Induced Systemic Resistance (ISR) by Pseudomonas spp. Impairs Pre- and Post-Infection Development of Pythium aphanidermatum on Cucumber Roots

The effect of induced systemic resistance (ISR) by Pseudomonas rhizobacteria on the pre- and post-infection development of Pythium aphanidermatum on cucumber roots was investigated. Cucumber plants (cv. Corona) were grown in vermiculite, roots were split with one side bacterized with Pseudomonas corrugata strain 13 or P. aureofaciens strain 63-28 (bacterized roots) and the other distant side was treated with water (distant, induced roots). For the non-induced control, roots on the bacterized side were treated with buffer instead of the bacterial treatment. Intact, non-split roots were also treated with the bacteria or buffer as a control. Cucumber root tissue from these treatments were harvested and incubated with a zoospore suspension of P. aphanidermatum for three hours. Most of the zoospores in the suspension were stimulated to encyst or germinate. The numbers of germinated zoospores were significantly decreased on distant induced cucumber roots in comparison to non-induced controls. Germination was also reduced on intact bacterized roots, compared to controls. There was less attachment, germ tube production and penetration on roots bacterized or induced by the rhizobacteria compared to non-induced roots. Effects were significantly greater on bacterized roots (roots colonized by bacteria) compared to distant induced roots (roots with the opposite side bacterized). Systemic resistance induced by the two Pseudomonas spp. also reduced pathogen spread on split cucumber roots in planta. Crown infection from induced or bacterized roots was delayed for four to six days in comparison to the non-induced control. Results indicated that Pseudomonas spp. can exert both an indirect influence on P. aphanidermatum zoospore behaviour and infection via induced systemic resistance (ISR) and a local influence via antibiosis or local induced resistance.     

Pseudomonas aeruginosa 7NSK2-induced Systemic Resistance in Tobacco Depends on in planta Salicylic Acid Accumulation but is not Associated with PR1a Expression

Root colonization by rhizobacteria can induce a systemic resistance in plants that is phenotypically similar to systemic acquired resistance induced by a localized pathogen infection. We used the tobacco–tobacco mosaic virus model to investigate whether the systemic resistance induced by the rhizobacterium Pseudomonas aeruginosa 7NSK2 is mediated by the systemic acquired resistance signal transduction pathway. Experiments with nahG-transformed tobacco revealed that Pseudomonas aeruginosa 7NSK2-induced resistance depended on in planta salicylic acid accumulation for its expression but not for its induction and is, in this respect, similar to systemic acquired resistance. However, Pseudomonas aeruginosa 7NSK2-induced resistance was, unlike systemic acquired resistance, not associated with PR1a expression at the time of challenge with tobacco mosaic virus. This suggests that Pseudomonas aeruginosa 7NSK2 treatment would only potentiate defense gene expression in systemic tissue, which would also explain why its level of resistance is lower than in case of systemic acquired resistance. Because we demonstrated that induced resistance by Pseudomonas aeruginosa 7NSK2 exclusively depends on the production of salicylic acid by this strain our conclusions might also account for other salicylic acid-producing and resistance-inducing rhizobacteria.     

Induction of Systemic Resistance in Cucumber against Several Diseases by Plant Growth-promoting Fungi: lignification and Superoxide Generation

Five fungal isolates (Trichoderma, Fusarium, Penicillium, Phoma and a sterile fungus) from zoysiagrass rhizosphere that promote plant growth were tested for their ability to induce systemic resistance in cucumber plants against Colletotrichum orbiculare. Roots of cucumber plants were treated with these fungal isolates using barley grain inocula (BGI), mycelial inocula (MI) or culture filtrate (CF). Most isolate/inoculum form combinations significantly reduced the disease except BGI of Trichoderma. These fungal isolates were also evaluated for induction of systemic resistance against bacterial angular leaf spot and Fusarium wilt by treatment with BGI. Penicillium, Phoma and the sterile fungus significantly reduced the disease incidence of bacterial angular leaf spot. Phoma and sterile fungus protected plants significantly against Fusarium wilt. Roots treated with CFs of these fungal isolates induced lignification at Colletotrichum penetration points indicating the presence of an elicitor in the CFs. The elicitor activity of CFs was evaluated by the chemiluminescence assay using tobacco callus and cucumber fruit disks. The CFs of all isolates elicited conspicuous superoxide generation. The chemiluminescence activity of the CF of Penicillium was extremely high, and its intensity was almost 100-fold higher than that of other isolates. The chemiluminescence activity was not lost following treatment with protease or autoclaving or after removal of lipid. The MW 12,000 dialyzed CF fraction was highly effective in eliciting chemiluminescence activity. Chemiluminescence emission from cucumber fruit disks treated with Penicillium was the same as that obtained from tobacco callus, except that the lipid fraction also showed a high activity. Both the MW 12,000 fraction and the lipid fraction induced lignification in the epidermal tissues of cucumber hypocotyls.     

Application of Rhizobacteria for Induced Resistance

This article provides a review of experiments conducted over a six-year period to develop a biological control system for insect-transmitted diseases in vegetables based on induced systemic resistance (ISR) mediated by plant growth-promoting rhizobacteria (PGPR). Initial experiments investigated the factors involved in treatment with PGPR led to ISR to bacterial wilt disease in cucumber caused by Erwinia tracheiphila. Results demonstrated that PGPR-ISR against bacterial wilt and feeding by the cucumber beetle vectors of E. trachiphiela were associated with reduced concentrations of cucurbitacin, a secondary plant metabolite and powerful beetle feeding stimulant. In other experiments, PGPR induced resistance against bacterial wilt in the absence of the beetle vectors, suggesting that PGPR-ISR protects cucumber against bacterial wilt not only by reducing beetle feeding and transmission of the pathogen, but also through the induction of other plant defense mechanisms after the pathogen has been introduced into the plant. Additional greenhouse and field experiments are described in which PGPR strains were selected for ISR against cucumber mosaic virus (CMV) and tomato mottle virus (ToMoV). Although results varied from year to year, field-grown tomatoes treated with PGPR demonstrated a reduction in the development of disease symptoms, and often a reduction in the incidence of viral infection and an increase in tomato yield. Recent efforts on commercial development of PGPR are described in which biological preparations containing industrial formulated spores of PGPR plus chitosan were formulated and evaluated for use in a transplant soil mix system for developing plants that can withstand disease attack after transplanting in the field.     

Study of Bacterial Determinants Involved in the Induction of Systemic Resistance in Bean by Pseudomonas putida BTP1

The ability of Pseudomonas putida BTP1 to induce resistance in bean to Botrytis cinerea was demonstrated in soil experiments on plants pre-inoculated at the root level with the bacteria before challenge with the leaf pathogen. As a first step to characterize the molecules from BTP1 responsible for induction of systemic resistance in bean, heat-killed cells and supernatant from culture in an iron-limited medium were tested for their protective effect. Most of the resistance-eliciting activity of the strain was retained in the crude cell-free culture fluid. In vivo assays with samples from successive fractionation steps of the BTP1 supernatant led, (i) to the conclusion that salicylic acid, pyochelin and pyoverdin, previously identified as Pseudomonas determinants for induced systemic resistance (ISR), were not involved in systemic resistance triggered by BTP1, and (ii) to the isolation of fractions containing one main metabolite that retained most of the resistance-inducing activity in bean. Although this molecule remains to be structurally characterized, its isolation is an addition to the range of determinants from plant growth-promoting rhizobacteria (PGPR) known to stimulate plant defences.     

Searching by Differential Display for New Genes Implicated in Induced Disease Resistance of Cucumber Treated with Acibenzolar-S-methyl

Acibenzolar-S-methyl (ASM) is a chemical activator of systemic disease resistance in plants. In this study, we used differential display to identify ASM-inducible defense response genes involved in induced disease resistance. As a result, we cloned three ASM-inducible genes from cucumber, encoding a chitinase, a putative protein disulfide isomerase and a putative mitochondrial-protein-like protein. Expression of these genes was induced within 24 hr after treatment of cucumber leaves with ASM. These results suggest that differential display is a useful tool for understanding the mode of action of ASM and defense responses. Received 6 September 2000/ Accepted in revised form 11 April 2001     

Induced Resistance in Hypocotyl of Cucumber by Infection with Colletotrichum lagenarium in Leaves

Localized infection of cucumber leaves with Colletotrichum lagenarium induced resistance against infection after challenge inoculation with pathogenic fungi, including C. lagenarium and Pythium ultimum var. ultimum. Systemic acquired resistance in the hypocotyl was observed when challenge inoculation was made 4 to 7 days after the first inoculation of cotyledons. Seven days after the first inoculation of a primary leaf, induced resistance against the challenge inoculation in the hypocotyl was also observed. However, the hypocotyl did not develop induced resistance when plants were challenged by a wound inoculation with P. ultimum. Received 9 June 1999/ Accepted in revised form 13 December 1999     

Reduction of Bacterial Speck (Pseudomonas syringae pv. tomato) of Tomato by Combined Treatments of Plant Growth-promoting Bacterium, Azospirillum brasilense, Streptomycin Sulfate, and Chemo-thermal Seed Treatment

Inoculation of tomato seeds with the plant growth-promoting bacterium Azospirillum brasilense, or spraying tomato foliage with A. brasilense, streptomycin sulfate, or commercial copper bactericides, separately, before or after inoculation with Pseudomonas syringae pv. tomato, the casual agent of bacterial speck of tomato, had no lasting effect on disease severity or on plant height and dry weight. Seed inoculation with A. brasilense combined with a single streptomycin foliar treatment and two foliar bactericide applications at 5-day intervals (a third or less of the recommended commercial dose) reduced disease severity in tomato seedlings by over 90% after 4 weeks, and significantly slowed disease development under mist conditions. A. brasilense did not induce significant systemic resistance against the pathogen although the level of salicylic acid increased in inoculated plants. Treatment of tomato seeds that were artificially inoculated with P. syringae pv. tomato, with a combination of mild chemo-thermal treatment, A. brasilense seed inoculation, and later, a single foliar application of a copper bactericide, nearly eliminated bacterial leaf speck even when the plants were grown under mist for 6 weeks. This study shows that a combination of otherwise ineffective disease management tactics, when applied in concert, can reduce bacterial speck intensity in tomatoes under mist conditions.