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Development of a Bioassay System for the Predator, Xylocoris Flavipes

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Development of a Bioassay System for the Predator, Xylocoris Flavipes Development of a Bioassay System for the Predator, Xylocoris flavipes (Heteroptera: Anthocoridae), and Its Use in Subchronic Toxicity/Pathogenicity Studies of Beauveria bassiana Strain GHA Author(s): Florence V. Dunkel and Stefan T. Jaronski Source: Journal of Economic Entomology, 96(4):1045-1053. 2003. Published By: Entomological Society of America DOI: http://dx.doi.org/10.1603/0022-0493-96.4.1045 URL: http://www.bioone.org/doi/full/10.1603/0022-0493-96.4.1045 BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the biological, ecological, and environmental sciences. BioOne provides a sustainable online platform for over 170 journals and books published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/terms_of_use. Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder. BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research. BIOLOGICAL AND MICROBIAL CONTROL Development of a Bioassay System for the Predator, Xylocoris flavipes (Heteroptera: Anthocoridae), and Its Use in Subchronic Toxicity/Pathogenicity Studies of Beauveria bassiana Strain GHA 1 2, 3 FLORENCE V. DUNKEL AND STEFAN T. JARONSKI Department of Entomology, Montana State University, Bozeman, MT 59717 J. Econ. Entomol. 96(4): 1045Ð1053 (2003) ABSTRACT Microbial biocontrol agents are useful commercially only if they do not harm other natural biocontrol organisms, at recommended use rate in the environment where the microorganism is being used. To test the hypothesis that the predatory warehouse pirate bug, Xylocoris flavipes (Reuter), is not adversely affected by the entomopathogen, Beauveria bassiana (Balsamo) Vuillemin strain GHA, we developed a method using individually conÞned bugs during a 10-d feeding regime. Three concentrations of a conidial suspension were applied to assay surfaces (Þlter paper disks) to achieve 2.7 ϫ 107, 2.6 ϫ 106, and 2.6 ϫ 105 conidia per cm2, representing 100X, 10X, and a Þeld rate of 2.6 ϫ 1013 conidia per ha (Ϯ10%), respectively. Fifth instar X. flavipes suffered 0% infection at the Þeld rate when conÞned to treated Þlter paper for 10 d (16% and 42% infection, respectively, at 10X and 100X the Þeld rate). Second instar migratory grasshoppers, Melanoplus sanguinipes (F.), exposed to the same doses suffered 97, 92, and 100% mortality at the three respective doses 10 d after exposure. These data indicate that B. bassiana can be used safely at recommended application levels without signiÞcant effect on Þfth instar populations of X. flavipes. KEY WORDS mycoinsecticide, nontarget organism, biological control, Melanoplus sanguinipes, stored product insects, predators Beauveria bassiana (BALSAMO) Vuillemin is one of cial scale by Emerald BioAgriculture (formerly three species of fungus with a broad host range that is Mycotech Corp., Butte, MT) and has been registered currently registered for use as insect control agents. by the U.S. EPA as a microbial insecticide for use An ideal biocontrol agent fungus is one with a re- against a variety of Orthoptera, thrips, aphids, white- stricted host range (Goettel et al. 1990). Because the ßies, and chrysomelid beetles. Strain GHA also has various isolates of B. bassiana have a broad host range some Þeld efÞcacy against the diamondback moth, of Ͼ700 species (Li 1987), it is possible that some Plutella xylostella (L.) (Shelton et al. 1998, Vanden- isolates of this fungus are infectious to nontarget in- burg et al. 1998). Effectiveness of this strain against vertebrates. After isolates of candidate fungi have Lygus hesperus Knight and Lygus lineolaris (Palisot been shown efÞcacious against a target insect, it is de Beauvois) in the laboratory and Þeld, respectively, important to test their safety for nontarget inverte- was demonstrated by Noma and Strickler (1999) brates. Coccinellid predators tested have shown only and others (Steinkraus 1996, Steinkraus and Tugwell slight susceptibility to B. bassiana under conditions 1997). reßective of Þeld exposure at recommended Þeld rates The Anthocoridae (Heteroptera) or ßowerbugs, for target insects (Magalha˜es et al. 1988, Inglis et al. are important beneÞcial insects found throughout the 1997, Jaronski and Goettel 1997). Some isolates of world, in temperate and tropical areas. This group of B. bassiana, however, have virulence for both pestif- small insects, usually Ͻ4 mm as adults, is predaceous erous and beneÞcial Heteroptera, for example, Redu- in all instars and as adults. Prey include such pests as viidae, speciÞcally Triatominae (Luzet al. 1998), Rho- aphids (Rakauskas 1984), European corn borer eggs palidae (Reinert et al. 1999), and Miridae (Bilewicz- (Reid 1991) and larvae (Isenhour et al. 1989), corn Pawinka and Bajan 1973, Bidochka et al. 1993, Jaronski earworm eggs (Reid 1991) and larvae (Isenhour et al. et al. 1998, Noma and Strickler 1999). Strain GHA of 1989), boll weevils (Wright 1993), psyllids (Herard B. bassiana is presently being produced on a commer- and Chen 1985, Barrion et al. 1987, Hagen and Dreistadt 1990), whiteßies (Dolling 1991), mites (Heitmans et al. 1986, McCaffrey and Horsburgh 1 E-mail: [email protected]. 2 Mycotech Corporation, 529 E. Front Street, Butte, MT 59701. 1986a), thrips (Kajita 1986, Saucedo-Gonzales and 3 Current address: USDAÐARS Northern Plains Agricultural Re- Reyes-Villanueva 1987), scales (Dolling 1991), and search Laboratory, 1500 N. Central Ave., Sidney MT 59270. pine bark beetle larvae (Schmitt and Boyer 1983). 0022-0493/03/1045Ð1053$04.00/0 ᭧ 2003 Entomological Society of America 1046 JOURNAL OF ECONOMIC ENTOMOLOGY Vol. 96, no. 4 These prey can cause serious economic damage and Goettel 1997) and one and two orders of magni- throughout the world by decreasing yield in many tude greater (concentrations fatal for the migratory crops such as: sorghum (Steward et al. 1991), apples grasshopper, Melanoplus sanguinipes [F.]). For the (McCaffrey and Horsburgh 1986b), pears (Vilajeliu test, we developed a Strain GHA-treated surface that 1989), berries (Rakauskas 1984), soybeans (Isenhour would mimic exposure to a leaf surface over a 10-d and Marston 1981), tea (Carayon 1982), corn (Reid period. This bioassay regime is considered subchronic 1991), and cotton (Elov 1984). Potential prey can also because exposure to the substance (B. bassiana) was cause economic loss in postharvest commodities such conducted over an extended period of time (Ϸ20% of as grain (Donnelly and Phillips 2001), dry edible beans the organismÕs lifetime 56 d [Sing 1997; EPA, personal (Sing 1997), almonds (Brower and Press 1988, Press communication]) and included two developmental 1989), peanuts (Press et al. 1979, Keever et al. 1986, stages, the last immature instar and the adult. These Brower and Mullen 1990) and processed food (Krasz- studies were used to support registration of this my- pulski and Davis 1988). coinsecticide. Some anthocorids are presently sold commercially, for example, the insidious ßower bug, Orius insidiosus Materials and Methods (Say), and the minute ßower bug O. tristicolor (White) (Planet Natural 2001), O. tristicolor and B. bassiana Strain GHA. Technical powder having Xylocoris flavipes (Reuter) (Biofac, Inc. 2001) to con- 1.43 ϫ 1011 viable Strain GHA conidia per g residual trol agricultural pests in alfalfa, greenhouses, and culture solids (Mycotech Lot #930210GHA.A) was stored products. X. flavipes has been approved for suspended in 0.05% Tween 80 in deionized water with application on stored grain in the U.S. (Anon. 1992). a Potter-Elvejhem cell homogenizer. This resulted in Montantoniola moraguezi (Puton) has been intro- a stock suspension of 1.77 ϫ 1010 conidia per ml, which duced from the Philippines and Hawaii to control was veriÞed by hemocytometer counts. This concen- thrips in California (Henry 1988). Tetraphleps abdul- tration was within 10% of the target concentration, ghani Ghauri and T. raoi Ghauri have been introduced 1.7 ϫ 1010 conidia per ml. from India and Pakistan into British Columbia and The entire stock solution was then serially diluted Nova Scotia without apparent establishment to con- with deionized water to yield suspensions with 1.77 ϫ trol aphids (Henry 1988). Some anthocorid species are 109, 1.72 ϫ 108, and 1.76 ϫ 107 conidia per ml (deter- cosmopolitan because of their relationship with post- mined by hemocytometer count). These dilutions harvest commodities (Miller 1971, LeCato and Davis were applied at the rate of 20 ␮l to each Þlter paper 1973, Dunkel and Ivie 1994, Yao et al. 1998). disk (1.18 cm in diameter). These inocula yielded The anthocorid we selected for our study is the 2.7 ϫ 107, 2.6 ϫ 106, and 2.6 ϫ 105 conidia per cm2, warehouse pirate bug, X. flavipes. This pirate bug is which are 100, 10, and 1X (Ϯ10%) a Þeld rate of 2.5 ϫ cosmopolitan in distribution, and found wherever 1013 conidia per ha, respectively. The recommended storage insects are located, in prairie areas, agricul- U.S. Þeld rate is 2.5 ϫ 1013 conidia per ha, (Inglis et al. tural areas, and in wooded areas. The speciÞc micro- 1997, Jaronski and Goettel 1997). habitat typical for this anthocorid is decaying vegeta- Heat-killed Strain GHA was prepared by autoclav- tion and rotting seeds. Although four of the Þve studies ing conidia at 121ЊC (103.4 kilopoise [KPa] ϭ 15 psi) of the impact of conventional residual pesticides for 25 min. The nonviability of the resulting conidia (Press et al. 1978, Niemycczyk 1973, Parker and Nila- was conÞrmed by plating diluted spore suspensions on khe 1990, Baker and Arbogast 1995) and fumigants Sabouraud Dextrose agar plus 1% yeast extract and (Press and Flaherty 1978) on anthocorids used Sabouraud Dextrose agar and incubating the plates at X.
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