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The stimulation exerted by the endophytic bacterium Bacillus pumilus strain SE34 in plant defense reactions was investigated at the ultrastructural level using an in vitro system in which root-inducing T-DNA pea (Pisum sativum L.) roots were infected with the pea root-rotting fungus Fusarium oxysporum f. sp. pisi. In nonbacterized roots, the pathogen multiplied abundantly through much of the tissue including the vascular stele, whereas in prebacterized roots, pathogen growth was restricted to the epidermis and the outer cortex In these prebacterized roots, typical host reactions included strengthening the epidermal and cortical cell walls and deposition of newly formed barriers beyond the infection sites. Wall appositions were found to contain large amounts of callose in addition to being infiltrated with phenolic compounds. The labeling pattern obtained with the gold-complexed laccase showed that phenolics were widely distributed in Fusarium-challenged, bacterized roots. Such compounds accumulated in the host cell walls and the intercellular spaces as well as at the surface or even inside of the invading hyphae of the pathogen. The wall-bound chitin component in Fusarium hyphae colonizing bacterized roots was preserved even when hyphae had undergone substantial degradation. These observations confirm that endophytic bacteria may function as potential inducers of plant disease resistance.

Experiments were designed to evaluate the effectiveness of several methods for delivering 15 endophytic bacteria into cotton stem and root tissues. The delivery methods included stab-inoculation of bacteria into stems, soaking seeds in bacterial suspensions, methyl cellulose seed coating, foliar spray, bacteria-impregnated granules applied in-furrow, vacuum infiltration and pruned-root dip. The success of delivery was gaged by recovery of the bacteria from internal plant tissues 2 weeks after the plants had been grown in a glasshouse potting mix. Following stab-inoculation into stems or radicles, 10 of the bacterial endophytes which previously exhibited biological control against fusarium wilt of cotton were successfully re-isolated from 50% of the plants inoculated; however, this method was labor-intensive, involved wounding the plant and sometimes reduced plant growth. Four of the other methods established from six to eight of the 15 strains, and, with some strains, all methods effectively established endophytic bacteria, based on re-isolation of strains from internal tissues 2 weeks after inoculation. A method was developed which allowed more convenient isolation of endophytes from a large number of plants. The results suggest that introduction of beneficial endophytic strains into cotton plants could be accomplished by practical methods chosen specifically for each strain.

The efficacy of 2-furfuraldehyde for control of Sclerotium rolfsii was studied in laboratory and greenhouse experiments. Mycelial growth of the fungus was reduced proportionally with concentrations of 0.1-0.5 ml furfuraldehyde l-1 agar medium, and viability of sclerotia diminished on exposure to 2-furfuraldehyde vapours. Detectable populations of bacteria and fungi, including Trichoderma spp., were reduced significantly (9=0.05) when furfuraldehyde was added to the agar used for soil dilution plates of untreated soil. Repeated treatments of natural soil with the fumigant significantly increased populations of Trichoderma spp. and bacteria, but diminished numbers of actinomycetes. Increasing dosages applied to soil artificially infested with S. rolfsii caused a reduction of disease on lentil, Lens culinaris. Results indicate that the compound, when applied to field soil, changes the composition of soil microflora and has potential for integrated control of S. rolfsii.

Citral (3 , 7 - dimethyl - 2 , 6 - octadienal) , furfural (2 - furaldehyde) and benzaldehyde (benzoic adel hyde) previously demonstrated control activity against Meloidogyne incognita and fungal diseases on cotton . Plant growth - promoting rhizobacteria (PGPR) applied to cotton were previously found to promote plant growth and reduce seedling disease . Studies were under taken to determine if these compounds were compatible with PGPR . In tests with 12 PGPR strains , vapor of citral inhibited in vitro growth of most strains , and vapor of furfural and benzaldehyde , with one exception , killed all but the Bacillus spp . tested . When 0 . 35 ml kg 1 soil of each compound were applied to the soil 9 - 10 days prior to planting the cotton cultivar Deltapine 51 , only furfural significantly reduced rhizosphere colonization across all strains from 4 . 70 colony - forming units (CFUs) / g of root to 4 . 42 CFUs / g root . In greenhouse studies , the low seed - surface pH (2 . 3) of commercial seed did not reduce root colonization , compared with colonization on roots from seed at pH 5 . 4 . There were no synergistic interactions between seed - surface pH and any of the compounds . Although previous research indicated that application of both furfural and benzaldehyde increased the proportion of Burkholderia spp . in the soil , there is no indication that they increased cotton root colonization by the B. cepacia strain tested . These results indicate PGPR can be combined with citral and benzaldehyde in integrated management systems and that the low seed - surface pH of acid - delinted cotton will not limit their application .

Ten genera from 9 families of dipterous insects collected in the field in the San Luis Valley of Colorado were contaminated withErwinia carotovoravar.atroseptica (Eca)andErwinia carotovoravar.carotovora (Ecc).Erwinia-contaminated insects were collected from settling ponds near potato warehouses, potato cull piles on growers’ farms, municipal dumps, lettuce and potato fields. The percentage of contaminated insects reached 14.5% in early spring but decreased as the season progressed. Insects were more commonly contaminated withEcathanEccearly in the season. Later the proportion of contamination byEccincreased while that ofEcadecreased. We suggest that insects may play a potentially important role in the epidemiology of potato blackleg especially in the reintroduction ofErwiniaintoErwiniafreepotato fields.

Adult fruit flies (Drosophila melanogasterMeig.) artificially contaminated withErwinia carotovoravar.carotovora(Jones) Dye (Ecc) and/orE. carotovoravar.atroseptica(van Hall) Dye (Eca) readily transmitted the bacteria to plants in the field that had been injured by crushing the stem. Injured, inoculated plants developed disease symptoms when maintained at high relative humidities.Erwiniawas transmitted to ten-hour-old injuries and they became infected as frequently as freshly made wounds. Insect transmission ofEca, Ecc, and mixtures was greatest during the afternoon, which was the warmest part of the day.Eccwas transmitted significantly less frequently during the cold morning than during afternoon or evening hours. A potato cull pile placed in a commercial potato field attracted a natural insect population which increased during the season. BothEccandEcawere isolated from uninoculated rotting tubers in the cull pile and from insects associated with the pile from May through September. These naturally infested insects transmittedEccand/orEcafrom the cull pile to artificially injured field plants during July and August at distances as great as 183 m from the cull pile. NoErwiniawas isolated from injured plants in a neighboring control field, which lacked a cull pile, further than 6 m upwind from the cull pile. We suggest that insects are important agents in the epidemiology of potato blackleg and soft rot even in areas with low relative humidities.

Recent works suggest that the combination of several PGPRs could be more effective than individual strains as a horticultural product. LS213 is a product formed by a combination of two PGPRs, Bacillus subtilisstrain GB03 (a growth-promoting agent), B. amyloliquefaciensstrain IN937a (an inducer of systemic resistance) and chitosan. The aim of this work is to establish if the combination of three PGPR, B. licheniformisCECT 5106, Pseudomonas fluorescensCECT 5398 and Chryseobacterium balustinumCECT 5399 with LS213 would have a synergistic effect on growth promotion and biocontrol on tomato and pepper against Fusariumwilt and Rhizoctoniadamping off. When individual rhizobacterium and the LS213 were put together, the biometric parameters were higher than with individual rhizobacterium both in tomato and pepper, revealing a synergistic effect on growth promotion, being the most effective combination that of B. licheniformisand LS213. When P. fluorescensCECT 5398 was applied alone, it gave good results, which could be due to the production of siderophores by this strain. Biocontrol results also indicate that those treatments that combined LS213 and each of the bacteria (Treatments: T7 and T8) gave significantly higher percentages of healthy plants for both tomato (T7: 65%) and pepper (T7: 75% and T8: 70%) than the LS213 alone (45% of healthy plants for tomato and 60% for pepper) three weeks after pathogen attack. The effects in pepper were more marked than in tomato. The best treatment in biocontrol was the combination of P. fluorescensand LS213. In summary, the combination of microorganisms gives better results probably due to the different mechanisms used.