Genes expressed in tissue-cultured seedlings of mountain laurel (Kalmia latifolia L.) with colonizing Streptomyces padanus AOK30

Abstract

An endophytic actinomycete, Streptomyces padanus AOK30, is capable of protecting mountain laurel against infection by Pestalotiopsis sydowiana, a causal agent of Pestalotia disease, when applied to the seedling of the plant. In this study, suppression subtractive hybridization was used to identify genes differentially expressed in seedlings of mountain laurel after application of S. padanus AOK30. Subsequent dot hybridization with independent RNA from S. padanus-colonized and control plants identified nonredundant 180 cDNAs involving 71 and 109 clones, which were up- and downregulated after inoculation with the bacteria, respectively. Comparison of the sequences with databases revealed that a number of transcripts encoding proteins or enzymes that function directly in defense or stress response and regulatory proteins were regulated differentially in the seedlings with colonizing S. padanus AOK30. Semi-quantitative RT-PCR analysis for the selected genes demonstrated that inoculation of mountain laurel seedlings with S. padanus AOK30 increased expression of defense-related genes as well as distinct classes of glutathione S-transferase, although endochitinases were exclusively suppressed. These results clearly indicate that the S. padanus-colonizing seedlings likely initiate or prime plant defense responses toward pathogen infection. Selected genes were also differentially expressed in S. padanus-colonized seedlings, compared to those solely challenged with the fungal pathogen P. sydowiana. This approach will assist in efforts not only to understand the molecular basis of the enhanced tolerance and/or enhanced disease resistance of mountain laurel, but also to define a core set of genes during colonization or association with S. padanus AOK30.