SYSTEMATICS OF EVANIIDAE, CHARISMATIC PREDATORS OF COCKROACH EGGS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 0213039
• Project Start Date: Sep 1, 2007
• Project End Date: Sep 1, 2012
• Program Code: [(N/A)]- (N/A)

Recipient Organization
NORTH CAROLINA STATE UNIV
(N/A)
RALEIGH,NC 27695

Performing Department
Entomology

Non Technical Summary
All verifiable rearing records indicate that ensign wasp larvae (Hymenoptera: Evaniidae) develop as solitary egg predators within the oothecae of cockroaches. Adult ensign wasps are often found searching inside buildings or through leaf litter for potential prey and are easily recognized by their flag-shaped metasomas, which are often pumped up and down in a chopping motion. Despite many fascinating biological attributes, including extreme wing venation pattern heterogeneity, mimicry complexes involving color and morphology, variable oviposition behaviors, impact on cockroach evolution, and their potential use as biological control agents of pestiferous cockroaches, this group has been somewhat neglected by biologists due to an absence of phylogenetic knowledge and a distressed taxonomy. There are now 435 valid extant species of ensign wasps worldwide assigned to 20 genera, with at least 800 spp. remaining to be described. We seek to improve the higher-level taxonomy in order to facilitate desperately needed species-level revisions. We are constructing a database that will be revealed through a website dedicated to ensign wasp biology. The current version (http://evaniid.tamu.edu/) will be migrated to NC State and upgraded to include images and maps for all species. Phylogenetic relationships will be estimated using morphological (this will allow us to include fossils and rare species) and molecular data. Upon securing a robust pattern of relationships we can then estimate divergence times of major lineages to answer questions about how these wasps have impacted the evolution of their prey (cockroaches) - i.e., when did cockroaches begin to protect their egg cases, and was that a response to predation by ensign wasps? Cockroaches employ numerous behavioral and physiological strategies for defending their eggs against predation and parasitism. Our hypothesis is that these strategies are driven in part by evaniid predation upon cockroach eggs. Upon completion of this project we will have a complete phylogeny for all genera, a website that informs interested parties about ensign wasps, a clearer picture of the coevolution between cockroaches and ensign wasps.

Animal Health Component

(N/A)

Research Effort Categories

Basic

(N/A)

Applied

(N/A)

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
136	3110	1130	60%
721	3110	1130	10%
901	3110	1130	30%

Knowledge Area
136 - Conservation of Biological Diversity; 901 - Program and Project Design, and Statistics; 721 - Insects and Other Pests Affecting Humans;

Subject Of Investigation
3110 - Insects;

Field Of Science
1130 - Entomology and acarology;

Keywords

cockroach

blattaria

blattodea

ensign wasp

evaniidae

hymenoptera

coevolution

phylogenetics

taxonomy

systematics

Goals / Objectives
A phylogeny and a well-defined taxonomy of Evaniidae are critical to further explore evaniid evolution and open paths to new lines of research. With revised genera and an understanding of how the major evaniid lineages relate to one another we can build a robust tribal taxonomy that refers to biologically and morphologically distinct lineages. Using a holistic approach we will incorporate information from fossil taxa into our revised generic concepts. Fifteen exclusively fossil genera have been described since 1975, mostly without considering the vast array of morphologies exhibited by extant genera. Extant genera, likewise, are based almost exclusively on observations of other extant taxa. The resulting confounded classification includes synonyms and overlapping concepts. A resolved, robust phylogeny will solidify taxonomic concepts and alleviate problems with the current classification. A revision of current generic concepts is required to sort the 15,000+ evaniid specimens assembled from these projects. Questions abound concerning prey specificity, predictions of cockroach faunal composition based on the presence of evaniid spp., habitat specificity, elevational and latitudinal turnover, and the extent of mimicry complexes. Solid generic and higher-level concepts based on morphology are desperately needed to address these biodiversity questions. Though tens of thousands of specimens have been collected, we currently have no means to identify them to species other than direct comparison to type specimens. Revised genera will facilitate desperately needed species-level revisions. Dictyoptera is known as one of the oldest hexapod lineages, with fossils going back to at least the Carboniferous. Evaniidae potentially radiated on their Blattaria prey before the breakup of Gondwana, and current distribution of ensign wasp taxa may perfectly reflect ancient vicariance events. An alternative is that there have been long-distance, post break-up dispersal events. By estimating divergence times and relationships and comparing apparent patterns to known continental positions we can begin to address questions about current distributions and radiation events. 4000 spp. of cockroaches (Blattaria) have been described, 99% of which stack their eggs together and encase them inside a protein envelope called an ootheca. Two hypotheses account for the evolution of egg encasement in Blattaria: 1) the ootheca appeared as an adaptive reaction to climatic changes in the Permian, 2) the ootheca evolved during the Jurassic to protect against predation. Blattarian lineages subsequently evolved behaviors thought to further protect their developing embryos - burying or disguising the ootheca, securing it in a hostile environment, retaining egg cases externally and then internally, and finally true viviparity, which are all considered to be continued adaptive responses to either hypothesized selection pressure scenarios. Age estimates of Evaniidae and Blattaria combined with well-resolved phylogenies of both Evaniidae and Blattaria and the extensive literature on ootheca physiology, would provide conclusive insights into coevolutionary events between these insects.

Project Methods
We anticipate assembling a dataset for all fossil evanioid taxa (n=~80) and at least 120 extant Evanioidea, including 18-19 of the 20 extant genera. Evaniella, Prosevania, and Semaeomyia account for 50% of evaniid diversity and exhibit a wide range of morphotypes. These genera are optimal candidates for splitting and will be represented by at least 20 exemplars each. At least 30 other examplars from Evanioidea, Evaniomorpha, and Apocrita, including fossils, will be used as outgroups. All 230 exemplars will be scored for morphology and non-fossil taxa sequenced for nuclear protein-encoding genes. The mx content management system will be used to manage character hypotheses, sequencing progress, and to database images prior to deposition into Morphbank. Historically within Hymenoptera the use of molecular markers has been heavily biased toward mitochondrial and ribosomal genes, which reflects in the data already available for Evaniidae and its outgroups. Poor resolving power of these genes resulted in almost no resolution among the evaniid genera. We seek to strengthen our data with the addition of nuclear protein-encoding genes. No comprehensive morphological datasets have yet been assembled for Evaniidae. Two small studies addressed relationships between higher-level fossil evanioids, but each used the same ~25 characters. We have a preliminary dataset of 33 non-wing characters (no overlap with earlier studies) and expect to expand this to at least 150 characters. Each exemplar will be dissected (extant taxa only) and imaged using standard views, with micrographs deposited in morphbank (http://morphbank.net/). Morphbank is developing annotation tools for scoring characters and exporting NEXUS-formatted datasets. Each character will be associated with a collection of images that documents the state(s) for each exemplar. Each image in the collection will be annotated with textual descriptions and arrows and/or boxes to justify our scoring of said character. This portion of the grant will be the main focus of the graduate student. As part of our involvement in HymAToL we are developing an ontology of morphological terms (spearheaded by Deans). This ontology is available currently as a glossary and markup tool in mx and will be implemented in Morphbank as a markup tool, an enhanced search algorithm, and to establish homologies. This process will allow others to more easily replicate or comment on our data/results. Morphological and molecular datasets will be analyzed individually and then together in total evidence parsimony and Bayesian approaches using mixed models. Divergence times will be estimated using parametric (Bayesian in BEAST or MrBayes, e.g.) and non-parametric (r8s, e.g.) approaches.

Progress 10/01/10 to 09/30/11

Outputs
OUTPUTS: The database at http://evanioidea.info continues to grow with respect to updated taxonomy and numbers of databased, georeferenced specimens (more than 32000 now). We also have extracted DNA from 275 specimens, performed almost 6000 polymerase chain reactions in order to amplify 16 gene fragments for sequencing. These DNA data, in combination with specimen data, are being used to address questions regarding ensign wasp biodiversity patterns and host associations. We are also using DNA (specifically a fragment of the mitochondrial cytochrome c oxidase gene) data as another line of evidence to assist in the delimitation of species, along with more traditional phenotypic characters (e.g., anatomy). Deans is mentoring three graduate students, a technician, and an undergraduate in the process of insect systematics and biodiversity informatics. Seven talks were given at scientific meetings. Two videos of evaniid oviposition, the first ever made, were published to the Web: http://www.youtube.com/watchv=sB2b7fiRJhI http://www.youtube.com/watchv=rQAa0VtD32U PARTICIPANTS: Trish Mullins, graduate student at NC State University, is revising several New World genera. Matthew Bertone, postdoc at NC State University, has contributed videos of the ensign wasp oviposition behaviors. Dr. Barb Sharanowski, who was a postdoc at NC State,spearheaded the development of new molecular markers and the collection of molecular data. Katja Seltmann, project manager at NC State, is examining ensign wasp fossils and is developing a morphological data set for estimating relations and divergence times between lineages. Amy Bader, who was a technician at NC State University this year, collected molecular data and is databasing specimens. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
We've discovered several new nuclear protein-encoding genetic markers (e.g., tango7, RPS23) that have utility across multiple evolutionary levels, from delimiting species to estimating relationships between genera. These new markers enable the community to move beyond the typical, oftentimes limiting mitochondrial markers. Using these markers in combination with morphological characters we have identified several new species that are queued to be described, as well as several new synonymies that need to be published. Most of this work is focused on the evaniid fauna of the New World. The project website has a growing array of diagnostic tools (dichotomous and multi-entry keys), which have drawn almost 2500 hits (i.e., uses) in the last year. These kinds of diagnostic tools are not available elsewhere.

Publications

• No publications reported this period

Progress 10/01/09 to 09/30/10

Outputs
OUTPUTS: The database at http://evanioidea.info continues to grow with respect to updated taxonomy and numbers of databased, georeferenced specimens (more than 5000 now). We also have extracted DNA from 250 specimens, performed almost 5000 polymerase chain reactions in order to amplify 16 gene fragments for sequencing. These DNA data, in combination with specimen data, are being used to address questions regarding ensign wasp biodiversity patterns and host associations. We are also using DNA (specifically a fragment of the mitochondrial cytochrome c oxidase gene) data as another line of evidence to assist in the delimitation of species, along with more traditional phenotypic characters (e.g., anatomy). Deans is mentoring three graduate students, a technician, and an undergraduate in the process of insect systematics and biodiversity informatics. Five talks were given at scientific meetings. Hundreds of specimens were sent to us for identification. Two videos of evaniid oviposition, the first ever made, were published to the Web: http://www.youtube.com/watch?v=sB2b7fiRJhI http://www.youtube.com/watch?v=rQAa0VtD32U PARTICIPANTS: Trish Mullins, graduate student at NC State University, is revising several New World genera. Matthew Bertone, postdoc at NC State University, has contributed videos of the ensign wasp oviposition behaviors. Dr. Barb Sharanowski, postdoc at NC State,spearheaded the development of new molecular markers and the collection of molecular data. Katja Seltmann, project manager at NC State, is examining ensign wasp fossils and is developing a morphological data set for estimating relations and divergence times between lineages. Amy Bader, technician at NC State University, is collecting molecular data and is databasing specimens. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
We've discovered several new nuclear protein-encoding genetic markers that have utility across multiple evolutionary levels, from delimiting species to estimating relationships between genera. These new markers enable the community to move beyond the typical, oftentimes limiting mitochondrial markers. Using these markers in combination with morphological characters we have identified several new species that are queued to be described, as well as several new synonymies that need to be published. Most of this work is focused on the evaniid fauna of the New World. The project website has a growing array of diagnostic tools (dichotomous and multi-entry keys), which have drawn almost 1400 hits (i.e., uses) in the last year. These kinds of diagnostic tools are not available elsewhere.

Publications

• Trish Mullins and Matt Bertone. 2010 Oviposition behaviors of Hyptia thoracica and Evania appendigaster. Hamuli 1(1):10-11 Trish Mullins. Hyptia floridana. electronic publication: http://blog.insectmuseum.org/?p=1907

Progress 10/01/08 to 09/30/09

Outputs
OUTPUTS: Activities: Services: Specimens were submitted to Deans by the natural history museum in Brussels. Products: Our molecular dataset continues to grow, with the efforts of Barb Sharanowski (NCSU postdoc), Matt Hill (honors undergraduate), Kate Muirhead (Adelaide postdoc) and John Jennings (Adelaide collaborator). We have partial sequences of the mitochondrial cytochrome oxidase 1, the nuclear 28S rRNA, and the nuclear protein encoding genes CAD and tango-7. The Evanioidea Online website (http://evanioidea.info/) continues to grow, with new taxa added this year, as well as more references. Emerging data were presented by Sharanowski & Deans and Rinchhuanawma at the Entomological Society of America's annual meeting. PARTICIPANTS: A postdoc, Barb Sharanowski, is working on the project as a molecular data expert. One graduate student, Awma Rinchhuanawma, is developing morphological characters and collecting molecular data in order to test taxon hypotheses. He is being trained in the latest morphological methods, including microdissection and imaging. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
We continue to use ensign wasps as a model for changing the way descriptive taxonomy is done, by infusing the manuscripts and database products with an unprecedented level of informatics and community standards (e.g., those standards advocated by TDWG). In terms of molecular data, early results indicate that the markers we chose to sequence yield topologically congruent trees and results that are more fully resolved than any previous attempts by my lab.

Publications

• No publications reported this period

Progress 10/01/07 to 09/30/08

Outputs
OUTPUTS: Activities: A collecting trip to Costa Rica was organized last January. The primary purpose of the trip was to secure fresh material for DNA extraction, amplification, and sequencing. We also targeted the species Evaniscus marginatus, which has only been collected during the dry season in Costa Rica. I am spearheading a revision of the ensign wasp fauna of Costa Rica, and this rare species is one of the few for which we do not yet have high quality DNA. Services: I was asked by three different labs (2 in the USA, 1 in China) to identify evaniid specimens involved in biocontrol and/or ecology research. Products: Our molecular dataset continues to grow, with the efforts of Matt Hill (honors undergraduate) and John Jennings at the University of Adelaide (Australia). We have partial sequences of the mirochondrial cytochrome oxidase 1, the nuclear 28S rRNA, and the nuclear protein encoding gene CAD. We expect this dataset to be ready for publication in 2009. The Evanioidea Online website (http://evanioidea.info/) continues to grow, with new taxa added this year, as well as more references. Perhaps the most striking new additions are the electronic keys (multi-entry and bifurcating) to Alobevania spp. and its associated species pages: http://purl.oclc.org/NET/evanioidea/alobevaniakey (bifurcating key) http://purl.oclc.org/NET/alobevaniamultikey (multi-entry key) http://purl.oclc.org/NET/alobevania (genus page) http://purl.oclc.org/NET/alobevaniagattiae (species page) http://purl.oclc.org/NET/alobevaniatavaresi (species page) http://purl.oclc.org/NET/alobevanialongisaeta (species page) Taxon pages were also created on the Tree of Life Web Project: http://tolweb.org/Alobevania_longisaeta http://tolweb.org/Alobevania_tavaresi http://tolweb.org/Alobevania_gattiae http://tolweb.org/Alobevania The webpages were published as part of an important manuscript on descriptive taxonomy, which I co-authored this year as part of this project: Deans, A. R. & R. Kawada. 2008. Alobevania, a new genus of neotropical ensign wasps (Hymenoptera: Evaniidae), with three new species: integrating taxonomy with the World Wide Web. Zootaxa 1787: 28-44 PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
We published one of the first insect papers (Deans & Kawada 2008) to exploit ZooBank's Life Science Identifiers (LSIDs), and we were the first authors to embed links in our taxon descriptions and bifurcating keys to annotations and images in Morphbank. We are using ensign wasps as a model for changing the way descriptive taxonomy is done, by infusing the manuscripts and database products with an unprecedented level of informatics and community standards (e.g., those standards advocated by TDWG). In terms of molecular data, early results indicate that the markers we chose to sequence yield topologically congruent trees and results that are more fully resolved than any previous attempts by my lab.

Publications

• Deans, A. R. & R. Kawada. 2008. Alobevania, a new genus of neotropical ensign wasps (Hymenoptera: Evaniidae), with three new species: integrating taxonomy with the World Wide Web. Zootaxa 1787: 28-44
• Deans, Andrew R. 2008. Alobevania Kawada & Deans 2008. Version 09 June 2008 (under construction). http://tolweb.org/Alobevania/120177/2008.06.09 in The Tree of Life Web Project, http://tolweb.org/