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Sites with smoke-stimulated germination of wild species . And the list of s

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1 : Smoke Emilio Ghisalberti (UWA) Kingsley Dixon (KPBG) Robert Trengove (Murdoch) Investigation of the germination promoting compounds in smoke Gavin Flematti The University of Western Australia
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3 : Solving the bushfire germination mystery……..
4 : Smoke as a germination cue In 1990, researchers in South Africa identified smoke as the key agent for stimulating germination of a threatened fynbos species Audouinia capitata (Bruniaceae) Researchers at Kings Park and Botanic Garden (Western Australia) have since identified smoke as the key element for promoting the germination of many Australian native seeds
5 : Smoke found to improve seed germination of many Australian native species
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7 : Sites with smoke-stimulated germination of wild species And the list of species includes…. lettuce, celery, parsley and others….
8 : Apparatus for generating Plant Derived Smoke ‘Smoke-water’ ‘Aerosol Smoke’
9 : Benefits for mining restoration Lateritic nickel project Mineral sands mining restoration - BUT… need 10 tonnes smoke water per hectare!
10 : Attempts to identify the active compound(s) Baldwin et al. (1994) identified 71 compounds and tested a total of 233 Dixon et al. (1997) identified a further 43 compounds Researchers in South Africa (1995) and California (1997) have also identified compounds in smoke Importantly, burning cellulose produces the active compound(s)
11 : Lettuce (Grand Rapids) The Needle in the Haystack - finding the chemical(s) in smoke responsible for germination Conostylis aculeata Stylidium affine Bioassay - Test that uses a biological indicator for tracking the active components through separation steps
12 : Isolation of Smoke Compounds Smoke Water
13 : Isolation of Smoke Compounds Ether extract Smoke Water Ether layer Aqueous layer
14 : Isolation of Smoke Compounds Ether extract Smoke Water Ether layer Aqueous layer
15 : Isolation of Smoke Compounds NaHCO3 soluble (stronger acids - COOH) Ether extract Smoke Water Ether layer NaHCO3 layer
16 : Isolation of Smoke Compounds NaHCO3 soluble (stronger acids - COOH) Ether extract NaOH soluble (weaker acids - phenols etc.) Smoke Water Ether layer NaOH layer -Activity found in the neutral fraction-
17 : The Haystack! GC-MS chromatogram of neutral fraction
18 : Lots of Chromatography! Chromatography - Physical method of separating compounds Classical Liquid Chromatography Stationary phase Mobile phase
19 : General separation scheme Alumina Neutral Fraction 15 Fractions
20 : Typical germination of Grand Rapids lettuce when tested with C18 fractions
21 : ~ 2 years later…Only 3 Compounds! All had molecular formula C8H6O3 (mw=150) Separated and tested for activity 1 2 3 UV recorded at 320nm 25% MeOH/water
22 : Testing of the 3 compounds
23 : Identification of the 3 compounds Chemical structure proposed based on Nuclear Magnetic Resonance (NMR) and mass spectrometry (MS) data
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25 : Germination of karrikinolide (2) with Grand Rapids Lettuce
26 : Germination of karrikinolide (2) with confirming native species
27 : Back to the haystack!
28 : Expanded CDS-neutral fraction
29 : Summary First chemical identified from smoke that promotes germination Novel structure – new to science Devised a method of synthesis Bioassay data demonstrating effect with species from Australia, South Africa and North America Potential uses in agriculture, horticulture, land restoration etc… Compared with smoke water, karrikinolide operates at < 1 g per hectare
30 : What other species…? What other activity…? Is there a better (simpler) synthesis…? Are there active analogues…? What is the mode of action…? Potent Germination Promoter
31 : Mode of Action?? Label = fluorescent Questions ??
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