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Molecular Determination of the Predator Community of a Cassava Whitefly in Colombia: Pest-Specific Primer Development and field Validation

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Molecular Determination of the Predator Community of a Cassava Whitefly in Colombia: Pest-Specific Primer Development and field Validation J Pest Sci (2014) 87:125–131 DOI 10.1007/s10340-013-0509-7 ORIGINAL PAPER Molecular determination of the predator community of a cassava whitefly in Colombia: pest-specific primer development and field validation Jonathan G. Lundgren • Luis Augusto Becerra Lo´pez-Lavalle • Soroush Parsa • Kris A. G. Wyckhuys Received: 19 February 2013 / Accepted: 24 May 2013 / Published online: 8 June 2013 Ó Springer-Verlag Berlin Heidelberg (outside the USA) 2013 Abstract In South America, the whitefly Aleurotrachelus Condylostylus sp. (Diptera: Dolichopodidae), were both socialis is one of the principal pests of cassava (Manihot positive for whitefly DNA, but did not have the strongest esculenta Crantz), reaching high population levels trophic linkage to the pest relative to other predators. This throughout the Andean region. Management of this species study shows that a diverse predator community affects is primarily based upon the use of insecticides, while cassava whitefly in southern Colombia, and provides the biological control has received limited attention. Till groundwork for the design of cassava production systems present, knowledge of A. socialis natural enemies is with minimal pesticide inputs. restricted to occasional records of predators and parasit- oids. In this study, we developed PCR primer sets specific Keywords Aleurotrachelus socialis Á Biological control Á for the cassava whitefly, A. socialis, to identify their Harmonia axyridis Á Molecular gut analysis Á PCR Á predator community in Colombian cassava. Eleven percent Predator of 586 predator specimens (representing 131 taxa from 29 families) tested positive for cassava whitefly DNA. Of the 21 predator taxa that consumed cassava whiteflies, an Introduction unidentified netwing beetle (Lycidae), an unidentified spi- der species (Araneae), Harmonia axyridis (Coleoptera: Cassava is a nutritionally important crop plant for subsis- Coccinellidae), a Cereaochrysa sp. (Neuroptera: Chrys- tence farmers in tropical regions (UNCTAD 2009). It opidae), and a Leucochrysa sp. (Chrysopidae) were the taxa supports a diversity of pests, including a complex of 11 that consumed cassava whiteflies most frequently under species of whiteflies (Hemiptera: Aleyrodidae) (Bellotti field conditions. Two abundant predators in the system, et al. 1999). Dominant species in this complex vary among Delphastus sp. (Coccinellidae) and the long-legged fly, regions, and include especially Aleurothrixus aepim (Goldi), Bemisia tabaci (Gennadius), and Trialeurodes variabilis (Quaintance) (Thresh et al. 1994; Bellotti et al. Communicated by M. Traugott. 1999; Calvert et al. 2001). In Colombia and northern South America, an important region for cassava production and J. G. Lundgren (&) USDA-ARS, North Central Agricultural Research Laboratory research, the dominant cassava whitefly is Aleurotrachelus (NCARL), 2923 Medary Avenue, Brookings, SD 57006, USA socialis Bondani (Bellotti et al. 1999). Whiteflies affect e-mail: [email protected] cassava by sucking the phloem of the plant, transmitting viruses to the plant, and by inhibition of photosynthesis L. A. B. Lo´pez-Lavalle Á S. Parsa Á K. A. G. Wyckhuys CIAT, Centro Internacional de Agricultura Tropical, through their production of honeydew and associated sooty Apartado Ae´reo, 6713 Cali, Colombia mold (Bellotti and Arias 2001; Akinbo et al. 2012). Rapid population growth rates and prolonged exposure can dec- Present Address: imate local cassava fields, reducing yields substantially (by K. A. G. Wyckhuys CIAT, Centro Internacional de Agricultura Tropical, up to 80 % in some research) (Bellotti et al. 1983; Gold Hanoi, Vietnam et al. 1989a). The dominant method for controlling whitefly 123 126 J Pest Sci (2014) 87:125–131 outbreaks in tropical regions involves the frequent appli- predator and pest, and identifying the diversity of predators cation of insecticides, which leads to environmental and consuming a pest (Lundgren and Fergen 2011). human health risks and the rapid evolution of pest resis- The current study is an initial description of the tance. Developing farm management systems that couple predator community of cassava whitefly (A. socialis) near host plant resistance (Bellotti and Arias 2001; Carabalı´ Calı´, Colombia. We developed the molecular tools nec- et al. 2010; Akinbo et al. 2012) or intercropping (Gold essary for conducting PCR-based gut analysis to establish et al. 1989a) with the use of natural enemies could offer an trophic connections between a predator community and alternative to current pesticide-based whitefly management cassava whitefly, and validated the method by describing practices (Bellotti et al. 1999). A crucial knowledge gap which species are consuming whiteflies under field con- that challenges the development of this type of system is ditions during a short snapshot of the field season. This that the predator community of cassava whitefly remains developmental research can now be used to help develop poorly described. sustainable pest management plans for cassava whiteflies Generalist predators are an abundant source of biotic that target conservation efforts at key predators of the pest mortality in cropland, but their identity and role are yet pest. to be characterized in cassava cropping systems. Much of the previous study on cassava whitefly natural enemies has focused on parasitoids that would be amenable to classical Methods biological control releases (Evans and Castillo 1998; Trujillo et al. 2004). Describing important components of Insect collection the predator community of cassava whiteflies is challeng- ing. Effective predation often leaves no trace of the pre- Arthropods (non-target herbivores and putative whitefly dation event (unlike parasitism or infection with predators) were collected from a cassava resistance entomopathogens), and estimating the impact of predators screening experiment at the Cenican˜a research station on a pest requires carefully controlled experiments that (3.34°, -76.31° latitude and longitude) between April 25 often disrupt normal predator behaviors. Previous study has and May 16, 2011. Here, approximately 80 varieties of first identified Delphastus pusillus (Le Conte) (Coleoptera: year cassava plants are produced on a 4-ha plot; these Coccinellidae), = D. catalinae (Hoelmer and Pickett cassava genotypes display varying levels of whitefly 2003), as a commonly encountered predator of cassava resistance, and are produced without the use of insecti- whitefly (Lo´pez-A´ vila et al. 2001), with questionable cides. To ensure uniformly high whitefly population pres- impact on the pest (Gold et al. 1989b; Bellotti et al. 1994; sure, experimental blocks were bordered by a row of the Bento et al. 1999), but efforts to determine relative impacts highly susceptible cultivar CMC 40, which was artificially of various predators on whitefly populations have been infested with A. socialis obtained from greenhouse colonies cursory at best. Nevertheless, generalist predators are well maintained at the International Center for Tropical Agri- documented as regulating other pest populations, including culture (CIAT). These whiteflies were originally collected a diverse predator community of whiteflies in other crops at the CIAT research station (3.50°, -76.36°) and kept in (Hagler and Naranjo 1994a, b; Gerling et al. 2001; Lin culture for over 10 years, with periodic infusions of wild et al. 2008; Hodek and Honeˇk 2009; Moreno-Ripoll et al. individuals from the same location. At the field site, all life 2012). stages of the whitefly were abundant, although the popu- An important step in developing predator conservation lation was patchy in distribution. There were no obvious techniques is establishing which predators have a trophic deterrents to predator establishment at the field site (e.g., linkage with the focal pest species. One method that specific predators were not seeded at the site, nor were facilitates the establishment of these trophic linkages is insecticides applied that might shape predator communi- PCR-based gut analysis of the predator community ties), which was surrounded by grassy margins and other (Fournier et al. 2008; Lundgren et al. 2009; King et al. crops. An array of adult arthropods (131 herbivorous and 2010; Sint et al. 2011; Opatovsky et al. 2012). This method predatory taxa from 29 families) was collected from the implements PCR primer sets that selectively amplify a cassava foliage using sweep nets, or directly from the DNA fragment from a focal pest to search predator stom- foliage with aspirators. The goal of the study was to collect achs for pest DNA (Sheppard and Harwood 2005; Weber a diversity of predators, along with greater numbers of and Lundgren 2009a). While this method does not con- some dominant species, to validate the primer sets. As sistently correlate well with relative impact of the predators such, the field was not systematically sampled, some on the pest (impact is also related to predator densities, predator groups are over-represented, and others may have foraging patterns, etc.), it is a very effective method for been overlooked. Individual predators were isolated in assessing the strength of the trophic linkage between a 1.5 ll microcentrifuge tubes, and placed on ice until they 123 J Pest Sci (2014) 87:125–131 127 could be frozen in the laboratory in 70 % ethanol at incubated for 3 h in proteinase K. Whitefly DNA in the -20 °C. Non-target herbivores were generally left as predator stomachs was amplified using qPCR reactions morphotaxa used in specificity testing of the whitefly pri-
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