Influence of Fungal Infection and Wounding on Contents and Enantiomeric Compositions of Monoterpenes in Phloem of Pinus sylvestris

To identify chemical resistant markers induced by fungal or mechanical injury, young trees of Scots pine (Pinus sylvestris) were subjected to inoculations of blue stain fungi associated with the pine shoot beetles Tomicus piniperda and T. minor. Among the 20 trees selected for chemical analyses, 16 were divided into four groups: one as control and three were pretreated by wounding only, or by inoculation with either the blue stain fungus Leptographium wingfieldii or Ophiostoma canum. Four wk after pretreatment, all 16 pretreated trees were mass-inoculated with L. wingfieldii. The absolute and relative amounts, as well as the enantiomeric compositions of monoterpene hydrocarbons in the phloem, were determined via a small sample of the phloem before and after the pretreatment and mass inoculation, by using two-dimensional gas chromatography (2D GC) and GC-mass spectrometry (MS). After mass inoculation, the absolute amounts of most of the monoterpenes decreased in the phloem sampled >20 cm from the fungal infection, and were higher in the phloem sampled within the infected reaction zone. The relative amounts of both (−)-β-pinene and (−)-limonene increased in phloem samples taken >20 cm above the fungal inoculation in the preinoculated trees compared with phloem sampled from the remaining four control trees. The enantiomeric compositions of β-pinene and limonene changed, after fungal growth, at defined distances from the inoculation site: the proportion of the (−)-enantiomers was highest in the phloem sampled >20 cm from the fungal inoculation. Four wk after pretreatment, monoterpene production in the phloem at the site of inoculation was more enhanced by L. wingfieldii than by O. canum. However, the different virulence levels of the fungi did not affect the enantiomeric composition of the monoterpenes. The biosynthesis of monoterpene enantiomers is discussed in relation to induced pathogen resistance.

[1]  F. Lieutier,et al.  Champignons phytopathogènes associés à deux coléoptères scolytidae du pin sylvestre (Pinus sylvestris L.) et étude préliminaire de leur agressivité envers l'hôte , 1989 .

[2]  F. Lieutier,et al.  Monoterpene composition of the preformed and induced resins of Scots pine, and their effect on bark beetles and associated fungi. , 1990 .

[3]  V. Kitunen,et al.  Induced responses in stilbenes and terpenes in fertilized Norway spruce after inoculation with blue-stain fungus, Ceratocystis polonica , 2001, Trees.

[4]  K. Raffa,et al.  Interaction of pre-attack and induced monoterpene concentrations in host conifer defense against bark beetle-fungal complexes , 1995, Oecologia.

[5]  A. Bakke,et al.  Semiochemicals in host selection and colonization of pine trees by the pine shoot beetleTomicus piniperda , 2005, Naturwissenschaften.

[6]  Phillips,et al.  Resin-based defenses in conifers. , 1999, Trends in plant science.

[7]  J. Bohlmann,et al.  Insect-Induced Conifer Defense. White Pine Weevil and Methyl Jasmonate Induce Traumatic Resinosis, de Novo Formed Volatile Emissions, and Accumulation of Terpenoid Synthase and Putative Octadecanoid Pathway Transcripts in Sitka Spruce1[w] , 2005, Plant Physiology.

[8]  E. Christiansen,et al.  Phloem parenchyma cells are involved in local and distant defense responses to fungal inoculation or bark-beetle attack in Norway spruce (Pinaceae). , 2000, American journal of botany.

[9]  A. Borg-Karlson,et al.  Enantiomeric composition of monoterpene hydrocarbons in different tissues of Norway spruce, Picea abies (L.) Karst. a multi-dimensional gas chromatography study , 1993 .

[10]  H. Solheim,et al.  Effects of growth and virulence of associated blue‐stain fungi on host colonization behaviour of the pine shoot beetles Tomicus minor and T. piniperda , 2001 .

[11]  B. Ellis,et al.  Induction of phenylalanine ammonia-lyase activity by tryptophan in Ustilago maydis. , 2001, Phytochemistry.

[12]  T. Reglinski,et al.  Induction of phenylalanine ammonia lyase activity and control of Sphaeropsis sapinea infection in Pinus radiata by 5‐chlorosalicylic acid , 1998 .

[13]  B. Långström.  Life cycles and shoot-feeding of the pine shoot beetles , 1983 .

[14]  T. Norin,et al.  Enantiomeric compositions of monoterpene hydrocarbons in the wood of healthy and top-cut Pinus sylvestris , 1992 .

[15]  E. Christiansen,et al.  Artificial inoculation with Ips typographus-associated blue-stain fungi can kill healthy Norway spruce trees [Ceratocystis polonica, Ceratocystis penicillata, water stress] , 1983 .

[16]  R. Croteau,et al.  Monoterpene synthases of Pinus contorta and related conifers. A new class of terpenoid cyclase. , 1994, The Journal of biological chemistry.

[17]  G. Nordlander.  Host finding in the pine weevil Hylobius abietis: effects of conifer volatiles and added limonene , 1991 .

[18]  K. Sturgeon.  MONOTERPENE VARIATION IN PONDEROSA PINE XYLEM RESIN RELATED TO WESTERN PINE BEETLE PREDATION , 1979, Evolution; international journal of organic evolution.

[19]  A. Borg-Karlson,et al.  Enantiomeric composition of six chiral monoterpene hydrocarbons in different tissues of Picea abies , 1993 .

[20]  A. Berryman,et al.  Mechanical injury and fungal infection induce acquired resistance in Norway spruce. , 1999, Tree physiology.

[21]  H. Francke-Grosmann.  CHAPTER 3 – Ectosymbiosis in Wood-Inhabiting Insects , 1967 .

[22]  A. Berryman,et al.  Effects of Grand Fir Monoterpenes on the Fir Engraver, Scolytus ventralis (Coleoptera: Scolytidae), and its Symbiotic Fungus , 1985 .

[23]  A. Borg-Karlson,et al.  Host Selection in Tomicus piniperda L.: Composition of Monoterpene Hydrocarbons in Relation to Attack Frequency in the Shoot Feeding Phase , 2006, Zeitschrift fur Naturforschung. C, Journal of biosciences.

[24]  Diane M. Martin,et al.  A novel sex-specific and inducible monoterpene synthase activity associated with a pine bark beetle, the pine engraver, Ips pini , 2003, Naturwissenschaften.

[25]  A. Berryman.  Resistance of Conifers to Invasion by Bark Beetle-Fungus Associations , 1972 .

[26]  A. Borg-Karlson,et al.  Relative amounts and enantiomeric compositions of monoterpene hydrocarbons in xylem and needles of Picea abies , 1996 .

[27]  B. Långström,et al.  Shoot damage and growth losses following three years of Tomicus-attacks in Scots pine stands close to a timber storage site. , 1991 .

[28]  Krokene,et al.  Induced resistance to pathogenic fungi in norway spruce , 1999, Plant physiology.

[29]  B. Långström,et al.  Induced and spontaneous attacks by pine shoot beetles on young Scots pine trees: tree mortality and beetle performance , 1993 .

[30]  Diane M. Martin,et al.  Functional Characterization of Nine Norway Spruce TPS Genes and Evolution of Gymnosperm Terpene Synthases of the TPS-d Subfamily1[w] , 2004, Plant Physiology.

[31]  R. Croteau,et al.  Defense mechanisms of conifers : differences in constitutive and wound-induced monoterpene biosynthesis among species. , 1991, Plant physiology.

[32]  I. Valterová,et al.  Enantiomeric Composition of Monoterpene Hydrocarbons from the Liverwort Conocephalum-Conicum , 1992 .

[33]  F. Lieutier,et al.  Wound reactions of Scots pine (Pinus sylvestris L) to attacks by Tomicus piniperda L. and Ips sexdentatus Boern. (Col., Scolytidae) , 1995 .

[34]  J. Weiner,et al.  Fundamentals and applications , 2003 .

[35]  A. Berryman,et al.  Specialized phloem parenchyma cells in Norway spruce (Pinaceae) bark are an important site of defense reactions. , 1998, American journal of botany.

[36]  J. Gibbs,et al.  The pine shoot beetle Tomicus piniperda as a vector of blue stain fungi to windblown pine. , 1991 .

[37]  J. Byers.  Host-Tree Chemistry Affecting Colonization in Bark Beetles , 1995 .

[38]  R. Croteau,et al.  Monoterpene synthases of loblolly pine (Pinus taeda) produce pinene isomers and enantiomers. , 1999, Archives of biochemistry and biophysics.

[39]  Diane M. Martin,et al.  Methyl Jasmonate Induces Traumatic Resin Ducts, Terpenoid Resin Biosynthesis, and Terpenoid Accumulation in Developing Xylem of Norway Spruce Stems1 , 2002, Plant Physiology.

[40]  E. Sjöström,et al.  Wood Chemistry: Fundamentals and Applications , 1981 .

[41]  H. Solheim,et al.  Fungal Infection and Mechanical Wounding Induce Disease Resistance in Scots Pine , 2000, European Journal of Plant Pathology.

[42]  H. Solheim,et al.  Blue-stain fungi associated with Tomicus piniperda in Sweden and preliminary observations on their pathogenicity , 1991 .

[43]  H. Solheim,et al.  Induced resistance of Norway spruce, variation of phenolic compounds and their effects on fungal pathogens , 2000 .

[44]  H. Solheim,et al.  Effects of pruning young Scots pines on host vigour and susceptibility to Leptographium wingfieldii and Ophiostoma minus, two blue‐stain fungi associated with Tomicus piniperda , 1993 .

[45]  J. Byers,et al.  Host tree unsuitability recognized by pine shoot beetles in flight , 1989, Experientia.

[46]  H. Solheim,et al.  Pathogenicity of four blue-stain fungi associated with aggressive and nonaggressive bark beetles. , 1998, Phytopathology.

[47]  A. Berryman,et al.  Traumatic resin duct formation in Norway spruce (Picea abies (L.) Karst.) after wounding or infection with a bark beetle-associated blue-stain fungus, Ceratocystis polonica Siem. , 1999 .

[48]  R. Waring,et al.  Resistance of conifers to bark beetle attack: Searching for general relationships , 1987 .

[49]  F. Lieutier,et al.  Bioassays on Leptographium wingfieldii, a Bark Beetle Associated Fungus, with Phenolic Compounds of Scots Pine Phloem , 2004, European Journal of Plant Pathology.

[50]  William J. Bell,et al.  Chemical Ecology of Insects , 1985, Springer US.

[51]  A. Borg-Karlson,et al.  Enantiomeric compositions of monoterpene hydrocarbons in different tissues of four individuals of Pinus sylvestris , 1996 .

[52]  K. Raffa,et al.  Combined chemical defenses against an insect-fungal complex , 1996, Journal of Chemical Ecology.

[53]  H. Solheim.  Fungal succession in sapwood of Norway spruce infested by the bark beetle Ips typographus , 1992 .

[54]  R. Gref,et al.  Induced defence reaction in Scots pine following stem attacks by Tomicus piniperda. , 1992 .

[55]  E. Christiansen,et al.  Wound-induced traumatic resin duct development in stems of Norway spruce (Pinaceae): anatomy and cytochemical traits. , 2000, American journal of botany.

[56]  J. Bohlmann,et al.  cDNA cloning, characterization, and functional expression of four new monoterpene synthase members of the Tpsd gene family from grand fir (Abies grandis). , 1999, Archives of biochemistry and biophysics.

[57]  A. Borg-Karlson,et al.  Occurrence and Correlations of Monoterpene Hydrocarbon Enantiomers in Pinus sylvestris and Picea abies , 2000, Journal of Chemical Ecology.

[58]  P. Niemelä,et al.  Susceptibility of defoliated Scots pine to spontaneous and induced attack by Tomicus piniperda and Tomicus minor. , 1999 .

[59]  A. Borg-Karlson,et al.  Correlations between selected monoterpene hydrocarbons in the xylem of six Pinus (Pinaceae) species , 2001, CHEMOECOLOGY.

[60]  J. Millstein,et al.  Preliminary study of the monoterpene response of three pines to Ophiostoma clavigerum (Ascomycetes : Ophiostomatales) and two chemical elicitors , 1991 .

[61]  L. E. Browne,et al.  In vivo conversion of a labelled host plant chemical to pheromones of the bark beetle Ips paraconfusus , 1980, Nature.

[62]  F. Lieutier,et al.  Preliminary investigations on phenolics as a response of Scots pine phloem to attacks by bark beetles and associated fungi , 1991 .

[63]  R. Firn,et al.  On the evolution of plant secondary chemical diversity , 1991 .

[64]  H. Solheim,et al.  Pathogenicity of the blue-stain fungi Leptographium wingfieldii and Ophiostoma minus to Scots pine : effect of tree pruning and inoculum density , 1993 .

[65]  Diane M. Martin,et al.  Traumatic resin defense in Norway spruce (Picea abies): Methyl jasmonate-induced terpene synthase gene expression, and cDNA cloning and functional characterization of (+)-3-carene synthase , 2004, Plant Molecular Biology.

[66]  W. J. Mattson,et al.  Physiology and Genetics of Tree-phytophage Interactions , 1999 .