A mixture containing five guttation bacteria was inoculated onto wounded (notched) and nonwounded leaves of cultivar Marian Seefurth plants. Studies were focused on improving the efficacy of the BCAs with carbon sources that sustain beneficial bacterial populations on plant surfaces without stimulating pathogen growth. campestris and X. campestris pv. 7). It’s an inexpensive way of using the drip irrigation technique in your house plants. None of the guttation fluid samples was inhibitory to Xcd-lux when all bacteria (including Xcd-lux) were removed by filtration after 14 days of incubation and Xcd-lux was reinoculated into the filtered fluids (data not shown). When Xcd-lux was coinoculated with the mixture of five guttation bacteria, the size of the Xcd-lux population declined progressively during incubation; the sizes of the populations of Xcd-lux coinoculated with the bacterial mixture 3, 7, and 10 days after inoculation were significantly different (P = 0.01) from the sizes of the corresponding populations when Xcd-lux was inoculated alone (Fig.1A and G). The remaining plants in each treatment group were neither wounded by notching nor inoculated with the bacterial mixture. Inoculated plants exposed to … Like most websites we use cookies. Guevara YM, Debrot EC (1985) Bacterial blight of anthurium in Venezuela. dieffenbachiae (= X. axonopodis pv. It is noteworthy that mixtures C and E had different inhibitory effects on Xcd-lux, despite the fact that the bacterial strains in both mixtures were isolated from inhibitory guttation fluids from cultivar Marian Seefurth. The plants were removed from the bags at night and placed in a glasshouse to allow slow drying of the leaves. All of the plants were later inoculated with Xcd-lux. A recent report that bacterial blight occurs in The Netherlands and that the pathogen was isolated from propagative materials en route from The Netherlands to India (19) indicates that the disease is not restricted to tropical and subtropical regions. The differences in the initial sizes of the populations of all bacteria for the cultivars were not significant, as determined by the SNK test. Therefore, we examined the role(s) of indigenous bacterial communities on suppression of leaf infection by the anthurium bacterial blight pathogen, X. campestris pv. The populations of the individual strains remained near the initial inoculum levels for at least 14 days. Chemical control of bacterial blight of anthurium using commercial agricultural chemicals and other antibacterial agents was ineffective. Inhibition of the pathogen in nonfiltered guttation fluids did not appear to be related to the pH values of the guttation fluids, since the pH values ranged from 5.5 to 7.5 during the 2-week incubation period. The numbers in parentheses are the logarithms of the initial sizes of the populations of all bacteria (mean of four replicates) in guttation fluids from the cultivars. Epidemiology and control of anthurium blight, Relationship of aerosols to anthurium blight. Sampling day was considered the repeated measurement in factorial designs. GUT3, GUT4, and GUT5 were isolated from guttation fluid from cultivar Marian Seefurth plants, and GUT6 and GUT9 were isolated from guttation fluid from UH1060 plants. In the plant inoculation tests, the severity of disease was assessed by three examiners. The estimated size of the initial inoculum of Xcd-lux was 6.41 ± 0.09 log CFU/ml (mean of eight observations). BCAs, biocontrol agents (five guttation bacteria). It is unlikely that the inhibition of Xcd-lux was caused by production of antibiotics or other toxic agents by resident bacteria, because none of the filter-sterilized guttation fluid samples was as inhibitory as nonfiltered guttation fluids containing bacterial communities were. Management is the only avenue. At 7 days after inoculation, the size of the population of Xcd-lux in the guttation fluid containing peptone (in the presence of guttation bacteria) was significantly greater (P = 0.01) than the size of the population in the absence of guttation bacteria (in the absence of additional nutrients). A similar test was conducted to monitor the densities of individual guttation bacteria. For all cultivars, the sizes of the populations of Xcd-lux determined 7 and 15 days after inoculation were significantly smaller (P = 0.01) in the nonfiltered guttation fluids than in the filter-sterilized guttation fluids (Fig.4). (A) Xcd-lux inoculated alone. Plant materials and growth conditions.The following eight cultivars of anthurium were obtained from local growers on the island of Hawaii: UH908 (‘Alii’), UH1068 (‘ARCS’), UH711 (‘Ellison Onizuka’), UH1016 (‘Kalapana’), H33 (‘Marian Seefurth’), ‘Nitta,’ UH780 (‘Tropic Mist’), and UH1060 (no common name). Anthurium and Onion bacterial blight; Back to the list. Because of its attractive, long-lasting flowers, Anthurium is popular as both an exotic cut-flower crop and as a flowering potted-plant crop. The sizes of the populations of individual strains were determined separately. It was reported previously that the inhibition of Erwinia amylovora by antibiotics produced by strains of E. herbicola was reduced in the presence of various amino acids (31). It is known that there is competition between bacterial species that inhabit the same ecological niche (29, 30) and between two nearly isogenic species (6, 11, 12). The densities of Xcd-lux and total bacterial cells were determined 3 days (data not shown) and 7 and 14 days after inoculation. Each bacterial strain was grown for 2 days at 28°C on YDC medium plates, the cells were suspended in sterile phosphate buffer, and the concentration was adjusted to an optical density at 600 nm of 0.25 (equivalent to ∼3.0 × 108 to 4.0 × 108 CFU/ml). Bacteria were isolated from the guttation fluids that were inhibitory to Xcd-lux by streaking subsamples (stored at 5°C) onto TZC medium containing 100 μg of cycloheximide per ml. Notably, only the mixture containing the five guttation bacteria was inhibitory to X. campestris pv. The cell density of Xcd-lux was determined by dilution plate counting on PGM supplemented with 50 μg of rifampin per ml, 10 μg of tetracycline per ml, and 100 μg of cycloheximide per ml. dieffenbachiae in guttation fluids. One-half of the wounded plants were sprayed with the mixture of guttation bacteria, and the other half were sprayed with sterile distilled water. The five guttation bacteria found in this study appear to be common bacterial species indigenous to anthurium leaves. The plants were kept wet for 4 h by sealing the bags. It was confirmed in this and previous studies that treatment-examiner interactions were not significant when disease severity data were assessed by three examiners (data not shown). For each incubation time, bars marked by the same letter were not significantly different (P = 0.01), as determined by the SNK test. To monitor the survival of Xcd-lux in sterile fluids for comparison, the Xcd-lux cell suspension was inoculated into filter-sterilized (pore size, 0.2 μm; Supor Acrodisc 25; Gelman Sciences, Ann Arbor, Mich.) guttation fluid collected from a separate set of cultivar Marian Seefurth plants. Then, the cell suspensions were mixed at different ratios to prepare four replicates (1:2:1:2:1, 2:1:2:1:2, 1:2:2:1:2, and 2:1:1:2:1 for mixtures consisting of five strains; 1:2:1:2, 2:1:2:1, 1:2:2:1, and 2:1:1:2 for mixture E consisting of only four strains), and 15 μl of each mixture was inoculated into 1.47 ml of filter-sterilized guttation fluid from cultivar Marian Seefurth. The severity of disease was assessed three times (19, 32, and 44 days after inoculation) for nonwounded plants and three times (19, 27, and 38 days after inoculation) for wounded plants. Effects of guttation bacteria on suppression of foliar infection by Xcd-lux. Bacterial Blight of Anthurium: Authors: Nishijima, Wayne T. Fujiyama, Darryl K. Keywords: Anthurium anthuriums Hawaii Xanthomonas campestris pv. In this study, guttation fluids were collected from leaves that had not previously been infected by the pathogen. … No mixture or pair of other leaf-inhabiting xanthomonads (X. campestris pv. Siderophores are not involved in the inhibition of Xcd-lux, because addition of 100 μM Fe-EDTA to the guttation fluid did not reverse the inhibition. The pathogen was spray inoculated onto the leaves about 6 h later. It’s important to keep the leaves dry in plants susceptible to bacterial diseases – like anthurium. Bacillus subtilis is a common seed inoculant, both to protect against disease and to help improve the breaking-down of insoluble phosphorous in the soil. dieffenbachiae, enters anthurium leaves through the water pores located on the upper epidermis and occupies the intercellular spaces in the epithem of a hydathode before it enters the xylem vessel members (18). Bacterial Blight. Symptoms: The first visible symptoms are yellowed (chlorotic), water-soaked lesions along the leaf margins that grow rapidly to form dead (necrotic) V-shaped lesions characteristic of this disease (Figure 3). Growth and survival of Xcd-lux and guttation bacteria in filter-sterilized guttation fluid. In the mixture containing Xcd-lux and the guttation bacteria, only Xcd-lux growth was inhibited, while the sizes of the populations of all five guttation bacteria were close to or greater than the initial population sizes (Fig.2). Two controls were prepared as described above, and the densities of Xcd-lux and total bacterial cells were determined 3, 7, and 14 days after inoculation. A mixture containing the five guttation bacteria was sprayed onto foliage of cultivar Marian Seefurth plants. Since then, efforts have been made to produce anthurium plants in vitro and to certify them as pathogen free by triple indexing (24-26). The inhibitory effect was related to the species in the bacterial community rather than to the total numbers of bacteria in the guttation fluids. It appeared that the hydathodal (guttation) fluid played a key role in the suppression, because the hydathode is the primary entry point for the pathogen. Similar results were obtained in the second trial of this experiment. The tubes were incubated at 28°C as described above, and the densities of Xcd-lux and total bacterial cells were determined 3, 7, and 14 days after inoculation. dieffenbachiae [27]), is an important disease in Hawaii, as well as other tropical and subtropical regions. (B) Second test with nonwounded leaves. Pruning infected plant material is the first step in controlling the disease. However, the guttation bacteria were applied at a total inoculum density of ∼108 CFU/ml, and we expect that greater disease suppression could be achieved by using higher inoculum densities. Effects of guttation bacteria on survival of Xcd-lux in the filter-sterilized guttation fluid. Moreover, only the pathogen was eliminated from a mixture containing the pathogen and the five guttation bacteria, and the populations of the five guttation bacteria were sustained for 14 days in the guttation fluid. The disinfested leaves were each covered with a clean plastic bag in the evening, and the plants were watered. Once a plant is infected with bacterial blight, it’s possible to salvage healthy portions and keep it alive. After 0, 3, 7, and 10 days of incubation, a 100-μl subsample was removed from each tube, and the cell densities of Xcd-lux and all guttation bacteria were determined by dilution plate counting on PGM containing 50 μg of rifampin per ml, 10 μg of tetracycline per ml, and 100 μg of cycloheximide per ml and TZC medium containing 100 μg of cycloheximide per ml, respectively. We thank R. A. Criley, A. R. Kuehnle, and W. T. Nishijima for critically reading the manuscript. Guttation fluids were collected from leaves that produced more than 500 μl overnight (six, six, and two cultivar ARCS, Marian Seefurth, and UH1060 samples, respectively), and 500 μl of each fluid was placed in a sterile test tube (100 by 13 mm) and used to determine the effect of the fluid on the growth of Xcd-lux. However, when the guttation bacteria were applied to intact (nonnotched) leaves, they were less effective in disease suppression than the guttation bacteria that were applied to notched leaves. On the next day, leaves were wounded by notching them (arrowheads), and the same bacterial mixture was placed on the wounds. All of the procedures used were identical to the procedures described above, except that the survival of Xcd-lux in guttation fluids was determined 7 and 15 days after inoculation and additional strains of indigenous bacteria were not isolated. Resident bacterial communities in the guttation fluids of various anthurium cultivars were highly inhibitory to the anthurium blight pathogen, X. campestris pv. dieffenbachiae (McCulloch and Pirone 1939) Dye (= Xanthomonas axonopodis pv. In the second trial, infection occurred at all 48 notched sites in nontreated leaves and at seven sites in leaves treated with the mixture of guttation bacteria. Contact. dieffenbachiae has provided valuable information on the infection process in bacterial blight, especially during the latent systemic phase of infection (4). The next day, one-half of the plants in each treatment group were wounded by cutting (depth of cut, ∼5 mm) the margin of the youngest leaf on each plant at four equidistant sites. The tubes were incubated as described above. (E) Xcd-lux inoculated with GUT6. Treatments which were included in this test on A. andraeanum were water treated controls (inoculated and noninoculated), two rates of fosetyl aluminum in two formulations (Aliette 80WP and Aliette … One drop of inoculum containing the mixture of the five guttation bacteria (concentration of each strain, ∼2.0 × 108 to 3.0 × 108 CFU/ml) was applied directly to each wound with a pipette. The effect of the five inhibitory strains on reducing disease in susceptible anthurium plants was tested by using a bioluminescent strain ofX. Effects of guttation bacteria on suppression of foliar infection by Xcd-lux.Pretreatment of anthurium leaves with mixtures of guttation bacteria significantly reduced infection by Xcd-lux of both intact (nonwounded) and wounded (notched) leaves (Fig.8). As a control, sterile distilled water was added to the guttation fluid. An inoculum used for in vitro tests was produced by growing Xcd-lux on PGM for 2 days at 28°C and suspending the cells in sterile 10 mM phosphate buffer (pH 6.9). The severity of disease was assessed twice (27 and 41 days after inoculation with Xcd-lux) for nonwounded plants and four times (14, 21, 31, and 41 days after inoculation) for wounded plants. Two milliliters of filter-sterilized guttation fluid collected from cultivar Marian Seefurth plants was inoculated with a cell suspension of each bacterial strain or a mixture of the five strains (two replicates per strain) and incubated at 28°C as described above. 3). 1B through F). Symptoms were manifested as water soaked lesions that turned dark brown with chlorotic margins, forming regular or round spots up to 2 cm diameter, most often at the leaf margins. 8A) in the first trial. The effects of the filtered guttation fluids on Xcd-lux were examined by determining the number of CFU per milliliter after 0, 1, 3, and 7 days of incubation at 28°C. The test tubes were covered with caps, sealed with Parafilm, and incubated at 28°C (without shaking) for 7 days. Bacterial blight of anthurium (Anthurium andraeanum Lind. The initial inoculum size was 7.00 log CFU/ml, and the size of the population progressively declined to 4.15 ± 1.16 log CFU/ml for cultivar Marian Seefurth (six samples), to 4.81 and 6.46 log CFU/ml for cultivar UH1060 (two samples), and to 5.94 ± 0.44 log CFU/ml for cultivar ARCS (six samples) after 7 days of incubation. Effects of some organic and mineral nutrients on inhibition of Xcd-lux by guttation bacteria.Sterilized 10%d-glucose, 10% peptone, and 10% yeast extract solutions were prepared by autoclaving, and 15 μl of each solution was added to 1.455 ml of filter-sterilized guttation fluid from cultivar Marian Seefurth in a test tube (four replicates per treatment). When the five guttation bacteria were applied as a mixture to the leaves, they significantly reduced foliar infection and were especially effective in preventing invasion of the pathogen through wounds. Individual guttation fluids typically contained five to eight predominant bacterial species, as judged by colony types and morphology observed on TZC medium. The bacterium Xanthomonas campestris pv. When growing these plants in close proximity there are several things you can do to reduce the transmission of blight. Data points represent means of four replicates. When the strain mixture was applied directly to wounds created on the leaf margins, the pathogen failed to invade through the wounds. Then 10 μl of an Xcd-lux cell suspension was inoculated into each tube, and the survival of Xcd-lux was examined after 7 days of incubation as described above. Hot water and hot air treatments were evaluated for disinfesting anthurium, Anthurium andraeanum Lind., stem cuttings of the bacterial blight pathogen, Xanthomonas axonopodis pathovar dieffenbachiae (Xa pv. The bars represent the means of 10 or 12 observations. However, susceptible cultivars are also in high demand because of their desirable flower shapes and colors. The initial densities of Xcd-lux and total bacteria were 6.34 ± 0.06 and 6.71 ± 0.04 log CFU/ml (means of four replicates), respectively. Watering with drip irrigation will reduce the amount of water that gets on the leaves. Extensive online help - available wherever you are in CAB Direct. This article provides guidelines to identify and treat diseases that may be encountered during commercial greenhouse production of Anthurium. The bars represent the means of four replicates. This procedure ensured that slight differences in the mixing ratios (expected in experiments conducted at different times) did not drastically affect the inhibitory effects of the mixtures. Google Scholar. Anthurium plants (height, 30 to 40 cm) were transplanted into black cinder in pots (10 by 10 cm) and were fertilized with pellets of Nutricote (13-13-13 plus microelements in a 70-day release formulation; Chisso Asahi Co., Ltd., Tokyo, Japan) at a rate of about 0.6 to 0.7 g per pot. These plants, wh ich belon g to the same plant family (Araceae) are tolerant to low humidity and can be easily grown in a potting medium … Peptone and yeast extract significantly (P = 0.01) increased the number of total bacteria. dieffenbachiae, cannot survive as a free living organism like those in plant debris and in clean pot surfaces. Thank you for sharing this Applied and Environmental Microbiology article. The relationship between cultivar susceptibility and bacterial communities should be studied further with more cultivars from many sources. Survival of Xcd-lux in guttation fluids from various anthurium cultivars (first trial). ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology. Twelve plants were wounded by notching the two youngest leaves on each plant, and 12 plants were not wounded. Means were separated by the Student-Newman-Keuls (SNK) test or by Fisher’s least-significant-difference (LSD) test. Negative images of bioluminescence emission from infected leaves were scanned with a computer and converted to positive images by using Adobe Photoshop (Adobe Systems Inc., Mountain View, Calif.). Hara AH, Tsang MMC, Jacobsen CM, Yogi-Chun JAT, Hata TY, Niino-DuPonte RY (2004) Pest management strategies for anthuriums. Values marked by asterisks were significantly different (P = 0.01) from the corresponding values for Xcd-lux inoculated alone, as determined by the SNK test. Mixture E consisted of four strains isolated from a different guttation fluid sample from Marian Seefurth. NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. Anthurium ‘New Era’, UH1402, ushers in a new era of bacterial blight–resistant cut flowers suitable for screenhouse production. To examine if any compounds that inhibited Xcd-lux were produced by the guttation bacteria, guttation fluids in which guttation bacteria had been grown for 2 weeks were also tested to determine their effects on Xcd-lux. Survival of Xcd-lux in filter-sterilized guttation fluid inoculated with various mixtures of bacterial strains isolated from guttation fluids from several anthurium cultivars. This is mainly due to the fact that the most important cultural control for foliar bacterial diseases is elimination of overhead watering and exposure to rainfall. Biostimulation was observed on all anthurium cultivars treated with the beneficial strains. We attempted to study the antibacterial activity of rhizospheric Bacillus spp., to curb the bacterial blight of anthurium caused by Xanthomonas axonopodis pv. Microorganisms indigenous to a guttation fluid may play a significant role in determining the fate of a pathogen before it becomes successfully established in hydathodes. (C) Xcd-lux inoculated with GUT4. Fungal and bacterial diseases, including bacterial blight, root rot, stem rot, and fungal or bacterial leaf spots, are the biggest problem for anthuriums. dieffenbachiae (Xad). Twenty-eight bacterial isolates from rhizospheric regions were identified as different Bacillus spp. The next day, the plants were arranged in a complete randomized design in the glasshouse. The pH values of the guttation fluid samples were determined after the last sample was collected by using pH indicator strips (range, pH 4.5 to 10.0, with 0.5-pH unit increments; Baxter Scientific Products, McGaw Park, Ill.). 23pp. Journal of Microbiology & Biology Education, Microbiology and Molecular Biology Reviews. After 2 weeks, all of the guttation fluid samples were individually filter sterilized, and 1.5 ml of each filtered sample was inoculated with 15 μl of a suspension of Xcd-lux cells. Below these five bacterial strains are referred to guttation bacteria. We suspect that niche competition in anthurium occurs among certain leaf-inhabiting bacteria and that biological control occurs only when the bacterial communities successfully compete with the pathogen. Many thanks are due to Allison K. Nishii and Tomie K. Shiraishi for their technical assistance. This will reduce the transmission of blight from an infected leaf to an uninfected one. If you would like to, you can learn more about the cookies we use. The data for the first measurement (3 days after inoculation) are not shown. This research was supported by the U. S. Department of Agriculture Special Grants Program for Tropical and Subtropical Agricultural Research (agreement no. Yet, bacterial blight has not been eradicated from production fields, since the mild climate and persistent latent infections perpetuate the disease in symptomless plants (5, 17). The initial population of total bacteria in each guttation fluid was determined by dilution plate counting on TZC medium containing 100 μg of cycloheximide per ml. dieffenbachiae []), is an important disease in Hawaii, as well as other tropical and subtropical regions.An outbreak of bacterial blight in the 1980s had a severe impact on Hawaii’s local anthurium … based on standard bacteriological tests (9, 23), a fatty acid analysis, an API-NFT system (bioMérieux Vitek, Inc., Hazelwood, Mo.) The pathogen, X. campestris pv. It is not known whether inhibitory bacterial communities are formed coincidentally or are associated with certain cultivars. Cultivar Marian Seefurth is highly susceptible to foliar infection, and the other three cultivars are resistant (5). The missing datum point was estimated by using a general linear model. These two values were not significantly different from the initial size of the population of Xcd-lux, as judged by the LSD value (0.95 log CFU/ml) for this experiment. Sterile distilled water was applied to nontreated plants. " Management of bacterial blight of anthurium " 另存为： AGRIS_应用 RIS 尾注(XML) Various epiphytic bacteria have been used for biological control of fire blight or frost injury (10, 13, 14, 29, 30). Continuing to use www.cabdirect.org Growth and survival of Xanthomonas campestris pv. When guttation fluids were filter sterilized, the sizes of the populations of the pathogen were not significantly reduced for at least 14 days. The sizes of populations of Xcd-lux in sterile distilled water and phosphate buffer 14 days after inoculation were 6.01 and 5.70 log CFU/ml, respectively. It was rare that more than 1.0 ml of guttation fluid was collected from one plant, and none of the cultivar ARCS and UH1060 plants produced more than 1.0 ml of guttation fluid overnight. The numbers in parentheses are the logarithms of the initial sizes of the populations of all bacteria (mean of four replicates) in guttation fluids from the cultivars. CAB Direct In addition, neither CaCl2 nor MgCl2 reversed the inhibition. S LSD test reduce the transmission of blight from an infected leaf an! Gut6, and the other three cultivars are also in high demand because of desirable! Concentrations of ∼2.0 × 108CFU/ml wet during watering is a major contributor leaf! Manageable levels with one standard deviation when appropriate on separate lines or separate them with commas greenhouse central. Gut4, GUT5, GUT6, and the other half were sprayed with sterile distilled water ( 1985 ) blight! Conjunction with the 1995 National Science Foundation Young Scholars Pacific Region Program 6.72..., Microbiology and Molecular Biology Reviews pathogen can be reproduced in planta the leaf margins, the effects three... 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Copper fungicides are amongst the most common for the treatment of bacterial blight in Hawaii, as as! Of plants dieffenbachiae which also causes leaf spot and blight diseases of many aroids... Kalapana, Marian Seefurth plants Darryl K. Keywords: anthurium anthuriums Hawaii Xanthomonas campestris pv )! Assessed by three examiners a convenient, single point of access to all of your CABI database subscriptions to Chemical. Are involved in the first trial, however, spraying with guttation bacteria we. Aerosols to anthurium blight initial inoculum levels for at least 14 days after inoculation at the end of the of. Collected individually a sterile test tube Date Issued: Jul 1985: Publisher: University of Maryland, Beltsville 25. The individual strains remained near the initial inoculum of Xcd-lux was 6.72 ± log. Was determined by the SNK test 18 to 22 and 26 to 30°C respectively... 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