Journal of Insect Science, (2018) 18(3): 11; 1–8 doi: 10.1093/jisesa/iey051 Research Screening for Resistance Against the Sugarbeet Root Maggot, Tetanops myopaeformis (Diptera: Ulidiidae), Using a Greenhouse Bioassay Tucker B. Daley and Erik J. Wenninger1 Department of Entomology, Plant Pathology, and Nematology, University of Idaho, Kimberly Research & Extension Center, Kimberly, ID 83341, and 1Corresponding author, e-mail: [email protected] Subject Editor: Junwei (Jerry) Zhu Received 7 March 2018; Editorial decision 7 May 2018 Abstract The sugarbeet root maggot, Tetanops myopaeformis (von Röder) (Diptera: Ulidiidae), is a major pest of sugar beet Beta vulgaris L. (Carophyllales: Amaranthaceae)in the United States and Canada. Larval feeding on roots can reduce both stand and yield. Current management practices are heavily reliant on chemical control. However, the carbamate and organophosphate insecticides that are commonly used against T. myopaeformis are being phased out of use. Host plant resistance against this pest shows promise, but difficulties with maintainingT. myopaeformis in culture have largely limited such studies to the field. A primary objective of this study was to develop protocols for rearing a laboratory colony of T. myopaeformis that would expedite assays aimed at screening for host plant resistance. Third (final) instar larvae were collected from the field and reared to the adult stage. These laboratory-reared adults laid eggs and ultimately produced a second generation of third-instar larvae in the lab. Adult flies reared from field-collected larvae were used to examine the modality of resistance of a known resistant variety by performing no-choice and paired-choice experiments alongside a susceptible variety in the greenhouse. Paired-choice tests showed no difference in oviposition rates between the two varieties, whereas no-choice tests showed significantly greater feeding damage and abundance of larvae on the susceptible variety. For the resistant variety examined here, we observed evidence of antibiosis, not antixenosis, as the putative modality of resistance. Our laboratory and greenhouse protocols can be used to expedite the process of developing varieties with resistance to this key pest of sugar beet. Key words: host plant resistance, antibiosis, antixenosis, Beta vulgaris Sugar beet (Beta vulgaris L. [Carophyllales: Amaranthaceae]) is (Boetel et al. 2009) and in certain parts of Idaho (Bechinski et al. an economically important crop in the United States that accounts 1989, Wenninger et al. 2018). for about 55% of U.S. sugar production (McConnell 2017). Since T. myopaeformis is native to North America (von Röder 1881, 2015, approximately 56% (257,000 hectares) of the sugar beet Hawley 1922, Jones et al. 1952). American colonists brought acreage in the United States has been grown in the Red River Valley B. vulgaris to North America (Harveson et al. 2009), which ulti- of North Dakota and Minnesota (USDA-NASS 2017), while Idaho mately resulted in a new encounter between T. myopaeformis and has ranked third among U.S. states for sugar beet production with sugar beet. Hosts that support larval development include different an average of approximately 69,200 hectares planted each year cultivated forms of B. vulgaris as well as spinach (Spinacia olera- (USDA-NASS 2017). cea L. [Caryophyllales: Amaranthaceae]), spear saltbush (Atriplex Sugarbeet root maggot, [Tetanops myopaeformis (von Röder) patula L. [Caryophyllales: Amaranthaceae]), and garden orache (Diptera: Ulidiidae)], is a major pest of sugar beet in the United States (Atriplex hortensis L. [Caryophyllales: Amaranthaceae])(Mahrt and and Canada. These insects can be found nearly anywhere in North Blickenstaff 1979, Msangosoko 2012). Eggs have been found on America that sugar beet is grown and have been reported in Alberta common lambsquarters (Chenopodium album L. [Caryophyllales: and Manitoba, Canada, as well as in the U.S. states of Washington, Amaranthaceae]), redroot pigweed (Amaranthus retroflexus L. Oregon, California, Colorado, Idaho, Montana, Minnesota, North [Caryophyllales: Amaranthaceae]), prostrate pigweed (Amaranthus Dakota, Nebraska, Utah, and Wyoming (Mahrt and Blickenstaff blitoides Watson [Caryophyllales: Amaranthaceae]), black night- 1979, Lange 1987, Hein 1995). Although T. myopaeformis is wide- shade (Solanum nigrum L. [Solanales: Solanaceae]), and curly dock spread, it is a serious pest of sugar beet only in the Red River Valley (Rumex crispus L. [Caryophyllales: Polygonaceae]) (Hawley 1922, © The Author(s) 2018. Published by Oxford University Press on behalf of Entomological Society of America. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/ 1 licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact [email protected] 2 Journal of Insect Science, 2018, Vol. 18, No. 3 Mahrt and Blickenstaff 1979, Msangosoko 2012); however, these successfully reared beyond the second instar in laboratory cultures have been deemed unsuitable hosts (Mahrt and Blickenstaff 1979). (Ure 1966). Here, we describe a protocol for rearing T. myopae- All known hosts are not native to North America, so the natural host formis in a laboratory setting to generate mature, third-instar larvae. plant of T. myopaeformis remains unknown (Mahrt and Blickenstaff Further, we compare oviposition, larval development, and feeding 1979). damage between a resistant and susceptible variety in no-choice and T. myopaeformis overwinter as third-instar larvae and move paired choice experiments. The results presented here contribute toward the surface to pupate when soil temperatures rise in the to our understanding of the modality of resistance for the resistant spring (Harper 1962). Adults emerge starting in late spring, then variety tested and outline a greenhouse protocol for screening ger- mate and lay eggs near the base of young sugar beet seedlings mplasm with putative resistance to T. myopaeformis. (Harper 1962). Male flies may live for an average of 6 d, whereas females live an average of 14 d (Ure 1966). Females lay 1 to 31 eggs Materials and Methods at a time and individuals have been reported to lay an average of 120 eggs during their lifespan (Harper 1962, Wenninger et al. 2018). Collection and Rearing of Flies Eggs begin hatching within 3 to 5 d after being laid and progress Third-instar maggots were collected in the Red River Valley of through three larval instars as they feed on the roots of sugar beet North Dakota in portions of commercial sugar beet fields in which plants (Gojmerac 1956, Ure 1966). After reaching the third instar, insecticides were not used. Collections were performed in July dur- usually during late summer, larvae tunnel deeper into the soil to ing the 2013, 2014, and 2015 growing seasons, after maggots had begin diapause (Harper 1962). The winter diapause period is obliga- reached maturity. T. myopaeformis typically reach the third instar tory for successful maggot pupation in the spring and typically lasts and stop feeding by late June to early July and tunnel deeper in the 6 mo (Chirumamilla et al. 2008). However, flies have been observed soil as temperatures decrease at the end of the summer (Hein 1995). to successfully pupate after 4 mo in diapause (S. D. Eigenbrode, per- Maggots were then transported to the University of Idaho Kimberly sonal communication). Research and Extension Center, in Kimberly, ID. T. myopaeformis larvae feed by rasping the root surface with Maggots were stored together in plastic vials filled with soil and their mouth hooks and consuming beet juices (Anderson et al. 1975, held in an environmental chamber (Percival Scientific, Inc., Perry, Wenninger et al. 2018). Feeding damage on older roots produces IA) until needed for use in experiments. The environmental chamber black scarring on the root surface and reduces yield. However, yield was held at 4°C with 70% RH and no light. Maggots were held at reductions can be even more severe if T. myopaeformis feeding sev- 4°C for a minimum of 6 mo to allow the insects to go through the ers the tap root of sugar beet seedlings, resulting in stand reduction. diapause period needed for pupation (Harper 1962). An organic pot- Such reductions in stand can be especially severe if beets are planted ting soil (“The EarthWorks”; Brandtastic Soil, LLC., Twin Falls, ID) later than usual (Campbell et al. 1998). Yield losses can range from was used for storing maggots and for all experiments. The soil mix 10 to 100% in areas where T. myopaeformis pressure is high (Cooke was comprised of equal parts peat moss, composted cow manure, 1993). perlite, and coconut coir. Potting soil was autoclaved at 121°C Cultural and biological control practices have been pursued for and 137,895 Pa for 30 min to reduce mortality of maggots from management of T. myopaeformis. However, these control meth- soil-borne pathogens. After autoclaving soil, deionized water was ods are not as effective as chemical control in areas of high pest added to achieve gravimetric soil moisture content of 30 to 40%. pressure (Campbell et al. 2000, Majumdar et al. 2008, Dunkel Rearing containers consisted of clear plastic 30-ml cups (Sovereign et al. 2010, Wenninger et al. 2018); thus, insecticides remain the Drinkware, Federalsburg, MD) partially filled with soil into which a cornerstone for management of this pest. The two groups of insec- maggot was gently placed using sterilized forceps and then covered ticides most commonly used against T. myopaeformis, carbamates with a ca. 6-mm layer of soil. After fitting each cup with a lid (Dixie, and organophosphates, are being phased out of use (Anonymous Easton, PA), cups were transferred to clear, plastic trays. Each tray 2010), and application of these pesticides is opposed by advocates was then placed inside a two-gallon plastic freezer bag. A 3 × 5 × for farm workers, child-safety, and the environment (Donley 2016). 0.5 cm cutting of a sponge moistened with ca.