Agriculture and Agri-Food Canada Agriculture et agroalimentaire Canada From the molecule to the organism: Physiology of the insect to help plant protection Desde la molécula al organismo: Fisiologia del insecto en ayuda de la protección Yvan Pelletier Potato Research Centre, AAFC Canada XXIII Congreso de la ALAP´08 VI Seminario Latinoamericano de Uso y C omercialización de la Papa Potato Research Centre Agriculture and Agri-Food Canada Potato Research Centre Agriculture and Agri-Food Canada 30% Production in 2007 25% 158,966 ha (4,969,506 t) 20% 15% 10% 5% 0% C.-B. Alb. Sask. Man. Ont. Qué. N.-B. N.-E I.-P.-E. T.-N. Potato Research Centre Agriculture and Agri-Food Canada Fredericton Potato Research Centre Agriculture and Agri-Food Canada Fredericton Main site (administrative and scientific) 17 scientists 20 laboratories and 300 hectares Benton Ridge Breeding sub-station 350 hectares Potato Research Centre Agriculture and Agri-Food Canada Insect Physiology Laboratory Potato Research Centre Mandate: Study potato insect pests Insect-plant interactions Potato Research Centre Agriculture and Agri-Food Canada Insect Physiology Laboratory Insect Physiology and Biomechanics Insecticide Resistance Potato Resistance to Insects PVY Transmission by Aphids Agriculture and Agri-Food Canada Agriculture et agroalimentaire Canada Green Peach Aphid Potato Aphid Colorado Potato Potato Beetle Potato Tuber Moth fleabeetle Agriculture and Agri-Food Canada Agriculture et agroalimentaire Canada Leptinotarsa decemlineata Colorado Potato Beetle Agriculture and Agri-Food Canada Agriculture et agroalimentaire Canada Agriculture and Agri-Food Canada Agriculture et agroalimentaire Canada The Research and Development Continuum Research Development Application Control Method How it works How to make it work Work with it Haemolymph Plasma Ions Macrosiphum albifrons Aphid blood collection: 1 student, 2 weeks, 70 hours 4,000 aphids 0.4 ml of blood Haemolymph Plasma Ions Haemolymph Plasma Ions Development of physiological saline solutions CPB anti-diuretic hormone aDH collection: 25,000 CPB heads CPB anti-diuretic hormone First insect anti-diuretic hormone active on Malpighian tubules CPB water loss Conditions: 30°C, 5% RH Plastic lined trench CPB biomechanics Walking on slope CPB biomechanics Walking on slope First description of the biomechanics of an insect walking on slope CPB biomechanics Obstacle perception First time antennae were associated with terrestrial locomotion Physical control method Flamer Physical control method Thermal injuries Physical control method Thermal injuries Injury level of adult CPB dipped in water of different temperature for one second. Physical control method Thermal injuries Physical control method Lethal temperature Physical control method Lethal temperature Physical control method CPB scorcher Insecticide resistance Colorado potato beetle Whalon, M., D. Mota-Sanchez, and L. Duynslager. 2004. CPB has The database of arthropod resistance to pesticides. developed http://www.pesticideresistance.org/DB/index.html. acetamiprid resistance to 49 aldicarb dioxacarb oxamyl aldrin endosulfan parathion insecticides azinphos-methyl endrin parathion-methyl Bt CryIIIA esfenvalerate permethrin carbaryl fenvalerate phorate carbofuran hydrogen cyanide phosmet cartap imidacloprid phoxim chlordane lindane/BHC propoxur chlorfenvinphos malathion quinalphos cloethocarb methamidophos rotenone clothianidin methidathion Spinosad cypermethrin methoxychlor tetrachlorvinphos DDT monocrotophos thiacloprid deltamethrin N-Desmethylthiamethoxam thiamethoxam Dieldrin N-Methylimidacloprid toxaphene dinotefuran nitenpyram trichlorfon Insecticide resistance Molecular tools Low Density Reverse BAC libraries Northern Array Identification of ~70 detoxification genes Sequencing of 3 nAChE receptor genes Dr. Jianhua Zhang Method for quantification of point mutation in populations Insecticide resistance Cuticular proteins Insecticide resistance Cuticular proteins First demonstration of expression of cuticular protein genes at adult stage Response to environmental stresses (insecticide and dehydration) Provide protection against environmental stresses? Cuticular protein genes over expressed after treatment with insecticide,Cuticular and more protein so in genes insecticide over expressed resistant CinPB adults Expression of cuticular protein in the epidermis of CPB adults maintained in dry conditions and adult CPB when starved Resistance to insects Wild Solanum Resistance to insects Research approach The approach Performance on wild Solanum Behaviour on wild Solanum Identification of the mode of resistance Can the insects adapt to the resistant potato? Resistance to insects Solanum tarijense Resistance to insects Solanum tarijense Resistance to insects Solanum tarijense Effect of trichomes on CPB behavior n % Fell Time to fall tar with trichomes 30 60% 2 min 23s tar without trichome 20 0% tbr 30 0% Resistance to insects Solanum tarijense Effect of trichomes on the feeding of adult CPB Feeding (g) tar with trichomes 8.62 " 2.2 ac tar without trichome 7.69 " 1.5 a tbr with trichomes 20.61 " 2.1 b tbr without trichomes 15.34 " 2.1 bc Resistance to insects Solanum tarijense Consumption (mm 2) of potato leaf disk treated with leaf surface extract from S. tarijense or potato Feeding (mm 2) tar 5.9 " 0.55 a tbr 8.3 " 0.53 b Solvent 8.6 " 0.50 b Resistance to insects Potato Tuber Moth Dr. Finbarr Horgan Resistance to insects Potato Tuber Moth S. tarijense adaxial abaxial trichone A hairs trichone A hairs S. berthaultii adaxial abaxial trichone A trichome B trichone A trichome B Proportion of eggs on eggs of leaf Proportion Trichome density (cm -2) Resistance to insects Potato Tuber Moth S. tarijense adaxial abaxial trichone A hairs trichone A hairs S. berthaultii adaxial abaxial trichone A trichome B trichone A trichome B Proportion of neonates established of Proportion neonates Trichome density (cm -2) Resistance to insects Potato Tuber Moth Resistance in tubers of S. berthaultii and S. tarijense as determined from laboratory challenges with PTM neonate larvae. Species Accession Total Resistance Proportion of Resistance b Resistance a after Perforation Periderm Cortex Ber HAM 141 0.47 ± 0.10a 0.07 ± 0.04 0.81 0.19 ± 0.13 Ber HCH 4422 0.70 ± 0.13a 0.39 ± 0.11 0.48 0.52 ± 0.06 Ber HHCH 4562 0.33 ± 0.15a 0.07 ± 0.07 0.78 0.22 ± 0.33 Ber HHCH 4728 0.33 ± 0.11a 0.07 ± 0.07 0.75 0.25 ± 0.25 Ber HHCH 4746 0.53 ± 0.06a 0.17 ± 0.07 0.67 0.33 ± 0.11 Ber OCH 12008 0.33 ± 0.00a 0.20 ± 0.08 0.4 0.60 ± 0.24 Ber OCHS 15599 0.40 ± 0.12a 0.07 ± 0.07 0.75 0.25 ± 0.25 Ber UGN 4561 0.67 ± 0.09a 0.17 ± 0.05 0.72 0.28 ± 0.09 Tar HOFF 2017 0.43 ± 0.06a 0.14 ± 0.07 0.64 0.36 ± 0.18 Tar OCH 11993 0.58 ± 0.12a 0.25 ± 0.06 0.55 0.46 ± 0.16 Tar OCH 12001 0.60 ± 0.16a 0.07 ± 0.07 0.9 0.10 ± 0.10 Tar OCHS 15561 0.70 ± 0.06ab 0.40 ± 0.08 0.38 0.62 ± 0.12 Tar OCHS 15584 0.76 ± 0.08ab 0.33 ± 0.08 0.52 0.48 ± 0.11 Tar OCHS 15592 0.67 ± 0.00a 0.33 ± 0.11 0.5 0.50 ± 0.16 Tar OCHS 15593 0.67 ± 0.08a 0.26 ± 0.07 0.61 0.39 ± 0.11 Tar OCHS 15596 0.78 ± 0.08b 0.37 ± 0.04 0.48 0.52 ± 0.07 Tbr c ‘Peruanita’ 0.00 ± 0.00 0.00 ± 0.00 0 0.00 ± 0.00 Resistance to insects Aphids Macrosiphum euphorbiae Potato Aphid Resistance to insects Electric Penetration Graph Mesophyll Xylem Phloem Resistance to insects Aphid probing 0.5 0.4 0.3 0.2 Xylem /(xylem+phloem) Xylem 0.1 0.0 0 1 2 3 6 8 1012 aphid age (day) Resistance to insects Aphid probing fecundity Xylem proportion 2.5 0.5 Xylem/xylem+phloem) 2.0 0.4 1.5 0.3 1.0 0.2 Nymphs/female 0.5 0.1 0.0 0.0 0 1 2 3 4 5 6 7 8 9101112 aphid age (day) Resistance to insects Aphid probing flight capability fecundity Xylem proportion 100% 3.00 2.50 80% Nymphs/female 2.00 60% 1.50 40% Aphids flying Aphids 1.00 20% 0.50 0% 0.00 0 1 2 3 4 5 6 7 8 9101112 aphid age (day) Resistance to insects Aphid probing fecundity Xylem proportion 1.0 0.9 4 0.8 0.7 Nymphs/female 3 0.6 0.5 2 0.4 0.3 Xylem/xylem+phloem) 0.2 1 0.1 0.0 0 0 1 2 3 4 5 6 7 8 9 10111213141516 aphid age (day) Resistance to insects Aphid probing Xylem consumption is modulated by the physiological state of the aphid and linked to fecundity and/or flight. Xylem consumption is part of the host selection process, maintaining the aphid ability to flight (light weight). Factor of plant resistance in the xylem might contribute to the defense of a plant towards a specific aphid species. Potato Research Centre Agriculture and Agri-Food Canada Insect Physiology Laboratory Insect Physiology and Biomechanics Potato Resistance to Insects Insecticide Resistance.