Holy Guacamole! Insights into the laurel wilt pandemic

Transcription

1 Holy Guacamole! Insights into the laurel wilt pandemic JASON A. SMITH 1, T. DREADEN, M. HUGHES, L. SOBEL, J. RIGGINS, J. ROLLINS, K. SMITH, A. BLACK, R. PLOETZ & C. HARMON 1 ASSOCIATE PROFESSOR, EMERGING THREATS TO FORESTS RESEARCH TEAM, SCHOOL OF FOREST RESOURCES AND CONSERVATION, UNIVERSITY OF FLORIDA, GAINESVILLE, FL JASONS@UFL.EDU

2 Ambrosia beetles are typically harmless But, some are causing mass tree mortality

3 Photo: Chip Bates

4 Xyleborus glabratus redbay ambrosia beetle Raffaelea lauricola - Ophiostomatales Clonal symbiosis!

5 0.1 Ambrosiella_ips_CBS Ambrosiella_xylebori_CBS S22 Raffaelea_ambrosiae_CBS Raffaelea_ellipticospora S21 Ambrosiella_ferruginea_JB13 Ophiostoma_stenoceras_CMW3202 Ophiostoma_quercus Ceratocystiopsis_minuta_bicolor_CBS Sporothrix_humicola_CMW7618 Grosmannia_piceiperda Claviceps_fusiformis_ATCC_26019 Ambrosiella_macrospora_CBS Grosmannia_penicillata Ophiostoma_montium_CBS Ceratocystis_adiposa_CBS Daldinia_concentrica Leptographium_abietinum_DAOM60343 Ophiostoma_floccosum Esteya_vermicola_CBS S32 Sporothrix_schenckii_CMW7614 Leptographium_lundbergii_UAMH9584 Raffaelea_montetyi Ophiostoma_ips Grosmannia_serpens Raffaelea_amasae_CBS Ophiostoma_abietinum Fragosphaeria_purpurea_CBS Raffaelea_canadensis_CBS Xylaria_sp. Raffaelea_sulphurea_CBS Ophiostomataceae_sp._TR25 Raffaelea_subalba_C2401 Ceratocystiopsis_manitobensis_UM237 Ambrosiella_tingens_CBS Raffaelea_brunnea_CBS Penicillium_expansum Epichloe_typhina Raffaelea_subfusca_C2335 Raffaelea_quercus-mongolicae_KACC44405 PL4 PL1 Raffaelea_tritirachium_CBS Ceratocystiopsis_minuta_CBS Ceratocystis_moniliformis_CBS Leptographium_fruticetum_DAOM Fragosphaeria_reniformis_CBS S31 Raffaelea_gnathotrichi_CBS Raffaelea_quercivora_MAFF Raffaelea_fusca_C2394 Ophiostoma_setosum Sporothrix_schenckii Grosmannia_cucullata Ophiostoma_pulvinisporum_CMW9022 Raffaelea_arxii_CBS Ophiostoma_ulmi Raffaelea_montetyi_PC Leptographium_terebrantis_UAMH9722 Ophiostoma_bicolor Microascus_cirrosus_CBS Raffaelea_lauricola_PL159 Ophiostoma_piceae Grosmannia_clavigera_ATCC18086 Raffaelea_santoroi_CBS Ambrosiella_hartigii_CBS Raffaelea_sulcati_CBS Petriella_setifera_CBS Taphrina_populina_CBS Leptographium_longiclavatum Grosmannia_abiocarpa_MUCL18351 S28 Raffaelea_albimanens_CBS PL1635 Ophiostoma_canum Raffaelea_lauricola Ceratocystis_coerulescens_CL13_12 Raffaelea_canadensis_CBS Ophiostoma_novo_ulmi_CMW10573 Raffaelea_scolytodis_CCF3572 Ambrosiella_ferruginea_CBS

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7 Known hosts in the USA Many more hosts may remain! Persea borbonia Redbay Persea palustris Swamp bay Persea humilis Silkbay Persea americana Avocado Persea indica* Cinnamomum camphora Camphortree Sassafras albidum Sassafras Umbellularia californica California bay laurel* Laurus nobilis European bay laurel Lindera benzoin Northern spicebush* Lindera melissifolia Pondberry Litsea aestivalis Pondspice Licaria triandra* Gulf licaria Ocotea coriacea* Lancewood Persea mexicana* Mexican redbay * = Artificial inoculation

8 Global Lauraceae Forests

9 Lateral transfer to additional vectors * * * * * * Transfer to at least 8 additional species complicates management and regulatory control. * Carrillo et al., 2014

10 426,988,027. and counting!!! 30% of all redbay lost 90-95% stand mortality in 3 years.

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12 4,925 to 133,740 ha in 26 months Rodgers et al., 2014

13 Photo: LeRoy Rodgers

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15 Photo: Randy Ploetz

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17 Simmonds, 5 cm + inoc pt A. Mock inoculated. External symptoms (es) = 1, internal symptoms (is) = 1 B. 3 days after inoculation (dai), es = 1, is=1 C. 7 dai, es=1, is=2 D. 14 dai, es = 2, is=3 E. 21 dai, es = 3, is = 5 F. 21 dai, es = 5, is = 6 G. 42 dai es = 8, is = 9 H. 42 dai, es = 9, is = 9 Scale bar = 0.5 cm. Arrows denote side of stem that was inoculated. Inch et al., 2012

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19 Disease symptoms co occur with tylosis

20 Genome comparisons R. lauricola, R. sp. 272 and several close relatives sequenced with both Ion Torrent and Illumina Transcriptomes RNAseq underway Cerato platanin homolog exists in RL, but not nonpathogenic species (potential effector?) Transformed isolates have been made and will be tested for function

21 Native Plants Journal, 2014

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24 FGG-L FGG-leaves - 30Jan14 XM D-limonene 36 leaves - spot A, 02-Mar :32:59 Scan EI+ TIC 1.25e8 Camphor % Eucalyptol FGG Time α- pinene FCD-L FCD-leaves - 30Jan14 36 leaves - spot B, 02-Mar :53:26 XM-005 Scan EI+ TIC 1.48e8 FCD % Time

25 Experimental protocol Fungus manipulation of plant odor (Martini and Hughes) Baseline µl of water 3 DAI 10 DAI 20 DAI CONTROL INFECTION µl of x 10 5 spores/ml Infected with fungus Volatile collection Volatile collection Volatile collection Volatile collection Behavioral assays Behavioral assays Behavioral assays Behavioral assays

26 Behavioral assays Beetle trap The olfactometer was covered with a black cover 4-way olfactometer Beetles caught in traps Leaf volatiles collected from a redbay

27 Disease progression Baseline INFECTION 3 DAI 10 DAI 20 DAI Asymptomatic First wilt symptoms Wilt/Death

28 Results Behavioral assays -1 DAI Baseline control Infected 3 DAI Asymptomatic 10 DAI First wilt symptoms 20 DAI Wilt/Death ** ** ** * % 50% 0% 50% % Percent beetles response

29 Results Volatile collection: potential repellents raf-infection proj: d3-raf1 15-sept-14 XM ul of 0.2ul/ug nonyl acetate + 0.2ug/u, 01-Oct :48: Scan EI+ TIC 1.15e7 % Control 0 XM-066 % 0 XM-068 % (E)-4,8-dimethyl-1,3,7,-nonatriene (DMNT) Methyl Salicylate inoculation Time Days after fungal inoculation 10 Days after fungal Scan EI+ TIC 2.68e8 Scan EI+ TIC 1.29e7

30 Trap Logs A. Fresh redbay bolt B. SPLAT repellent C.Sticky Card (x 2) D.Metal Pole (1 dia.) Trap Approx. 4-5 ft. tall To refresh odors, a cork-borer was used to remove 2 circular bark sections every 2 weeks

31 Potential Repellents of Xyleborus glabratus Mean # X. glabratus beetles caught X. glabratus Trap Catches Weeks after Repellent Deployment Mean Total X. glabratus per Trap Over 10 weeks A C B CD D (+) Redbay ( ) Poplar Rb + MeSA Rb + MeSA/Verb Rb + Verb 0 (+) Redbay ( ) Poplar Rb + MeSA Rb + MeSA/Verb Rb + Verb

32 Did the Repellents Stop Boring Activity? Refrigerate logs De-bark Count entrance holes No. 1 paperclip Exposed bark Under bark 140 Beetle Boring redbay poplar (Haile)/laurel oak (Itch) MeSA MeSA + Verb Verb positive Negative haile ichetucknee Positive control redbay log

33 Research questions to be addressed in near future What are the mechanisms of resistance? Genetic mechanism? How do implement long term management and carry out restoration? What is the potential host range expansion and is there tolerance in other hosts? How does pathogen kill hosts? Are other Raffaelea killers out there?

34 Research Team Thank You Tyler Dreaden, Marc Hughes, Fred Beckman, Don Spence, Keumchul Shin; Randy Ploetz, Lukasz Stelinski, Jiri Hulcr, Sharon Inch, Bud Mayfield, Jeff Eickwort, Alina Campbell, John Riggins, Steve Fraedrich, Grechen Pruett, Ben Held, Robert Blanchette, Tom Harrington, Adam Black Funding USDA (AFRI, Critical Issues and SCRI), UF/IFAS Research, USDA Forest Service, FHP, SRS/R8

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