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Eucalyptus grandis  (W. Hill ex Maiden) is an Australian Myrtaceae tree grown for timber in many parts of the world and for which the annotated genome sequence is available. Known to be susceptible to a number of pests and diseases,  E. grandis  is a useful study organism for investigating defense responses in woody plants. Chitinases are widespread in plants and cleave glycosidic bonds of chitin, the major structural component of fungal cell walls and arthropod exoskeletons. They are encoded by an important class of genes known to be up-regulated in plants in response to pathogens. The current study identified 67 chitinase gene models from two families known as glycosyl hydrolase 18 and 19 (36 GH18 and 31 GH19) within the  E. grandis  genome assembly (v1.1), indicating a recent gene expansion. Sequences were aligned and analyzed as conforming to currently recognized plant chitinase classes (I–V). Unlike other woody species investigated to date,  E. grandis  has a single gene encoding a putative vacuolar targeted Class I chitinase. In response to  Leptocybe invasa  (Fisher &amp; La Salle) (the eucalypt gall wasp) and  Chrysoporthe austroafricana  (Gryzenhout &amp; M.J. Wingf. 2004) (causal agent of fungal stem canker), this  Class IA chitinase  is strongly up-regulated in both resistant and susceptible plants. Resistant plants, however, indicate greater constitutive expression and increased up-regulation than susceptible plants following fungal challenge. Up-regulation within fungal resistant clones was further confirmed with protein data. Clusters of putative chitinase genes, particularly on chromosomes 3 and 8, are significantly up-regulated in response to fungal challenge, while a cluster on chromosome 1 is significantly down-regulated in response to gall wasp. The results of this study show that the  E. grandis  genome has an expanded group of chitinase genes, compared with other plants. Despite this expansion, only a single  Class I chitinase  is present and this gene is highly up-regulated within diverse biotic stress conditions. Our research provides insight into a major class of defense genes within  E. grandis  and indicates the importance of the  Class I chitinase .

Identification of the Eucalyptus grandis chitinase gene family and expression characterization under different biotic stress challenges