Functional Characterization of Tomato ShROP7 in Regulating Resistance against Oidium neolycopersici.

ROPs (Rho-like GTPases from plants) are a unique family of small GTP-binding proteins in plants and play vital roles in numerous cellular processes, including growth and development, abiotic stress signaling, and plant defense. In the case of the latter, the role of ROPs as response regulators to obligate parasitism remains largely enigmatic. Herein, we isolated and identified ShROP7 and show that it plays a critical role in plant immune response to pathogen infection. Real-time quantitative PCR analysis revealed that the expression of ShROP7 was significantly increased during incompatible interactions. To establish its requirement for resistance, we demonstrate that virus-induced gene silencing (VIGS) of ShROP7 resulted in increased susceptibility of tomato to Oidium neolycopersici (On) Lanzhou strain (On-Lz). Downstream resistance signaling through H2O2 and the induction of the hypersensitive response (HR) in ShROP7-silenced plants were significantly reduced after inoculating with On-Lz. Taken together, with the identification of ShROP7-interacting candidates, including ShSOBIR1, we demonstrate that ShROP7 plays a positive regulatory role in tomato powdery mildew resistance.

The tomato Arp2/3 complex is required for resistance to the powdery mildew fungus Oidium neolycopersici.

The actin-related protein 2/3 complex (Arp2/3 complex), a key regulator of actin cytoskeletal dynamics, has been linked to multiple cellular processes, including those associated with response to stress. Herein, the Solanum habrochaites ARPC3 gene, encoding a subunit protein of the Arp2/3 complex, was identified and characterized. ShARPC3 encodes a 174-amino acid protein possessing a conserved P21-Arc domain. Silencing of ShARPC3 resulted in enhanced susceptibility to the powdery mildew pathogen Oidium neolycopersici (On-Lz), demonstrating a role for ShARPC3 in defence signalling. Interestingly, a loss of ShARPC3 coincided with enhanced susceptibility to On-Lz, a process that we hypothesize is the result of a block in the activity of SA-mediated defence signalling. Conversely, overexpression of ShARPC3 in Arabidopsis thaliana, followed by inoculation with On-Lz, showed enhanced resistance, including the rapid induction of hypersensitive cell death and the generation of reactive oxygen. Heterologous expression of ShARPC3 in the arc18 mutant of Saccharomyces cerevisiae (i.e., ∆arc18) resulted in complementation of stress-induced phenotypes, including high-temperature tolerance. Taken together, these data support a role for ShARPC3 in tomato through positive regulation of plant immunity in response to O. neolycopersici pathogenesis.

The dual role of oxalic acid on the resistance of tomato against Botrytis cinerea.

In order to define the role of oxalic acid (OA) in the invasion of Botrytis cinerea in tomato plants, the OA induction of resistance related to oxalate oxidase (O×O) and germin was examined. In greenhouse experiments, OA at 3 mmol/L significantly induced resistance in tomato plants against B. cinerea strains B05.10 and T4, reducing lesion size of 37.55% and 24.91% by compared with distilled water control, respectively, while 20 mmol/L OA increasing by 36.14% and 41.48%. OA contents were 98 and 46 µg/mL when tomato plants were infected by B. cinerea strains B05.10 and T4, respectively. To define the molecular-genetic mechanisms, we compared the gene expression under four different conditions: 3 mmol/L OA-treated plants, 20 mmol/L OA-treated plants, B. cinerea strain B05.10-infected plants (B05.10 Inf plants) and B. cinerea strain T4-infected plants (T4 Inf plants). In 3 mmol/L OA-treated plants, the expressions of O×O and Germin peaked at 48 h after spraying, with approximate threefold and 18-fold increase compared with the control expression, respectively. In T4 Inf plants, the expression (mRNA accumulation) of O×O and Germin reached the highest levels at 24 h after inoculation, with 3- and 13-times that immediately after inoculation, respectively. In total, these findings suggest that elevated levels of OA correlated with increased fungal invasion and lower OA induced resistance in tomato plants by increasing expressions of O×O and Germin.

Synergistic effect of the combined bio-fungicides ε-poly-l-lysine and chitooligosaccharide in controlling grey mould (Botrytis cinerea) in tomatoes.

The antifungal properties and the induction of resistance by ε-poly-l-lysine (ε-PL) and chitooligosaccharide (COS) were examined to find an alternative to synthetic fungicides currently used in the control of the devastating fungal pathogen Botrytis cinerea, the causal agent of grey mould disease of tomatoes. As presented herein, this combined treatment (200 mg/L ε-PL + 400 mg/L COS) was found to have optimal in vitro antifungal activities, achieving an inhibition rate of 90.22%. In vivo assays with these combined bio-fungicides, under greenhouse conditions using susceptible tomato plants, demonstrated good protection against severe grey mould. In field tests, the combined bio-fungicides had a control effect of up to 66.67% against tomato grey mould. To elucidate the mechanisms of the combined bio-fungicide-induced resistance in the tomato, plants were subjected to three treatments: 1) inoculation with B. cinerea after spraying with 200 mg/L ε-PL alone, 2) inoculation with the combined bio-fungicides, and 3) inoculation with 400 mg/L COS alone. Compared to the control (sterile water), increases in salicylic acid (SA) and jasmonic acid (JA) levels and increased phenylalanine ammonia lyase (PAL), peroxidase (POD), and superoxide dismutase (SOD) activities were observed. Catalase (CAT) activity and abscisic acid (ABA) and gibberellin (GA) levels decreased, particularly in the combined bio-fungicide-treated plants. Altogether, these findings reveal that the combined bio-fungicides (200 mg/L ε-PL + 400 mg/L COS) should be an excellent biocontrol agent candidate that combines direct antifungal activity against B. cinerea with plant resistance.