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Microbes Associated with Hylobius abietis: A Chemical and Behavioral Study
KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis is based on three inter-related studies: the first part deals with the microbial consortium, the identification of microbes and their volatiles, the second part deals with the study of bio-chemical control methods of two conifer pests; the pine weevil Hylobius abietis (L.) and the root rot fungi Heterobasidion spp., and the third part describes the production of styrene by a fungus using forest waste.The large pine weevil (Hylobius abietis L.) is an economically important pest insect of conifers in reforestation areas of Europe and Asia. The female weevils protect their eggs from feeding conspecifics by adding frass (mixture of weevil feces and chewed bark) along with the eggs. In order to understand the mechanism behind frass deposition at the egg laying site and to find repellents/antifeedants for pine weevils, microbes were isolated from the aseptically collected pine weevil frass. Microbial produced volatile organic compounds (VOCs) were collected by solid phase micro extraction and analyzed by GC-MS after cultivating them on weevil frass broth. The major VOCs were tested against pine weevils using a multi-choice olfactometer. Ewingella sp., Mucor racemosus, Penicillium solitum, P. expansum, Ophiostoma piceae, O. pluriannulatum, Debaryomyces hansenii and Candida sequanensis were identified as abundant microbes. Styrene, 6-protoilludene, 1-octene-3-ol, 3-methylanisole, methyl salicylate, 2-methoxyphenol and 2-methoxy-4-vinylphenol were the VOCs of persistently isolated microbes. In behavioral bioassay, methyl salicylate, 3-methylanisole and styrene significantly reduced the attraction of pine weevils to their host plant volatiles. Heterobasidion spp. are severe pathogenic fungi of conifers that cause root and butt rot in plants. Bacterial isolates were tested for the antagonistic activity against fungi on potato dextrose agar. Bacillus subtilis strains significantly inhibited the growth of H. annosum and H. parviporum. Styrene is an industrial chemical used for making polymeric products, currently produced from fossil fuel. A strain of Penicillium expansum isolated from pine weevil frass was investigated for the production of styrene using forest waste. Grated pine stem bark and mature oak bark supplemented with yeast extract produced greater amounts of styrene compared to potato dextrose broth.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2013. , 66 p.
Series
Trita-CHE-Report, ISSN 1654-1081 ; 2013:23
Keyword [en]
Hylobius, Ewingella, Penicillium, Heterobasidion, Bacteria, Fungi, Bark, Forest waste, Metabolites, Styrene, Methyl salicylate.
National Category
Natural Sciences Organic Chemistry
Identifiers
URN: urn:nbn:se:kth:diva-121887ISBN: 978-91-7501-727-3 (print)OAI: oai:DiVA.org:kth-121887DiVA: diva2:619700
Public defence
2013-05-30, F3, Lindstedtsvägen 26, KTH, Stockholm, 10:00 (English)
Opponent
Supervisors
Note

QC 20130507

Available from: 2013-05-07 Created: 2013-05-06 Last updated: 2013-05-07Bibliographically approved
List of papers
1. Chemo- and biodiversity of microbes associated with pine weevil (Hylobius abietis)
Open this publication in new window or tab >>Chemo- and biodiversity of microbes associated with pine weevil (Hylobius abietis)
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(English)Manuscript (preprint) (Other academic)
National Category
Chemical Sciences
Identifiers
urn:nbn:se:kth:diva-121988 (URN)
Note

QS 2013

Available from: 2013-05-07 Created: 2013-05-07 Last updated: 2013-05-07Bibliographically approved
2. Fungal metabolite mask the host plant odor of the pine weevil (Hylobius abietis)
Open this publication in new window or tab >>Fungal metabolite mask the host plant odor of the pine weevil (Hylobius abietis)
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(English)Article in journal (Other academic) Submitted
National Category
Chemical Sciences
Identifiers
urn:nbn:se:kth:diva-121992 (URN)
Note

QS 2013

Available from: 2013-05-07 Created: 2013-05-07 Last updated: 2013-05-07Bibliographically approved
3. Penicillium expansum Volatiles Reduce Pine Weevil Attraction to Host Plants
Open this publication in new window or tab >>Penicillium expansum Volatiles Reduce Pine Weevil Attraction to Host Plants
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2013 (English)In: Journal of Chemical Ecology, ISSN 0098-0331, E-ISSN 1573-1561, Vol. 39, no 1, 120-128 p.Article in journal (Refereed) Published
Abstract [en]

The pine weevil Hylobius abietis (L.) is a severe pest of conifer seedlings in reforested areas of Europe and Asia. To identify minimally toxic and ecologically sustainable compounds for protecting newly planted seedlings, we evaluated the volatile metabolites produced by microbes isolated from H. abietis feces and frass. Female weevils deposit feces and chew bark at oviposition sites, presumably thus protecting eggs from feeding conspecifics. We hypothesize that microbes present in feces/frass are responsible for producing compounds that deter weevils. Here, we describe the isolation of a fungus from feces and frass of H. abietis and the biological activity of its volatile metabolites. The fungus was identified by morphological and molecular methods as Penicillium expansum Link ex. Thom. It was cultured on sterilized H. abietis frass medium in glass flasks, and volatiles were collected by SPME and analyzed by GC-MS. The major volatiles of the fungus were styrene and 3-methylanisole. The nutrient conditions for maximum production of styrene and 3-methylanisole were examined. Large quantities of styrene were produced when the fungus was cultured on grated pine bark with yeast extract. In a multi-choice arena test, styrene significantly reduced male and female pine weevils' attraction to cut pieces of Scots pine twigs, whereas 3-methylanisole only reduced male weevil attraction to pine twigs. These studies suggest that metabolites produced by microbes may be useful as compounds for controlling insects, and could serve as sustainable alternatives to synthetic insecticides.

Keyword
Penicillium expansum, Hylobius, Styrene, 3-Methylanisole, Frass, Feces, Fungal volatiles, "Green" chemistry
National Category
Biochemistry and Molecular Biology Ecology
Identifiers
urn:nbn:se:kth:diva-119064 (URN)10.1007/s10886-012-0232-5 (DOI)000314363000012 ()2-s2.0-84873525306 (Scopus ID)
Funder
Formas
Note

QC 20130306

Available from: 2013-03-06 Created: 2013-03-05 Last updated: 2017-12-06Bibliographically approved
4. Antagonistic activity of Bacillus subtilis A18 – A19 against Heterobasidion species
Open this publication in new window or tab >>Antagonistic activity of Bacillus subtilis A18 – A19 against Heterobasidion species
(English)Article in journal (Other academic) Submitted
National Category
Chemical Sciences
Identifiers
urn:nbn:se:kth:diva-121993 (URN)
Note

QS 2013

Available from: 2013-05-07 Created: 2013-05-07 Last updated: 2013-05-07Bibliographically approved
5. Sustainable bio-production of styrene from forest waste
Open this publication in new window or tab >>Sustainable bio-production of styrene from forest waste
2013 (English)In: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 144, 684-688 p.Article in journal (Refereed) Published
Abstract [en]

A strain of Penicillium expansum was studied for the production of styrene using forest waste biomass as a feeding substrate. The fungal strain was cultivated on bark of various trees supplemented with yeast extract and the volatiles produced were collected on Tenax TA and analyzed by gas chromatography-mass spectrometry. Fungus cultured on grated soft bark of pine (Pinus sylvestris) stems (GPB) and mature bark of oak (Quercus robur) supplemented with yeast extract produced relatively the highest amounts of styrene. The maximum styrene production rate was 52.5 mu g/h, 41 mu g/h and 27 mu g/h from fungus cultivated on 50 mL liquid media with 10 g GPB or mature bark of oak and potato dextrose broth respectively. These promising results suggest that the fungal strain could be used to produce "green" styrene plastics using renewable forest waste biomass.

Keyword
Forest biomass, Penicillium expansum, Styrene, Bark, Fungal volatiles
National Category
Other Chemistry Topics Energy Engineering
Identifiers
urn:nbn:se:kth:diva-121994 (URN)10.1016/j.biortech.2013.07.033 (DOI)000324280100093 ()2-s2.0-84881558867 (Scopus ID)
Funder
Formas
Note

QC 20131017. Updated from submitted to published.

Available from: 2013-05-07 Created: 2013-05-07 Last updated: 2017-12-06Bibliographically approved

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