snail genus Fontigens (Prosobranchia: Hydrobiidae). Smithsonian Contributions to Zoology 509: 1-49.

1990. Hershler, R. Pyrgulopsis bruneauensis, a new springsnail (Gastropoda: Hydrobiidae) from the Snake River Plain, southern Idaho. Proceedings of the Biological Society of Washington 103: 803-814.

1990. Hershler, R., J.M. Pierson, and R.S. Krotzer. Rediscovery of Tulotoma magnifica (Conrad)(Gastropoda: Viviparidae). Proceedings of the Biological Society of Washington 103: 815-824.

1991. Thompson, F.G. and R. Hershler. Two new hydrobiid snails (Amnicolinae) from Florida and Georgia, with a discussion of the biogeography of freshwater gastropods of south Georgia streams. Malacological Review 24: 55-72.

1991. Thompson, F.G. and R. Hershler. New hydrobiid snails (Mollusca: Gastropoda: Prosobranchia: Truncatelloidea) from North America. Proceedings of the Biological Society of Washington 104: 669-683.

29 1992. Hershler, R. and F.G. Thompson. A review of the aquatic gastropod subfamily Cochliopinae (Prosobranchia: Hydrobiidae). Malacological Review, Supplement 5: 1- 140.

1993. Hershler, R. and F. Velkovrh. A new genus of hydrobiid snails (Prosobranchia: Rissooidea) from northern South America. Proceedings of the Biological Society of Washington 106: 182-189.

1993. Kabat, A. and R. Hershler. The prosobranch snail family Hydrobiidae (Gastropoda: Rissooidea): review of classification and supraspecific taxa. Smithsonian Contributions to Zoology 547: 1-94.

1994. Hershler, R., T.J. Frest, E.J. Johannes, P.A. Bowler, and F.G. Thompson. Two new genera of hydrobiid snails (Prosobranchia: Rissooidea) from the northwestern United States. Veliger 37: 221-243.

1994. Hershler, R. A review of the North American freshwater snail genus Pyrgulopsis. Smithsonian Contributions to Zoology 554: 1-115.

1995. Hershler, R. New freshwater snails of the genus Pyrgulopsis (Rissooidea: Hydrobiidae) from California. Veliger 38: 343-373.

1996. Hershler, R. and F.G. Thompson. Redescription of Paludina integra Say, 1821, type species of genus Cincinnatia (Gastropoda: Hydrobiidae). Journal of Molluscan Studies 62: 33-55.

1996. Hershler, R. Review of the North American aquatic snail genus Probythinella. Invertebrate Biology 115: 120-144.

1996. Hershler, R. and T.J. Frest. A review of the North American freshwater snail genus Fluminicola (Hydrobiidae). Smithsonian Contributions to Zoology 583: 1-41.

1998. Hershler, R. A systematic review of the hydrobiid snails (Gastropoda: Rissooidea) of the Great Basin, western United States. Part I. Genus Pyrgulopsis. Veliger 41: 1-132.

1998. Hershler, R. and W.F. Ponder. A review of morphological characters of hydrobioid snails. Smithsonian Contributions to Zoology 600. In press.

1998. Hershler, R., M. Mulvey, and H.-P. Liu. Biogeography in the Death Valley region: evidence from springsnails (Hydrobiidae: nyonia). Zoological Journal of the Linnean Society, London. Accepted, with revision. 40 mss pages.

1998. Hershler, R. and D.W. Sada. Biogeography of Great Basin freshwater snails of the genus Pyrgulopsis. Smithsonian Contributions to Earth Sciences. In review. 59 mss pages.

30 1998. Hershler, R. A systematic review of the hydrobiid snails (Gastropoda: Rissooidea) of the Great Basin, western United States. Part II. Genera Colligyrus, Fluminicola, Pristinicola, and Tryonia. The Veliger. In review. 71 mss pages.

Other publications (e.g., reviews, contract reports, scientific editorials, articles for a lay audience).

1985. Hershler, R. Status of phreatic hydrobiid snails of the Edwards (Balcones Fault Zone) Aquifer, south-central Texas. Report to the U.S. Fish and Wildlife Service (Albuquerque, NM). 30 pages.

1986. Hershler, R. Survey of the rissoacean snails (Gastropoda: Prosobranchia) of the Death Valley drainage system, California. Report to the U.S. Fish and Wildlife Service (Great Basin Complex, Reno, NV) and Bureau of Land Management (Riverside, CA). 39 pages.

1988. Hershler, R. Status survey of Hydrobiidae in Owens River drainage. Report to California Department of Fish and Game (Rancho Cordova, CA). 29 pages.

1989. Hershler, R. Status survey of hydrobiid snails in the Great Basin of northern California. Report to California Department of Fish and Game (Rancho Cordova, CA). 21 pages.

1989. Hershler, R. Literature review of California springsnails. Report to California Department of Fish and Game (Rancho Cordova, CA). 30 pages.

1989. Hershler, R. Status survey of Tulotoma magnifica (Conrad). Report to U.S. Fish and Wildlife Service (Jackson, MS). 20 pages.

1990. Hershler, R. Status survey of Hydrobiidae in Mojave Desert and selected areas adjacent to Death Valley hydrographic system. Report to California Department of Fish and Game (Rancho Cordova, CA). 27 pages.

1991. Hershler, R. Status survey of the Grapevine Creek Snail. Report to California Department of Parks and Recreation (Lodi, CA). 10 pages.

1992. Hershler, R. Taxonomic status of Tryonia in Long Valley, Mono County, California. Report to California Department of Fish and Game (Bishop, CA). 7 pages.

1994. Hershler, R. Taxonomic validation of federal candidate snails of the family Hydrobiidae. Report to the National Biological Service (Washington, D.C.). 11 pages.

1994. Hershler, R. Field survey and preliminary taxonomy of Great Basin springsnails. Report to the Bureau of Land Management (Washington, D.C). 112 pages.

1995. Hershler, R. Status survey of three species of hydrobiid snails in California. Report to California Department of Fish and Game (Rancho Cordova, CA). 7 pages.

31 Previous Awards to Hershler from the Scholarly Studies Program (with resulting publications)

1988-1989. Generic review of aquatic snails of the subfamily Littoridininae (Gastropoda: Hydrobiidae). $42,977.

Thompson, F.G. and R. Hershler. 1991. New hydrobiid snails (Mollusca: Gastropoda: Prosobranchia: Truncatelloidea) from North America. Proceedings of the Biological Society of Washington 104:669-683.

Hershler, R. and F.G. Thompson. 1992. A review of the aquatic gastropod subfamily Cochliopinae (Prosobranchia: Hydrobiidae). Malacological Review, Supplement 5:140 pages.

1991-1993. Biogeography of Great Basin springsnails. $78,500 (two separate awards).

Hershler, R. 1998. A systematic review of the hydrobiid snails (Gastropoda: Rissooidea) of the Great Basin, western United States. Part I. Genus Pyrgulopsis. The Veliger 41:132 pages.

Hershler, R. 1998. A systematic review of the hydrobiid snails (Gastropoda: Rissooidea) of the Great Basin, western United States. Part II. Genera Colligyrus, Fluminicola, Pristinicola, and Tryonia. Submitted to The Veliger. 71 mss pages.

Hershler, R. and D.W. Sada. 1998. Biogeography of Great Basin freshwater snails of the genus Pyrgulopsis. Submitted to Smithsonian Contributions to Earth Sciences. 59 mss pages.

1994-1995. Evolution of the North American springsnail genus Tryonia. $25,000.

Hershler, R., M. Mulvey, and H.-P. Liu. 1998. Biogeography in the Death Valley region: evidence from springsnails (Hydrobiidae: Tryonia). Zoological Journal of the Linnean Society, London. Accepted, with revision. 40 mss pages.

Hershler, R., H.-P. Liu, and M. Mulvey. 1998. Phylogenetic relationships within the North American hydrobiid snail genus Tryonia: taxonomic and biogeographic implications. In preparation for Molecular Biology and Evolution.

32 CURRICULUM VITAE Hsiu-Ping Liu Biology Department Montclair State University Upper Montclair, NJ 07043 Personal Information Born April 17, 1965, Keelung, Taiwan, Female, U.S. citizen. Education Ph. D., Department of Environmental, Population, and Organismic Biology, University of Colorado at Boulder, 1994. M.S., Institute of Fisheries Science, National Taiwan University, Taipei, Taiwan, 1988. B.S., with Honors, Department of Aquaculture, National Taiwan University of Marine Science and Technology, Keelung, Taiwan, 1986. Workshop attended Recent Advances in Conservation Genetics at NOAHS-Conservation Research Center, Front Royal, VA 1996 Employment Assistant Professor, Montclair State University, 1998-present. Visiting Assistant Professor, Colgate University, 1997-1998. Postdoctoral Fellow, University of Georgia, Savannah River Ecology Laboratory, 1995 - 1997. Field Zoologist, Colorado Heritage Program, Nature Conservancy, 1994. Graduate Assistant, University of Colorado Museum, 1988 - 1994. Teaching Assistant, Department of Environmental, Population, and Organismic Biology, University of Colorado, 1989 - 1990; 1992. Graduate Assistant, Institute of Zoology, Academia Sinica, Taipei, 1986 - 1988. Awards and Funding Smithsonian Institution Biodiversity Surveys and Inventories Program, Biodiversity in northern Mexico: springsnails of the genus Tryonia (Hydrobiidae) (pending) California Game and Fish, Taxonomic clarification of springsnails (Tryonia variegata complex), Amargosa River Basin. (pending) Short-term Visitor Fellowship, Smithsonian Institution, 1998. Colgate Student Wage Grant, Colgate University, 1997. Savannah River National Environmental Research Park Grant, Conservation genetic study of freshwater Lampsiline mussels, co-principal investigator, 1996. Smithsonian Institution Biodiversity Initiative Grant, Deep Phylogeny of Hydrobiid Gastropods, Postdoctoral Fellow, 1995-1996. Best Student Paper Award, American Malacological Union, 1994.

33 Research Grant in Malacology, the Western Society of Malacologists, the Southwest Shell Club, the Santa Barbara Shell Club and the San Diego Shell Club, 1992. Environmental, Population, and Organismic Biology Graduate Fellowship, University of Colorado, Colorado, 1992. Graduate School Dean's Small Grant Award, University of Colorado, Colorado, 1992. Walker Van Riper Award, University of Colorado Museum, Colorado, 1992. Scholarship Award, Hawaiian Malacological Society, Hawaii, 1991. Graduate Fellowship, National Taiwan University, Taipei, 1986-1988. Scholarship, the Ministry of Education, Republic of China, 1986-1987. Scholarship Award, Chun-Kuan Company, Taipei, 1985. Scholarship Award, Chia-Shin Company, Taipei, 1983, 1984.

Publications (Published) Hershler, R., Mulvey, M., and Liu, H.-P. 1998. Biogeography in the Death Valley region: Evidence from springsnails (Hydrobiidae: Tryonia). Accepted with revision in Zoological Journal of Linnaean Society). Mulvey, M., Liu, H.-P., and Kandl, K. L. 1998. Application of molecular genetic markers to conservation of freshwater mussels (in press in Symposium on Genetics of Freshwater Mussels, National Shellfisheries Association) Liu, H.-P., Mitton, J. B., and Wu, S.-K. 1996a. Paternal mitochondrial DNA differentiation far exceeds maternal mitochondrial DNA and allozyme differentiation in the freshwater mussel, Anodonta grandis grandis. Evolution 50(2): 952-957. Liu, H.-P., Mitton, J.B., and Herrmann, S.J. 1996b. Genetic differentiation and management recommendations for the freshwater mussel, Pyganodon grandis Say 1829. American Malacological Bulletin 13(1/2): 117-125. Liu, H.-P., and Mitton, J.B. 1996. Tissue-specific maternal and paternal mitochondrial DNA in the freshwater mussel, Anodonta grandis grandis Say 1829. The Journal of Molluscan Studies 62: 393-394. Liu, H.-P. 1993. Diagnostic genetic loci for species in the genus Physella. Malacological Review, 26: 1-8. Liu, H.-P., and Mitton, J. B. 1993. Techniques to reveal restriction fragment length polymorphisms (RFLPs) in the genus Physa. Malacological Review, 26: 89-90. Liu, H.-P., Liao, K.-Y., and Wu, W.-L. 1993. Comparison of the reproductive ecology of Meretrix lusoria from Tam-Shui and Lu-Kang area. Bulletin of Malacology, Taipei, 17: 69- 77. Wu, W.-L. and Liu, H.-P. 1993. Studies on the gonad development of Meretrix lusoria from Taiwan (Bivalvia: Veneridae). Bulletin of Malacology, Taipei, 17: 79-86. Wu, W.-L., Liu, H.-P. and Liao, K.-Y. 1993. Morphological and anatomical studies of Meretrix lusoria from Taiwan (Bivalvia: Veneridae). Bulletin of Malacology, Taipei, 17: 91-98. Wu, W.-L., and Liu, H.-P. 1989. Malacological research on Meretrix resources in Taiwan. II. History review and evaluation on the studies of the Taiwan Meretrix. Bulletin of Malacology, Taipei, 14: 49-61. Wu, W.-L., and Liu, H.-P. 1988. Malacological research on Meretrix resources in Taiwan. I. Preliminary report on isozyme study. Bulletin of Malacology, Taipei, 13: 1-7.

34 Publications (submitted, and in preparation) Liu, H.-P. and Mulvey, M. 1998. The utility of different regions of rDNA sequences in inferring phylogenetic relationships in freshwater mussels. (submitted to Molecular Phylogenetics and Evolution) Aars, J., Ims, R. A., Liu, H.-P., Mulvey, M., Smith, M. 1998. Bank voles in linear habitats shows restricted gene flow as revealed by mtDNA sequences (submitted to Molecular Ecology). Kandl, K. L., Liu, H.-P., Butler, R., Hoeh, R. W., and Mulvey, M. 1998. Clarification of Pleurobema pyriforme as a species or species-complex and implications for the conservation of rare freshwater mussels (submitted to Malacologia). Liu, H.-P., and Mulvey, M., and Mitton, J. B. 1998. Novel mtDNA variation in Pyganodon gibbosa provides insight to the Dual Uniparental Inheritance (DUI) of freshwater mussels (Unionidae) (in preparation for Evolution). Hershler, R., Liu, H.-P., and Mulvey, M. 1998. Phylogenetic relationships within the North American hydrobiid snail genus Tryonia: taxonomic and biogeographic implications. (in preparation for Molecular Biology and Evolution). Theses Liu, H.-P. 1994. Evolutionary Studies of Freshwater Mussels and Systematic Studies of Species of the Family Physidae. Doctoral dissertation. Hsiu-Ping Liu. 1988. Population Analysis and Reproductive Ecology of Meretrix in Tam-Shui and Lu-Kang Area. Master's Thesis, National Taiwan University, Taipei. Abstracts Liu, H.-P., and Mulvey, M. 1996. Genetic relationships among some Elliptio species: RFLP's analysis of amplified ITS region. Triannual Unionid Report 9: 10. Liu, H.-P., Mitton, J. B., and Wu S.-K. 1994. Mitochondrial DNA variation of Anodonta grandis grandis Say in Colorado. Triannual Unionid Report 3, January 1994. Liu, H.-P. 1992. The preliminary report on the Physa species by using isozyme technique (Gastropoda: Physidae). The Western Society of Malacologists, Annual Report 24: 35. Hsiu-Ping Liu. 1987. Application of electrophoresis on Meretrix study. Symposium on Marine Sciences, 1987: 20-21, Taipei, Taiwan. Research Experience Postdoctoral research, with Dr. M. Mulvey, Savannah River Ecology Laboratory, University of Georgia. Evolutionary studies of hydrobiid snails, genetic survey of rare freshwater mussels, and systematic studies of doubly uniparental inheritance in freshwater mussels. 1995 - 1997. Doctoral dissertation, with Dr. J. B. Mitton, University of Colorado at Boulder. Systematic studies of species of the Family Physidae. 1988 - 1994. Evolutionary studies of freshwater mussels. 1993 - 1994. Graduate Assistant, with Dr. S.-K. Wu, University of Colorado Museum. Colorado freshwater mollusks and using isozyme techniques to determine Physa and Catinella species. 1988 - 1994. Graduate independent research, with Dr. J. B. Mitton and Dr. K. Armitage, University of Colorado at Boulder. The relationship between shell length, shell weight, oxygen consumption and heterozygosity on Anguispira alternata. 1988 - 1989. Research Assistant, with Dr. W.-L. Wu, Institute of Zoology, Academia Sinica, Taipei. Natural resources studies on genus Meretrix. 1986 - 1988. Undergraduate honor study with Dr. C.-L. Liu, National Taiwan College of Marine Science and technology, Keelung. Studies on essential nutrients in shrimp diet. 1985 - 1986. Internship with Dr. Y.-L. Yu, Institute of Zoology, Academia Sinica, Taipei. Using isotope technique to study shrimp physiology. 1984. Papers presented 1997 Molecular phylogeny of hydrobiid gastropods, in the 63"' Annual Meeting of the American Malacological Union. 1997 Evolutionary patterns of Doubly Uniparental Inheritance in freshwater mussels, in 1997 Annual Meeting of the Society for the Study of Evolution. 1997 Molecular phylogenetic relationships among freshwater mussels of the subfamily Anodontinae: Conservation implication, in the 1997 Annual Meeting of the National Shellfisheries Association. 1996 Genetic relationships among Atlantic lanceolate Elliptio species: RFLP's analysis of amplified ITS region, in the 62" Annual Meeting of the American Malacological Union. 1995 Genetic data and management recommendations for the freshwater mussel, Anodonta grandis grandis Say 1829, in 9th Annual Meeting of the Society for Conservation Biology. 1995 Paternal mitochondrial DNA differentiation far exceeds maternal mitochondrial DNA and allozyme differentiation in the freshwater mussel, Anodonta grandis grandis, in 1995 Annual Meeting of the Society for the Study of Evolution. 1994 Gender-specific mitochondrial DNA within and among populations of the freshwater mussel, Anodonta grandis grandis, in Colorado in 60th Annual Meeting of the American Malacological Union. 1993 Molecular method for species identification in the Family Physidae in 26th Annual Meeting of the Western Society of Malacologists. 1991 The preliminary report on the Physa species by using isozyme technique (Gastropoda: Physidae) in 17th Annual Meeting of the Guild of Rocky Mountain Population Biologists. 1991 The preliminary report on the Physa species by using isozyme technique (Gastropoda: Physidae) in the 1991 meeting of American Malacological Union Poster Section. 1987 Natural resources studies on Genus Meretrix by using isozyme techniques in Symposium on Marine Science.