Fern (Non-Idiindigenous Vi Ne) Light Green Ornamental # 4 Categgyory 1 Invasive (Langeland, K.A
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Potential for Use of Native Phytopathogens as Biocontrol Agents for Invasive Plant Species K. Jayachandran, K. G. Shetty, Tainya Clarke, LeRoy Rodgers III, ShiLi Miao, and Robert Johnson 1. Theoretical basis or Rationale 2. Discovery – Survey, collection and isolation 3Elti3. Evaluation – PthPathogen iiticity testing Invasive Exotic Species Biological invasions increasingly threaten the biologgyggyical diversity and ecological integrity of natural ecosystems. Invasive species have contributed to the decline of endangered and threatened species in the United States. Non-indigenous weeds alone cost U.S. agriculture $7-27 billion per year. Global warming/climate change effect on the invasive species spread is a serious concern. Natural and agricultural ecosystems in Flor ida are be ing threat ened b y th e continuing exotic aquatic, wetland, and ttilltiiterrestrial plant invasions: Problems with utilization and management of water resources Disturb and displace the native flora and fauna itlin natural areas Negative effects on productivity of crop and pastures lands Managing invasive plants in natural areas is challenging: Economic, environmental health risks with herbicides. Certain weeds are not controlled effectively by availa ble meth od s. Discovery and development of novel weed- maaageetagetsadtecoogesaeonagement agents and technologies are of greater importance now than ever before. Problem of Invasive exotic plants in Florida There are about 1000 non-native plant species in Florida There are more than 50 exotic invasive plant species listed under the category-1 Florida's Most Invasive Species (Florida Exotic Pest Plant Council, www.fleppc.org) One of alternative weed management tools is biologgygpical control by using plant pathogens: Cause significant tissue damage, Retard plant growth, Hinder or Suppress reproduction ↓ Leading to reduction in population sizes Overall ecological impacts in natural area are reduced Classical Biological control Introduction of non -natives Insects Plant pathogens NtiNative PhtPhytopath ogens Bio herbi cid es Avoid potential hazards of introduced biocontrol agggents from other regions Locally available and locally adapted Examples of biocontrol projects utilizing pathogens a lread y present i n H awaii are 1. Kahili ginger (Hedychium gardnerianum) host-specific strain of the common plant pathogenic bacterium Pseudomonas solanacearum (Anderson and Gardner, 1996, 1997). 2. The presence of Botryitis cinerea attacking fruit of Myrica faya. 3. Controlling effect of a combination of a rust fungus and an eriophyid mite for introduced fuschia in Hawaii Volcanoes National Park ((,)Gardner, 1987). 4. The attack of ivy gourd (Coccinia grandis), an aggressive introduced vine invading lower -elevation forests, by a powdery mildew fungus, Oidium sp., (Gardner, 1994) It may be possible to use native phytopathogens for biocontrol of Schinus aadnd Lygodiu m, but it will be necessary to first acquire more basic knowledge about the pathosystem before this idea can be developed further. Brazilian Pepper (Schinus terebinthifolius Raddi) Family - Anacardiaceae Everg ree n inv asiv e ex oti c h ar dw ood tr ee species Native to Brazil, Argentina and Paraguay Brazilian pepper is the most widespread exotic plant in the state – occupying more than 700,,(,, 000 acres (Ferriter, SFWD, unpublished). Dense grow th o f Braz ilian P epper t ree EtExtens ive leaf spot and c hloros is of leaves Extensive foliar disease lesions Comm onl y en coun ter ed f un gal i sol ates from infected Brazilian pepper leaf Endophytes Pestaloptiopsis sp. Phoma sp. Alternaria sp. Exserohilum sp. Cylindrocladium sp. Neofusicoccum sp Inoculation Studies In greenhouse and in field In the greeenhouse by circular surface incision (< 1mm depth) at the base of sapling with sterile Sharp scalpel blade. Mycelial plug was placed on the incision and sealed with parafilm. In the field (FIU preserve), similar procedure but the incision was made on a small terminal branch 1 – 2 ft below the terminal bud. Wilting o f Bra zilia n peppe r sap lin gs fo llo wing BPS PF-1 inoculat io n Wilting and defoliation of BPSPF-1 inoculated Brazilian ppppepper branch What is Lygodium microphyllum? F(Fern (non-idiindigenous v ine) Light green ornamental # 4 Categgyory 1 invasive (Langeland, K.A. 1998; Van Driesche et al 2002) Why is Lygodium a problem? Blocks light from the understory plants Forms thick mats on ground Does not degrade easily Acts as a fire ladde r Jonathan Dickinson State Park Diseases with severe symptoms covering large areas of Lygodium BtbtktBrown spots, brown streak spots, chlorosis, vein necrosis, leaf curling, blight and defoliation Leaf spots with grey center on fertile frond Fertile frond showing brown – dark brown necrotic patches with chlorosis Among the 78 fungal isolates tested 12 iltisolates causedtd extens ive necrosi ifs of detached leaflets and 9 of the isolates were able cause lflttileaflet tissue necrosi s in less than 72 hours after inoculation. Detached leaf assay Control leaflets Tissue necrosis of leaflets fllfollow ing i nocul ltiation with C-7 After completion of the first greenhouse experiment, based on the results 12 isolates were selected for the liquid based inoculation study Characteristics of the fungal isolates selected for inoculation studies Fungal isolate Hyphae/reproductive Host/Source ID structures C-2 Septate, microsclerotia L. microphyllum/Field Unidentified C-7 Sepp,tate, microsclerotia L. microppyhyllum/Field Unidentified C-15 Septate, melanised L. microphyllum/Field Unidentified C-22 Septate, conidia L. microphyllum/Field Alternaria sp. D-18 Septate L. microphyllum/Field Unidentified J-5 Septate, mealanised L. microphyllum/Field Unidentified J-20 Septate, conidia L. microphyllum/Field Curvularia sp. J-33 Septate L. microphyllum/Field Unidentified J-34 Septate LiL. microphyll hllum/Field UidUnident ifidified GL3 Septate, conidia L. microphyllum/Greenhouse Alternaria sp. N-1 Septate, conidia Decaying L. microphyllum plant Cylindrocladium sp. debris F-1 Septate, melanised Brazilian pepper seed. Unidentified Effect of native fungal isolates on disease incidence (%) on L. microphyllum 100.00 90.00 80.00 70.00 nce (%) nce 60.00 ee 50.00 40.00 ease incid ss 30. 00 Di 20.00 10.00 0.00 C2 C7 C15 C22 GL3 D18 J5 J20 J33 N1 F1 Fungal isolates Effect of native fungal isolates on disease severity in L. microphyllum 4.00 3.50 3.00 2.50 erity vv 2.00 1.50 Disease se 1.00 0.50 0000.00 C2 C7 C15 C22 GL3 D18 J5 J20 J33 N1 F1 Fungal isolates Die-back, leaflet lesions, leaf blight and defoliation on N-1 inoculated plant Leaf blight, defoliation and death of F-1 inoculated plant Host-range study Anemia adiantifolia (Pine fern) Nephrolepsis exaltata (Boston fern) Polypodium polypodioides (Resurrection fern) Adiantum tenerum (Mainden hair fern) Pteris bahamensis (Bahama ladder brake) Thelypteris cf. kunthii (Southern shield fern) Pathogenicity of selected fungal isolates on native ferns (% disease incidence) Fungal isolates Native fern species Number of control C7 J5 F1 N1 plants/treat. Anemia adiantifolia 300000 Adiantum tenerum 200000 Polypodium p olyp odioides 2004000 Nephrolepsis exaltata 200000 Thelypteris cf. kunthii 300000 Pteris bahamensis 300000 Scale insect infestation and suppression of Lygodium Initial scale population adapted to the host through selection? Over the period the scale insect infestation became more severe. Infested plants developed severe disease symptoms and died. Native insect also can adapt to an exotic invasive plant species and suppress its growth There are reports of scale insects being used as biological control agents in the invasive plant management. Report of facilitation by scale insect of deeper fungal infection of host plant . Dead Lygodium plant rhizome, wax covered scale insect on the basal portion of stipe and on crozier Close-up of scale infestation Chlorosis, necrotic lesions on leaflet and on rachis due to scales Necrosis of rachis and gradual blighting of leaflets due to scales Conclusion Successful in documenting natural occurrence of diseases symptoms on Schinus and Lygodium in the South Florida natural areas and isolating native pathogens Native phytopathogens can be highly virulent and are capable of killing Schinus and Lygodium Native phytopathogens can do extensive damage to the vegetative (leaflet) as well as the reproductive tissues (fertile frond) of Lygodium Acknowledgements South Florida Water Management District The Everglades National Park HID Program Jennifer Possley Jose Pacheco.