Aims The goal of this study was to identify the induced resistance pathway mediated by biochar in the tomatoBotrytis cinerea pathosystem. Methods Tomato wild types and mutants modified in their salicylic acid (SA), ethylene (ET) or jasmonic acid (JA) metabolism were grown in a potting medium amended with biochar produced at 450ºC from greenhouse wastes, to identify the possible pathway(s) involved in biochar-mediated resistance to B. cinerea.Early cellular response of H 2 O 2 accumulation was biochemically tested, and the transcriptional changes of 12 defense-related genes upon B. cinerea challenge of detached leaflets were analyzed. Results Biochar amendment resulted in about 50 % reduction in B. cinerea disease severity in all tested genotypes with the exception of a JA deficient mutant, def1. Biochar amendment induced priming of early as well as late-acting defense responses particularly in the genes Pti5 (ET-related) and Pi2 (JA-related), which are known to be crucial in resistance against B. cinerea. Stronger and earlier H 2 O 2 accumulation subsequent to B. cinerea inoculation in all genotypes was observed as a result of biochar amendment, with the exception of the def1 mutation. Conclusion Biochar-mediated IR in the B. cinerea-tomato pathosystem involves the JA pathway.

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Gray mold (Botrytis cinerea) is an important disease of tomato (Solanum lycopersicum). This study examined defense-related gene expression involved in the resistance to B. cinerea that is induced in tomato plants by benzothiadiazole and Trichoderma harzianum T39 soil drench. In whole plants, transcriptional changes related to salicylic acid and ethylene were induced by the application of a 0.01% benzothiadiazole solution, whereas changes related to jasmonic acid were induced by the application of a 0.4% T39 suspension. On detached leaves, soil treatment by T39 led to enhanced resistance to B. cinerea infection that was proportional to the concentration of the T39 suspension. By 5 days after pathogen inoculation, the plants that had received the 0.04% T39 drench exhibited 62% less severe disease than the untreated plants. The 0.4% T39 drench led to an 84% reduction in disease severity. Observations of B. cinerea infection in leaves harvested from plants grown in the treated soils revealed that drenching with a T39 suspension induces systemic resistance against B. cinerea and primes salicylic acid- and ethylene-related gene expression in a manner proportional to the concentration of the biocontrol agent. Benzothiadiazole treatment induced resistance to gray mold independently of salicylic acid and led to strong priming of two genes known to be involved in defense against B. cinerea, Pti5 and PI2.

Powdery mildew of strawberry is caused by the obligate pathogenic fungus Sphaerotheca macularis f. sp. fragariae. The disease affects the leaves, flowers and fruit of this crop. This study examined the effects of different environmental factors on disease cycle components (germination, conidiation and survival) in strawberry to determine which conditions limit the progress of the disease. The optimal environmental conditions for conidial germination and conidial germ tube length ranged between 15 and 25°C with relative humidity (RH) higher than 75%, but less than 98%. High light intensity reduced germination and hyphal growth. The viability of conidia on infected leaves was examined at temperatures ranging from 15 to 35°C at 80-85% RH. Conidia survival declined over time, but a certain percentage of conidia remained active after 5 months incubation. The rate of conidial germination was significantly higher on young leaves than on older leaves. This observation was consistent across all four tested cultivars. Conidiation at 70-75% RH was similar to that at 80-85%, but greater than that at ‡95% RH. The shortest time from inoculation to appearance of the first disease symptoms was 4 days, at 20 and 30°C with RH above 75%. In growth chambers, temperatures of 10 and 30°C, RH above 95%, radiation of 7000 lux and the use of a more tolerant cultivar were all detrimental to disease development. In general, the environmental conditions required for germination and dispersal of powdery mildew are conducive to disease progress under strawberry production conditions in Israel. Furthermore, viability and survival of the pathogen during and between seasons appears to be dependent on asexual inoculum production.

Botrytis cinerea is a non-specific, necrotrophic pathogen that attacks many plant species, including Arabidopsis and tomato. Since senescing leaves are particularly susceptible to infection by B. cinerea, we hypothesized that the fungus might induce senescence as part of its mode of action and that delaying leaf senescence might reduce the severity of B. cinerea infections. To examine these hypotheses, we followed the expression of Arabidopsis SAG12, a senescence-specific gene, upon infection with B. cinerea. Expression of SAG12 is induced by B. cinerea infection, indicating that B. cinerea induces senescence. The promoter of SAG12, as well as that of a second Arabidopsis senescence-associated gene, SAG13, whose expression is not specific to senescence, were previously analyzed in tomato plants and were found to be expressed in a similar manner in the two species. These promoters were previously used in tomato plants to drive the expression of isopentenyl transferase (IPT) from Agrobacterium to suppress leaf senescence (Swartzberg et al. in Plant Biology 8:579-586, 2006). In this study, we examined the expression of these promoters following infection of tomato plants with B. cinerea. Both promoters exhibit high expression levels upon B. cinerea infection of non-senescing leaves of tomato plants, supporting our conclusion that B. cinerea induces senescence as part of its mode of action. In contrast to B. cinerea, Trichoderma harzianum T39, a saprophytic fungus that is used as a biocontrol agent against B. cinerea, induces expression of SAG13 only. Expression of IPT, under the control of the SAG12 and SAG13 promoters in response to infection with B. cinerea resulted in suppression of B. cinerea-induced disease symptoms, substantiating our prediction that delaying leaf senescence might reduce susceptibility to B. cinerea.

Downy mildew (Peronospora belbahrii) is a major disease of sweet basil (Ocimum basilicum). We examined the effects of potassium, calcium and magnesium, individually and in combination, on sweet basil downy mildew (SBDM) in potted plants and under commercial-greenhouse conditions over six growing seasons. An increased K concentration in the fertigation solution increased SBDM severity, whereas foliar-applied KCl and K2SO4 suppressed SBDM. The application of higher concentrations of those salts increased the K concentrations in the shoots and significantly alleviated SBDM. Increased concentrations of Ca or Mg in the fertigation solution decreased SBDM severity, as did foliar-applied CaCl2. However, the combination of Ca and Mg did not have any synergistic effect. Foliar-applied K2SO4 provided better disease suppression than some of these treatments. The 3.3 mM Mg + fungicide treatment and the 5.0 mM Mg + fungicide treatment each provided synergistic disease control in one of two experiments. SBDM severity was significantly reduced by MgCl2 and MgSO4 (both 3.3 mM Mg), as compared with the basic Mg fertigation (1.6 mM), with MgCl2 providing better control. The combined Mg salts + fungicide treatments reduced SBDM better than any of those treatments alone. These results demonstrate that macro-elements can contribute to SBDM control.

Biocontrol agents can control pathogens by re-enforcing systemic plant resistance through systemic acquired resistance (SAR) or induced systemic resistance (ISR). Trichoderma spp. can activate the plant immune system through ISR, priming molecular mechanisms of defense against pathogens. Entomopathogenic fungi (EPF) can infect a wide range of arthropod pests, and play an important role in reducing pests' population. Here, we investigated the mechanisms by which EPF control plant diseases. We tested two well studied EPF, Metarhizium brunneum isolate Mb7 and Beauveria bassiana as the commercial product Velifer, for their ability to induce systemic immunity and disease resistance against several fungal and bacterial phytopathogens, and their ability to promote plant growth. We compared the activity of these EPF to an established biocontrol agent, T. harzianum T39, a known inducer of systemic plant immunity and broad disease resistance. The three fungal agents were effective against several fungal and bacterial plant pathogens and arthropod pests. Our results indicate that EPF induce systemic plant immunity and disease resistance by activating the plant host defense machinery, as evidenced by increases in reactive oxygen species (ROS) production and defense gene expression, and that EPF promote plant growth. EPF should be considered as control means for Tuta absoluta. We demonstrate that, with some exceptions, biocontrol in tomato can be equally potent by the tested EPF and T. harzianum T39, against both insect pests and plant pathogens. Taken together, our findings suggest that EPF may find use in broad-spectrum pest and disease management and as plant growth promoting agents.