Insect Science (2011) 18, 473–483, DOI 10.1111/j.1744-7917.2010.01401.x ORIGINAL ARTICLE Characterization of three transcripts encoding small heat shock proteins expressed in the codling moth, Cydia pomonella (Lepidoptera: Tortricidae) Stephen F. Garczynski, Thomas R. Unruh, Christelle Gu ´ edot and Lisa G. Neven USDA-ARS, Yakima Agricultural Research Laboratory, Wapato, Washington, USA Abstract Codling moth is a major pest of apples and pears worldwide. Increasing knowledge of how this insect responds to environmental stress will improve field and post- harvest control measures used against it. The small heat shock proteins (sHsps) play a major role in cellular responses to environmental stressors. A degenerate oligonucleotide primer, designed against the conserved α-crystallin domain, was used in 3 ′ rapid amplification of complementary DNA (cDNA) ends reactions to amplify transcripts encoding sHsps expressed in the codling moth cell line, Cp169, subjected to heat shock. Three full-length cDNAs were cloned from Cp169 cells that contained open reading frames encoding sHsps. The cDNA for CpHsp19.8 was 795 bp encoding 177 amino acids. The cDNA for CpHsp19.9 was 749 bp encoding 175 amino acids. The cDNA for CpHsp22.2 was 737 bp encoding 192 amino acids. Analysis of the protein sequences of the three CpHsps indicated the presence of 83 amino acids with homology to the α-crystallin domain. For each of the CpHsps, the α-crystallin domain was surrounded by divergent N- and C-terminal regions, consistent with the conserved structural features of sHsps. Real-time polymerase chain reaction, used to determine the expression patterns of each of the sHsps in different developmental stages of codling moth revealed the presence of transcripts in all stages tested. Consistent with characteristics of other sHsps, expression of CpHsp transcripts were greatly enhanced when insects were subjected to heat shock. The results of this research can be used as a guide to study the roles of sHsps in codling moth control using various post-harvest treatments. Key words α-crystallin domain, heat shock response, qPCR Introduction Codling moth, Cydia pomonella (L.), is a major pest of apples and pears throughout most of the apple growing regions of the world (Barnes, 1991). Numerous field and post-harvest control measures have been implemented to reduce infestation levels in fruit and meet quarantine restrictions in countries where this pest is not present Correspondence: Stephen F. Garczynski, USDA-ARS, Yakima Agricultural Research Laboratory, 5230 Konnowac Pass Rd, Wapato, WA 98951, USA. Tel: 509 454 6572; email: steve.garczynski@ars.usda.gov (Hansen et al., 2000; Jang & Moffitt, 1994; Neven, 2005; Neven et al., 2006; Neven & Rehfield-Ray, 2006a,b). Many post-harvest quarantine procedures have been de- veloped against codling moth, including a combination cold storage followed by methyl bromide fumigation (Hansen et al., 2000), cold storage alone (Moffitt & Al- bano, 1972; Moffitt & Burdett, 1989), and heat plus con- trolled atmospheres (Neven, 2005; Neven et al., 2006; Neven & Rehfield-Ray, 2006a, 2006b). These procedures can be classified as environmental stresses and as such may elicit the production of a specific cadre of stress re- sponse proteins called heat shock proteins (Hsps), which can serve to protect the cells from damage incurred by exposure to environmental extremes (Feder & Hofmann, C  2011 The Authors Journal compilation C  Institute of Zoology, Chinese Academy of Sciences 473