SYSTEMATICS OF HEMIPTERA: BIODIVERSITY, PHYLOGENETICS AND EVOLUTIONARY HISTORY OF PLANT PESTS, POLLINATOR PREDATORS, BENEFICIAL INSECTS, AND DISEASE VECTORS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1016681
• Project Start Date: Jun 27, 2019
• Project End Date: Sep 30, 2023
• Program Code: [(N/A)]- (N/A)

Recipient Organization
UNIVERSITY OF CALIFORNIA, RIVERSIDE
(N/A)
RIVERSIDE,CA 92521

Performing Department
Entomology, Riverside

Non Technical Summary
Most Heteroptera, the true bugs, are plant feeding and many species are serious crop pests. Other Heteroptera are beneficial and includeimportant natural enemies. Others may cause damage to pollinators. Most fields of entomology, including applied areas such as biological control, rely on an accuratetaxonomic framework for the group in question. This framework includes reliable species diagnoses and knowledge on relatedspecies. Heteroptera including the two families that are targeted in this project are in severe need for systematic research on alltaxonomic levels. Descriptive and revisionary work for many groups is not up to current standards, hypotheses on relationshipsare nonexistent or have not been tested in a rigorous framework, and our understanding of the natural history of many true bugsis limited. This project will study systematics and evolution of two groups ofHeteroptera, predatory Reduviidae and mostly plant-feeding Miridae, but also exapnd to other heteropteran groups as well as their close relatives. We will improve systematics in the target groups. We will conduct revisionary work and produce online identification keysand dissemination of systematic and biological information. The research on gland systems and genitalia will provide newinsights into two aspects of the natural history of an insect that are central to integrated pest management. This research,integrating systematics and natural history, will make a significant contribution to our knowledge on the systematics ofpests and natural enemies, the evolution of prey specialization, and thost plant associations.

Animal Health Component

(N/A)

Research Effort Categories

Basic

100%

Applied

(N/A)

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
211	0120	1130	90%
722	0120	1130	10%

Knowledge Area
211 - Insects, Mites, and Other Arthropods Affecting Plants; 722 - Zoonotic Diseases and Parasites Affecting Humans;

Subject Of Investigation
0120 - Land;

Field Of Science
1130 - Entomology and acarology;

Keywords

hemiptera

biodiversity

evolutionary history

plant pests

natural enemies

Goals / Objectives
A) Improve systematics of Reduviidae that comprise natural enemies, pollinator predators, vectors of Chagas Disease providing a basis for the development of integrated pest management and vector control strategies.B) Collect biodiversity data on predators of pests and pollinators in California, e.g. Apiomerus and Phymata, to investigate their potential as natural enemies and assess their influence on pollinators.C) Advance taxonomy and phylogenetics of plant bugs, which include numerous crop pests, allowing for accurate species identification.D) Provide online identification tools and distribution maps of natural enemies (Reduviidae) and crop pests (Miridae) and online documentation of host plant preferences in phytophagous Hemiptera.E) Study comparative and functional morphology (e.g., gland, genitalic structures), basic biology, and ecology (e.g., insect-host plant associations, predator-prey, blood feeder-host, insect-microbial) of heteropteran pests, natural enemies, and disease vectors to provide the basis for their management.

Project Methods
Objective 1) Taxonomy and biodiversity of Reduviidae (natural enemies, pollinator predators, and human disease vectors).?Essential working plan, location, and analysis: We will continue to use established taxonomic protocols that present information in a highly structured way including species description and diagnosis, geographic distribution, identification keys, and illustrations (e.g., Berniker et al. 2011, Zhang and Weirauch 2011a, Zhang et al. 2016). Given the virtual absence of high-quality images (habitus, morphological details) for any of the taxa we propose to monograph, this will be a primary goal. Macrophotography and SEM documentation will allow for fast and consistent documentation, with plates being compiled in Photoshop or CorelDRAW. Specimens are derived from fieldwork in California and abroad and loaned from natural history museums. Research will be performed at UCR. Specimens will be databased and the PBI specimen database allows for download of coordinates that will be plotted resulting in distribution maps (Weirauch 2006e,f, 2007b). Keys will be made available as print keys in monographic treatments and online through Discover Life (see http://www.discoverlife.org/mp/20q?guide=Apiomerini for a model). Field work for assassin bugs in Southern California will continue, as will databasing of historic collections.2) Taxonomy, biodiversity, and phylogenetics of Miridae (agricultural pests and natural enemies). Essential working plan, location, and analysis: We will continue our fieldwork in Southern California and elsewhere. The target habitats will be chaparral and oak-woodlands. Miridae are collected by beating vegetation. Plants will be collected, prepared for deposition in a herbarium, and identified by specialists. Plant bugs will be mounted, sorted, and databased. Taxonomic revisionary work will follow the procedures published, e.g. Weirauch and Schuh (2011a) and other lab publications. These projects will taxonomically revise genera in the Western Nearctic and generate an overall phylogeny of Mirinae that will provide insights into the evolution of host plant associations within the group. We are also working towards a comprehensive phylogeny across the Miroidea, i.e. Miridae and the closely related families Tingidae and Thaumastocoridae, using long-range PCR and next-generation sequencing methods and phylogenomic approaches to optimize sampling of genetic loci.3) Higher-level phylogenetics of Heteroptera, Paraneoptera, and Reduviidae. Essential working plan, location, and analysis: In addition to traditional Sanger sequencing molecular methods, our lab now uses a combination of genomic, transcriptomic, and exon target enrichment (both AHE and UCE) datasets to inform evolutionary relationships at various taxonomic levels and to explore the evolution of proteins involved in predatory and blood-feeding behaviors and vision (opsin evolution). We continue to use morphological characters for phylpogenetic purposes that are critical to tie in fossil species and to translate phylogenetic results into classifications. Morphological characters are examined and documented through dissections, light and SEM imaging, and line drawings. Images will be vouchered using MorphBank [http://www.morphbank.net/]. Coding (binary and multistate, non-additive, ordered) will follow standard procedures as in Weirauch (2008). Research will be conducted in the Weirauch lab.4) Tri-trophic database. Essential working plan, location, and analysis: Procedures for estimating areas of endemism are established (Weirauch et al. 2016a), as are those for phylogenetic components (see above), ecological niche modeling procedures, and methods in historical biogeography. Research will be conducted in the Weirauch lab.5) Natural history and ecology of Heteroptera. Essential working plan, location, and analysis: We are using a range of methods from microdisscetions of glands (aspublished by us; see references above) to molecular approaches involved in documenting prey associations and microbial communities. We are using the full range of methods in molecular diet analyses, from using specific primers to amplify target taxa (Gordon and Weirauch, 2016; Georgieva et al., 2017), over the use of blocking primers to inhibit amplification of host DNA (Masonick et al., in prep), to metagenomic approaches. Harnessing online images documenting predator-prey associations for assassin bugs from the internet is another avenue we pursue to improve our knowledge bases of these interactions. We are also in the process of describing the natural history of the rarely collected Bactrodinae, where we show that species take advantage of arthropod carrion entrapped on sticky plants, while engaging in maternal care. In addition to interactions between true bugs and their prey (Reduviidae) or plant hosts (Miridae), we also conduct research on microbiomes of selected groups of true bugs. Building on expertise acquired during a project focusing on Pyrrhocoroidea (Gordon et al. 2016), we are now investigating microbiomes of sylvatic Triatominae and investigate evolutionary patterns of microbiomes across the Miroidea (Miridae and the closely related Tingidae and Thaumastocoridae). Research will be conducted in the Weirauch lab.6) "True Bugs of the World". Essential working plan, location, and analysis: Co-authors Schuh and Weirauch have divided the responsibility for all book chapters. Images plates have been assembled and chapters are being finalized and send for review. Work will be conducted in the Weirauch lab.

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:My lab's research output continues to target a scientific audience. Online research and outreach tools (e.g., online distribution maps, voucher images) and published identification keys allow a broader audience to reliably identify potential crop pests (mostly Miridae) and natural enemies (mostly Reduviidae). The recently published "True Bugs of the World" textbook (Schuh and Weirauch, 2020) aims on educating professionals and amateurs on the natural history and classification of Heteroptera. Basic biodiversity data (species distributions, phenology, host plant associations) are an important commodity for conservation scientists and policy makers. We also generate and disseminate data on trophic interactions and microbial associates. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the review period, three PhD students were based in the PI's lab and involved in studying the systematics of Reduviidae (Samantha Smith; Stephanie Castillo; Paul Masonick). One postdoc (Alex Knyshov) focused on advancing phylogenomic methods and in charge of the overarching phylogeny of Reduvioidea. Together with the Heraty lab, we organize reading groups (phylogenomics; systematic theory). Two research undergraduates (Tatiana Bush, Madison Hernandez) focused on taxonomic and phylogenetic research on Reduviidae. Tatiana Bush is now enrolled in the 4+1 program in Entomology, and her research will focus on two projects on Miridae and Reduviidae. A visiting Fulbright researcher (Dimitri Forero) worked towards revising species groups of Apiomerus. The pandemic led to several cancelled conferences that we had planned to attend. Frequent lab and individual meetings facilitated communication during the pandemic How have the results been disseminated to communities of interest?During this reporting period, we continued to rely on print publications, online databases, and our research webpage. What do you plan to do during the next reporting period to accomplish the goals? Higher-level phylogenetics of Heteroptera: a collaboration with the hemipteroid AToL project is ongoing that will resolve higher-level relationships of true bugs using combined transcriptomic and genomic datasets. These datasets will also allow us the provide first insights into the evolution of opsin and OR proteins across Heteroptera. Phylogenetic projects on Reduviidae: Most of the above-mentioned projects are ongoing. The manuscripts for the large reduvioid phylogeny are nearing completion and are expected to be submitted soon. In addition, a combined molecular and morphological dataset that will allow us to re-classify Reduviidae is in progress. Other projects with manuscripts nearing completion are those focused on Triatominae, Emesinae, and Stenopodainae. Taxonomic revisions on Pseudocetherinae, Ectrichodiella, Apiomerus, and Salyavatinae will continue. Miridae: We will focus on advancing phylogenetics of the plant bugs subfamily Phylinae, with emphasis on the Western Nearctic including California. . Some new field work combined with sorting of existing voucher specimen collections will form the basis for this study. As part of this study, we will also investigate the historical biogeography and host plant evolution. Reduviidae that show prey specializations or/and unusual predation techniques: We will finalize a manuscript documenting the intriguing natural history (sticky trap predation, maternal care) of Bactrodinae. Planned field work is postponed due to the pandemic. We also continue to work towards a better understanding of salivary proteins across Reduvioidea. Salivary proteins comprise defensive and offensive (used during predatory behaviors) compounds that are derived from separate parts of the salivary glands. The evolution of salivary proteins in the context of prey specialization (millipedes, ants, termites, etc.) and specialized predatory behaviors (e.g., sticky trap predation) is currently unknown. Reduviid transcriptomes and genomes generated by us offer the first glimpses into the evolution of salivary proteins across the group.

Impacts
What was accomplished under these goals? A) Improve systematics of Reduviidae that comprise natural enemies, pollinator predators, vectors of Chagas Disease providing a basis for the development of integrated pest management and vector control strategies. Revisionary taxonomic projects on Reduviidae focused on Phymata (Masonick et al., 2020), Pseudocetherinae (Castillo et al., in prep.), the maya group of Apiomerus (Masonick et al., under review), the remaining species groups of the large New World genus Apiomerus (Forero et al., in prep.), and the millipede assassin bug genus Ectrichodiella (Bush et al., in prep.). We also published a taxonomic revision of Camarochilus that belongs to the sister group of assassin bugs (Weirauch et al., 2020). Phylogenetic projects on assassin bugs continued to focus on a) the overarching phylogeny of Reduvioidea based on transcriptomic and genomic data (three manuscripts in process); b) a phylogeny of Triatominae, the kissing bugs, that vector the Chagas disease agent, based on Ultraconserved Element data (Kiernan et al., in prep.); c) phylogenetics and host associations of spider-web inhabiting Emesinae (Smith et al., in prep.); d) phylogenetics of the pirate assassins Peiratinae (two manuscripts; Castillo et al., in prep.); e) phylogenetics of Stenopodainae (Hernandez et al., in prep.), and f) phylogenetics of Harpactorinae (Weirauch et al., in prep.), the clade with the greatest number of actual and potential biocontrol agents. B) Collect biodiversity data on predators of pests and pollinators in California, e.g., Apiomerus and Phymata, to investigate their potential as natural enemies and assess their influence on pollinators. A study analyzing crowd sourced predator-prey images of commonly encountered assassin bugs in North America was published (Hernandez et al., 2019). C) Advance taxonomy and phylogenetics of plant bugs, which include numerous crop pests, allowing for accurate species identification. Pretrained convolutional networks are a powerful tool for insect identification. We conducted research on applying this method to notoriously difficult to identify plant bugs to advance use of this approach on insect systems (Knyshov et al., in prep.). D) Provide online identification tools and distribution maps of natural enemies (Reduviidae) and crop pests (Miridae) and online documentation of host plant preferences in phytophagous Hemiptera. Specimen databasing of Reduviidae and Miridae continued during the reporting period. Data are available through the Heteroptera Species Pages. Natural history, phylogenetics, and classification of Heteroptera: the second edition of the "True Bugs of the World" was published (Schuh and Weirauch, 2020) providing an up to date account of the classification and natural history of this insect group. We also published a transcriptome-based phylogeny of Heteroptera with improved taxon sampling and support (de Moya et al. 2019).

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: De Moya, R., Weirauch, C., Sweet, A. D., Skinner, R., Walden, K. K. O., Swanson, D. R., Dietrich, C., and Johnson, K. 2019. Deep instability in the phylogenetic backbone of Heteroptera is only partly overcome by transcriptome-based phylogenomics. Insect Systematics and Diversity, 3: 7; 1-14.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Hernandez, M., Masonick, P., and Weirauch, C. 2019. Crowdsourced online images provide insights into predator-prey interactions of putative natural enemies. Food Webs (Short Communication), 21 (2019), e00126.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Weirauch, Forero, and Schuh. 2020. Taxonomic revision of Camarochilus Harris (Hemiptera: Pachynomidae). American Museum Novitiates 2020: 1-31.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Masonick, P. and Weirauch, C. 2020. Taxonomic revision of the Nearctic erosa species group of Phymata Latreille, 1802 (Heteroptera: Reduviidae: Phymatinae). Canadian Journal of Arthropod Identification 41 https://cjai.biologicalsurvey.ca/mw_41/mw_41.html

Progress 06/27/19 to 09/30/19

Outputs
Target Audience:Most of my lab's research output continues to be targeted towards a scientific audience. Our online research and outreach tools (e.g., online distribution maps, voucher images), however, as well as published identification keys including those that are part of taxonomic revisions, allow a broader audience to reliably identify potential crop pests (mostly Miridae) and natural enemies (mostly Reduviidae). They provide valuable resources to those involved in biological control, integrated pest management and other applications. Basic biodiversity data (species distributions, phenology, host plant associations) are an important commodity for conservation scientists and policy makers. We also generate and disseminate data on trophic interactions and microbial associates. Our ongoing efforts to document true bug biodiversity in the biodiversity hotspot "California Floristic Province" thus contribute to a strong knowledge base for our natural resources. The "True Bugs of the World" textbook aims on educating professionals and amateurs on the natural history and classification of Heteroptera. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the review period, three PhD students were based in the PI's lab and involved in revisionary systematics of Reduviidae (Samantha Smith; Stephanie Castillo; Paul Masonick). One postdoc (Alex Knyshov) focused on advising phylogenomic methods and in charge of the overarching phylogeny of Reduvioidea. Together with the Heraty lab, we organize reading groups (phylogenomics; systematic theory). Smith and Knyshov presented at the Evolution meeting. Two research undergraduates (Tatiana Bush, Madison Hernandez) focused on taxonomic and phylogenetic research on Reduviidae. How have the results been disseminated to communities of interest?Our premier avenue for dissemination of results continues to be print publications, online databases, and our research webpage. Smith and Knyshov presented at the Evolution 2019 meeting, the PI at the Association for Tropical Biology and Conservation 201 meeting. What do you plan to do during the next reporting period to accomplish the goals?Higher-level phylogenetics of Heteroptera: a collaboration with the hemipteroid AToL project is ongoing that will resolve higher-level relationships of true bugs using data generated from transcriptomes. Other ongoing research continues to emphasize Dipsocoromorpha, both from a phylogenetic and integrative taxonomic revisionary perspective. Classification and natural history of Heteroptera: The second edition of ""True Bugs of the World" is at the proof stage. We expect to finalize and publish this book during the next reporting period. Reduviidae systematics: The focus in this area will continue to closely follow the goals of my current NSF project: 1) test phylogenetic hypotheses with emphasis on the poorly resolved Higher Reduviidae and contentious relationships of the hematophagous Chagas disease vectors Triatominae, while maximizing taxonomic coverage and integrating transcriptomic, anchored hybrid enrichment, traditional molecular, and morphological datasets; 2) describe and redescribe fossil reduvioids to bridge taxonomic and geological gaps in the fossil record and test hypotheses on the timing of key biological events; 3) study diversification across Reduvioidea, focusing on hypotheses of diversification rate shifts and correlated key innovations, and morphological, physiological, and ecological traits involved in the transition between predation and hematophagy; 4) use the phylogenetic information generated during this project to propose a new classification of Reduvioidea. We will also continue a number of revisionary projects (e.g., revisions of Camarochilus, Salyavatinae, Nearctic Phymata, Pseudocetherinae, Ectrichodiella). Miridae identification: Pretrained convolutional networks are a powerful tool for insect identification. We are planning on using a dataset notoriously difficult to identify plant bugs to advance use of this approach on our study systems. Reduviidae that show prey specializations or/and unusual predation techniques: We will finalize and publish our study using crowd sources images documenting predator-prey interactions of Reduviidae. We will also focus on advancing a manuscript documenting the intriguing natural history (sticky trap predation, maternal care) of Bactrodinae. Planned field work in Panama (November 2019) will allow us to improve our dataset on spider web-inhabiting Emesinae.

Impacts
What was accomplished under these goals? A) Improve systematics of Reduviidae that comprise natural enemies, pollinator predators, vectors of Chagas Disease providing a basis for the development of integrated pest management and vector control strategies. Revisionary taxonomic projects on Reduviidae focused on Emesinae (Smith et al. 2019), Pseudocetherinae (Castillo et al., in prep.), Phymatinae (Masonick et al., in prep), and Apiomerus (Masonick et al., in prep.). Phylogenetic projects on assassin bugs continued to focus on a) the overarching phylogeny of Reduvioidea based on transcriptomic and genomic data; b) a phylogeny of Triatominae, the kissing bugs, that vector the Chagas disease agent, based on Ultraconserved Element data; c) phylogenetics and host associations of spider-web inhabiting Emesinae; d) phylogenetics of the pirate assassins Peiratinae; e) phylogenetics of Stenopodainae, and f) phylogenetics of Harpactorinae, the clade with the greatest number of actual and potential biocontrol agents. B) Collect biodiversity data on predators of pests and pollinators in California, e.g., Apiomerus and Phymata, to investigate their potential as natural enemies and assess their influence on pollinators. A study documenting tropic interactions between ambush bugs and flower-visiting insects was published (Masonick et al., 2019). The analysis of crowd sourced predator-prey interactions of commonly encountered assassin bugs in North America was another major emphasis under this objective. C) Advance taxonomy and phylogenetics of plant bugs, which include numerous crop pests, allowing for accurate species identification. The graduate student researching the phylogenetics of the subfamily Mirinae continued to be on sick leave. A study investigating new molecular procedures to generate large datasets from dry museum specimens was published (Knyshov et al., 2019). This study used plant bugs as model system and significantly improved our understanding of evolutionary relationships and host use evolution of several lineages of plant bugs that are restricted (or almost so) to California. D) Provide online identification tools and distribution maps of natural enemies (Reduviidae) and crop pests (Miridae) and online documentation of host plant preferences in phytophagous Hemiptera. Specimen databasing of Reduviidae and Miridae continued during the reporting period. Data are available through the Heteroptera Species Pages. E) Study comparative and functional morphology (e.g., gland, genitalic structures), basic biology, and ecology (e.g., insect-host plant associations, predator-prey, blood feeder-host, insect-microbial) of heteropteran pests, natural enemies, and disease vectors to provide the basis for their management. A functional morphological study on male-specific glands in the small heteropteran infraorder Dipsocoromorpha was published (Knyshov et al., 2019). While this group is not of economic importance, this study serves as a model for future investigations into Reduviidae and Miridae.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Knyshov, A., Gordon, E.R.L., and Weirauch, C. 2019. Cost-efficient capture of historical arthropod specimen DNA using PCR-generated baits. Methods in Ecology and Evolution, 10: 841-852 https://doi.org/10.1111/2041-210X.13169
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Smith, S., Hwang, W. S., and Weirauch, C. 2019. Synonymy of Mangabea Villiers and Stenorhamphus Elkins, with the description of two new species (Hemiptera: Reduviidae: Emesinae: Collartidini). Raffles Bulletin of Zoology 67: 135-149.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Masonick, P., Hernandez, M., and Weirauch, C. 2019. No guts, no glory: Gut content metabarcoding unveils the diet of a flower?associated coastal sage scrub predator. Ecosphere 10: e02712, https://doi.org/10.1002/ecs2.2712