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Colorado potato beetles compensate for tomato cathepsin D inhibitor expressed in transgenic potato.
Arch Insect Biochem Physiol. 2004 Mar; 55(3):103-13.AI

Abstract

We reported earlier the importance of digestive cathepsin D-like activity for initiating dietary protein hydrolysis in Colorado potato beetle, Leptinotarsa decemlineata Say [Brunelle et al. (1999) Arch. Insect Biochem. Physiol. 42:88-98]. We assessed here whether transgenic lines of potato (Solanum tuberosum L.) expressing a cathepsin D inhibitor (CDI) from tomato would show resistance to the beetle, or if the insect would compensate for the loss of cathepsin D activity after ingesting the recombinant inhibitor. Transgenic potato lines expressing tomato CDI were developed by Agrobacterium tumefaciens genetic transformation, and selected based on their relative amount of CDI. After confirming the absence of detectable visible effects of CDI on the plant's phenotype, diet assays with control and transgenic lines were carried out to assess the impact of the inhibitor on growth and development of the insect. Leaf consumption, relative growth rate, molting incidence, and digestive protease activity were monitored at 12-h intervals over 132 h for 3rd-instar larvae provided with transgenic potato foliage. Leaf consumption and relative growth rate were slightly reduced during the first 12 h for larvae fed CDI, but no significant differences were observed thereafter. In contrast, time for molting to the 4th larval stage was significantly longer for larvae fed modified plants, with developmental delays of approximately 10 h (0.5 day) compared to control larvae. Recombinant CDI also had an impact on the insect's digestive physiology, readily inducing overproduction of digestive proteases (rubiscases), followed by a gradual decrease of total and pepstatin-sensitive activity. Overall, these observations show the ability of Colorado potato beetle to compensate for the loss of cathepsin D activity by modulating its digestive protease complement in response to aspartate-type inhibitors in the diet. From a practical viewpoint, these data stress the importance of devising improved strategies for the effective inhibition of insect digestive proteinases in vivo, based on the use of hybrid inhibitors active against different protease classes.

Authors+Show Affiliations

Brunelle F
Département de Phytologie, Centre de recherche en horticulture, Université Laval, Québec, Canada.
Cloutier C
No affiliation info available
Michaud D
No affiliation info available

MeSH

Animal FeedAnimalsColeopteraEndopeptidasesGerminationLarvaMetamorphosis, BiologicalPepstatinsPlant LeavesPlant ProteinsPlant StemsPlants, Genetically ModifiedProtease InhibitorsRecombinant ProteinsSolanum tuberosumTosyl CompoundsTransgenes

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

14981655

Citation

Brunelle, France, et al. "Colorado Potato Beetles Compensate for Tomato Cathepsin D Inhibitor Expressed in Transgenic Potato." Archives of Insect Biochemistry and Physiology, vol. 55, no. 3, 2004, pp. 103-13.
Brunelle F, Cloutier C, Michaud D. Colorado potato beetles compensate for tomato cathepsin D inhibitor expressed in transgenic potato. Arch Insect Biochem Physiol. 2004;55(3):103-13.
Brunelle, F., Cloutier, C., & Michaud, D. (2004). Colorado potato beetles compensate for tomato cathepsin D inhibitor expressed in transgenic potato. Archives of Insect Biochemistry and Physiology, 55(3), 103-13.
Brunelle F, Cloutier C, Michaud D. Colorado Potato Beetles Compensate for Tomato Cathepsin D Inhibitor Expressed in Transgenic Potato. Arch Insect Biochem Physiol. 2004;55(3):103-13. PubMed PMID: 14981655.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Colorado potato beetles compensate for tomato cathepsin D inhibitor expressed in transgenic potato. AU - Brunelle,France, AU - Cloutier,Conrad, AU - Michaud,Dominique, PY - 2004/2/26/pubmed PY - 2004/4/7/medline PY - 2004/2/26/entrez SP - 103 EP - 13 JF - Archives of insect biochemistry and physiology JO - Arch Insect Biochem Physiol VL - 55 IS - 3 N2 - We reported earlier the importance of digestive cathepsin D-like activity for initiating dietary protein hydrolysis in Colorado potato beetle, Leptinotarsa decemlineata Say [Brunelle et al. (1999) Arch. Insect Biochem. Physiol. 42:88-98]. We assessed here whether transgenic lines of potato (Solanum tuberosum L.) expressing a cathepsin D inhibitor (CDI) from tomato would show resistance to the beetle, or if the insect would compensate for the loss of cathepsin D activity after ingesting the recombinant inhibitor. Transgenic potato lines expressing tomato CDI were developed by Agrobacterium tumefaciens genetic transformation, and selected based on their relative amount of CDI. After confirming the absence of detectable visible effects of CDI on the plant's phenotype, diet assays with control and transgenic lines were carried out to assess the impact of the inhibitor on growth and development of the insect. Leaf consumption, relative growth rate, molting incidence, and digestive protease activity were monitored at 12-h intervals over 132 h for 3rd-instar larvae provided with transgenic potato foliage. Leaf consumption and relative growth rate were slightly reduced during the first 12 h for larvae fed CDI, but no significant differences were observed thereafter. In contrast, time for molting to the 4th larval stage was significantly longer for larvae fed modified plants, with developmental delays of approximately 10 h (0.5 day) compared to control larvae. Recombinant CDI also had an impact on the insect's digestive physiology, readily inducing overproduction of digestive proteases (rubiscases), followed by a gradual decrease of total and pepstatin-sensitive activity. Overall, these observations show the ability of Colorado potato beetle to compensate for the loss of cathepsin D activity by modulating its digestive protease complement in response to aspartate-type inhibitors in the diet. From a practical viewpoint, these data stress the importance of devising improved strategies for the effective inhibition of insect digestive proteinases in vivo, based on the use of hybrid inhibitors active against different protease classes. SN - 0739-4462 UR - https://www.unboundmedicine.com/medline/citation/14981655/Colorado_potato_beetles_compensate_for_tomato_cathepsin_D_inhibitor_expressed_in_transgenic_potato_ DB - PRIME DP - Unbound Medicine ER -