Dirigent proteins in conifer defense II: Extended gene discovery, phylogeny, and constitutive and stress-induced gene expression in spruce (Picea spp.)

Analysis of expressed sequence tags (ESTs) and full-length (FL)cDNAs from species of spruce (Picea spp.) revealed a family of 35 unique dirigent proteins (DIR) and DIR-like proteins. Phylogenetic analysis indicates the spruce DIR and DIR-like genes cluster into three distinct subfamilies, DIR-a, DIR-b/d, and DIR-f, of a larger plant DIR and DIR-like gene family. Gene-specific primers were designed for 31 unique spruce DIR family genes, and closely related isoforms, and used to evaluate patterns of constitutive expression, as well as responses to herbivory by stem-boring insects (i.e., white pine weevil, Pissodes strobi) in bark tissue and defoliating insects (i.e., western spruce budworm, Choristoneura occidentalis) in green apical shoots. Furthermore, meta-analysis of microarray gene expression data obtained from a series of independent experiments using the same 16.7K cDNA array platform identified several distinct expression clusters of the spruce DIR transcriptome closely matching phylogenetic clusters of sequence similarity. Members of the DIR-a family, which also contains functionally characterized DIR from other plant species, are most prominent for their induced response to feeding by weevils on Sitka spruce bark.