Florida Cooperative Fish and Wildlife Research Unit

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plants. The overall goal of the project is to develop a reliable genetic database used to delineate and determine thesource of sea oats to ensure ecologically sound beach and dune restoration. Ultimately, plant micropropagationtechnology and cryopreservation will be used to create a germplasm library of multiple genotypes from each majorsea oats population. This both ensures a long-term reserve of population specific genotypes for beach and dunerestoration.OBJECTIVES: To establish a germplasm library of sea oats genotypes from all major populations along Florida’s Atlanticand Gulf coasts. To determine the genetic diversity and distance of seedlings collected from the major sea oats populationsalong the Florida Gulf and Atlantic coasts using AFLP fingerprinting procedures. To evaluate use of the sea oats diversity genetic database as a tool to delineate sea oats provenance distancealong Florida’s Atlantic and Gulf coasts.PROGRESS:All immediate objectives have been accomplished. Sea oats seed was harvested from sixteen Florida State Parks orRecreation Areas: Perido Key, Navarre Beach, Henderson Beach, St. Andrews, St. George, Little Talbot Island,Anastasia, Gamble Rogers, Honeymoon Island, Sebastian Inlet, John D. MacArthur, Don Pedro Island, Delnor-Wiggins Pass, John U. Lloyd, and Bill Braggs Cape Florida. Park managers and biologists were consulted to assurethat seed was only harvested from areas that had not been replanted. Seed production varied significantly betweenpopulations. Seed was surfaced sterilized in a three-step process and germinated in vitro on Murashige & SkoogMedium in 150 X 25 mm glass culture tubes. We found that germination could be significantly enhanced bymaintaining the culture tubes on a heating mat at about 37 C. In vitro germination rates and seedling growth variedconsiderable between the sea oats populations and genotypes. We have now completed establishment of seedcultures from all sea oats the populations sampled. These data are being analyzed to determine if there are anypopulation specific differences in germination and initial seedling growth. To establish clonal lines, unbranched 7-week old seedlings were transferred to sea oats shoot multiplication medium.______________________________Determination of Population Diversity in the Florida EndangeredOrchid Cyrtopodium punctatumPrincipal Investigator: Michael E. KaneFunding Agencies: U.S. Department of Interior, USGSExpected Completion: 9/30/2010 (RWO 251, UF#77491)Cytopodium punctatum, the cigar orchid, is an endangered plant in the state of Florida. The species distributionranges from Florida and the West Indies. The genus Cyrtopodium comprises about 35 species, with C. punctatumbeing the only epiphytic member and northernmost ranging species. Cyrtopodium punctatum is a very large showyorchid that bears showy flowers. Due to its appeal, the species was over collected during the past century and todayonly a few plants still exist in inaccessible and protected areas. Three distinct populations are located in Unit 51 (ca.7 plants), 54 ca. 14 plants) and an 3 plants in Unit 38 at the Florida Panther National Wildlife Refuge (FPNWR;Collier Co., FL). With previous funding from the FPNWR, a seed propagation protocol has been developed for thefuture reintroduction of C. punctatum. Breeding system type is one of the most important determinants of the geneticcomposition of plant populations. Consequently, pollination biology and breeding system studies are beingcompleted in two FPNWR C. punctatum populations to understand the ecology and population genetics of thisspecies in situ. However, the current genetic diversity and structure in the FPNWR C. punctatum populations is notknown. This information is critical for development of ecologically sound integrated conservation plans.OBJECTIVES OF PROJECT: Determine genetic diversity of C. punctatum populations in the FPNWR. Compare genetic diversity between and within C. punctatum populations Interpret results in light of ongoing reintroduction efforts with this species.34
OBJECTIVES YR 1: Determine genetic diversity of C. punctatum populations in the FPNWR.TASKS: Leaf samples from newly developed leaves will be collected from all known plants throughoutthe FPNWF (totaling about 20 plants). DNA will be extracted using DNeasy Plant Mini Kits. PurifiedDNA will be subjected to Amplified Fragment Polymorphism (AFLP)analysis.OBJECTIVES YR 2: Compare genetic analysis between and within C. punctatum populations.TASKS: Analyze AFLP data using GeneMarker software, Use POPGENE software to estimate fixationindexes (F IS , F IT , and F ST ), effective population size (N m ), H O , and expected Nei’s and Shannon’sheterozygosity estimates (H E ). Use the program STRUCTURE v 2.2 for population assignment andprinciple coordinate analysis of data. Intepret results with respect to development of a ecologically-soundre-introduction program.PROGRESS 2009:A site visit was made to the FPNWR on December 2, 2008 to discuss sampling procedures with Larry Richardson.Since newly developed leaves of C. punctatum are required for “clean” DNA extraction, sampling will not beginuntil late March 2009 when new leaves on the donor plants will available. A site visitation with Larry Richardsonwas made on March 17, 2009 to identify the locations of the individual plants from which tissue samples will betaken for DNA analysis. Plants were just beginning to flower. The DNA extraction kits have been ordered andarrangements were made to work in the laboratory of Dr. Charles Guy to perform the DNA extractions. Variousmethods are being compared to decide which method to use to store the plant tissues prior to DNA extraction.______________________________Techniques for Field Establishment and Reintroduction ofCalopogon tuberosus Var. tuberosusPrincipal Investigator: Michael E. KaneCo-Principal Investigator: Philip J KauthFunding Agencies: U.S. Department of Interior, FWSExpected Completion: 8/30/09 (UF#69941)While much of the literature regarding orchids focuses on propagationtechniques, little information exists on reintroduction, trans-location, andfield transplanting. Habitat destruction or degradation is responsible forloss of orchid habitat and orchid populations. Reintroduction of seedlingsinto natural habitats is becoming a popular technique for conservation, butfield-establishment often fails. Much of the information regarding reestablishmentfocuses on seedlings. A major obstacle to fieldestablishment is initial survival. Only a few articles discuss techniques forincreasing survival of orchid seedlings. One technique used to increasesurvival is planting dormant storage organs such as tubers or corms.Calopgon tuberosus in the FL PantherNational Wildlife RefugeCalopogon tuberosus var. tuberosus is a cormforming species found throughout eastern NorthAmerica includingsouthwest Florida. In south Florida, this species has up to ten magenta flowers that open in succession. Theflowering season begins in April and continues through the end of May in south Florida. Seed capsules are fullyripe approximately 6-8 weeks after pollination. Reintroduction of Calopogon tuberosus to suitable habitats is thenext logical step in a propagation experiment. We have successfully germinated seeds of C. tuberosus to theseedling stage, and have successfully grown seedlings from other populations under greenhouse conditions.35
Page 1 and 2: Florida Cooperative Fishand Wildlif
Page 3: COOPERATING AGENCIES:FLORIDA FISH A
Page 6 and 7: RESEARCH MISSION STATEMENTThe St. J
Page 8 and 9: Elizabeth Martin - NBII Bird Conser
Page 10 and 11: Students and PersonnelFull Name: Ad
Page 12 and 13: Full Name: Brian E. ReichertDegree
Page 14 and 15: Strategic Habitat Conservation for
Page 16 and 17: PROGRESS:The pilot phase of the pro
Page 18 and 19: droughts) disproportionally impact
Page 20 and 21: system. Cape San Blas experiences t
Page 22 and 23: 2009 PROGRESS:Four Nova 2 small unm
Page 24 and 25: ______________________________Exper
Page 26 and 27: OBJECTIVES: Annually document numbe
Page 28 and 29: Following the successes of this spr
Page 30 and 31: 1) Excavate a minimum of three hist
Page 32 and 33: Directing Succession Through Adapti
Page 36 and 37: OBJECTIVES: Reintroduce Calopogon t
Page 38 and 39: This project is a continuation of e
Page 40 and 41: Babcock/Webb WMA has been an import
Page 42 and 43: Everglades. Recon flights for Sawgr
Page 44 and 45: Beach nourishment is used to repair
Page 46 and 47: ______________________________Struc
Page 48 and 49: Gulf Sturgeon from the Apalachicola
Page 50 and 51: those released under Control condit
Page 52 and 53: Eglin Air Force Base (EAFB) owns ap
Page 54 and 55: of those habitats (changes in hydro
Page 56 and 57: COMPLETED PROJECTS of Florida Unit
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Page 60 and 61: Channel (RWO62) PI: A.B. Bolten, K.
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Page 64 and 65: Kappes, Jr.; Completion Date: Augus
Page 66 and 67: 172. Biological Diversity in Florid
Page 68 and 69: R.R. Carthy, J.D. Nichols; Personne
Page 70 and 71: A. Schwarzer; Completed: June 20072
Page 72: small unmanned aircraft system for

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