Functional Genomics of Conifer Defense and Resistance Mechanisms

Conifers display a very large array of defenses against insects and insect-associated fungal pathogens. These defenses include a myriad of combinations of constitutive and induced, chemical and physical, direct and indirect, as well as local and systemic defenses. Among some of the most prominent defenses in conifers are terpenoid (e.g. oleoresin) and phenolic secondary metabolites. Traumatic resinosis involves methyl jasmonate or ethylene-inducible de novo differentiation of specialized anatomical structures (traumatic resin ducts) for induced terpenoid accumulation in the developing xylem. Similarly, phenolic defenses may involve the induction of specialized phloem parenchyma (PP) cells. Insect induced volatile emissions are based on passive release of resin terpenoids and on the active de novo formation and emission of non-resin monoterpenes and sesquiterpenes.

We are investigating the defense and resistance mechanism in weevil resistant and susceptible genotypes of Sitka spruce and in interior spruce. For example, in our genomics-based biochemical characterization of insect-induced terpenoid defenses, we are functionally characterizing a very large family of terpenoid synthases (TPS) and analyze their expression in response to insect attack. Other work is targeting cytochrome P450 genes and other insect-induced defense gene families.

Our research on the large and rapidly evolving TPS gene family and on terpenoid metabolite profiles has revealed molecular and biochemical mechanisms that underlie plasticity and diversity of conifer chemical defenses.  The diversity of terpenoid and other chemical defenses contributed at least in part, to the successful evolution of long-lived conifer trees, which often survive for several hundred years in the same location defeating many generations of faster evolving insect pests and pathogens.

Large-scale genomics resources for species of spruce as well as optimized proteomics and metabolite profiling tools have been developed in our conifer genomics program and have been applied for a comprehensive analysis of conifer chemical defense against insect attack. In parallel, we are studying the possible counterattack mechanisms of insects (e.g. bark beetles) and their associated fungal pathogens which may help these organisms to overcome the defenses of conifers.

Among the goals we would like to achieve is the identification of constitutive and insect-induced defense mechanisms which will be pursued by:

  1. transcript-, proteome-, and metabolite profiling;
  2. characterization of cell- or tissue-specific defense mechanisms based on tissue micro-dissection;
  3. functional identification of defense genes and gene families using in vivo and in vitro assay systems;
  4. investigation of the size and diversification of defense gene families in spruce;
  5. identification of regulatory systems for selected defense genes and mechanisms.