Abstract

Plants respond to various stress stimuli by activating broad-spectrum defense responses both locally as well as systemically. As such, identification of expressed genes represents an important step towards understanding inducible defense responses and assists in designing appropriate intervention strategies for disease management. Genes differentially expressed in tobacco cell suspensions following elicitation with isonitrosoacetophenone (INAP) were identified using mRNA differential display and pyro-sequencing. Sequencing data produced 14579 reads, which resulted in 198 contigs and 1758 singletons. Following BLAST analyses, several inducible plant defense genes of interest were identified and classified into functional categories including signal transduction, transcription activation, transcription and protein synthesis, protein degradation and ubiquitination, stress-responsive, defense-related, metabolism and energy, regulation, transportation, cytoskeleton and cell wall-related. Quantitative PCR was used to investigate the expression of 17 selected target genes within these categories. Results indicate that INAP has a sensitising or priming effect through activation of salicylic acid-, jasmonic acid- and ethylene pathways that result in an altered transcriptome, with the expression of genes involved in perception of pathogens and associated cellular re-programming in support of defense. Furthermore, infection assays with the pathogen Pseudomonas syringae pv. tabaci confirmed the establishment of a functional anti-microbial environment in planta.

Highlights

  • Due to the lack of a circulative adaptive immune system, plants have adapted to biotic stressors by developing resistance mechanisms to recognize and counter-attack prospective colonists and pathogens [1]

  • This study aimed to assess changes in gene expression associated with isonitrosoacetophenone-associated defense induction in Nicotiana tabacum. mRNA differential display was successfully used for detection and recovery of up-regulated PCR amplicons as previously used to identify and isolate genes involved in plant innate immunity [7,13]

  • Based on the ratios obtained for A260/280 and A260/230, as well as the integrity, it was concluded that the RNA was of high quality and free from protein, polyphenol and polysaccharide contamination

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Summary

Introduction

Due to the lack of a circulative adaptive immune system, plants have adapted to biotic stressors by developing resistance mechanisms to recognize and counter-attack prospective colonists and pathogens [1]. Resistance involves several pre-formed and inducible mechanisms that inhibit pathogen growth and can be dependent on host resistance- as well as pathogen avirulent genes [2]. Plant resistance is considered effective if it remains operative even in an environment.

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