Mayra Rodríguez

NmDef02, a novel antimicrobial gene isolated from Nicotiana megalosiphon confers high‐level pathogen resistance under greenhouse and field conditions

Plant defensins are small cysteine-rich peptides that inhibit the growth of a broad range of microbes. In this article, we describe NmDef02, a novel cDNA encoding a putative defensin isolated from Nicotiana megalosiphon upon inoculation with the tobacco blue mould pathogen Peronospora hyoscyami f.sp. tabacina. NmDef02 was heterologously expressed in the yeast Pichia pastoris, and the purified recombinant protein was found to display antimicrobial activity in vitro against important plant pathogens. Constitutive expression of NmDef02 gene in transgenic tobacco and potato plants enhanced resistance against various plant microbial pathogens, including the oomycete Phytophthora infestans, causal agent of the economically important potato late blight disease, under greenhouse and field conditions.

Expression of a Nicotiana tabacum pathogen-induced gene is involved in the susceptibility to black shank

Many host genes induced during compatible plant-pathogen interactions constitute targets of pathogen virulence factors that act to suppress host defenses. In order to identify Nicotiana tabacum L. genes for pathogen-induced proteins involved in susceptibility to the oomycete Phytophthora parasitica var. nicotianae, we used SuperSAGE technology combined with next-generation sequencing to identify transcripts that were differentially upregulated during a compatible interaction. We identified a pathogen-induced gene (NtPIP) that was rapidly induced only during the compatible interaction. Virus-induced gene silencing of NtPIP reduced the susceptibility of N. tabacum to P. parasitica var. nicotianae. Additionally, transient expression of NtPIP in the resistant species Nicotiana megalosiphon Van Heurck & Mull. Arg. compromised the resistance to P. parasitica var. nicotianae. This pathogen-induced protein is therefore a positive regulator of the susceptibility response against an oomycete pathogen in tobacco.

NmEXT Extensin Gene: a Positive Regulator of Resistance Response Against the Oomycete Phytophthora nicotianae

The oomycete pathogens produce important diseases in many plant species. To identify extensin genes expressed during the oomycete Phytophthora nicotianae-Nicotiana megalosiphon interaction, we used the SuperSAGE technology. Using this approach, we detected a N. megalosiphon extensin gene (NmEXT) triggered during the interaction. The extensin gene accumulation induced by the pathogen correlated with disease resistance in different Nicotiana species. Transient expression of NmEXT gene in susceptible Nicotiana tabacum enhanced the resistance to P. nicotianae. Our date indicated that NmEXT gene served a positive role in N. tabacum resistance against P. nicotianae.

High-throughput SuperSAGE for gene expression analysis of Nicotiana tabacum – Rhizoctonia solani interaction

ObjectiveThe ubiquitous soil pathogen Rhizoctonia solani causes serious diseases in different plant species. Despite the importance of this disease, little is known regarding the molecular basis of susceptibility. SuperSAGE technology and next-generation sequencing were used to generate transcript libraries during the compatible Nicotiana tabacum–R. solani interaction. Also, we used the post-transcriptional silencing to evaluate the function of a group of important genes.ResultsA total of 8960 and 8221 unique Tag sequences identified as differentially up- and down-regulated were obtained. Based on gene ontology classification, several annotated UniTags corresponded to defense response, metabolism and signal transduction. Analysis of the N. tabacum transcriptome during infection identified regulatory genes implicated in a number of hormone pathways. Silencing of an mRNA induced by salicylic acid reduced the susceptibility of N. tabacum to R. solani. We provide evidence that the salicylic acid pathway was involved in disease development. This is important for further development of disease management strategies caused by this pathogen.

Cellular target of helminthosporium sacchari Toxin in sugarcane tissue

Spindles from sugarcane (Saccharum spp. hybrid) tissue were challenged using labeledHelminthosporium (Bipolaris) sacchari toxin to determine the general effect on different organelles and structures. Toxin labeling was characterized by biochemical and physiological behaviour of the fungal pathogen (dry weight, pH, refractive index, invertase and β -galactosidase activity and the biological activity of the toxin), allowing to improve the metabolic uptake of a labeled structural precursor of the toxin. A more efficient toxin labeling than previously reported was obtained. The labeled toxin was used as a tracer for determination of interactions at mitochondrial level. Binding activity reached saturation in time, confirming respiratory and ultrastructural analysis of the toxin effect.