Molecular characterization of HEXOKINASE1 in plant innate immunity

Hexokinase1 (HXK1) is an Arabidopsis glucose sensor that has a variety of roles during plant growth and devlopment, including during germination, flowering, and senescence. HXK1 also acts as a positive regulator of plant immune responses. Previous research suggested that HXK1 might influence plant immune responses via responses to glucose. Plant immune responses are governed by two main pathways: PAMP-triggered immunity (PTI) and effector-triggered immunity (ETI). PTI involves the recognition of Pathogen-Associated Molecular Patterns (PAMPs) and leads to increased callose formation and accumulation of pathogenesis response (PR) proteins. ETI acts in response to effectors secreted by Gram-negative bacteria. During ETI, the membrane-localized protein RPM1-interacting protein 4 (RIN4) becomes phosphorylated in reponse to interactions with effectors and mediates the downstream response. In this study, the effects of glucose on plant immune responses against infection with Pseudomonas syringae pv. tomato DC3000 and other P. syringae strains were investigated in the presence and absence of HXK1. Infiltration of leaves with glucose prior to infection led to decreases in bacterial populations and reductions in disease symptoms in wild-type Arabidopsis plants, indicating that glucose plays a role in plant immunity. Both PTI and ETI responses were affected. However, these effects were not observed in a hxk1 mutant, indicating that the effects of glucose on plant immune responses were mediated by HXK1-related pathways.

HCPro Suppression of Callose Deposition Contributes to Strain-Specific Resistance Against Potato Virus Y

Potato virus Y (PVY; Potyviridae) is a continuing challenge for potato production owing to the increasing popularity of strain-specific resistant cultivars. Hypersensitive resistance (HR) is one type of plant defense responses to restrict virus spread. In many potato cultivars, such as cultivar Premier Russet (PR), local necrosis at the site of infection protects against the most common PVYO strain, but the HR often fails to restrain necrotic strains, which spread systemically. Here, we established the role of callose accumulation in the strain-specific resistance responses to PVY infection. We first uncovered that PVY, independent of the strain, is naturally capable of suppressing pathogenesis-related callose formation in a susceptible host. Such activity can be dissociated from viral replication by the transient expression of the viral-encoded helper component proteinase (HCPro) protein, identifying it as the pathogen elicitor. However, unlike the necrotic strain, PVYO and its corresponding HCPro are unable to block callose accumulation in resistant PR potatoes, in which we observed an abundance of callose deposition and the inability of the virus to spread. The substitution of eight amino acid residues within the HCPro C-terminal region that differ between PVYO and PVYN strains and were previously shown to be responsible for eliciting the HR response, are sufficient to restore the ability of HCProO to suppress callose accumulation, despite the resistant host background, in line with a new viral function in pathogenicity.

Active defence by an Australian native host, Lomandra longifolia, provides resistance against Phytophthora cinnamomi

Resistance is rare against the oomycete plant pathogen Phytophthora cinnamomi Rands. Only a limited number of species have been recorded as field-resistant species in Australia. However, understanding the nature of resistance of those species when grown under controlled conditions is challenging because of their slow growth and the inherent difficulties of working with a root pathogen. We assessed the Australian native species, Lomandra longifolia Labill., as a resistant species by analysing in detail the response of roots to infection by P. cinnamomi in a series of comparative tests with Lupinus angustifolius L., a highly susceptible species. Following inoculation of L. longifolia roots, lesion length and colonisation percentage were significantly less than in roots of the susceptible species. Moreover, there was no statistical difference in root growth rate, whole-plant FW and leaf relative chlorophyll content between controls and inoculated L. longifolia. We then examined three key cellular responses that are related to resistance: the production of the reactive oxygen species, H2O2, callose formation and lignin deposition in L. longifolia roots following inoculation with P. cinnamomi. The upregulation of these resistance-related components in the early hours after inoculation suggested their involvement in resistance and that this is controlled by the coordinated response of multiple components. Resistance assessment and a detailed investigation of cellular resistance components along with gene expression analysis provides a platform for further understanding of the mechanisms of resistance against this generalist pathogen and presents opportunities for manipulating susceptible species for disease resistance.

Studies on cell wall regeneration in protoplast culture of legumes – the effect of organic medium additives on cell wall components

The cell wall regeneration in mesophyll protoplasts of yellow lupin and grass pea was studied. The occurrence of cell wall components: cellulose, callose and arabinogalactan proteins was analysed during 15 days of culture. Protoplasts were cultured in different media to test the effect of culture environment on the cell wall regeneration. Medium supplementation with 2 mg/l chitosan resulted in prolonged viability, more balanced cellulose resynthesis, increased callose formation and induction of mitotic divisions in protoplast-derived cells of both examined legumes. In chitosan-enriched medium arabinogalactan proteins were detected in cell plates of divided cells. Medium rich in additional organic compounds, i.e. free amino acids, organic acids and monosaccharides, was inferior to media of simpler composition. In both species the relatively quick cellulose resynthesis negatively affected the viability of protoplast-derived cells. In grass pea cellulose appeared during 24 h of culture. In yellow lupin the process started significantly later and after 10 days the frequency of walled cells did not exceed 50%. Callose was detected in cultures of both species and its pattern suggested that the synthesis was unlikely to be a result of protoplast wounding. Arabinogalactan proteins were localized in cell walls of different types of cells: dividing, elongating, but predominantly in degenerating ones. Occurrence and organization of the cell wall components studied were discussed in relation to recalcitrance of grass pea and yellow lupin protoplasts.

Callose formation in injured cells of the vegetative and generative thallus of Chara vulgaris L. Absence of callose in the process of cytodifferentiation

In an alga <em>Chara vulgaris</em> L. the processes of differentiation of vegetative system cells of the thallus, and initiation and development of generative organs are not associated with callose formation. It was demonstrated that damage to any of the somatic cells and also generative and nongenerative cells of the antheridium and oogonium are capable of callose formation independently of their developmental stage. The localisation and thickness of these layers depend on the way the cells are injured and on their size. The protective role of callose in such cells may consist, beside strengthening the damaged walls, in protection of the symplast by formation of callose deposits on the walls with plasmodesmata; it may also consist in increasing the water potential of the cells. Experiments in which callose deposition was provoked by pressing of the cells or damage leading to a sudden increase of the water potential of the extracellular environment suggest that a sudden increase of tension in the cells may be a factor triggering the "callose effect".

Salicylic Acid Is Important for Basal Defense of Solanum tuberosum Against Phytophthora infestans

The importance of the signaling compound salicylic acid for basal defense of potato (Solanum tuberosum L. cv. Désirée) against Phytophthora infestans, the causal agent of late blight disease, was assessed using transgenic NahG potato plants which are unable to accumulate salicylic acid. Although the size of lesions caused by P. infestans was not significantly different in wild-type and transgenic NahG plants, real-time polymerase chain reaction analyses revealed a drastic enhancement of pathogen growth in potato plants depleted of salicylic acid. Increased susceptibility of NahG plants correlated with compromised callose formation and reduced early defense gene expression. NahG plants pretreated with the salicylic acid analog 2,6-dichloro-isonicotinic acid allowed pathogen growth to a similar extent as did wild-type plants, indicating that salicylic acid is an important compound required for basal defense of potato against P. infestans.