Resistance of Arabidopsis thaliana to the green peach aphid, Myzus persicae, involves camalexin and is regulated by microRNAs.

· Small RNAs play important roles in resistance to plant viruses and the complex responses against pathogens and leaf-chewing insects. · We investigated whether small RNA pathways are involved in Arabidopsis resistance against a phloem-feeding insect, the green peach aphid (Myzus persicae). We used a 2-wk fecundity assay to assess aphid performance on Arabidopsis RNA silencing and defence pathway mutants. Quantitative real-time polymerase chain reaction was used to monitor the transcriptional activity of defence-related genes in plants of varying aphid susceptibility. High-performance liquid chromatography-mass spectrometry was employed to measure the accumulation of the antimicrobial compound camalexin. Artificial diet assays allowed the assessment of the effect of camalexin on aphid performance. · Myzus persicae produces significantly less progeny on Arabidopsis microRNA (miRNA) pathway mutants. Plants unable to process miRNAs respond to aphid infestation with increased induction of PHYTOALEXIN DEFICIENT3 (PAD3) and production of camalexin. Aphids ingest camalexin when feeding on Arabidopsis and are more successful on pad3 and cyp79b2/cyp79b3 mutants defective in camalexin production. Aphids produce less progeny on artificial diets containing camalexin. · Our data indicate that camalexin functions beyond antimicrobial defence to also include hemipteran insects. This work also highlights the extensive role of the miRNA-mediated regulation of secondary metabolic defence pathways with relevance to resistance against a hemipteran pest.

Control of Arabidopsis meristem development by thioredoxin-dependent regulation of intercellular transport.

Cell-to-cell transport in plants occurs through cytoplasmic channels called "plasmodesmata" and is regulated by developmental and environmental factors. Callose deposition modulates plasmodesmal transport in vivo, but little is known about the mechanisms that regulate this process. Here we report a genetic approach to identify mutants affecting plasmodesmal transport. We isolated 5 mutants, named gfp arrested trafficking (gat), affected in GFP unloading from the phloem into the meristem. gat1 mutants were seedling lethal and carried lesions in an m-type thioredoxin that is expressed in non-green plastids of meristems and organ primordia. Callose and hydrogen peroxide accumulated in gat1 mutants, and WT plants subjected to oxidative conditions phenocopied the gat1 trafficking defects. Ectopic expression of GAT1 in mature leaves increased plasmodesmal permeability and led to a delay in senescence and flowering time. We propose a role for the GAT1 thioredoxin in the redox regulation of callose deposition and symplastic permeability that is essential for meristem maintenance in Arabidopsis.

Plasmodesmata "in Communicado".

Cell-to-cell communication is fundamental to multicellular life. For this to occur effectively there must be pathways and dynamic networks for communication. These might depend upon electrical or chemical signals or the mass transfer of molecules between adjacent cells. Molecular communication occurs either via an extra-cellular pathway or through physical structures, called plasmodesmata, that connect the cytoplasm of neighboring cells. Plasmodesmata bridge the rigid physical barrier presented by the cell wall to extend the symplasm from single cells to tissue domains that have functional importance for tissue growth, development, and defense. Although recent years have seen advances in our knowledge of the physical nature of PD, the trafficked molecules, and of the wider processes they affect, our knowledge of PD structure and function is still relatively rudimentary. This article will consider the technical/experimental difficulties hindering PD research and suggest priorities in the future research effort that might advance the field at a significantly faster rate.

Opportunities and successes in the search for plasmodesmal proteins.

The proteinaceous composition of plasmodesmata (PDs) is a puzzle for which pieces have proven particularly difficult to find. This review describes the numerous approaches that have been undertaken in the search for PD-associated proteins and what each has contributed to our understanding of PD structure and function. These approaches include immunolocalisation of known proteins, proteomic characterisation of PD-enriched tissue fractions, high-throughput screens of random cDNAs and mutant screens. In addition to components of the cytoskeleton, novel proteins with predicted or unknown functions have been identified. Many of these have properties that relate to the symplastic and/or apoplastic faces of the plasma membrane. Mutant screens have identified proteins involved in previously unconnected cell pathways such as ROS signalling, implicating ROS in PD formation and regulation. Proteins associated with callose synthesis and degradation have also been identified and characterised, providing considerable weight to the hypothesis that callose deposition around the neck of the PD pore is one mechanism by which the PD aperture is regulated. The techniques described in this review have been developed such that it is to be expected that a considerable number of new PD proteins will be identified in coming years to fill in further detail of the structure and functional mechanisms of these dynamic pores.

Redox regulation of intercellular transport.

Plant cells communicate with each other via plasmodesmata (PDs) in order to orchestrate specific responses to environmental and developmental cues. At the same time, environmental signals regulate this communication by promoting changes in PD structure that modify symplastic permeability and, in extreme cases, isolate damaged cells. Reactive oxygen species (ROS) are key messengers in plant responses to a range of biotic and abiotic stresses. They are also generated during normal metabolism, and mediate signaling pathways that modulate plant growth and developmental transitions. Recent research has suggested the participation of ROS in the regulation of PD transport. The study of several developmental and stress-induced processes revealed a co-regulation of ROS and callose (a cell wall polymer that regulates molecular flux through PDs). The identification of Arabidopsis mutants simultaneously affected in cell redox homeostasis and PD transport, and the histological detection of hydrogen peroxide and peroxidases in the PDs of the tomato vascular cambium provide new information in support of this novel regulatory mechanism. Here, we describe the evidence that supports a role for ROS in the regulation of callose deposition and/or in the formation of secondary PD, and discuss the potential importance of this mechanism during plant growth or defense against environmental stresses.

Symplastic domains in the Arabidopsis shoot apical meristem correlate with PDLP1 expression patterns.

Symplastic domains in plants are defined by spatial limitations on cell-to-cell communication through plasmodesmata (Pds) and establish tissue boundaries necessary for metabolic and developmental programming. With the exception of the physical closure of Pds by callose, the cues and the processes for creating symplastic domains remain poorly understood. Recently, we identified a novel family of eight proteins, called Pd-located protein 1 (PDLP1). These proteins span the plasma membrane within Pds and likely form part of a signal transduction system that perceives external signals to regulate molecular trafficking between cells. For two members of this family that have high expression in the shoot apex we show that they have defined and partially overlapping tissue-specific expression patterns that correlate in part with previously defined symplastic domains. The importance of non-cell-autonomous proteins in shoot development and of the spatial rules that govern leaf and floral development highlight the need to have a clearer understanding of symplastic domains.