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Prospects for biological control of serrated tussock and Chilean needle grass

Prospects for biological control of serrated tussock and Chilean needle grass. Seona Casonato 1,3,4 , Freda Anderson 5 , Ann Lawrie 1 and David McLaren 2,3 1 Biotechnology & Environmental Biology, RMIT, Bundoora, Vic. 3083 2 Department of Primary Industries, Frankston Centre

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Prospects for biological control of serrated tussock and Chilean needle grass

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  1. Prospects for biological control of serrated tussockand Chilean needle grass Seona Casonato1,3,4, Freda Anderson5, Ann Lawrie1 and David McLaren2,3 1Biotechnology & Environmental Biology, RMIT, Bundoora, Vic. 3083 2Department of Primary Industries, Frankston Centre 3CRC for Australian Weed Management 4Current address: HortResearch, Auckland, NZ 5CERZOS-UNS, Bahía Blanca, Argentina TT biocontrol talk

  2. Prospects for biological control of serrated tussockand Chilean needle grass 1. Biological control – Argentina 1.1 Chilean needle grass (Nassella neesiana) 1.2 Serrated tussock (Nassella trichotoma) 2. Biological control – endemic 2.1 Serrated tussock (Nassella trichotoma) TT biocontrol talk

  3. Chilean needle grass http://www.weeds.crc.org.au/main/wom_chilean_needle_grass_0905.html# http://www.humeshire.nsw.gov.au/files/1733/File/CNGWeed2.jpg TT biocontrol talk

  4. Chilean needle grass http://www.weeds.org.au/cgi-bin/weedident.cgi?tpl=plant.tpl&state=&s=&ibra=all&card=G12 TT biocontrol talk

  5. Serrated tussock (Nassella trichotoma) NRE 1998 TT biocontrol talk

  6. Seed spread – inflorescences break off and blow away http://www.weeds.crc.org.au/documents/wmg_serrated_tussock.pdf 140,000 seeds plant-1 yr-1 TT biocontrol talk

  7. Prospects for biological control of serrated tussockand Chilean needle grass 1. Biological control – Argentina 1.1 Chilean needle grass (Nassella neesiana) 1.2 Serrated tussock (Nassella trichotoma) 2. Biological control – endemic 2.1 Serrated tussock (Nassella trichotoma) TT biocontrol talk

  8. 1. BIOLOGICAL CONTROL - ARGENTINA • Nassella spp. native to Peru, Chile, Uruguay and Argentina (Healy, 1945; Campbell, 1982) • Epicentre Argentina • Look for classical biological control organisms in these regions • Import to Australia • Distribute and leave to establish • Natural spread and control of Nassella spp. TT biocontrol talk

  9. Pathogens collected from Nassella spp. in Argentina during 1988-99 TT biocontrol talk Briese and Evans, 1998; Briese et al., 1999)

  10. search for biological control pathogens in Argentina • Begun in 1998 through CERZOS-UNS • Funded by • NSW Agriculture • RIRDC • Consortium of local government and community groups in SE Australia • Serrated Tussock Working Party in NSW • Serrated Tussock Taskforce in Victoria • Latterly WONS TT biocontrol talk

  11. Areas surveyed for pathogens in Argentina Plant pathologist in Argentina: Dr Freda Anderson, CERZOS-UNS Australian scientist in Argentina: Dr Bill Pettit, CSIRO http://www.rirdc.gov.au/reports/Ras/01-27.pdf TT biocontrol talk

  12. Microorganisms collected from grasses of the genus Nassella in Argentina in 1999 TT biocontrol talk http://www.rirdc.gov.au/reports/Ras/01-27.pdf

  13. Microorganisms collected from grasses of the genus Nassella in Argentina in 1999 (cont.) TT biocontrol talk http://www.rirdc.gov.au/reports/Ras/01-27.pdf; Anderson 2004a

  14. heteroecious species – alternate host RUST LIFE CYCLES • Rust needs to be autoecious (have all life stages on host plant) I aecia macrocyclic e.g. P. graminis 0 pycnia demicyclic e.g. P. lagenophora microcyclic e.g. P. heterospora IV II basidia uredinia leptoforms e.g. P. malvacearum III telia TT biocontrol talk

  15. Rust on ST and CNG • ST • Puccinia nassellae abundant at several sites • CNG • 3 rust species associated with severe leaf damage • Sphaerellopsis filum (mycoparasite) • frequently associated with P. nassellae pustules • ‘significant controlling effect on rust populations’ TT biocontrol talk Briese et al. (2001); Anderson (2004a)

  16. Sphaerellopsis (Darluca) filum TT biocontrol talk http://www.niaes.affrc.go.jp/inventry/microorg/eng/z65e-Sphaer-fil.html

  17. Prospects for biological control of serrated tussockand Chilean needle grass 1. Biological control – Argentina 1.1 Chilean needle grass (Nassella neesiana) 1.2 Serrated tussock (Nassella trichotoma) 2. Biological control – endemic 2.1 Serrated tussock (Nassella trichotoma) TT biocontrol talk

  18. CNG - rust fungi in Argentina • 3 spp. of rust fungi found • P. nassellae most widespread and damaging • Uredinia and telia in field (aecia on alternate host?) • P. graminella at only one site • Telia, pycnia and aecia in field (no urediniospores) • P. aff. avocensis no good • x heteroecious (aecia on Malvaceae) • Uromyces pencanus very damaging • Uredinia and telia in field (aecia on alternate host?) TT biocontrol talk Briese et al. (2001)

  19. Uredinia on N. neesiana (site NT 27) Telia on N. neesiana (site NT 16) Puccinia nassellae - CNG TT biocontrol talk Anderson (2005a)

  20. Germinating basidiospores of Puccinia nassellae Puccinia nassellae - CNG http://www.rirdc.gov.au/reports/Ras/01-27.pdf TT biocontrol talk Anderson (2005a)

  21. P. nassellae - CNG • Infection of glasshouse plants with urediniospores • No infection with teliospores • Problems with germinating teliospores (cold treatment needed) • No cross-infection with ST (different) • Possibly heteroecious? • Aecia ± pycnia on plants close to CNG in field • Clematis montevidensis (P. recondita?) • Solidago chilensis (no pycnia)(P. stipae?) • Cestrum parquii (immature, only one site) • Verbena sp. (P. verbeniphila?) TT biocontrol talk Anderson (2004a)

  22. P. nassellae - CNG • Plants of all 7 Australian provenances inoculated with urediniospores • All 7 infected with isolate Pn27 • None infected with isolate Pn52 • Host-strain specificity • Subspecies known in CNG TT biocontrol talk Anderson (2004a)

  23. Puccinia graminella - CNG • Autoecious • Aecia and telia always present in field • No urediniospores known from life cycle • Germination of freshly collected • Aeciospores • Teliospores • Glasshouse infection trials under way TT biocontrol talk Anderson (2005a); Anderson (2005c)

  24. P. graminella. Aecia on N. neesiana P. graminella. Aecia and telia on N. neesiana Puccinia graminella - CNG TT biocontrol talk Anderson (2005a)

  25. Puccinia graminella - CNG Aecia on Chilean needle grass at site NN 8 Germinating teliospores with basidiospores TT biocontrol talk Anderson (2005c)

  26. Uromyces pencanus - CNG • Trap plants of Australian CNG heavily infected • known on 2 accessions of N. neesiana in Argentina (Lindquist, 1982) • Autoecious • No paraphyses in uredinia • No germination in teliospores • Infection of glasshouse plants with urediniospores • No aecia yet in field or glasshouse • on ‘Stipa setigera’ in Chile by Holway, 1919-1920 (Arthur, 1925) • Aecial description matches P. graminella – mistaken identity? TT biocontrol talk Anderson (2004); Anderson (2005)

  27. Telia on Nassella neesiana (site NN3) Uromyces pencanus - CNG Uredinia on inoculated plant Uredinia on inoculated plant (site NN3) TT biocontrol talk Anderson (2005a)

  28. Uromyces pencanus - CNG Appresorium over stoma Germinating urediniospore on leaf TT biocontrol talk Anderson (2005c)

  29. Uromyces pencanus - CNG • Isolates (urediniospores) tested on: • 7 provenances from Australia • 2 provenances from NZ • Australian 6/7 susceptible (not Ballarat) • Neither from NZ infected • 2 Austrostipa spp. • Neither infected • ST (ACT) • No infection TT biocontrol talk Anderson (2005a)

  30. Pathogenicity and virulence of Argentinian fungi on CNG - promising • Puccinia nassellae (rust) • Autoecious? (teliospore infection not shown) • Much obvious damage in field • urediospores infect but host-strain specificity • Puccinia graminella • Autoecious, demicyclic • Much obvious damage in field • plants currently being tested • Uromyces pencanus • Autoecious? (no teliospore infection yet) • Much obvious damage in field • urediospores infect but host-strain specificity TT biocontrol talk

  31. Prospects for biological control of serrated tussockand Chilean needle grass 1. Biological control – Argentina 1.1 Chilean needle grass (Nassella neesiana) 1.2 Serrated tussock (Nassella trichotoma) 2. Biological control – endemic 2.1 Serrated tussock (Nassella trichotoma) TT biocontrol talk

  32. Urediniospores on site http://www.rirdc.gov.au/reports/Ras/01-27.pdf Puccinia nassellae - ST • Only uredinia and telia in literature (Cummins, 1971; Lindquist, 1982) • Only uredospores found at sites • Trap plants transplanted into infected sites • Most plants infected during autumn, not winter • Uredinia do not burst open easily • Urediniospores collected by suction • Used to infect ST and CNG in glasshouse TT biocontrol talk Briese et al. (2001)

  33. Puccinia nassellae (from ST) specificity • Inoculation trials • ST vs CNG • ST - uredinia on all inoculated Argentinian plants • CNG - no cross-infection Argentinian ST • Rust strains species-specific • Argentinian vs Australian ST • Argentinian ST – uredinia • Australian ST – not infected • Rust site-specific TT biocontrol talk Anderson (2004a); Anderson (2005b)

  34. Ustilago sp. • Ustilago sp. on one population of ST • Totally prevented seed production • Important effect on plant populations? TT biocontrol talk Briese et al. (2001)

  35. http://www.rirdc.gov.au/reports/Ras/01-27.pdf Ustilago sp. - ST • Might not be U. hypodytes • Glasshouse experiments • Inoculated germinating seeds • No infected plant produced seed (controls did) • Little fusion of hyphae in germinating spores TT biocontrol talk Briese et al. (2001)

  36. Corticium • Corticium ubiquitous on ST and CNG etc. • Associated with tussock decline • Heavy sporulation (mats) observed at crown on dead and dying culms (pull out easily) • Isolated on to PDA – hyphae only • Transplant experiment set up on site – ST • Under way TT biocontrol talk Briese et al. (2001)

  37. Pathogenicity and virulence of Argentinian fungi on ST – much less promising • Puccinia nassellae (rust) • Some obvious damage in field • Problems infecting plants in glasshouse • Ustilago sp. • As for rust • Corticium sp. • Dieback in field, plants in field pull out easily • Roots covered with white layer of fungus • Could not grow easily in media or in glasshouse • Only hyphae on PDA (may not be infective) TT biocontrol talk

  38. Prospects for biological control of serrated tussockand Chilean needle grass 1. Biological control – Argentina 1.1 Chilean needle grass (Nassella neesiana) 1.2 Serrated tussock (Nassella trichotoma) 2. Biological control – endemic 2.1 Serrated tussock (Nassella trichotoma) TT biocontrol talk

  39. Search for local pathogens for inundative biocontrol of ST • Plants of serrated tussock surveyed within 50 km radius of Melbourne, Victoria during 1996-7 and 1998-9 • 6 fungi found • Arthrinium sp.(seed epiphyte) • Ascochyta sp. (leaf spot) • Dinemasporium sp. (leaf spot) • Fusariumoxysporum (crown rot) • Pleospora sp.(leaf spot) • Zinzipegasa argentinensis (leaf spot)* • *new record for Australia • Previously recorded only on one ‘Stipa sp.’ in Argentina Hussaini, I.P., Lawrie, A.C. and McLaren, D.A. (1998). Fungi in Victoria with biological control potential for Nassella trichotoma (serrated tussock). Plant Protection Quarterly 13, 99-101. TT biocontrol talk

  40. Fruiting bodies on leaf (incurved hairs) Fruiting body inside stem and wall detail Zinzipegasa argentinensis Conidia (spores) with tails Hussaini, I.P., Lawrie, A.C. and McLaren, D.A. (1998). Fungi in Victoria with biological control potential for Nassella trichotoma (serrated tussock). Plant Protection Quarterly 13, 99-101. TT biocontrol talk

  41. Effects on seedlings of ST Germination (n=20) of Nassella trichotoma seeds inoculated with fungi at 3x106 CFU. Data are meanSE. Hussaini, I.P., Lawrie, A.C. and McLaren, D.A. (2000). Pathogens on and variation in Nassella trichtoma (Poales: Poaceae) in Australia. In: Proceedings of the X International Symposium on Biological Control of Weeds 4-14 July 1999 (ed N. R. Spencer), pp.269-280. (MontanaState University, Bozeman). TT biocontrol talk

  42. Effectiveness of endemic fungi on ST • Host specificity and pathogenicity tested on mature plants for 3 spp.: • Arthrinium sp. • Dinemasporium sp. • Fusarium oxysporum • Pot trial in glasshouse • 5-6-month-old serrated tussock • 30 other grass species incl. Austrostipa Casonato, S.G. (2003). An investigation into the biology and natural enemies of serrated tussock in Australia. Ph.D. thesis, RMIT. TT biocontrol talk

  43. Effectiveness of endemic fungi on ST • Dinemasporium sp. most effective/specific • fruiting bodies, but little necrosis around them • leaf blades died from the tip if fruiting bodies on the same blade • no plant died Casonato, S.G. (2003). An investigation into the biology and natural enemies of serrated tussock in Australia. Ph.D. thesis, RMIT. TT biocontrol talk

  44. Specificity of endemic fungi on ST • Arthrinium sp. also infected: • 2/47 plants of other species • 1/2 Dichelachne crinita, 1/3 Poa poiformis • none died • Dinemasporium sp. also infected: • 2/53 plants ofother species • 1/2 A. exilis, 1/3 A. mollis • none died • Fusarium oxysporum also infected: • 26/29 plants from 16 Austrostipa spp. • 5/21 plants from 11 other native grass species • 5 Austrostipa species had ≥50% plants die (too wet?) Casonato, S.G. (2003). An investigation into the biology and natural enemies of serrated tussock in Australia. Ph.D. thesis, RMIT. TT biocontrol talk

  45. Effectiveness and specificity of endemic fungi on ST • fungi not host-specific • 2-18 other species infected too • Dinemasporium best for specificity • did not kill serrated tussock plants • But 6 plants of 4 Austrostipa species died with Fusarium oxysporum (not forma specialis) • unlikely that any could be used ‘as is’ for inundative biological control • Mutate (irradiate spores with UV)? Casonato, S.G. (2003). An investigation into the biology and natural enemies of serrated tussock in Australia. Ph.D. thesis, RMIT. TT biocontrol talk

  46. Seed burial trial - ST Effects of main factors and significant interactions on proportion of Nassella trichotoma seed infected after recovery from burial at 6 sites for 3 burial durations. • Hypothesis: • Serrated tussock infestation not uniform in area • Some areas not/less infested than others • Possible soil microbe action on seeds? • Bury seeds - see if infected differently at different sites • Infection did vary with site, duration and provenance Casonato, S.G. (2003). An investigation into the biology and natural enemies of serrated tussock in Australia. Ph.D. thesis, RMIT. TT biocontrol talk

  47. Infection (%) of retained serrated tussockseed buried in soil for 7 months by the major types of microorganisms. TT biocontrol talk Casonato, S.G. (2003). An investigation into the biology and natural enemies of serrated tussock in Australia. Ph.D. thesis, RMIT.

  48. Germination (%) of serrated tussockseed in the presence of five fungi used as the inoculum (sed for transformed data=5.08) • Possible use in biological control? Casonato, S.G. (2003). An investigation into the biology and natural enemies of serrated tussock in Australia. Ph.D. thesis, RMIT. TT biocontrol talk

  49. Search for more biological control organisms for ST • looked in areas with established serrated tussock • Bacchus Marsh • Class 5 infestation TT biocontrol talk

  50. Property near Bacchus Marsh Class 5 tussock TT biocontrol talk

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