Source: UNIV OF MARYLAND submitted to
STYLET MORPHOLOGY OF THE INVASIVE SPOTTED LANTERNFLY: IMPLICATIONS FOR HOST TREE ASSOCIATIONS AND TREE DAMAGE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
1017864
Grant No.
(N/A)
Project No.
MD-ENTM-1808
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 17, 2018
Project End Date
Sep 30, 2019
Grant Year
(N/A)
Project Director
Lamp, WI, .
Recipient Organization
UNIV OF MARYLAND
(N/A)
COLLEGE PARK,MD 20742
Performing Department
Entomology
Non Technical Summary
The invasive spotted lanternfly, Lycorma delicatula (Hemiptera: Fulgoridae) is an economically important invasive insect pest that is rapidly spreading in the eastern US. It poses a significant economic threat to many woody tree species in MD including native and economically-important trees and woody plants. Nymphs and adults cause substantial damage by sucking phloem sap and subsequently reducing photosynthesis, causing weeping wounds, and creating conditions for sooty mold. Lanternfly nymphs switch host plants during their development. However, little is known about the relationship between the lanternfly and its tree hosts. Information is particularly needed regarding the association between the lanternfly mouthparts and its feeding behavior. Our objectives include (1) investigating how the lanternfly utilizes its host plants by assessing its stylet morphology (the piercing-sucking mouthpart which allows it to penetrate bark), (2) predicting potential tree host-associations at each developmental stage, and (3) providing information to forest managers for effective monitoring of the lanternfly on tree hosts throughout the season. We expect the lanternfly mouthparts to be highly adapted to various host trees and to undergo structural changes throughout the lanternfly development. This information will be used to help predict lanternfly dispersal to new host trees and prevent rapid infestation of trees.
Animal Health Component
0%
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2110613107020%
2110613113080%
Goals / Objectives
The invasive spotted lanternfly, Lycorma delicatula (Hemiptera: Fulgoridae) is an economically important invasive insect pest that is rapidly spreading in the eastern US. It poses a significant economic threat to many woody tree species in MD including native and economically-important trees and woody plants. Nymphs and adults cause substantial damage by sucking phloem sap and subsequently reducing photosynthesis, causing weeping wounds, and creating conditions for sooty mold. Lanternfly nymphs switch host plants during their development. However, little is known about the relationship between the lanternfly and its tree hosts. Information is particularly needed regarding the association between the lanternfly mouthparts and its feeding behavior.To address these issues, in this project we focus on exploring and predicting lanternfly host plant usage by assessing changes in its stylet morphology at each developmental stage.Our specific objectives are:Objective 1. To assess the changes in morphology and morphometric characteristics of L. delicatula stylets during the lanternfly development and their potential effect on tree host plants.Objective 2. To estimate the depth of the lanternfly stylet penetration and stylet movement within plant tissues and predict potential tree host-associations at each developmental stage of the lanternfly.Objective 3. To provide information about the lanternfly stylet morphology and susceptible host trees to forest managers and other stakeholders - to help them effectively scout and monitor the lanternfly nymphs and adults on potential tree host plants.We expect the lanternfly mouthparts to be highly adapted to various host trees and to undergo structural changes throughout the lanternfly development. This information will be used to help predict lanternfly dispersal to new host trees and prevent rapid infestation of trees.
Project Methods
This one-year project will be conducted at the Department of Entomology, University of Maryland, and will include a laboratory study, as well as several collection trips to Pennsylvania (or in Maryland once the spotted lanternfly invades Maryland). During this year we will (a) perform a morphological analysis of the lanternfly mouthparts based on measurements and observations of stylet morphology at each insect developmental stage (Objective 1); and (b) propose potential tree host-associations at each developmental stage based on established characteristics of tree bark (Objective 2). Based on these findings we will then provide information to forest managers for effective scouting and monitoring of the spotted lanternfly throughout the season (Objective 3).Our specific methods for each objective include:Objective 1- assess changes in morphology and morphometric characteristics of L. delicatula stylets. Developing a step-by-step protocol for insect dissection, isolating stylets, and tissue preparation for morphological analysis. Individual insects at each developmental stage will then be placed in a Petri dish with a standard saline. Under the dissecting microscope the stylets will be isolated, and then transferred to a microscope slide (for light microscopy) and/or fixation solution (for scanning electron microscopy). Using resources for light and scanning electron microscopy located in the Department of Entomology and the Laboratory for Biological Ultrastructure, the morphology of the stylets will be investigated, and all the morphological structures will be photographed. These photographs will serve as a reference for searching same morphological structures at each developmental stage.Identifying morphometric characteristics used in a comparative analysis of the stylet morphology at each developmental stage. We will specifically focus on (a) determining the length of the stylet, and (b) describing the stylet morphology (for the first time) for each developmental stage of the lanternfly. Both the stylet length and stylet morphology will allow us to pinpoint a type of host plants the lanternfly is associated with at each developmental stage; specifically, we will be able to determine relative bark thickness and plant tissue density. Based on the results from this morphological analysis we will identify a number of morphometric characteristics which will be used in the comparative analysis of the stylet morphology at each developmental stage.Morphometric comparative analysis of the lanternfly stylets among developmental stages. For this comparative analysis, the lanternfly nymphs and adults will be collected from different host plants at several field sites in PA during summer 2019. We will collect specimens at five locations in PA from up to 10 different host trees creating a range of host plant sizes and bark structure. We are especially interested in comparing early nymphs which are broad generalists and have the potential to impact a wide range of trees, late nymphs with narrower host plant range, and adults which tend to attack only a few preferred host plants. Difference in the stylet length and other characteristics among nymphs and adults may indicate the difference in distance from the bark surface to the phloem tissue of the lanternfly host trees; which, in turn, may reflect the pattern of switching host trees during the lanternfly development.Objective 2 - estimate stylet penetration and movement:Morphometric comparative analysis of the lanternfly stylet penetration and investigating plant tissue samples. We will set up a series of host preference feeding experiments which will be conducted either in University of Maryland environmental chambers (if the lanternfly invades MD before Summer 2019) or at field sites in PA (the experimental sites and potential collaborators in PA will be determined during Spring 2019). For these feeding experiments, lanternfly nymphs and adults will be allowed to feed on different host trees. Following techniques used by Freeman et al. (2001), insects and plant tissues from the feeding site will be frozen with dry ice cooled 2,2-dimethoxypropane. Using rapid methods of killing and fixing adults and nymphs as they feed on host branches and leaves will allow us to obtain more accurate measurements of stylet lengths and the mechanism of stylet insertion, before the insects could retract their stylets. We will collect and fix at least 10 individuals at each developmental stage. Based on the comparative analysis of stylet length, we will estimate the depth of stylet penetration in various host plants which the lanternfly needs to overcome to reach the phloem. We will also identify probe sites in plant tissue samples (leaves and branches) and examine them under light and scanning electron microscopy. Following Freeman et al. (2001), we will determine whether the lanternfly probed and fed on host plant or abandoned the feeding site after probing. Based on these results as well as known characteristics of bark from woody plants, we will then propose a range of host trees which are most susceptible to the lanternfly feeding at each developmental stage of the pest.Objective 3 - provide information to forest managers and other stakeholders:Develop recommendations and share intermediate results at forestry meetings. During this project, all the findings will be distributed to forest managers, researchers, regional specialists, growers, and other MD stakeholders through extension events and publications. We will especially focus on sharing the information about the ecology and sustainable management of the lanternfly, such as: (1) the lanternfly life cycle; (2) the lanternfly host range; (3) tree species which are most susceptible to infestation by the lanternfly at each developmental stage; and (4) offer recommendations for effective scouting and monitoring of the lanternfly throughout the year.

Progress 10/17/18 to 09/30/19

Outputs
Target Audience:The proposed activities will benefit many MD stakeholders, and especially forest managers and fruit growers. Findings from this project will be distributed through field day events, commodity and in-service meetings, local conferences, and extension publications. Our findings will help the forest managers and fruit growers (particularly, apple and grape growers) monitor and scout the insect, and as a result, reduce impact from the lanternfly dispersal on woody plants. In addition, through the distributed detailed information on the lanternfly ecology and its host plants, as well as our recommendations, forest managers and fruit growers will become more knowledgeable on (1) the ecology of the lanternfly; (2) the lanternfly host range; and (3) woody plants which are most susceptible to different developmental stages of the lanternfly. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Our work on this project during the past year has resulted in the following important outcomes: (a) it has substantially increased our knowledge on the morphology of the spotted lanternfly mouthparts; (b) it has provided the opportunity to develop our skills in the insect dissection, tissue preparation, and scanning electron microscopy. As a result, we have developed a step-by-step dissection protocol (manuscript in review) for the spotted lanternfly, available here: http://dx.doi.org/10.17504/protocols.io.8tthwnn How have the results been disseminated to communities of interest?(a) The results were described and submitted for publication in PLOSOne. The manuscript is currently in review. (b) Additionally, we have presented our results at the Eastern Branch ESA meeting (March 2019), and we are presenting them at the Annual ESA meeting (November 2019). (c) Intermediate results of our work on this grant were presented at the MOFFA Winter Meeting 2019 (Maryland Organic Food & Farming Association meeting; 02/23/2019). What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective 1 has been fully completed: (a) A step-by-step protocol has been developed for insect dissection, insect dissection, isolating stylets, and tissue preparation for morphological analysis. (b) Changes in morphology of the lanternfly mouthparts (labium and stylets) at each developmental stage have been assessed using both scanning electron microscopy and morphometric analysis (c) Morphometric characteristics have been identified and a comparative analysis of the stylet morphology at each developmental stage has been conducted Objective 2. (has not been completed, but preliminary work has been done to facilitate further studies) (a) Preliminary testing of the lanternfly fixation in dry ice cooled 2,2-dimethoxypropane has been initiated. However, fixation of leaf tissues and estimation of the depth of the stylet penetration was not possible because of active moving of the lanternfly nymphs on the host trees. Consequently, scanning electron microscopy (using the proposed technique) has not been conducted. (b) We were able, however, to estimate the length of the exposed stylet part in 3rd instar and 4th instar nymphs. These morphometric characteristics can be further used as a proxy for the depth of the stylet penetration in the host plant tissue. Objective 3 has been fully completed: Intermediate results of our work on this grant were presented at the MOFFA Winter Meeting 2019 (Maryland Organic Food & Farming Association meeting; 02/23/2019). During this presentation, we shared the information about the lanternfly life cycle, and its host plant usage at each developmental stage. We had also provided recommendations for growers to effectively monitor and scout the lanternfly during each developmental stage, to prevent its dispersal from non-crop host plants to cropping systems.

Publications

  • Type: Journal Articles Status: Under Review Year Published: 2020 Citation: Avenesyan, A., and W. Lamp. External morphology and developmental changes of tarsal tips and mouthparts of the invasive spotted lanternfly, Lycorma delicatula (Hemiptera: Fulgoridae. PLOS ONE (in review)