Species Delimitation and Host Specialization of Ceratocystis laricicola and C. polonica to Larch and Spruce

Abstract
Ceratocystis laricicola and C. polonica are fungal symbionts of bark beetle species of the genus Ips that attack species of Larix and Picea, respectively, across Eurasia. Earlier studies found that these fungal species were morphologically identical, had similar isozymes patterns, and had identical internal transcribed spacer (ITS) sequences of the rDNA operon. We analyzed 27 isolates from Europe, southwestern Siberia (Russia) and Japan, representing the known geographic ranges of the two species. Phylogenetic analysis of the DNA sequences of a portion of the MAT-2 idiomorph showed these species to be distinct, with the Japanese isolates of C. laricicola having a sequence slightly different (5 bp) from those of the Russian and European isolates of C. laricicola. Sexual compatibility tests showed full interfertility among isolates of C. polonica from Europe, Russia and Japan, but isolates of C. polonica were not fully interfertile with isolates of C. laricicola. A Russian and a European isolate of C. laricicola mated with each other but not with the Japanese isolates of C. laricicola. Mature L. sibirica and P. obovata were inoculated with isolates of C. laricicola and C. polonica from Europe, Russia, and Japan, and measurement of lesions in the inner bark/cambium region demonstrated strong host specialization. The data suggest that the two fungal species are very closely related and are distinguished primarily by their physiological specialization to the hosts of their bark beetle vectors.