Canadian Forest Service Publications

Establishment of a cell line from the ash and privet borer beetle Tylonotus bimaculatus Haldemand and assessment of its sensitivity to diacylhyrazine insecticides. 2015. Wen, F.; Caputo, G.; Hooey, S.; Bowman,S.; Pinkney, K.; Krell, P.J.; Arif, B.; Doucet, D. In Vitro Cellular and Developmental Biology 51:905-914.

Year: 2015

Issued by: Great Lakes Forestry Centre

Catalog ID: 36140

Language: English

Availability: PDF (request by e-mail)

Available from the Journal's Web site.
DOI: 10.1007/s112626-015-9917-8

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A novel cell line, NRCAN-Tb521, was developed from larvae of the longhorn beetle Tylonotus bimaculatus (Coleoptera: Cerambycidae), a pest of North American ash trees. The cell line has been successfully passaged more than 50 times and displayed very strong attachment to the substrate and a modal chromosomal count distribution of 19. Sequencing of a 649 bp fragment of the mitochondrial cytochrome oxidase I gene confirmed the identity of NRCANTb521 as T. bimaculatus. The response of the cell line to 20- hydroxyecdysone and diacylhydrazine ecdysone agonist insecticides was also studied. At 10−6 M, 20-hydroxyecdysone, tebufenozide, methoxyfenozide and halofenozide triggered the production of numerous filamentous cytoplasmic extensions, and the cells tended to form aggregates, indicative of a cell differentiation response. This response was followed by a strong decrease in viability after 4 d. Reverse transcription polymerase chain reaction (PCR) experiments and sequencing of PCR fragments showed that the 20E receptor gene EcR is expressed in the cells and that 20E, tebufenozide, methoxyfenozide and halofenozide also induce the expression of the nuclear hormone receptor gene HR3. This report establishes that NRCAN-Tb521 is a valuable in vitro model to study effects of ecdysone agonists in wood-boring cerambycids.

Plain Language Summary

Wood-boring insects can pose threats to tree health and reduce the value of timber. In some cases these insects can be managed with insecticides. However, the testing of novel insecticide formulations for wood-boring beetles requires a large number of larvae for assays which can be cost prohibitive. We have developed a cell line from a wood boring beetle (the ash and privet borer) that can be propagated almost indefinitely under laboratory conditions and can be used to test the efficacy of insecticides. This article first presents a detailed description of the morphological and molecular characteristics of the cell line. Three different insecticide molecules that interfere with insect molting were also tested on the cell line. The insecticides triggered changes in cell morphology and caused cell death in a dose-dependent manner. The compounds also interfered with the expression of molting genes. However, the effects were the same irrespective of the insecticide used. The results demonstrate that the ash privet borer cell line can be used to test the activity of multiple insecticide compounds in an efficient manner.