Transcriptome analysis of WIPK/SIPK-suppressed plants reveals induction by wounding of disease resistance-related genes prior to the accumulation of salicylic acid

Abstract

Salicylic acid (SA) plays a key role in plant resistance to pathogens. Accumulation of SA is induced by wounding in tobacco plants in which the expression of WIPK and SIPK, two mitogen-activated protein kinases, is suppressed. Here, the mechanisms underlying the abnormal accumulation of SA in WIPK/SIPK-suppressed plants have been characterized. SA accumulation started around 12 h after wounding and was inhibited by cycloheximide (CHX), a protein synthesis inhibitor. SA accumulation, however, was enhanced several fold when leaf discs were transferred onto CHX after floating on water for ≥6 h. Temporal and spatial analyses of wound-induced and CHX-enhanced SA accumulation suggested that wounding induces activators for SA accumulation followed by the generation of repressors, and late CHX treatment inhibits the production of repressors more efficiently than that of activators. Microarray analysis revealed that the expression of many disease resistance-related genes, including N, a Resistance (R) gene for Tobacco mosaic virus and R gene-like genes, was up-regulated in wounded WIPK/SIPK-suppressed plants. Expression of the N gene and R gene-like genes peaked earlier than that of most other genes as well as SA accumulation, and was mainly induced in those parts of leaf discs where SA was highly accumulated. Moreover, wound-induced SA accumulation was decreased by the treatments which compromise the function of R proteins. These results indicate that signaling leading to the expression of disease resistance-related genes is activated by wounding in WIPK/SIPK-suppressed plants, and induction of R gene and R gene-like genes might lead to the biosynthesis of SA.

Keywords: MAPK; Nicotiana tabacum; R gene; Salicylate; Wound.