Diagnosis of Lobesia botrana (Lepidoptera: Tortricidae) Using Real-Time PCR.
A real-time PCR assay is reported for identification of Lobesia botrana (Denis and Schiffermüller) collected in California. This assay multiplexes two independent TaqMan probe systems in a single reaction tube to reduce handling time and sample exposure to environmental contaminants. One probe system targets a segment of DNA located in the internal transcribed spacer region 2 (ITS2) that is present in the L. botrana genome but absent in native North American Tortricidae. The second probe system serves as a control for DNA quality by targeting a segment of the 18S rDNA gene that is conserved in L. botrana and all of the tested nontarget species. The assay successfully diagnosed 70 Lobesia botrana specimens and 95 nontarget specimens. No false-positive or false-negative results were observed supporting its application for identification of this pest in California.
Center for Plant Health Science and Technology, Mission Laboratory, USDA-APHIS-PPQ-S&T, Moore Air Base, Edinburg, TX 78541 (lisa.a.ledezma@aphis.usda.gov; Norman.B.Barr@aphis.usda.gov).
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Ledezma
Center for Plant Health Science and Technology, Mission Laboratory, USDA-APHIS-PPQ-S&T, Moore Air Base, Edinburg, TX 78541 (lisa.a.ledezma@aphis.usda.gov; Norman.B.Barr@aphis.usda.gov) Norman.B.Barr@aphis.usda.gov.
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Barr
California Department of Food and Agriculture, Plant Pest Diagnostics Branch, 3294 Meadowview Rd., Sacramento, CA 95832 (marc.epstein@cdfa.ca.gov).
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Epstein
Identification Technology Program, USDA-APHIS-PPQ-S&T, Fort Collins, CO 80526 (Todd.M.Gilligan@aphis.usda.gov).
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Gilligan
D000818:Animals; D002140:California; D021903:DNA, Ribosomal Spacer / Q000032:analysis; D007300:Insect Control / Q000379:methods*; D007814:Larva / Q000145:classification / Q000235:genetics; D009036:Moths / Q000145:classification* / Q000235:genetics; D011679:Pupa / Q000145:classification / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
D021903:DNA, Ribosomal Spacer; D012337:RNA, Ribosomal, 18S
European grape vine moth; Paralobesia; internal transcribed spacer