Induction of pigmentation of Thielaviopsis basicola and other fungi by a bacterium isolated from soil

1970 ◽  
Vol 48 (10) ◽  
pp. 1879-1886 ◽  
Author(s):  
Z. A. Patrick ◽  
Maury Schlifer

A blue-pigmented bacterium was isolated from soil and as a contaminant in cultures of Thielaviopsis basicola isolated from diseased tobacco roots. When the bacterium, a species of Pseudomonas, was grown in culture in association with T. basicola, a blue pigmentation was observed in the fungus. In addition to pigmentation of hyphae and conidia, other deleterious effects were induced in the fungus, including cytoplasmic granulation, vacuolation, contraction of cellular contents, reduced growth, and subsequent death of the affected cells. Other fungi, including Fusarium oxysporum f. sp. lycopersici, Botrytis convoluta, Rhizopus sp., and Mucor sp., were similarly affected. Hyphal pigmentation and other effects were observed only when viable bacteria were growing in intimate contact with the fungus mycelium. The pigment-inducing substance was not secreted into the culture media and was not obtained from lysed bacteria. Pigmentation and the other characteristic effects on T. basicola were not observed when the fungus and bacteria were added to soil. Similarly no significant reduction in severity of black root rot of tobacco seedlings was obtained when the blue bacteria were added to T. basicola infested soil.

Plant Disease ◽  
2009 ◽  
Vol 93 (4) ◽  
pp. 354-362 ◽  
Author(s):  
Harun Toksoz ◽  
Craig S. Rothrock ◽  
Terrence L. Kirkpatrick

The efficacy of triazole and host resistance–inducing seed treatment chemicals was examined for black root rot on cotton caused by Thielaviopsis basicola in both artificially and naturally infested soils with and without nematodes. In naturally infested soil, myclobutanil was effective in reducing root and hypocotyl discoloration over a wide range of soil population densities. Treatments containing high rates (42 g a.i./100 kg seed) of myclobutanil provided greater reductions in disease than low rates (21 g a.i./100 kg seed) in some experiments. Acibenzolar-S-methyl applied to the seed reduced black root rot or colonization by T. basicola on seedlings in artificially infested soils. Rates of acibenzolar-S-methyl did not differ in efficacy. In controlled studies, root colonization by T. basicola was significantly lower when seeds were treated with both myclobutanil and acibenzolar-S-methyl than with either chemical alone. In naturally infested soil under low (24 CFU/g soil) and high (154 CFU/g soil) populations of T. basicola, a combination of myclobutanil and acibenzolar-S-methyl at the high rate resulted in the lowest root discoloration and colonization. The nematicide seed treatment abamectin improved the control of black root rot in the presence of Meloidogyne incognita. The semi-selective medium TB-CEN allowed the importance of T. basicola to be evaluated in the presence of other pathogens that contribute to the seedling disease complex on cotton by quantifying the isolation frequency and percent colonization of T. basicola.


1936 ◽  
Vol 14c (1) ◽  
pp. 11-26 ◽  
Author(s):  
A. A. Hildebrand ◽  
L. W. Koch

Similarity of organisms encountered in studies of black root of strawberry and of tobacco, respectively, carried out contemporaneously but independently, suggested the co-operative investigation, the results of which are embodied in this paper. Strawberry and tobacco seedlings growing (i) in seed-bed muck heavily infested with Thielaviopsis basicola (Berk.) Ferraris, and other organisms known to be pathogenic on tobacco, (ii) in soil from a commercial plantation where strawberry root rot had occurred in severe and typical form, and (iii) in greenhouse compost soil, were examined microscopically daily, commencing a few hours after germination and continuing throughout a period of four weeks. Organisms observed definitely within root tissues of both hosts included the "phycomycetous mycorrhizal" fungus, T. basicola (observed in plants grown in muck only), Rhizoctonia (Solani and endophytic orchid types), forms of Pythium, Asterocystis (Olpidiaster), certain unidentified fungi, a minute filamentous alga and nematodes. Organisms observed on the surface of roots included representatives of the genera Cylindrocarpon (Ramularia), Fusarium, Helminthosporium, Sphaeropsis, and Cephalothecium. The sequence of appearance, percentage occurrence, and parasitic capabilities of certain of the organisms varied in roots grown in the different soils. Because of early infection by, and ultimate almost universal occurrence of, the phycomycetous mycorrhizal fungus this organism received especial attention. Evidence based on certain morphological differences suggests the occurrence of strains of this organism. Of interest, too, is an alga invading living root tissue.From observations not limited alone to the examination of diseased roots of strawberry and tobacco, the authors are led to conclude, (1) that a root rot as it occurs in nature is extremely complex even in cases where a primary causal agent is recognized, and (2) that fungi representative of comparatively few groups or genera are "common factors" in root-rot complexes of different host plants.The technique described offers distinct advantages in that it permits a study of the sequence and severity of infection by the organisms involved in a root-rot complex; it reveals the occurrence of obligate parasites the presence of which would never be detected by the soil-plating, the Cholodny, or the tissue-isolation methods; and it is readily adaptable to the study of other root-rot complexes.


Plant Disease ◽  
2010 ◽  
Vol 94 (9) ◽  
pp. 1168-1168 ◽  
Author(s):  
W. S. Monfort ◽  
A. G. Carroll ◽  
M. J. Emerson ◽  
J. Fortner ◽  
C. S. Rothrock

Thielaviopsis basicola (Berk. & Broome) Ferraris (synonym Chalara elegans Nag Raj & Kendrick) is a soilborne plant-pathogenic fungus reported in many parts of the world. In Arkansas, T. basicola is found commonly in cotton fields (4). This fungus colonizes cortical tissue of seedlings under cool wet conditions, causing a dark brown or black discoloration of the roots and hypocotyls, resulting in stunted, slow-developing plants (4). In 2008, large areas of stunted soybean plants with shortened internodes were reported in a field in Phillips County, AR, where cotton had previously been produced. Soybean was planted in this field in early April when cool soil temperatures (~21 to 24°C) prevailed. Soybean plants at the v3 to v5 growth stages were observed to have extensive areas of black cortical root necrosis. Plant samples were collected and roots were excised, washed, and surface disinfested in a 10% NaOCl solution. Root segments were incubated on the carrot-based selective medium TB-CEN (3). T. basicola was isolated from incubated segments after 2 weeks at 21°C in the dark. Chlamydospore chains (44.8 to 56.0 × 8.4 to 11.2 μm) consisting of an average of six spores and endoconidia (8 to 30 × 3 to 5 μm) were observed with a compound microscope. In addition to plant tissue, soil was assayed and confirmed to be positive for T. basicola by the pour plate technique (3) with the medium TB-CEN. Greenhouse trials were conducted to confirm field observations. Soil from the Phillips County field was sterilized and reinfested with 100 CFU of chlaymdospore suspension per gram (dry weight) of soil. Fifty soybean seeds (cv. Schillinger 457) were planted in infested and sterilized soil and grown for 29 days. Results showed that 38% of plants germinated and survived in the T. basicola-infested soil compared with 71% in the sterile soil treatment. Fifteen of the nineteen plants that survived in the infested soil were positive for T. basicola, while all plants in the sterilized soil were negative for the fungus. Soybean has previously been reported to be a host of T. basicola worldwide, but North American reports have been confined to Canada and Michigan, where cool soil temperatures persist for longer periods during the early part of the growing season (1,2). To our knowledge, this is the first report of T. basicola being important in the growth of soybean in warmer latitudes where the pathogen has been observed frequently on cotton and tobacco. In areas where cotton has historically suffered seedling damage from T. basicola, black root rot may become important on soybean as production of the latter crop increases. Since the initial field observation and confirmation in 2008, multiple soybean fields in 10 Arkansas counties have been documented with black root rot, with an estimated 5 to 30% of plants in each field infected. References: (1) T. R. Anderson. Can. J. Plant Pathol. 6:71, 1984. (2) J. L. Lockwood et al. Plant Dis. Rep. 54:849, 1970. (3) L. P. Specht and G. J. Griffin. Can. J. Plant Pathol. 7:438, 1985. (4) N. R. Walker et al. Phytopathology 89:613, 1999.


2011 ◽  
Vol 101 (2) ◽  
pp. 223-230 ◽  
Author(s):  
Brantlee Spakes Richter ◽  
Kelly Ivors ◽  
Wei Shi ◽  
D. M. Benson

Wood-based mulches are used in avocado production and are being tested on Fraser fir for reduction of Phytophthora root rot, caused by Phytophthora cinnamomi. Research with avocado has suggested a role of microbial cellulase enzymes in pathogen suppression through effects on the cellulosic cell walls of Phytophthora. This work was conducted to determine whether cellulase activity could account for disease suppression in mulch systems. A standard curve was developed to correlate cellulase activity in mulches with concentrations of a cellulase product. Based on this curve, cellulase activity in mulch samples was equivalent to a cellulase enzyme concentration of 25 U ml–1 or greater of product. Sustained exposure of P. cinnamomi to cellulase at 10 to 50 U ml–1 significantly reduced sporangia production, but biomass was only reduced with concentrations over 100 U ml–1. In a lupine bioassay, cellulase was applied to infested soil at 100 or 1,000 U ml–1 with three timings. Cellulase activity diminished by 47% between 1 and 15 days after application. Cellulase applied at 100 U ml–1 2 weeks before planting yielded activity of 20.08 μmol glucose equivalents per gram of soil water (GE g–1 aq) at planting, a level equivalent to mulch samples. Cellulase activity at planting ranged from 3.35 to 48.67 μmol GE g–1 aq, but no treatment significantly affected disease progress. Based on in vitro assays, cellulase activity in mulch was sufficient to impair sporangia production of P. cinnamomi, but not always sufficient to impact vegetative biomass.


2013 ◽  
Vol 2013 ◽  
pp. 1-13 ◽  
Author(s):  
Kamel Kamal Sabet ◽  
Magdy Mohamed Saber ◽  
Mohamed Adel-Aziz El-Naggar ◽  
Nehal Samy El-Mougy ◽  
Hatem Mohamed El-Deeb ◽  
...  

Five commercial composts were evaluated to suppress the root-rot pathogens (Fusarium solani (Mart.) App. and Wr, Pythium ultimum Trow, Rhizoctonia solani Kuhn, and Sclerotium rolfsii Sacc.) of cucumber plants under in vitro and greenhouse conditions. In vitro tests showed that all tested unautoclaved and unfiltrated composts water extracts (CWEs) had inhibitor effect against pathogenic fungi, compared to autoclaved and filtrated ones. Also, the inhibitor effects of 40 bacteria and 15 fungi isolated from composts were tested against the mycelial growth of cucumber root-rot pathogens. Twenty two bacteria and twelve fungal isolates had antagonistic effect against root-rot pathogens. The antagonistic fungal isolates were identified as 6 isolates belong to the genus Aspergillus spp., 5 isolates belong to the genus Penicillium spp. and one isolate belong to the genus Chaetomium spp. Under greenhouse conditions, the obtained results in pot experiment using artificial infested soil with cucumber root-rot pathogens showed that the compost amended soil reduced the percentage of disease incidence, pathogenic fungi population, and improved the cucumber vegetative parameters as shoot length, root length, fresh weight, and dry weight. These results suggested that composts are consequently considered as control measure against cucumber root-rot pathogens.


Plant Disease ◽  
2008 ◽  
Vol 92 (9) ◽  
pp. 1368-1368 ◽  
Author(s):  
S. T. Koike

In 2005 and 2006, field-grown iceberg lettuce (Lactuca sativa) in California's coastal Salinas Valley (Monterey County) was affected by a previously unreported disease. Symptoms were observed on iceberg lettuce at the post-thin rosette stage (8 to 12 leaves). Plants were stunted and slightly chlorotic. Fine feeder roots had numerous, small (4 to 8 mm long), elongated, dark brown-to-black lesions. Larger secondary roots and taproots lacked lesions. No vascular discoloration was present. Isolations from root lesions consistently resulted in gray fungal colonies that formed catenulate, cylindrical, thin-walled, hyaline endoconidia and catenulate, subrectangular, thick-walled, dark aleuriospores. The fungus was identified as Thielaviopsis basicola (2). Conidial suspensions (5.0 × 105) of eight isolates from iceberg lettuce were used for pathogenicity tests. Iceberg cv. Ponderosa and romaine cv. Winchester were grown for 3 weeks in soilless peat moss rooting mix. Roots of 20 plants per cultivar were washed free of the rooting mix and soaked in conidial suspensions for 5 min. Plants were repotted and grown in a greenhouse. Control plant roots were soaked in sterile distilled water (SDW). After 3 weeks, inoculated iceberg exhibited slight chlorosis in comparison with control plants. Feeder roots of all iceberg plants inoculated with the eight isolates exhibited numerous black lesions and T. basicola was reisolated from these roots. Romaine lettuce, however, did not show any foliar symptoms. Small segments of roots had tan-to-light brown discoloration and T. basicola was occasionally reisolated (approximately 40% recovery). Roots of control iceberg and romaine showed no symptoms. Results were similar when this experiment was repeated. To explore the host range of T. basicola recovered from lettuce, two isolates were prepared and inoculated as described above onto 12 plants each of the following: iceberg lettuce (cv. Ponderosa), bean (cv. Blue Lake), broccoli (cv. Patriot), carrot (cv. Long Imperator #58), celery (cv. Conquistador), cotton (cv. Phy-72 Acala), cucumber (cv. Marketmore 76), green bunching onion (cv. Evergreen Bunching), parsley (cv. Moss Curled), pepper (cv. California Wonder 300 TMR), radish (cv. Champion), spinach (cvs. Bolero and Bossanova), and tomato (cv. Beefsteak). Control plant roots of all cultivars were soaked in SDW. After 4 weeks, only lettuce and bean roots had extensive brown-to-black lesions, from which the pathogen was consistently resiolated. Roots of cotton, pepper, spinach, and tomato had sections of light brown-to-orange discoloration; the pathogen was not consistently recovered from these sections. All other species and the control plants were symptomless. This experiment was repeated with similar results except that inoculated peppers were distinctly stunted compared with control plants. To my knowledge, this is the first report of black root rot caused by T. basicola on lettuce in California. Disease was limited to patches along edges of iceberg lettuce fields; disease incidence in these discrete patches reached as high as 35%. Affected plants continued to grow but remained stunted in relation to unaffected plants and were not harvested. Black root rot of lettuce has been reported in Australia (1); that report also showed that lettuce cultivars vary in susceptibility to T. basicola and isolates from lettuce were highly aggressive on bean but not on many other reported hosts of this pathogen. References: (1) R. G. O'Brien and R. D. Davis. Australas. Plant Pathol. 23:106, 1994. (2) C. V. Subramanian. No. 170 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1968.


Author(s):  
Haque ME ◽  
◽  
Parvin MS ◽  

Rhizoctonia solani causes pre-emergence and post-emergence damping-off, as well as crown and root rot of sugar beet (Beta vulgaris L.), which significantly affects the yield returns in the USA and Europe. The pathogen overwinters as sclerotia or melanized mycelium. Traditionally, the resistance of cultivars to R. solani is evaluated by scoring disease reactions at the crowns and roots of older seedlings, thus resistance is not evaluated during seed germination. Moreover, earlier studies evaluated cultivars resistance to R. solani using colonized whole barley or wheat grains which, unlike sclerotia, are artificial inocula of the pathogen that require time, space and technical know-how to produce. Moreover, colonized grains are prone to contamination with other pathogens, consumed by rodents/birds while applied in the field, and are often uneconomic. Considering those limitations, a study was undertaken (1) to develop in vitro methods to generate large-scale sclerotia, (2) to compare pathogenic potentials of sclerotia, mycelia, and colonized barley grains for optimization of dampingoff assays, and (3) to evaluate Rhizoctonia resistance of selected commercial cultivars during the seed germination phase. Comparing six different culture media, we found that R. solani had the highest radial growth (8.9 ± 0.04, cm³) at 8-days and the maximum number of sclerotia produced (203 ± 4.6) at 28-days in CV8 medium. We demonstrated significant differences in pathogenicity of the three different forms of R. solani inocula and susceptibility of cultivars to preand post-emergence damping-off. The highest pre-emergence damping-off and root rot were observed with sclerotia, and the highest post-emergence dampingoff was recorded with both sclerotial and colonized barley inocula. In addition, varietal differences in susceptibility to pre- and post-emergence damping-off were noted. The highest pre-emergence damping-off was recorded on cv Crystal 101RR and lowest in Maribo MA 504. The highest post-emergence damping-off was recorded on BTS 8500 and the lowest in Crystal 467. The maximum mean root rot was observed in BTS 8500, BTS 8606, and Crystal 101R. Our studies demonstrated that sclerotia serve as efficient natural inocula, reemphasized that host-pathogen interactions differ at the early vs. late stages of sugar beet growth, and highlighted the need to reevaluate commercial sugar beet cultivars for resistance at the seed germination stage.


1999 ◽  
Vol 89 (8) ◽  
pp. 613-617 ◽  
Author(s):  
N. R. Walker ◽  
T. L. Kirkpatrick ◽  
C. S. Rothrock

Controlled environments were used to study the relationship between the root-knot nematode (Meloidogyne incognita) and Thielaviopsis basicola on cotton. Temperature treatments were continuous 20, 24, and 28°C or two cyclic linear regimes with ranges of 14 to 32 or 18 to 28°C over 24 h. Cotton seeds were planted in fumigated soil infested with T. basicola, M. incognita, or both. After 42 days, pathogen effects on plant growth and pathogen development were evaluated. Histology was conducted on roots collected 14, 28, and 42 days after planting in the continuous 24°C treatment. Reductions in plant height-to-node ratio and total fresh weight were observed for soils infested with both pathogens compared with the control or with soils infested with either pathogen, except for M. incognita-infested soil at 28°C. T. basicola reduced root galling and reproduction of the nematode at all temperatures. Vascular discoloration caused by T. basicola was greater in the presence of M. incognita compared with that by T. basicola alone. At 2 and 4 weeks, histological studies showed that plants grown in all T. basicola-infested soils contained chlamydospore chains on the root surface and in cortical cells. The fungus was not observed inside the vascular cylinder. Roots from 4-week-old plants from soils infested with T. basicola and M. incognita showed fungal sporulation in vascular tissue and localized necrosis of vascular tissue adjacent to the nematodes. At 6 weeks, plants grown in soil infested with T. basicola alone exhibited no remaining cortical tissue and no evidence of vascular colonization by the fungus. Six-week-old plants grown in T. basicola + M. incognita-infested soils exhibited extensive vascular necrosis and sporulation within vascular tissue. These studies suggest that coinfection expands the temperature ranges at which the pathogens are able to cause plant damage. Further, M. incognita greatly increases the access of T. basicola to vascular tissue.


2020 ◽  
Vol 21 (1) ◽  
pp. 21-25
Author(s):  
Lindsey D. Thiessen ◽  
Grant H. Ellington ◽  
Justin A. Macialek ◽  
Chuck S. Johnson ◽  
David T. Reed

Pythium root rot is an economically important disease threatening greenhouse production of tobacco seedlings. Although methyl bromide was historically used for tray sanitation, the phase-out of the fumigant from agricultural use has left few options for growers to produce disease-free transplants. Steam sanitation at 80°C for 30 min has shown control of disease caused by Rhizoctonia solani and has been adopted for use to manage Pythium spp. This study evaluates other steam temperatures and time durations to effectively manage Pythium spp. in float-tray systems. Naturally infested trays steamed at 63, 71, and 77°C for 30 min significantly reduced Pythium spp. from trays compared with TriSan wash and CC-15 dip treatments. Float trays inoculated with Pythium spp. that were steamed at 70 and 80°C for 2 h 30 min, respectively, also significantly reduced Pythium spp. survival. Other fungi, likely saprophytic or beneficial organisms, were not significantly impacted by any steaming treatment.


Plant Disease ◽  
2019 ◽  
Vol 103 (12) ◽  
pp. 3234-3243
Author(s):  
David R. Cruz ◽  
Leonor F. S. Leandro ◽  
Gary P. Munkvold

Fusarium oxysporum (Fo) is an important pathogen that reduces soybean yield by causing seedling disease and root rot. This study assessed the effects of pH and temperature on Fo fungal growth and seedling disease. In an in vitro assay, 14 Fo isolates collected from symptomatic soybean roots across Iowa in 2007 were grown on artificial culture media at five pH levels (4, 5, 6, 7, and 8) and incubated at four temperatures (15, 20, 25, or 30°C). In a rolled-towel assay, soybean seeds from Fo-susceptible cultivar Jack were inoculated with a suspension of a pathogenic or a nonpathogenic Fo isolate; both isolates were previously designated for their relative aggressiveness in causing root rot at 25°C. The seeds were placed in rolled germination paper, and the rolls were incubated in all combinations of buffer solutions at four pH levels (4, 5, 6, and 7), and four temperatures (15, 20, 25, or 30°C). There was a significant interaction between temperature and pH (P < 0.05) for in vitro radial growth and root rot severity. Isolates showed the most in vitro radial growth after incubation at pH 6 and 25°C. For the rolled-towel assay, the pathogenic isolate caused the most severe root rot at pH 6 and 30°C. Gaussian regression analysis estimates for optimal conditions were pH 6.3 at 27.1°C for maximal fungal growth and pH 5.9 at 30°C for maximal root rot severity. These results indicate that optimal pH and temperature conditions are similar for Fo growth and disease in soybean seedlings and suggest that Fo may be a more important seedling pathogen when soybeans are planted under warm conditions in moderately acidic soils.


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