Alina Avanesyan
Among the redwoods -- Muir Woods

Ecology and evolution of species interactions: molecular, morphological, and behavioral perspectives

In my current postdoctoral work at the University of Maryland (Department of Entomology; Dr. Bill Lamp’s lab) I’m interested in ecology and evolution of species interactions. Using arthropods and plant-insect study systems I’m curious what mechanisms (behavioral, molecular, physiological, etc.) drive species interactions, and especially interactions between native and introduced species.

I’m working on a variety of projects that involve primarily molecular biology techniques, microscopy, and morphometric analysis, but also field and greenhouse work. My primary projects include (a) using molecular biology tools to assess wetland-stream connectivity; (b) external morphology and host plant usage of the invasive spotted lanternfly using molecular approach, light microscopy and scanning electron microscopy; (c) plant DNA detection from gut contents of sap-feeders; (d) molecular identification of prey from predator diet; (e) plant resistance and tolerance to herbivores. A brief description for some of my projects is below. I will be adding more details as the projects grow.


The spotted lanternfly: host plants, external morphology, molecular confirmation of diet

I’ve been working on the spotted lanternfly (SLF) since April 2018. This emerging invasive insect pest is one of the most aggressive leaf-hopping pests in Mid-Atlantic region: it is extremely polyphagous and can utilize over 70 host plants, primarily trees. Since its introduction in Pennsylvania in 2014, the spotted lanternfly has rapidly spread in the eastern US, and it was finally detected in Maryland in October 2018. Nymphs and adults cause substantial plant damage by sucking phloem sap. Lanternfly nymphs switch host plants during their development; however, little is known about the relationship between the lanternfly and its tree hosts, as well as about the mechanisms of its unusual use of plant hosts during development (i.e. polyphagous behavior of early instars and nearly monophagous behavior of adults). We also don’t really know which plants the lanternfly utilize for feeding and which plants it uses for resting, migration, laying eggs, etc.

To address these issues in my research on the spotted lanternfly, I largely utilizes microscopy and molecular biology tools to (1) explore changes in external morphology of the lanternfly during its development, and (2) to confirm the utilization of host plants. Our two grant proposals on studying SLF external morphology and conducting molecular gut content analysis were funded by MDA and MAES in August 2018. Since then, I’ve been spending almost half of my time in the microscope room and the SEM laboratory. I’m curious to see how the lanternfy’s mouthparts and tarsi are so well adapted to feeding and moving on its host trees. Also, I’m applying our DNA barcoding protocol we’ve developed for another sap-feeder, the potato leafhopper, to confirm the lanternfly’s host plants. Almost two years later, since I started studying this species, I can’t stop being amazed by SLF unique ecology and behavior.

The spotted lanternfly, Lycorma delicatula; 4th instars (PA, 2018)
The spotted lanternfly, Lycorma delicatula; 4th instars (PA, 2018)
First amplification of the chloroplast trnL gene from the lanternfly body ('head+mouthparts' and 'gut content'-samples)

Utilization of molecular biology tools. To confirm host plant consumption and explore the potential host plant range of the lanternfly, I use molecular biology techniques to detect host plant DNA within the lanternfly gut contents. Plant DNA detection in insect gut contents has been one of the most accurate ways to confirm host plant utilization, however this method becomes challenging when dealing with sap-feeding insects: (a) the host plants are difficult to discern because piercing-sucking insects, such as the spotted lanternfly, often cause no direct symptoms to plants; (b) the lack of plant tissue in the gut of a piercing-sucking insect that feeds on phloem makes gut contents difficult to discern. Currently, I utilize Sanger sequencing to obtain a portion of plant chloroplast DNA region within the lanternfly gut contents. I successfully isolate and identify plant DNA for ‘single-DNA’-samples (i.e. when there is only one ingested host plant species in the lanternfly gut), and we are planning to use NGS technology to explore mixed DNA samples and compose a host plant range of the spotted lanternfly.

Scanning electron microscopy. To explore the morphological adaptations of the lanternfly to host plant utilization, I focus on the following two objectives: (a) to assess changes in morphology of the lanternfly mouthparts (stylets and labium), and (b) to assess changes in morphology of the lanternfly tarsal tips (arolia and tarsal claws) at each developmental stage. The labium, stylets, and tarsal tips are the structures which are associated with primary contact of the lanternfly with its host plant, and which potentially facilitate the lanternfly successful host plant use. I assess the developmental changes in these structures using both scanning electron microscopy (SEM) and morphometric analysis. We expect these structures to undergo substantial morphological and morphometric changes throughout the lanternfly development which could potentially indicates the lanternfly association with certain host trees at each developmental stage.

Tarsal tip of the 3rd instar: tarsal claws + arolium
Labium tip of the 3rd instar with numerous sensilla
Labrum and stylets of the 4th instar

Update – 01/25/19: I’m presenting my intermediate results at the at the Entomological Society of America Annual Meeting, Eastern Branch, March 9-11,2019. Blacksburg, VA.

Update - 02/23/19: I’ve presented at the Annual Meeting of the Maryland Organic Food & Farming Association (Maryland Dept. of Agriculture, Annapolis, MD). My talk was on the lanternfly biology, behavior, and host plant usage.

Update - 02/28/19: I’ve finished the 2nd round of scanning electron microscopy of the lanternfly mouthparts and tarsi. Most of the beautiful SEM images are included in my poster for the upcoming EB-ESA meeting.

Update - 06/06/19: Our paper on external morphology of the spotted lanternfly has been submitted to PLOS ONE.

Update - 11/14/19: I’ve obtained all the sequences from the lanternfly gut content samples we used and finished data analysis. We’ve got some interesting results, and I’m currently working on getting the paper done (Tentative title: “Molecular gut content analysis reveals the host plant range of the invasive spotted lanternfly, Lycorma delicatula”). We are planning to submit it to Insects, the special issue on molecular gut content analysis of insect herbivores. Bill and I are co-editing this special issue and we hope to see many interesting studies there.

Update - 11/18/19: I’ve presented our updated results on the lanternfly external morphology at the ESA meeting. Here is my poster.


Plant DNA detection within gut contents in sap-feeding insects

Plant DNA detection in insect gut contents is one of the most accurate ways to confirm host plant utilization, determine insect diet and interactions with other organisms. Most of the previous studies that involved molecular analysis of insect gut contents were primarily conducted on leaf-chewing insects, such as beetles, moths, or grasshoppers. Sap-feeders could be more challenging for molecular analysis of their gut contents because phloem sap presumably doesn’t contain plant DNA. However, this method was shown to be effective for potato psyllids (Cooper et al 2016) - apparently, while feeding an insect stylet (a piercing mouthpart) can consume not only phloem sap but can also pick up some of the surrounding plant cells which allows DNA to be detected in the insect gut contents. In this project, we would like to investigate whether (and how) we can apply this method for yet another sap-feeder, the potato leafhopper, to explore its host plant usage.

Experimental set up and detection of DNA of consumed fava beans in the guts of potato leafhoppers. Additionally, we've found that contamination of the insect surface with plant DNA is not significant.

Update - 07/07/18: we are making some good progress, and we’ve submitted our oral presentation (A. Avanesyan and W. O. Lamp, “Use of molecular markers for plant DNA to determine host plant usage for potato leafhopper, Empoasca fabae”) also to the ESA meeting in Vancouver, BC, Canada.

Update – 11/15/18: presented!

Update - 06/10/19: the manuscript is in preparation.

Update - 09/10/19: the manuscript has been submitted to Environmental Entomology.


Wetland-stream connectivity: DNA barcoding of native isopods as a tool for assessing watershed connectivity

This is an ongoing project in Dr. Lamp’s lab. Using isopods from multiple wetland sites and streams in Maryland Delmarva Bays Wetlands we explore the potential connectivity between wetland and stream communities. Morphological identification of isopod species is tricky, and DNA barcoding, which I’m focusing on, is very helpful for estimating how ‘close’ wetland isopod species are to the isopod species inhabiting streams. I’m currently mentoring three students in this DNA barcoding work, and together with my mentees we isolate and sequence a portion of mitochondrial gene, cytochrome oxidase 1 (CO1), create a reference library of these sequences, and using a BLAST search engine in the NCBI GenBank database we assess species and genera identity of isopods.

Maryland Delmarva Bays (May, 2018)
Collecting isopods
Developing PCR protocol with a black fly sample (BF) as a control
First successful amplification!

Update - 02/22/19: Nina, my mentee, a high school student, who was working on this project since September 2018, presented her research project earlier this week at her school’s science fair and got 3rd prize! So exciting!

Update - 04/26/19: Nina presented her final poster at the ERHS Research Symposium.

Update - 06/06/19: We are currently exploring phylogenetic relationships between stream and wetland isopod species.


Molecular identification of prey in predator gut contents and feces

This is a new lab project I’m involved in and it is also a new challenge in my DNA barcoding work. I amplify DNA from dragonfly prey items. In addition to a gut content analysis, I’m working with degraded DNA from dragonfly feces. So far, I successfully amplified and sequenced COI partial gene from dragonfly feces produced during the first 2 hours post ingestion. The sequence analysis with subsequent BLAST results showed that the prey item was a crane fly. Surprisingly enough the DNA from the crane fly wasn’t degraded and an isolated fragment of 667 bp showed good sequence quality. These exciting results can make identification of dragonfly prey possible and accurately confirm (or even point to new) trophic interactions in dragonfly natural habitats.

Obtaining the DNA of a crane fly from dragonfly feces.

Update - 06/10/19: DNA from mayfly was detected in feces produced during first 7 hours post ingestion by the same dragonfly individual. The sequence had lower quality but it was still readable.


Should I eat or should I go: systematic review and meta-analysis

This is a systematic review I was working on last year (manuscript submitted). The review aimed to identify patterns of grasshopper feeding preferences for native versus introduced plants and, consequently, grasshopper potential for biotic resistance of native communities, that (as the review has shown) is surprisingly overlooked in experimental studies on invasion ecology.

My table pet for inspiration..
Literature search and data colelction: PRISMA flowchart

Update – 07/07/18: I’ve submitted a poster presentation on this review to the upcoming ESA meeting in Vancouver, BC, Canada. I’m also going to present it here, at the University of Maryland research symposium organized by the Office of Postdoctoral Affairs.

Update – 05/18/18: the manuscript has been submitted!

Update – 10/07/18: the manuscript has been published in Plants! I was also featured in our departmental news.


Other Projects


Herbivore resistance and tolerance in Miscanthus sinensis cultivars

Although exotic chinese silver grass, a gorgeous ornamental plant and important biofuel source, can be highly invasive in some states, not all of its cultivars are invasive. I’ve been conducting field and greenhouse experiments to explore plant resistance and tolerance to grasshopper herbivory, how these responses differ among Miscanthus cultivars, and whether the initially introduced wild type demonstrates the highest level of herbivore resistance.

Miscanthus sinensis cultivars I use in my experiments (from left to right): ‘Dixieland’, ‘Autumn Anthem’,‘Zebrinus’,‘Morning Light’,‘Gracillimus’

Update – 09/20/19: I’ve finished the analysis of the data we collected in 2018 and 2019 summer seasons; we see some interesting patterns in plant tolerance to herbivory and their association with plant morphological appearance; and I’m about to start working on the manuscript.


Biodiversity of agricultural drainage ditches

When I started working in Dr. Lamp’s lab I joined this ongoing project, and I spent some time ‘torturing’ herbarium plant specimens hoping that they will speak and tell me their taxonomic identity. I then analyzed plant diversity across several farms exploring whether and how plant diversity corresponds with the diversity of natural enemies, such as spiders.

Some of my new plant friends

Update - 12/17/18: I’m done with ‘torturing’ plant specimens. Plant diversity across different sites in Maryland has been analyzed, the plant data have been ‘matched’ to the spider diversity data, and my labmate, Dylan Kutz, will present the results at the Entomological Society of America Annual Meeting, Eastern Branch, March 9-11,2019. Blacksburg, VA.

Update - 05/24/19: Yet another two presentations with our updated results were submitted to the Annual meeting ‘Entomology-2019’.


Reproductive biology and distribution of invasive agricultural pests (previous postdoctoral work)

In my postdoctoral work at the University of Wisconsin-Madison I studied one of the emerging insect pests – the spotted wing drosophila, Drosophila suzukii Matsumura (Diptera: Drosophilidae). This is a highly invasive insect species, which attacks undamaged ripening fruit of a wide variety of soft-skinned fruits and berries. Native to Southeast Asia, currently it is observed in Europe, North America, and South America. Drosophila suzukii has demonstrated a very high dispersal capacity and remarkable phenotypic plasticity – during only a couple of decades since its first introduction in Hawaii, D. suzukii invaded different temperate regions and now is being monitored in many northern and eastern states, as well as Canada.

I work on several projects on different aspects of D. suzukii biology and population distribution. I coordinated a multi-state bait comparison project for determining optimal attractants for D. suzukii. This project was conducted in Minnesota; we set up fly traps with eight different baits and conducted monitoring of D. suzukii in raspberry during several weeks. I also developed experimental design for the spatial and temporal distribution project which was supposed to be conducted later in the season when population of D. suzukii could be established.

I was also actively involved in a project on D. suzukii seasonal phenology focused on overwintering of D. suzukii and the effect of temperature and humidity on D. suzukii seasonal abundance. We are currently writing a paper on D. suzukii seasonal phenology which includes an analysis of the interactions between D. suzukii seasonal abundance and temperature and humidity dynamics during the collecting seasons in 2014-2015.

Additionally, I developed a protocol for tissue preparation, isolating spermathecae, and determining mating status of D. suzukii, which we have applied in our bait comparison and phenology studies. This protocol has been recently published in Insects (Special issue on invasive species). In this paper, we also demonstrated how this protocol can be applied for both field collected flies and flies reared in the lab, including fly specimens stored on a long-term basis.


Summer and fall raspberry varieties at West Madison
Agricultural Research Station on May 19, 2016.
Plastic high tunnel (A, B) with raspberry plants (C) at Hoch Orchard & Gardens farm (La Crescent, MN)

Peer-Reviewed Publications

Journal Articles

  1. Avanesyan, A., and W.O. Lamp. (2020) Use of molecular gut content analysis to decipher the range of food plants of the invasive spotted lanternfly, Lycorma delicatula. Insects: Special Issue " Molecular Gut Content Analysis: Deciphering Trophic Interactions of Insects", 11(4), 215, doi.org/10.3390/insects11040215. file_downloadfull text
  2. Avanesyan, A., Maugel T.K., and W. Lamp (2019) External morphology and developmental changes of tarsal tips and mouthparts of the invasive spotted lanternfly, Lycorma delicatula. PLOS ONE, doi.org/10.1371/journal.pone.0226995 file_downloadfull text
  3. Avanesyan, A., Lamp, W., Snook, K., and P. Follett. (2019) Short-term physiological response of a native Hawaiian plant, Hibiscus arnottianus, to injury by the exotic leafhopper, Sophonia orientalis (Hemiptera: Cicadellidae). Environmental Entomology 48(2): 363-369. file_downloadfull text
  4. Avanesyan, A. (2018) Should I eat or should I go? Acridid grasshoppers and their novel host plants: implications for biotic resistance. Plants: Special Issue "Plants Interacting with other Organisms: Insects", 7(4), 83, doi.org/10.3390/plants7040083. Invited paper. file_downloadfull text
  5. Guédot, C., Avanesyan, A., and K. Hietala-Henschell. (2018) Effect of temperature and humidity on the seasonal phenology of Drosophila suzukii (Diptera: Drosophilidae) in Wisconsin. Environmental Entomology 47(6): 1365–1375. file_downloadfull text
  6. Jaffe, B.D., Avanesyan, A., Bal, H. K., Grant, J., Grieshop, M.J., Lee, J.C., Liburd, O.E., Rhodes, E., Rodriguez-Saona, C., Sial, A.A., Zhang, A., and C. Guédot (2018) Multistate comparison of attractants and the impact of fruit development stage on trapping Drosophila suzukii (Diptera: Drosophilidae) in raspberry and blueberry. Environmental Entomology, 47(4): 935–945. exit_to_applink to publication
  7. Avanesyan, A., Jaffe, B.D., and C. Guédot (2017) Isolating spermatheca and determining mating status of the invasive spotted wing drosophila, Drosophila suzukii: a protocol for tissue dissection and its applications. Insects: Special issue “Invasive Insect Species”. 8(1), 32; doi:10.3390/insects8010032. Invited paper. file_downloadfull text

Manuscripts in preparation

  1. Avanesyan, A., and W.O. Lamp. Molecular approaches to diet analysis of insect herbivores: a systematic review.
  2. Avanesyan, A., and W.O. Lamp. Use of molecular markers for gut content analysis of potato leafhopper, Empoasca fabae.

Conference Presentations

  1. Avanesyan, A., Maugel, T., and W. Lamp. (2019) External morphology and developmental changes of tarsal tips and mouthparts of the invasive spotted lanternfly, Lycorma delicatula. Annual Meeting of the Entomological Society of America, St. Lois, MO. Poster presentation. file_downloadfull text
  2. Smith, D., Avanesyan, A., and W. Lamp. (2019) Are natural enemies related to plant diversity in agricultural drainage ditches? Annual Meeting of the Entomological Society of America, St. Lois, MO. Poster presentation.
  3. Kutz, D., Avanesyan, A., and W. Lamp. (2019) Drainage ditches as sources of beneficial spiders on farms to enhance conservation biological control. Annual Meeting of the Entomological Society of America, St. Lois, MO. Oral presentation.
  4. Avanesyan, A., and W. Lamp (2019) External morphology of the spotted lanternfly, Lycorma delicatula, and its association with insect host plants. Postdoctoral Research Symposium. University of Maryland, College Park, MD. Poster presentation. file_downloadfull text
  5. Avanesyan, A., and W. Lamp (2019) External morphology of the spotted lanternfly, Lycorma delicatula, and its association with insect host plants. Entomological Society of America Annual Meeting, Eastern Branch. Blacksburg, VA. Poster presentation. file_downloadfull text
  6. Avanesyan, A., and W. Lamp (2019) Feeding preferences of native acridid grasshoppers for novel host plants: a case study of biotic resistance. Entomological Society of America Annual Meeting, Eastern Branch. Blacksburg, VA. Oral presentation. file_downloadfull text
  7. Kutz, D., Avanesyan, A., and W. Lamp (2019) Drainage ditches as sources of beneficial spiders on farms: A closer look at plant-spider community associations. Entomological Society of America Annual Meeting, Eastern Branch. Blacksburg, VA. Oral presentation.
  8. Avanesyan, A., and W. Lamp (2018) Use of molecular markers for plant DNA to determine host plant usage for potato leafhopper, Empoasca fabae. Annual Meeting of the Entomological Society of America: 2018 ESA, ESC, and ESBC Joint Annual Meeting, Vancouver, BC, Canada. Oral presentation. file_downloadfull text
  9. Avanesyan, A. (2018) Should I eat or should I go? Acridid grasshoppers and their novel host plants: implications for biotic resistance. Annual Meeting of the Entomological Society of America: 2018 ESA, ESC, and ESBC Joint Annual Meeting, Vancouver, BC, Canada. Poster presentation. file_downloadfull text
  10. Avanesyan, A. (2018) Should I eat or should I go? Acridid grasshoppers and their novel host plants: implications for biotic resistance. Postdoctoral Research Symposium. University of Maryland, College Park, MD. Poster presentation. file_downloadfull text

Upcoming Presentations

  1. Lamp W.O, and A. Avanesyan, A. (2020) Molecular gut content analysis reveals the host plant range of the invasive spotted lanternfly, Lycorma delicatula. Joint Eastern and Southeastern Branch Meeting of the Entomological Society of America, Atlanta, GA. Oral presentation.

Invited Talks

  1. Avanesyan, A. (2019) Spotted lanternfly: information and update. Maryland Organic Food & Farming Association, Maryland Dept. of Agriculture, Annapolis, MD. file_downloadfull text
  2. Avanesyan, A. (2018) Novel plant-insect associations: implications of the lack of coevolution. Department of Entomology, University of Maryland, College Park, MD; weekly seminar series. Seminar speaker. file_downloadfull text
  3. Avanesyan A. (2016). Identifying and controlling spotted wing drosophila. Berry Field Day organized by Wisconsin Berry Growers Association. River Falls, WI.

GenBank Submissions

  1. Smith, D.K., Avanesyan, A. and W. O. Lamp. (2019) Eupatorium serotinum tRNA-Leu (trnL) gene, partial sequence; chloroplast. Direct Submission, GenBank Accession no. MN395725
  2. Smith, D.K., Avanesyan, A. and W. O. Lamp. (2019) Lonicera maackii tRNA-Leu (trnL) gene, partial sequence; chloroplast. Direct Submission, GenBank Accession no. MN365276
  3. Smith, D.K., Avanesyan, A. and W. O. Lamp. (2019) Pisum sativum isolate slf-2 tRNA-Leu (trnL) gene, partial sequence; chloroplast. Direct Submission, GenBank Accession no. MN335637
  4. Stancliff, B., Avanesyan, A. and W. Lamp. (2019) Vicia faba var. major tRNA-Leu (trnL) gene, partial sequence; chloroplast. Direct Submission, GenBank Accession no. MK934667 exit_to_applink to publication
  5. Stancliff, B., Smith, D., Avanesyan, A. and W. Lamp. (2019) Pisum sativum tRNA-Leu (trnL) gene, partial sequence; chloroplast. Direct Submission, GenBank Accession no. MK919208 exit_to_applink to publication
  6. Stancliff, B., Abdelwahab, O., Avanesyan, A. and W. Lamp. (2019) Vigna unguiculata tRNA- Leu (trnL) gene, partial sequence; chloroplast. Direct Submission, GenBank Accession no. MK883492 exit_to_applink to publication
  7. Stancliff, B., Ho, J., Avanesyan, A. and W. Lamp. (2019) Helianthus annuus tRNA-Leu (trnL) gene, partial sequence; chloroplast. Direct Submission, GenBank Accession no. MK875279 exit_to_applink to publication
  8. Avanesyan, A., and W. Lamp. (2019) Vicia faba var. major isolate PLH_fb tRNA-Leu (trnL) gene, partial sequence; chloroplast. Direct Submission, GenBank Accession no. MK837073 exit_to_applink to publication

Non Peer-Reviewed Publications

Newsletters

  1. Avanesyan, A. and C. Guédot. (2016) Exclusion barriers as a sustainable strategy for management of Spotted Wing Drosophila. Wisconsin Fruit News, 1(6). file_downloadfull text
  2. Avanesyan, A. and C. Guédot. (2016) Raspberry varieties and their infestation by Drosophila suzukii. Wisconsin Fruit News, 1(4). file_downloadfull text