We DNA barcoded 2,597 parasitoid wasps belonging to 6 microgastrine braconid genera reared from parapatric tropical dry forest, cloud forest, and rain forest in Area de Conservació n Guanacaste (ACG) in northwestern Costa Rica and combined these data with records of caterpillar hosts and morphological analyses. We asked whether barcoding and morphology discover the same provisional species and whether the biological entities revealed by our analysis are congruent with wasp host specificity. Morphological analysis revealed 171 provisional species, but barcoding exposed an additional 142 provisional species; 95% of the total is likely to be undescribed. These 313 provisional species are extraordinarily host specific; more than 90% attack only 1 or 2 species of caterpillars out of more than 3,500 species sampled. The most extreme case of overlooked diversity is the morphospecies Apanteles leucostigmus. This minute black wasp with a distinctive white wing stigma was thought to parasitize 32 species of ACG hesperiid caterpillars, but barcoding revealed 36 provisional species, each attacking one or a very few closely related species of caterpillars. When host records and/or within-ACG distributions suggested that DNA barcoding had missed a species-pair, or when provisional species were separated only by slight differences in their barcodes, we examined nuclear sequences to test hypotheses of presumptive species boundaries and to further probe host specificity. Our iterative process of combining morphological analysis, ecology, and DNA barcoding and reiteratively using specimens maintained in permanent collections has resulted in a much more fine-scaled understanding of parasitoid diversity and host specificity than any one of these elements could have produced on its own.Area de Conservació n Guanacaste ͉ Costa Rica ͉ caterpillar ͉ Braconidae ͉ host specificity

Insect parasitoids are a major component of global biodiversity and affect the population dynamics of their hosts. However, identification of insect parasitoids is often difficult, and they are suspected to contain many cryptic species. Here, we ask whether the cytochrome c oxidase I DNA barcode could function as a tool for species identification and discovery for the 20 morphospecies of Belvosia parasitoid flies (Diptera: Tachinidae) that have been reared from caterpillars (Lepidoptera) in Area de Conservació n Guanacaste (ACG), northwestern Costa Rica. Barcoding not only discriminates among all 17 highly host-specific morphospecies of ACG Belvosia, but it also raises the species count to 32 by revealing that each of the three generalist species are actually arrays of highly hostspecific cryptic species. We also identified likely hybridization among Belvosia by using a variable internal transcribed spacer region 1 nuclear rDNA sequence as a genetic covariate in addition to the strategy of overlaying barcode clusters with ecological information. If general, these results will increase estimates of global species richness and imply that tropical conservation and host-parasite interactions may be more complex than expected.Area de Conservació n Guanacaste ͉ Belvosia ͉ cytochrome c oxidase I ͉ internal transcribed spacer region 1 ͉ species richness

A DNA barcode based on 650 bp of mitochondrial gene cytochrome c oxidase I is proving to be highly functional in species identification for various animal groups. However, DNA degradation complicates the recovery of a full-length barcode from many museum specimens. Here we explore the use of shorter barcode sequences for identification of such specimens. We recovered short sequences -i.e. ∼ ∼ ∼ ∼ 100 bp -with a single PCR pass from more than 90% of the specimens in assemblages of moth and wasp museum specimens from which full barcode recovery was only 50%, and the latter were usually less than 8 years old. Short barcodes were effective in identifying specimens, confirming their utility in circumstances where full barcodes are too expensive to obtain and the identification comparisons are within a confined taxonomic group.

Many species of tachinid flies are viewed as generalist parasitoids because what is apparently a single species of fly has been reared from many species of caterpillars. However, an ongoing inventory of the tachinid flies parasitizing thousands of species of caterpillars in Area de Conservació n Guanacaste, northwestern Costa Rica, has encountered >400 species of specialist tachinids with only a few generalists. We DNA-barcoded 2,134 flies belonging to what appeared to be the 16 most generalist of the reared tachinid morphospecies and encountered 73 mitochondrial lineages separated by an average of 4% sequence divergence. These lineages are supported by collateral ecological information and, where tested, by independent nuclear markers (28S and ITS1), and we therefore view these lineages as provisional species. Each of the 16 apparently generalist species dissolved into one of four patterns: (i) a single generalist species, (ii) a pair of morphologically cryptic generalist species, (iii) a complex of specialist species plus a generalist, or (iv) a complex of specialists with no remaining generalist. In sum, there remained 9 generalist species among the 73 mitochondrial lineages we analyzed, demonstrating that a generalist lifestyle is possible for a tropical caterpillar parasitoid fly. These results reinforce the emerging suspicion that estimates of global species richness are likely underestimates for parasitoids (which may constitute as much as 20% of all animal life) and that the strategy of being a tropical generalist parasitic fly may be yet more unusual than has been envisioned for tachinids.

The role of DNA barcoding as a tool to accelerate the inventory and analysis of diversity for hyperdiverse arthropods is tested using ants in Madagascar. We demonstrate how DNA barcoding helps address the failure of current inventory methods to rapidly respond to pressing biodiversity needs, specifically in the assessment of richness and turnover across landscapes with hyperdiverse taxa. In a comparison of inventories at four localities in northern Madagascar, patterns of richness were not significantly different when richness was determined using morphological taxonomy (morphospecies) or sequence divergence thresholds (Molecular Operational Taxonomic Unit(s); MOTU). However, sequence-based methods tended to yield greater richness and significantly lower indices of similarity than morphological taxonomy. MOTU determined using our molecular technique were a remarkably local phenomenon-indicative of highly restricted dispersal and/or long-term isolation. In cases where molecular and morphological methods differed in their assignment of individuals to categories, the morphological estimate was always more conservative than the molecular estimate. In those cases where morphospecies descriptions collapsed distinct molecular groups, sequence divergences of 16% (on average) were contained within the same morphospecies. Such high divergences highlight taxa for further detailed genetic, morphological, life history, and behavioral studies.

Inventory of the caterpillars, their food plants and parasitoids began in 1978 for today's Area de Conservacion Guanacaste (ACG), in northwestern Costa Rica. This complex mosaic of 120 000 ha of conserved and regenerating dry, cloud and rain forest over 0-2000 m elevation contains at least 10 000 species of non-leaf-mining caterpillars used by more than 5000 species of parasitoids. Several hundred thousand specimens of ACG-reared adult Lepidoptera and parasitoids have been intensively and extensively studied morphologically by many taxonomists, including most of the co-authors. DNA barcoding -the use of a standardized short mitochondrial DNA sequence to identify specimens and flush out undisclosed species -was added to the taxonomic identification process in 2003.

Seasonally dry tropical forests are distributed across Latin America and the Caribbean and are highly threatened, with less than 10% of their original extent remaining in many countries. Using 835 inventories covering 4660 species of woody plants, we show marked floristic turnover among inventories and regions, which may be higher than in other neotropical biomes, such as savanna. Such high floristic turnover indicates that numerous conservation areas across many countries will be needed to protect the full diversity of tropical dry forests. Our results provide a scientific framework within which national decision-makers can contextualize the floristic significance of their dry forest at a regional and continental scale. N eotropical seasonally dry forest (dry forest) is a biome with a wide and fragmented distribution, found from Mexico to Argentina and throughout the Caribbean (1, 2) ( Fig. 1). It is one of the most threatened tropical forests in the world (3), with less than 10% of its original extent remaining in many countries (4).Following other authors (5, 6), we define dry forest as having a closed canopy, distinguishing it from more open, grass-rich savanna. It occurs on fertile soils where the rainfall is less thañ 1800 mm per year, with a period of 3 to 6 months receiving less than 100 mm per month (5-7), during which the vegetation is mostly deciduous. Seasonally dry areas, especially in Peru and Mexico, were home to pre-Columbian civilizations, so human interaction with dry forest has a long history (8). The climates and fertile soils of dry forest regions have led to higher human population densities and an increasing demand for energy and land, enhancing degradation (9). More recently, destruction of dry forest has been accelerated by intensive cultivation of crops, such as sugar cane, rice and soy, or by conversion to pasture for cattle.Dry forest is in a critical state because so little of it is intact, and of the remnant areas, little is protected (3). For example, only 1.2% of the total Caatinga region of dry forest in Brazil is fully protected compared with 9.9% of the Brazilian Amazon (10). Conservation actions are urgently needed to protect dry forest's unique biodiversity-many plant species and even genera are restricted to it and reflect an evolutionary history confined to this biome (1).We evaluate the floristic relationships of the disjunct areas of neotropical dry forest and highlight those that contain the highest diversity and endemism of woody plant species. We also explore woody plant species turnover across geographic space among dry forests. Our results provide a framework to allow the conservation significance of each separate major region of dry forest to be assessed at a continental scale. Our analyses are based on a subset of a data set of 1602 inventories made in dry forest and related semi-deciduous forests from Mexico and the Caribbean to Argentina and Paraguay that covers 6958 woody species, which has been compiled by the Latin American and Caribbean Seasonally Dry Tropica...

Three new genera are described: Michener (Proteropinae), Bioalfa (Rogadinae), and Hermosomastax (Rogadinae). Keys are given for the New World genera of the following braconid subfamilies: Agathidinae, Braconinae, Cheloninae, Homolobinae, Hormiinae, Ichneutinae, Macrocentrinae, Orgilinae, Proteropinae, Rhysipolinae, and Rogadinae. In these subfamilies 416 species are described or redescribed. Most of the species have been reared and all but 13 are new to science. A consensus sequence of the COI barcodes possessed by each species is employed to diagnose the species, and this approach is justified in the introduction. Most descriptions consist of a lateral or dorsal image of the holotype, a diagnostic COI consensus barcode, the Barcode Index Number (BIN) code with a link to the Barcode of Life Database (BOLD), and the holotype specimen information required by the International Code of Zoological Nomenclature. The following species are treated and those lacking authorship are newly described here with authorship attributable to Sharkey except for the new species of Macrocentrinae which are by Sharkey & van Achterberg: AGATHIDINAE: Aerophilus paulmarshi, Mesocoelus davidsmithi, Neothlipsis bobkulai, Plesiocoelus vanachterbergi, Pneumagathis erythrogastra (Cameron, 1905), Therophilus bobwhartoni, T. donaldquickei, T. gracewoodae, T. maetoi, T. montywoodi, T. penteadodiasae, Zacremnops brianbrowni, Z. coatlicue Sharkey, 1990, Zacremnops cressoni (Cameron, 1887), Z. ekchuah Sharkey, 1990, Z. josefernandezi, Zelomorpha sarahmeierottoae. BRACONINAE: Bracon alejandromarini, B. alejandromasisi, B. alexamasisae, B. andresmarini, B. andrewwalshi, B. anniapicadoae, B. anniemoriceae, B. barryhammeli, B. bernardoespinozai, B. carlossanabriai, B. chanchini, B. christophervallei, B. erasmocoronadoi, B. eugeniephillipsae, B. federicomatarritai, B. frankjoycei, B. gerardovegai, B. germanvegai, B. isidrochaconi, B. jimlewisi, B. josejaramilloi, B. juanjoseoviedoi, B. juliodiazi, B. luzmariaromeroae, B. manuelzumbadoi, B. marialuisariasae, B. mariamartachavarriae, B. mariorivasi, B. melissaespinozae, B. nelsonzamorai, B. nicklaphami, B. ninamasisae, B. oliverwalshi, B. paulamarinae, B. rafamoralesi, B. robertofernandezi, B. rogerblancoi, B. ronaldzunigai, B. sigifredomarini, B. tihisiaboshartae, B. wilberthbrizuelai, Digonogastra montylloydi, D. montywoodi, D. motohasegawai, D. natwheelwrighti, D. nickgrishini. CHELONINAE: Adelius adrianguadamuzi, A. gauldi Shimbori & Shaw, 2019, A. janzeni Shimbori & Shaw, 2019, Ascogaster gloriasihezarae, A. grettelvegae, A. guillermopereirai, A. gustavoecheverrii, A. katyvandusenae, A. luisdiegogomezi, Chelonus alejandrozaldivari, C. gustavogutierrezi, C. gustavoinduni, C. harryramirezi, C. hartmanguidoi, C. hazelcambroneroae, C. iangauldi, C. isidrochaconi, C. janecheverriae, C. jeffmilleri, C. jennyphillipsae, C. jeremydewaardi, C. jessiehillae, C. jesusugaldei, C. jimlewisi, C. jimmilleri, C. jimwhitfieldi, C. johanvalerioi, C. johnburnsi, C. johnnoyesi, C. jorgebaltodanoi, C. jorgehernandezi, C. josealfredohernandezi, C. josefernandeztrianai, C. josehernandezcortesi, C. josemanuelperezi, C. josephinerodriguezae, C. juanmatai, C. junkoshimurae, C. kateperezae, C. luciariosae, C. luzmariaromeroae, C. manuelpereirai, C. manuelzumbadoi, C. marianopereirai, C. maribellealvarezae, C. markmetzi, C. markshawi, C. martajimenezae, C. mayrabonillae, C. meganmiltonae, C. melaniamunozae, C. michaelstroudi, C. michellevanderbankae, C. mingfangi, C. minorcarmonai, C. monikaspringerae, C. moniquegilbertae, C. motohasegawai, C. nataliaivanovae, C. nelsonzamorai, C. normwoodleyi, C. osvaldoespinozai, C. pamelacastilloae, C. paulgoldsteini, C. paulhansoni, C. paulheberti, C. petronariosae, C. ramyamanjunathae, C. randallgarciai, C. rebeccakittelae, C. robertoespinozai, C. robertofernandezi, C. rocioecheverriae, C. rodrigogamezi, C. ronaldzunigai, C. rosibelelizondoae, C. rostermoragai, C. ruthfrancoae, C. scottmilleri, C. scottshawi, C. sergioriosi, C. sigifredomarini, C. stevearonsoni, C. stevestroudi, C. sujeevanratnasinghami, C. sureshnaiki, C. torbjornekremi, C. yeimycedenoae, Leptodrepana alexisae, L. erasmocoronadoi, L. felipechavarriai, L. freddyquesadai, L. gilbertfuentesi, L. manuelriosi, Phanerotoma almasolisae, P. alvaroherrerai, P. anacordobae, P. anamariamongeae, P. andydeansi, P. angelagonzalezae, P. angelsolisi, P. barryhammeli, P. bernardoespinozai, P. calixtomoragai, P. carolinacanoae, P. christerhanssoni, P. christhompsoni, P. davesmithi, P. davidduthiei, P. dirksteinkei, P. donquickei, P. duniagarciae, P. duvalierbricenoi, P. eddysanchezi, P. eldarayae, P. eliethcantillanoae, P. jenopappi, Pseudophanerotoma alanflemingi, Ps. albanjimenezi, Ps. alejandromarini, Ps. alexsmithi, Ps. allisonbrownae, Ps. bobrobbinsi. HOMOLOBINAE: Exasticolus jennyphillipsae, E. randallgarciai, E. robertofernandezi, E. sigifredomarini, E. tomlewinsoni. HORMIINAE: Hormius anamariamongeae, H. angelsolisi, H. anniapicadoae, H. arthurchapmani, H. barryhammeli, H. carmenretanae, H. carloswalkeri, H. cesarsuarezi, H. danbrooksi, H. eddysanchezi, H. erikframstadi, H. georgedavisi, H. grettelvegae, H. gustavoinduni, H. hartmanguidoi, H. hectoraritai, H. hesiquiobenitezi, H. irenecanasae, H. isidrochaconi, H. jaygallegosi, H. jimbeachi, H. jimlewisi, H. joelcracrafti, H. johanvalerioi, H. johnburleyi, H. joncoddingtoni, H. jorgecarvajali, H. juanmatai, H. manuelzumbadoi, H. mercedesfosterae, H. modonnellyae, H. nelsonzamorai, H. pamelacastilloae, H. raycypessi, H. ritacolwellae, H. robcolwelli, H. rogerblancosegurai, H. ronaldzunigai, H. russchapmani, H. virginiaferrisae, H. warrenbrighami, H. willsflowersi. ICHNEUTINAE: Oligoneurus kriskrishtalkai, O. jorgejimenezi, Paroligoneurus elainehoaglandae, P. julianhumphriesi, P. mikeiviei. MACROCENTRINAE: Austrozele jorgecampabadali, A. jorgesoberoni, Dolichozele gravitarsis (Muesebeck, 1938), D. josefernandeztrianai, D. josephinerodriguezae, Hymenochaonia kalevikulli, H. kateperezae, H. katherinebaillieae, H. katherineellisonae, H. katyvandusenae, H. kazumifukunagae, H. keithlangdoni, H. keithwillmotti, H. kenjinishidai, H. kimberleysheldonae, H. krisnorvigae, H. lilianamadrigalae, H. lizlangleyae, Macrocentrus fredsingeri, M. geoffbarnardi, M. gregburtoni, M. gretchendailyae, M. grettelvegae, M. gustavogutierrezi, M. hannahjamesae, M. harisridhari, M. hillaryrosnerae, M. hiroshikidonoi, M. iangauldi, M. jennyphillipsae, M. jesseausubeli, M. jessemaysharkae, M. jimwhitfieldi, M. johnbrowni, M. johnburnsi, M. jonathanfranzeni, M. jonathanrosenbergi, M. jorgebaltodanoi, M. lucianocapelli. ORGILINAE: Orgilus amyrossmanae, O. carrolyoonae, O. christhompsoni, O. christinemcmahonae, O. dianalipscombae, O. ebbenielsoni, O. elizabethpennisiae, O. evertlindquisti, O. genestoermeri, O. jamesriegeri, O. jeanmillerae, O. jeffmilleri, O. jerrypowelli, O. jimtiedjei, O. johnlundbergi, O. johnpipolyi, O. jorgellorentei, O. larryspearsi, O. marlinricei, O. mellissaespinozae, O. mikesmithi, O. normplatnicki, O. peterrauchi, O. richardprimacki, O. sandraberriosae, O. sarahmirandae, O. scottmilleri, O. scottmorii, Stantonia billalleni, S. brookejarvisae, S. donwilsoni, S. erikabjorstromae, S. garywolfi, S. henrikekmani, S. luismirandai, S. miriamzunzae, S. quentinwheeleri, S. robinkazmierae, S. ruthtifferae. PROTEROPINAE: Hebichneutes tricolor Sharkey & Wharton, 1994, Proterops iangauldi, P. vickifunkae, Michener charlesi. RHYSIPOLINAE: Pseudorhysipolis luisfonsecai, P. mailyngonzalezaeRhysipolis julioquirosi. ROGADINAE: Aleiodes adrianaradulovae, A. adrianforsythi, A. agnespeelleae, A. alaneaglei, A. alanflemingi, A. alanhalevii, A. alejandromasisi, A. alessandracallejae, A. alexsmithi, A. alfonsopescadori, A. alisundermieri, A. almasolisae, A. alvarougaldei, A. alvaroumanai, A. angelsolisi, A. annhowdenae, A. bobandersoni, A. carolinagodoyae, A. charlieobrieni, A. davefurthi, A. donwhiteheadi, A. doylemckeyi, A. frankhovorei, A. henryhowdeni, A. inga Shimbori & Shaw, 2020, A. johnchemsaki, A. johnkingsolveri, A. gonodontovorus Shimbori & Shaw, 2020, A. manuelzumbadoi, A. mayrabonillae, A. michelledsouzae, A. mikeiviei, A. normwoodleyi, A. pammitchellae, A. pauljohnsoni, A. rosewarnerae, A. steveashei, A. terryerwini, A. willsflowersi, Bioalfa pedroleoni, B. alvarougaldei, B. rodrigogamezi, Choreborogas andydeansi, C. eladiocastroi, C. felipechavarriai, C. frankjoycei, Clinocentrus andywarreni, Cl. angelsolisi, Cystomastax alexhausmanni, Cy. angelagonzalezae, Cy. ayaigarashiae, Hermosomastax clavifemorus Quicke sp. nov., Heterogamus donstonei, Pseudoyelicones bernsweeneyi, Stiropius bencrairi, S. berndkerni, S. edgargutierrezi, S. edwilsoni, S. ehakernae, Triraphis billfreelandi, T. billmclarneyi, T. billripplei, T. bobandersoni, T. bobrobbinsi, T. bradzlotnicki, T. brianbrowni, T. brianlaueri, T. briannestjacquesae, T. camilocamargoi, T. carlosherrerai, T. carolinepalmerae, T. charlesmorrisi, T. chigiybinellae, T. christerhanssoni, T. christhompsoni, T. conniebarlowae, T. craigsimonsi, T. defectus Valerio, 2015, T. danielhubi, T. davidduthiei, T. davidwahli, T. federicomatarritai, T. ferrisjabri, T. mariobozai, T. martindohrni, T. matssegnestami, T. mehrdadhajibabaei, T. ollieflinti, T. tildalauerae, Yelicones dirksteinkei, Y. markmetzi, Y. monserrathvargasae, Y. tricolor Quicke, 1996. Y. woldai Quicke, 1996. The following new combinations are proposed: Neothlipsis smithi (Ashmead), new combination for Microdus smithi Ashmead, 1894; Neothlipsis pygmaeus (Enderlein), new combination for Microdus pygmaeus Enderlein, 1920; Neothlipsis unicinctus (Ashmead), new combination for Microdus unicinctus Ashmead, 1894; Therophilus anomalus (Bortoni and Penteado-Dias) new combination for Plesiocoelus anomalus Bortoni and Penteado-Dias, 2015; Aerophilus areolatus (Bortoni and Penteado-Dias) new combination for Plesiocoelus areolatus Bortoni and Penteado-Dias, 2015; Pneumagathis erythrogastra (Cameron) new combination for Agathis erythrogastra Cameron, 1905. Dolichozele citreitarsis (Enderlein), new combination for Paniscozele citreitarsis Enderlein, 1920. Dolichozele fuscivertex (Enderlein) new combination for Paniscozele fuscivertex Enderlein, 1920. Finally, Bassus brooksi Sharkey, 1998 is synonymized with Agathis erythrogastra Cameron, 1905; Paniscozele griseipes Enderlein, 1920 is synonymized with Dolichozele koebelei Viereck, 1911; Paniscozele carinifrons Enderlein, 1920 is synonymized with Dolichozele fuscivertex (Enderlein, 1920); and Paniscozele nigricauda Enderlein,1920 is synonymized with Dolichozele quaestor (Fabricius, 1804). (originally described as Ophion quaestor Fabricius, 1804).