Yigal Elad

Biochar impact on development and productivity of pepper and tomato grown in fertigated soilless media

The impact of additions (1–5% by weight) of a nutrient-poor, wood-derived biochar on pepper (Capsicum annuum L.) and tomato (Lycopersicum esculentum Mill.) plant development and productivity in a coconut fiber:tuff growing mix under optimal fertigation conditions was examined. Pepper plant development in the biochar-treated pots was significantly enhanced as compared with the unamended controls. This was reflected by a system-wide increase in most measured plant parameters: leaf area, canopy dry weight, number of nodes, and yields of buds, flowers and fruit. In addition to the observed increases in plant growth and productivity, the rhizosphere of biochar-amended pepper plants had significantly greater abundances of culturable microbes belonging to prominent soil-associated groups. Phylogenetic characterization of unique bacterial isolates based on 16S rRNA gene analysis demonstrated that of the 20 unique identified isolates from roots and bulk soil from the char-amended growing mix, 16 were affiliated with previously described plant growth promoting and/or biocontrol agents. In tomato, biochar treatments positively enhanced plant height and leaf size, but had no effect on flower and fruit yield. The positive impacts of biochar on plant response were not due to direct or indirect effects on plant nutrition, as there were no differences between control and treatments in leaf nutrient content. Nor did biochar affect the field capacity of the soilless mixture. A number of organic compounds belonging to various chemical classes, including n-alkanoic acids, hydroxy and acetoxy acids, benzoic acids, diols, triols, and phenols were identified in organic solvent extracts of the biochar. We conjecture two related alternatives to explain the improved plant performance under biochar treatment: (i) the biochar stimulated shifts in microbial populations towards beneficial plant growth promoting rhizobacteria or fungi, due to either chemical or physical attributes of the biochar; or (ii) low doses of biochar chemicals, many of which are phytotoxic or biocidal at high concentrations, stimulated plant growth at low doses (hormesis).

Induced systemic resistance in Trichoderma harzianum T39 biocontrol of Botrytis cinerea

Biocontrol of Botrytis cinerea with Trichoderma spp. is generally believed to result from direct interaction of the biocontrol agent with the pathogen or from a Trichoderma-induced change in environmental conditions that affects B. cinerea development. In this work we provide arguments for the participation of induced plant defence in T. harzianum T39 control of B. cinerea. In tomato, lettuce, pepper, bean and tobacco, T. harzianum T39 application at sites spatially separated from the B. cinerea inoculation resulted in a 25–100%percnt; reduction of grey mould symptoms, caused by a delay or suppression of spreading lesion formation. Given the spatial separation of both micro-organisms, this effect was attributed to the induction of systemic resistance by T. harzianum T39. The observation that in bean the effect of T. harzianum T39 was similar to that of the rhizobacterium Pseudomonas aeruginosa KMPCH, a reference strain for the induction of systemic resistance, confirmed this hypothesis. Since B. cinerea control on tobacco leaves sprayed with T. harzianum T39 was similar to the control on leaves from T. harzianum T39 soil-treated plants, induction of plant defence might also participate in biocontrol on treated leaves.

Impact of Biochar Application to Soil on the Root-Associated Bacterial Community Structure of Fully Developed Greenhouse Pepper Plants

ABSTRACT Adding biochar to soil has environmental and agricultural potential due to its long-term carbon sequestration capacity and its ability to improve crop productivity. Recent studies have demonstrated that soil-applied biochar promotes the systemic resistance of plants to several prominent foliar pathogens. One potential mechanism for this phenomenon is root-associated microbial elicitors whose presence is somehow augmented in the biochar-amended soils. The objective of this study was to assess the effect of biochar amendment on the root-associated bacterial community composition of mature sweet pepper (Capsicum annuum L.) plants. Molecular fingerprinting (denaturing gradient gel electrophoresis and terminal restriction fragment length polymorphism) of 16S rRNA gene fragments showed a clear differentiation between the root-associated bacterial community structures of biochar-amended and control plants. The pyrosequencing of 16S rRNA amplicons from the rhizoplane of both treatments generated a total of 20,142 sequences, 92 to 95% of which were affiliated with the Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes phyla. The relative abundance of members of the Bacteroidetes phylum increased from 12 to 30% as a result of biochar amendment, while that of the Proteobacteria decreased from 71 to 47%. The Bacteroidetes-affiliated Flavobacterium was the strongest biochar-induced genus. The relative abundance of this group increased from 4.2% of total root-associated operational taxonomic units (OTUs) in control samples to 19.6% in biochar-amended samples. Additional biochar-induced genera included chitin and cellulose degraders (Chitinophaga and Cellvibrio, respectively) and aromatic compound degraders (Hydrogenophaga and Dechloromonas). We hypothesize that these biochar-augmented genera may be at least partially responsible for the beneficial effect of biochar amendment on plant growth and viability.

Induction of Systemic Resistance in Plants by Biochar, a Soil-Applied Carbon Sequestering Agent

Biochar is the solid coproduct of biomass pyrolysis, a technique used for carbon-negative production of second-generation biofuels. The biochar can be applied as a soil amendment, where it permanently sequesters carbon from the atmosphere as well as improves soil tilth, nutrient retention, and crop productivity. In addition to its other benefits in soil, we found that soil-applied biochar induces systemic resistance to the foliar fungal pathogens Botrytis cinerea (gray mold) and Leveillula taurica (powdery mildew) on pepper and tomato and to the broad mite pest (Polyphagotarsonemus latus Banks) on pepper. Levels of 1 to 5% biochar in a soil and a coconut fiber-tuff potting medium were found to be significantly effective at suppressing both diseases in leaves of different ages. In long-term tests (105 days), pepper powdery mildew was significantly less severe in the biochar-treated plants than in the plants from the unamended controls although, during the final 25 days, the rate of disease development in the treatments and controls was similar. Possible biochar-related elicitors of systemic induced resistance are discussed.

Mechanisms involved in the biological control of Botrytis cinerea incited diseases

Mechanisms involved in the biological suppression of infection and inoculum potential ofBotrytis cinerea are numerous and variable and the involvement of two or more mechanisms has been demonstrated in several systems. Reported combinations include antibiosis with enzyme degradation ofB. cinerea cell walls; competition for nutrients followed by interference with pathogenicity enzymes of the pathogen or with induced resistance; and alteration of plant surface wettability combined with antibiosis. Since germinatingB. cinerea conidia are dependent on the presence of nutrients, competition for nutrients is regarded as important in systems where biocontrol is involved. Conidial viability and germination capacity are also potentially affected by the presence of antibiotics produced by biocontrol agents and present in the phyllosphere. Slower in action are mechanisms involving induced resistance in the host plant and production of hydrolytic enzymes that degradeB. cinerea cell walls. The latter has been demonstrated much more convincinglyin vitro than in the phyllosphere. Biocontrol in established lesions and reduction of sporulation on necrotic plant tissues is a means to minimize the pathogen inoculum.

Trichoderma Biocontrol of Colletotrichum acutatum and Botrytis cinerea and Survival in Strawberry

Trichoderma isolates are known for their ability to control plant pathogens. It has been shown that various isolates of Trichoderma, including T. harzianum isolate T-39 from the commercial biological control product TRICHODEX, were effective in controlling anthracnose (Colletotrichum acutatum) and grey mould (Botrytis cinerea) in strawberry, under controlled and greenhouse conditions. Three selected Trichoderma strains, namely T-39, T-161 and T-166, were evaluated in large-scale experiments using different timing application and dosage rates for reduction of strawberry anthracnose and grey mould. All possible combinations of single, double or triple mixtures of Trichoderma strains, applied at 0.4% and 0.8% concentrations, and at 7 or 10 day intervals, resulted in reduction of anthracnose severity; the higher concentration (0.8%) was superior in control whether used with single isolates or as a result of combined application of two isolates, each at 0.4%. Only a few treatments resulted in significant control of grey mould. Isolates T-39 applied at 0.4% at 2 day intervals, T-166 at 0.4%, or T-161 combined with T-39 at 0.4% were as effective as the chemical fungicide fenhexamide. The survival dynamics of populations of the Trichoderma isolates (T-39, T-105, T-161 and T-166) applied separately was determined by dilution plating and isolates in the mixtures calculated according to the polymerase chain reaction (PCR) using repeat motif primers. The biocontrol isolates were identified to the respective species T. harzianum (T-39), T. hamatum (T-105), T. atroviride (T-161) and T. longibrachiatum (T-166), according to internal transcribed spacer sequence analysis.

Mycorrhiza-Induced Changes in Disease Severity and PR Protein Expression in Tobacco Leaves

The development of leaf disease symptoms and the accumulation of pathogenesis-related (PR) proteins were monitored in leaves of tobacco (Nicotiana tabacum cv. Xanthinc) plants colonized by the arbuscular mycorrhizal fungus Glomus intraradices. Leaves of mycorrhizal plants infected with the leaf pathogens Botrytis cinerea or tobacco mosaic virus showed a higher incidence and severity of necrotic lesions than those of nonmycorrhizal controls. Similar plant responses were obtained at both low (0.1 mM) and high (1.0 mM) nutritional P levels and with mutant plants (NahG) that are unable to accumulate salicylic acid. Application of PR-protein activators induced PR-1 and PR-3 expression in leaves of both nonmycorrhizal and mycorrhizal plants; however, accumulation and mRNA steady-site levels of these proteins were lower, and their appearance delayed, in leaves of the mycorrhizal plants. Application of 0.3 mM phosphate to the plants did not mimic the delay in PR expression observed in the mycorrhizal tobacco. Together, these data strongly support the existence of regulatory processes, initiated in the roots of mycorrhizal plants, that modify disease-symptom development and gene expression in their leaves.

Biological control of Botrytis-incited diseases and powdery mildews in greenhouse crops

Abstract The greenhouse environment is very conducive to several diseases, among which grey mould and powdery mildews are the most important ones. Currently, much attention is given worldwide to the biological and integrated means of control of these two diseases, yielding reports on many potential antagonists. The two diseases differ greatly with respect to their epidemiology and the physiology of the pathogens. Hence, different approaches to their biocontrol are adopted. Biocontrol of powdery mildews is mostly based on the application of hyperparasitic fungi. In order to achieve a high level of parasitism, the biocontrol agents need a low vapour pressure deficit (VPD), compared to the VPD needed for the development of powdery mildews. Therefore, measures are necessary to overcome this problem. On the other hand, biocontrol of the high humidity-promoted Botrytis cinerea , is mostly based on saprophytes. The efficacy of biocontrol agents and their survival are dependent on biotic and abiotic factors. So far, results obtained under commercial conditions arc rather moderate, but even so there are biocontrol agents suitable for integration with other control measures in order to achieve a control level acceptable to growers. Formulations of biocontrol agents against B. cinerea and powdery mildews have already been released and registered in some countries. They are only moderately effective but are suitable for use in integrated control of the target diseases.

The role of phytohormones in basal resistance and Trichoderma-induced systemic resistance to Botrytis cinerea in Arabidopsis thaliana

Thirty-six phytohormone-affected mutants of Arabidopsis thaliana (L.) Heynh. and their parental ecotypes were tested for resistance/susceptibility to Botrytis cinerea Pers.; Fr. and ability to develop Trichoderma-mediated induced systemic resistance (ISR). Ecotype Colombia-0 (Col-0) was relatively resistant to B. cinerea, and Trichoderma harzianum Rifai T39 application at sites spatially separated (roots) from the B. cinerea inoculation (leaves) resulted in reduction of grey mold symptoms. Ecotypes Wassilewskija-4, Nossen-0 and Landsberg-0 had low levels of basal resistance to B. cinerea and were unable to express ISR. Mutants derived from ISR-non-inducible ecotypes displayed ISR-non-inducible phenotypes, whereas the ISR inducibility of mutants derived from the ISR-inducible genotype Col-0 varied according to the type of mutant. Thus, salicylic acid (SA)-impaired mutants derived from Col-0 were ISR-inducible, while ethylene/jasmonic acid (ethylene/JA)-impaired mutants of the same origin were ISR-non-inducible. SA-impaired mutants retained basal level of resistance to B. cinerea, while most ethylene/JA-impaired mutants were highly susceptible. Abscisic acid- and gibberellin-impaired mutants were highly susceptible to B. cinerea and showed ISR-non-inducible phenotypes irrespective of their lines of origin. Auxin-resistant mutants derived from Col-0 were ISR-inducible; mutant originating from Landsberg-0 and mutants which were resistant to both auxin and ethylene were ISR-non-inducible. Most of the arabidopsis genotypes which were unable to express Trichoderma-mediated ISR against B. cinerea exhibited enhanced susceptibility to this pathogen. T. harzianum treatments enhanced the growth of arabidopsis plants regardless of genotype or ISR inducibility.

Biological control of grape grey mould by Trichoderma harzianum

Abstract Biological and chemical control of grey mould was tested in vineyards of table and wine grapes. Treatments with Trichoderma harzianum (0.5-1.0 g 1 −1 ), dicarboximide fungicides (vinclozolin or iprodione) (0.5 g 1 −1 ) or diethofencarb plus carbendazim (0.25 g 1 −1 ) resulted in up to 78% disease reduction. A tank mix of the biocontrol agent with a dicarboximide fungicide was not superior to either treatment alone. It is suggested that alternate sprays of the biocontrol preparation with a fungicide should be employed in vineyards in order to reduce the use of chemicals. Populations of Trichoderma on grapes treated with the biocontrol agent were 4.5 × 10 5 per berry compared with 400–2000 per berry on untreated bunches. The application of T. harzianum in the vineyard did not interfere with processes involved in wine production from the grapes. T. harzianum and iprodione applied alone in the vineyard reduced the postharvest rot of grapes in one of two experiments. Alternation of T. harzianum with diethofencarb plus carbendazim, or its mixture with iprodione in the vineyard, resulted in a 64–68% reduction in postharvest rot caused by Botrytis cinerea .