Aphanomyces euteiches Race 2 in Central Illinois Alfalfa Fields

Approximately 260,000 ha of alfalfa is grown in Illinois. Two soil samples were collected randomly from slowly drained thin patches in each of four established alfalfa fields near Urbana in 2001. Plants in the thin patches were asymptomatic. Aphanomyces euteiches Drechs. was baited from the soil with cv. Saranac alfalfa seedlings and was isolated from 3- to 4-week-old infected seedlings using a medium containing metalaxyl and benomyl (1,2). It is difficult to isolate A. euteiches from field-grown roots. One to seven isolates were obtained per field, and all were identified as A. euteiches based on morphology (1,2). A. euteiches (races R1 and R2) causes root rot of alfalfa in slowly drained fields in Iowa, Kentucky, and Wisconsin (1,2). The race of 13 isolates was determined in tests repeated once with alfalfa populations Saranac (susceptible to R1 and R2), WAPH-1 (resistant only to R1), and WAPH-5 (resistant to R1 and R2) (1). Twelve 7-day-old seedlings in each of three pots per population were inoculated with 103 zoospores per seedling in a growth chamber (25°C). A disease index (DI) was determined 12 days later by scoring plants on a 1 to 5 scale, where 5 is a dead plant (1). Race was based on DI, R1: DI ≥3 for Saranac and <3 for WAPH-1, and R2: DI > 3 for Saranac and WAPH-1. The DI was 1.0 for noninoculated plants. All isolates were R2; the DI was >3.0 for inoculated Saranac and WAPH-1 and <3.0 for WAPH-5. To our knowledge, this is the first report of A. euteiches races in Illinois, and this pathogen was reported previously only from northwest Illinois. Control of Aphanomyces root rot is based on resistance; however, few alfalfa cultivars are resistant to R2. References: (1) D. Malvick and C. Grau. Plant Dis. 85:740, 2001. (2) G. Munkvold and W. Carlton. Plant Dis. 79:1251,1995.

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First Report of Pythium torulosum Causing Corn Root Rot in Northeastern China

Plant Disease ◽

10.1094/pdis-08-20-1679-pdn ◽

2020 ◽

Author(s):

Xiujun Tang ◽

Shuning Chen ◽

Xiaojing Yan ◽

Huizhu Yuan ◽

Daibin Yang

Keyword(s):

Root Rot ◽

Petri Dish ◽

Soil Samples ◽

Northeastern China ◽

Growth Chamber ◽

Corn Root ◽

First Report ◽

Corn Root Rot ◽

Soil Mix ◽

Pythium Torulosum

In October 2017, we collected five soil samples from each of several fields with a history of severe corn (Zea mays) seedling disease in Heilongjiang province of China. Affected seedlings were wilted with severe root rot, and a high incidence of seedling death was observed in the fields. Corn seeds were seeded in the collected soil samples and grown in a growth chamber for 21 days set at the following incubation temperatures: 21℃/7℃ for 6 days, 10℃/3℃ for 4 days, 16℃/7℃ for 5 days, 20℃/20℃ for 6 days (16 h/8 h, light/dark) (Tang et al. 2019). The corn seedlings in the growth chamber showed the same symptoms observed in the field as mentioned above. Corn root rot samples were collected from several symptomatic plants in the growth chamber to isolate the possible pathogen. Symptomatic roots were washed in 0.5% NaOCl for 2 min, rinsed in sterile water and cut into 1-2 mm segments and then plated on corn meal agar amended with pimaricin (5 μg/ml), ampicillin (250 μg/ml), rifampicin (10 μg/ml), pentachloronitrobenzene (50 μg/ml), and benomyl (10 μg/ml) (PARP+B), which is selective for oomycetes (Jeffers and Martin 1986). After 3 days of incubation in the dark at 25℃, colonies were transferred to 10% V8 juice agar and incubated at 25℃ for 2 weeks. Six isolates were identified as Pythium torulosum based on the morphology of sexual and asexual structures following van der Plaats-Niterink’s key (van der Plaats-Niterink 1981). On 10% V8 juice agar, the hypha were aseptate and colonies had filamentous sporangia with a dendroid or globose structure. The oogonia were globose or subglobose, laevis, terminal, rarely intercalary, ranging from 12-19 (average 16) μm. Antheridia were mostly sessile or brachypodous, and each oogonium was supplied by 1-2 antheridia cells. Oospores were globose, plerotic, ranging from 9-16 (average 13) μm. For the molecular identification, two molecular targets, the internal transcribed spacer (ITS) region of ribosomal DNA and cytochrome c oxidase subunit II (CoII), were amplified and sequenced using universal primer sets DC6/ITS4 (Cooke et al. 2000) and FM58/FM66 (Villa et al. 2006), respectively for one isolate, “copt”. BLAST analyses of a 971 bp ITS segment amplified from copt (GenBank Accession No. MT830918) showed 99.79% identity with a P. torulosum isolate (GenBank Accession No. AY598624.2). For the COⅡ gene of copt, BLAST analyses of a 553 bp segment (GenBank Accession MT843570) showed 98.37% identity with P. torulosum isolate (GenBank Accession No. AB095065.1). Thus, the isolate, copt, was identified as P. torulosum based on morphological characteristics and molecular analysis. To confirm pathogenicity and Koch’s postulates, a pathogenicity test was conducted as described by Zhang et al. (2000). Briefly, a 5 mm culture plug from the P. torulosum isolate, copt, was transferred to a 9-cm petri dish containing 20mL 10% V8 juice agar and incubated in the dark at 25℃ for 7 days. The culture was cut into small pieces and mixed with a sterilized soil mix (40% organic peat substrate, 40% perlite, and 20% soil) at a ratio of one petri dish per 100 g soil mix. Ten corn seeds were planted at a depth of 2 cm in a 500-mL pot containing the inoculated soil mix. The control pots were mock inoculated with plain 10% V8 juice agar. Pots were incubated in a greenhouse at temperatures ranging from 21 to 23℃. There were four replications. After 14 days, corn roots brown and rotted were observed, which was similar to those observed in the field and growth chamber. Control plants remained symptomless and healthy. P. torulosum copt was consistently re-isolated from the symptomatic roots. To our knowledge, this is the first report of P. torulosum causing root rot of corn in Northeastern China. Corn is an important crop in Heilongjiang and the occurrence of root rot caused by this pathogen may be a new threat to corn plants. There is a need to develop management measures to control the disease.

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First Report of Aphanomyces euteiches Race 1 and Race 2 Causing Aphanomyces Root Rot on Alfalfa (Medicago sativa) in South Dakota

Plant Disease ◽

10.1094/pdis-08-21-1733-pdn ◽

2021 ◽

Author(s):

Conner L. Tordsen ◽

Jennifer M. Giles ◽

Andrew Edward Sathoff

Keyword(s):

South Dakota ◽

Medicago Sativa ◽

Root Rot ◽

Severity Index ◽

The United States ◽

Soil Samples ◽

Aphanomyces Euteiches ◽

Specific Primers ◽

Lake County ◽

First Report

Aphanomyces euteiches causes Aphanomyces root rot (ARR) in alfalfa (Medicago sativa), along with root rot on many other legumes, including pea, clover, and lentil (Malvick et al., 2009). In 2020, South Dakota (SD) planted the most acres of alfalfa in the United States, which demonstrates the importance of alfalfa to the state. Several SD growers reported alfalfa establishment problems likely to be associated with ARR. Soil samples were collected from 16 fields under commercial alfalfa production in Lake County, SD in June 2020. Composite soil samples based on 24 subsamples were collected in a W-shaped pattern at a depth of 15 cm. Collected soil was sieved, and 80 cm3 was placed in plastic pots (6 cm x 6 cm). Each pot was planted with 25 seeds, covered with an additional 15 cm3 soil, and placed in a growth chamber with a 16-hour photoperiod at temperatures of 24 and 19 ℃ (day and night). Alfalfa seedlings, including Saranac (susceptible to races R1 and R2), WAPH-1 (resistant only to R1), WAPH-5 (resistant to both R1 and R2), and Mustang 625 (resistant to both R1 and R2 and coated with mefenoxam) grew in collected soil for 7 days, followed by 4 days under flooded conditions. Trays were drained, and at 21 days after planting (DAP), roots were removed from soil, washed in distilled water, and rated to measure severity of disease symptoms (Samac et al., 2015). The average severity index (ASI) used a 1-5 disease severity scale, 5 being a dead plant and 1 being no symptoms present (http://www.naaic.org/stdtests/Aphano.html). Race was based on ASI where R1 included an ASI of ≥3 for Saranac and <3 for WAPH-1, and R2 included an ASI of >3.0 for Saranac and WAPH-1 and <3.0 for WAPH-5 (Malvick and Grau, 2001). Race-typing experiments were repeated twice with six replicate pots per alfalfa cultivar per experiment and determined the presence of both R1 and R2 in Lake County, SD. ASI values for Mustang 625 and WAPH-5 were similar across all fields evaluated, which indicates limited confounding effects of other root rotting pathogens. DNA was extracted from three symptomatic roots from each field and was PCR amplified using A. euteiches specific primers (Vandemark et al., 2002). A PCR product was observed in all 16 fields evaluated, and the absence of a product was observed when DNA was extracted from alfalfa roots grown in vermiculite. Following race-typing, infected alfalfa roots were surfaced sterilized and placed on Aphanomyces selective media consisting of mefenoxam and benomyl in cornmeal agar (CMA) (Pfender et al., 1984). Isolates were identified as A. euteiches based on hyphal morphology (Malvick and Grau, 2001). Alfalfa seedlings (Saranac) were grown in vermiculite under growth conditions used for the race-typing assay and inoculated 6 DAP with two isolates of A. euteiches. Inoculation was completed using half plates of one week old A. euteiches mycelium on CMA blended with one liter of water (Samac et al., 2015). At 35 DAP, control alfalfa seedlings inoculated with blended CMA and water remained asymptomatic, and alfalfa infected with A. euteiches displayed symptoms including honey-brown colored lesions. For confirmation of Koch’s postulates, DNA from three re-infected seedlings was again PCR amplified using A. euteiches specific primers and confirmed our previous work. This is the first report of either R1 or R2 of A. euteiches causing ARR on alfalfa in SD. To avoid future yield loss, SD growers should consider planting available alfalfa cultivars that have resistance to both races of A. euteiches.

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A Rapid Method Using PCR-Based SCAR Markers for the Detection and Identification of Phoma sclerotioides: The Cause of Brown Root Rot Disease of Alfalfa

Plant Disease ◽

10.1094/pdis.2002.86.9.928 ◽

2002 ◽

Vol 86 (9) ◽

pp. 928-932 ◽

Cited By ~ 21

Author(s):

R. C. Larsen ◽

C. R. Hollingsworth ◽

G. J. Vandemark ◽

M. A. Gritsenko ◽

F. A. Gray

Keyword(s):

Root Rot ◽

Random Amplified Polymorphic Dna ◽

Dna Amplification ◽

Pythium Ultimum ◽

Soil Samples ◽

Root Tissue ◽

Aphanomyces Euteiches ◽

Amplification Product ◽

Phoma Sclerotioides ◽

Brown Root Rot

A rapid technique for identification and detection of Phoma sclerotioides, the causal agent of brown root rot of alfalfa, has been developed using polymerase chain reaction (PCR). Amplification products obtained from random amplified polymorphic DNA (RAPD) reactions were cloned and sequenced, and two extended primer sets were designed from the resulting data that were used to detect sequence-characterized DNA markers. A single 499-bp DNA amplification product was consistently obtained from primers PSB12499 that was specific for 19 isolates of P.sclerotioides but was not produced from Phoma medicaginis or Phoma betae, or from other soilborne pathogens including Aphanomyces euteiches, Rhizoctonia solani, Fusarium oxysporum, Pythium ultimum, or Phytophthora infestans. A 499-bp amplification product was also produced from root tissue known to be infected with the fungus as verified by microscopic examination. A similar PCR product was obtained from soil samples collected from fields with an established infection of P. sclerotioides on alfalfa. This PCR-based assay enables detection of P. sclerotioides from alfalfa root tissue and in soil samples in a single day, including extraction of DNA, compared with standard methods that require up to 100 days for identification using agar media.

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Bioassay and baiting methods for determining occurrence of races of Phytophthora clandestina in soil

Australian Journal of Experimental Agriculture ◽

10.1071/ea9960815 ◽

1996 ◽

Vol 36 (7) ◽

pp. 815 ◽

Cited By ~ 1

Author(s):

A Purwantara ◽

SP Flett ◽

PJ Keane

Keyword(s):

Disease Severity ◽

Root Rot ◽

Subterranean Clover ◽

Soil Samples ◽

Field Soil ◽

Flooded Soil ◽

Aphanomyces Euteiches ◽

Differential Cultivars ◽

Pure Cultures ◽

Race 1

The method currently used for determining races of Phytophthora clandestina requires isolation of pure cultures of the pathogen and testing of their pathogenicity on a range of differential cultivars. To date, the pathogen has not been isolated directly from soil and isolation of the pathogen from naturally infected seedlings is laborious. A bioassay involving the planting of differential cultivars in soil samples in small planting trays was developed to identify races of P. clandestina in soil. The specific races of the pathogen in the soil were determined by assessing the disease severity and the extent of sporulation of the pathogen on the roots of the differential cultivars. A more rapid baiting method using cotyledons of differential cultivars in flooded soil samples was also developed to determine the presence of different races in the samples. Both bioassays were used to confirm the presence of race 0 and race 1 in separate paddocks at Rutherglen, northern Victoria, over 4 seasons. The presence in field soil of another root rot pathogen of subterranean clover, Aphanomyces euteiches, was also detected using these techniques.

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First Report of Root Rot Caused by Pythium aphanidermatum on Bell Pepper (Capsicum annuum L.) in Italy

Plant Disease ◽

10.1094/pdis-11-13-1148-pdn ◽

2014 ◽

Vol 98 (6) ◽

pp. 854-854 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

G. Ortu ◽

M. L. Gullino

Keyword(s):

Capsicum Annuum ◽

Root Rot ◽

Agar Medium ◽

The United States ◽

Pythium Aphanidermatum ◽

Northern Italy ◽

Bell Pepper ◽

Growth Chamber ◽

Sodium Hypochlorite Solution ◽

First Report

During July 2012, symptoms of root rot were observed on bell pepper (Capsicum annuum) grown in 2,000 m2 of commercial greenhouses near Cuneo in northern Italy. Symptoms first developed 30 to 40 days after transplanting, when greenhouse temperatures ranged from 25 to 30°C, and 10% of the plants were affected. Affected plants were stunted with leaf chlorosis, reduced growth, and sudden wilting. Roots were severely affected with a brown discoloration, water-soaking, and soft rot. Eventually, affected plants collapsed. Tissue fragments of 1 mm2 were excised from symptomatic roots, dipped in a 1% sodium hypochlorite solution, and placed on potato dextrose agar (PDA) and an agar medium selective for oomycetes (3). Plates were incubated under constant fluorescent light at 22 ± 1°C for 5 days. An isolate grown for 12 days on V8 agar medium (200 ml V8 Campbell Soup, 15 g agar, 0.5 g CaCO3, and 1 liter distilled water) showed aseptate hyphae that were 3.5 to 6.3 μm (avg. 5.2 μm) wide. Oogonia were globose, smooth, and 24.3 to 29.0 (avg. 25.1) μm in diameter. Antheridia were barrel-shaped, while oospores were globose, and 17.3 to 23.5 μm (avg. 21.2 μm) in diameter. These morphological characters identified the microorganism as a Pythium sp. (4). The ITS region of rDNA of a single isolate was amplified using the primers ITS1/ITS4 and sequenced. BLAST analysis (1) of the 781-bp segment (GenBank Accession KF840479) showed 100% homology with the ITS sequence of an isolate of Pythium aphanidermatum in GenBank (AY598622.2). Pathogenicity tests were performed twice on 30-day-old plants of C. annuum cv. Cuneo grown in 2-L pots (4 plants/pot), containing a steam-disinfested, organic peat substrate (70% black peat and 30% white peat, pH 5.5 to 6.0, N 110 to 190 mg/liter, P2O5 140 to 230 mg/liter, K2O 170 to 280 mg/liter) that was infested with wheat and hemp kernels colonized by the isolate of P. aphanidermatum, at a rate of 1 g colonized kernels/liter potting medium. The inoculum was prepared by autoclaving at 121°C for 30 min a mixture of wheat-hemp kernels (2:1 v/v) in a 1-liter flask, to which the bell pepper isolate of P. aphanidermatum was added in the form of colonized agar medium selective for oomycetes plugs. Before use, the inoculated flask was incubated for 10 days at 22°C in the dark. Four plants/pot were transplanted into each of four pots filled with the infested medium/growth chamber, while the same number of plants were grown in non-infested substrate in pots in each growth chamber. Plants were kept in two growth chambers, one set at 20°C and the other at 28°C. Symptoms first developed 7 days after inoculation. After 30 days, 50% of inoculated plants showed brown roots and died in the growth chamber set at 28°C, while only 10% of the plants were symptomatic at 20°C. Control plants remained asymptomatic at both temperatures. P. aphanidermatum was re-isolated consistently from the symptomatic roots of plants grown in the infested soil by using the same protocol as the original isolations, while no fungal colonies were obtained from asymptomatic roots of the non-inoculated control plants. To our knowledge, this is the first report of the presence of P. aphanidermatum on C. annuum in Italy. The same disease was reported in the United States (2). The importance of the disease, although limited in distribution at present to the greenhouses surveyed in northern Italy, could increase in areas where sweet pepper is grown intensively. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. O. Chellemi et al. Plant Dis. 84:1271, 2000. (3) H. Masago et al. Phytopathology 67:425, 1977. (4) T. Watanabe. Pictorial Atlas of Soil and Seed Fungi. CRC Press, Boca Raton, FL, 2002.

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First Report of Aphanomyces euteiches Causing Aphanomyces Root Rot on Lentil in North Dakota

Plant Disease ◽

10.1094/pdis-11-19-2422-pdn ◽

2020 ◽

Vol 104 (6) ◽

pp. 1876

Author(s):

K. K. Zitnick-Anderson ◽

J. S. Pasche ◽

T. Gargouri-Jbir ◽

A. Kalil

Keyword(s):

North Dakota ◽

Root Rot ◽

Aphanomyces Euteiches ◽

First Report

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Characteristics and Frequency of Aphanomyces euteiches Races 1 and 2 Associated with Alfalfa in the Midwestern United States

Plant Disease ◽

10.1094/pdis.2001.85.7.740 ◽

2001 ◽

Vol 85 (7) ◽

pp. 740-744 ◽

Cited By ~ 21

Author(s):

D. K. Malvick ◽

C. R. Grau

Keyword(s):

Root Rot ◽

High Frequency ◽

Rapd Analysis ◽

Random Amplified Polymorphic Dna ◽

Aphanomyces Euteiches ◽

History Of ◽

Race 1 ◽

Race 2 ◽

Genotypic Characteristics ◽

Distribution Frequency

Aphanomyces root rot of alfalfa, caused by Aphanomyces euteiches, kills seedlings and causes decline of established plants in slowly drained soils. Two races of A. euteiches that are pathogenic to alfalfa have been identified. Despite the contribution of race 1 resistance to establishment and yield of alfalfa, race 1-resistant alfalfa cultivars perform poorly in some fields infested with A. euteiches. Many isolates of A. euteiches obtained from the soils of problematic fields are of a race 2 phenotype. The purpose of this study was to determine distribution, frequency, and pathogenic and genotypic characteristics of race 1 (R1) and race 2 (R2) isolates from 21 fields: 13 in Wisconsin, 7 in Minnesota, and 1 in Kentucky. A. euteiches was successfully isolated from the soil of 16 of the 21 fields; 405 isolates were obtained from Wisconsin, 4 from Minnesota, and 48 from Kentucky. Pathogenicity and race phenotype of isolates were characterized on Saranac (susceptible to R1 and R2 isolates) and WAPH-1 (resistant to R1 and susceptible to R2 isolates) alfalfa populations. One Wisconsin field with no recent history of alfalfa production had a high frequency (51%) of R2 isolates, and 43% of all isolates were R2 from fields with a history of alfalfa production. In a location that was planted continuously to pea for 30 years, 27% of the isolates were R2. Random amplified polymorphic DNA (RAPD) analysis of three R1 and three R2 isolates with eight primers generated 43 total polymorphic bands; however, none of the bands were uniquely associated with race phenotype. Cluster analysis based on RAPD bands revealed no consistent genotypic distinctions between R1 and R2 isolates of A. euteiches. Evaluation of eight commercial alfalfa cultivars for resistance to two R1 and two R2 isolates demonstrated that most are susceptible to R2 isolates; however, those selected for R2 resistance express resistance to R2 isolates. The results suggest that R2 isolates represent a widespread risk to alfalfa cultivars having resistance only to R1 isolates in fields with varied cropping histories.

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First Report of Brown Root Rot of Alfalfa Caused by Phoma sclerotioides in Wisconsin

Plant Disease ◽

10.1094/pdis.2004.88.7.769c ◽

2004 ◽

Vol 88 (7) ◽

pp. 769-769 ◽

Cited By ~ 4

Author(s):

R. C. Larsen ◽

C. R. Grau ◽

G. J. Vandemark ◽

T. J. Hughes ◽

B. D. Hudelson

Keyword(s):

United States ◽

Root Rot ◽

Lateral Roots ◽

The United States ◽

Soil Samples ◽

Soil Extraction ◽

Soil Dna ◽

First Report ◽

Phoma Sclerotioides ◽

Brown Root Rot

Brown root rot (BRR) has been associated with winterkill of alfalfa (Medicago sativa L.) in the temperate regions of North America where winters are severe (1). Although suspected, BRR has not been associated with winterkill of alfalfa in the upper Midwestern United States. Alfalfa plants exhibiting symptoms resembling those induced by the causal agent Phoma sclerotioides G. Preuss ex Sacc. were collected from fields in Marinette, Pierce, and Marathon counties in Wisconsin during the spring and early summer of 2003. Symptoms included stunting and decline in 1- to 3-year-old plants that were slow to break dormancy in the early spring. Roots frequently exhibited dark brown lesions or were entirely decayed. Advanced lesions often formed dark bands around the circumference of tap and secondary roots. Beaked pycnidial structures typical of P. sclerotioides were also observed on many samples with advanced lesions. Plants with symptoms of BRR were also observed in Clark, Langlade, Lincoln, Oconto, Shawno, Taylor, and Wood counties. Several lesion areas of tissue on the tap and lateral roots of each sample were excised with a sterile scalpel. Total DNA was extracted using the Fast DNA kit (Bio 101, Carlsbad, CA). In addition, soil samples were collected in the root rhizosphere of symptomatic plants from four fields in two counties. Soil DNA was extracted with the Ultra-Clean DNA soil extraction kit (Mo Bio, Solana Beach, CA). DNA extractions were diluted 1:10 or 1:50, and samples were evaluated for the presence of P. sclerotioides using polymerase chain reaction (PCR)-based sequence-characterized amplified region (SCAR) markers according to the method described previously (4). Amplicons of the expected size (499 bp) were detected from alfalfa roots sampled from Marathon (4 of 4), Marinette (4 of 5), and Pierce (4 of 4) counties but not in roots from healthy controls produced in the greenhouse at Prosser, WA. PCR amplicons were also produced from all field soil samples in Marathon and Marinette counties. Proof of pathogenicity via Koch's postulates for this host-pathogen system was not attempted because of the extensive time period required (1). However, characteristic beaked pycnidia were present, and the pathogen was identified using PCR on DNA from roots symptomatic of BRR. Detection of BRR has been limited in the United States to Wyoming (2), but has been thought to occur in other states with severe winters (3). To our knowledge, this is the first report of P. sclerotioides in Wisconsin. References: (1) J. G. N. Davidson. Brown root rot. Pages 29–31 in: Compendium of Alfalfa Diseases. 2nd ed. D. L. Stuteville and D. C. Erwin, eds. The American Phytopathological Society, St. Paul, MN, 1990. (2) F. A. Gray et al. Pages 27–28 in: Proc. 10th Western Alfalfa Improv. Conf., 1997. (3) C. R. Hollingsworth et al. Can. J Plant Pathol. 25:215, 2003. (4) R. C. Larsen et al. Plant Dis. 86:928, 2002.

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Comparison of Soil Receptivity to Thielaviopsis basicola, Aphanomyces euteiches, and Fusarium solani f. sp. pisi Causing Root Rot in Pea

Phytopathology ◽

10.1094/phyto.1997.87.5.534 ◽

1997 ◽

Vol 87 (5) ◽

pp. 534-541 ◽

Cited By ~ 16

Author(s):

P. J. Oyarzun ◽

G. Dijst ◽

F. C. Zoon ◽

P. W. Th. Maas

Keyword(s):

Fusarium Solani ◽

Root Rot ◽

Weibull Model ◽

Soil Samples ◽

Thielaviopsis Basicola ◽

Aphanomyces Euteiches ◽

Health Index ◽

Weibull Parameters ◽

Maximum Root ◽

Index Curve

Soil receptivity as a quantifiable characteristic ranging from conduciveness to suppressiveness to soilborne pea pathogens Thielaviopsis basicola and Aphanomyces euteiches was determined by analysis of differences in disease response curves obtained by artificial introduction of inoculum into natural field soil samples. Several parameters, including maximum root rot severity, the area under the health index curve, scores on the first axis of a principal component analysis (PCA) on dose responses, and Weibull model fitting were used to describe the disease responses. In all cases, the Weibull model gave satisfactory fits. PCA yielded a first axis that comprised 86% of the variance found when using Weibull predicted responses for T. basicola and 74% of the variance found for A. euteiches. This PCA axis essentially represented the average increase in disease severity due to the addition of increasing doses of inoculum to the soil. The Weibull scale parameter B, which represents the amount of inoculum necessary to increase root rot severity by 63% with respect to the level caused by pathogens naturally present in the soil, is another means of quantifying the receptivity of soils to these plant pathogens. Weibull parameter B, maximum root rot severity, the areaunder the health index curve, and the scores on the first PCA axis were strongly correlated for each of the pathogens tested individually. To compare the extent and behavior of soil receptivity responses to different pathogens, Weibull parameters B and C (slope at dose B) were chosen because of their universal definition, in contrast to PCA scores. Comparison of the average levels of Weibull parameters B and C indicated significant differences between the pathogens. Yet, no significant similarity in the ranking of the soils was found for the three pathogens, demonstrating that individual soils may interact with different pathogens in totally different ways. In general, soils were suppressive to T. basicola but conducive to A. euteiches, whereas their response to Fusarium solani f. sp. pisi ranged from conducive to suppressive. Therefore, risk assessment of soils prior to planting may require different strategies for each pathogen. Bioassays with soil samples taken before the last pea crop in 1987 and 1991 revealed a significant increase in the natural inoculum potential of soils that mainly was accounted for by A. euteiches and Pythium spp. These results strongly indicate that A. euteiches must be considered one of the most threatening pathogens to pea crops in the Netherlands.

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Isolation, identification, and assessment of soil bacteria as biocontrol agents of pea root rot caused by Aphanomyces euteiches

Canadian Journal of Soil Science ◽

10.1139/cjss-2019-0133 ◽

2020 ◽

Vol 100 (3) ◽

pp. 206-216

Author(s):

Ashebir T. Godebo ◽

James J. Germida ◽

Fran L. Walley

Keyword(s):

Root Rot ◽

Soil Bacteria ◽

Developmental Stages ◽

Antagonistic Activity ◽

Culture Collection ◽

Field Pea ◽

Biocontrol Agents ◽

Growth Chamber ◽

Aphanomyces Euteiches

Aphanomyces euteiches is a soil-borne pathogen that causes root rot of pea and can significantly affect pea production in western Canada. This study aimed to isolate and identify soil bacteria with antagonistic activity towards A. euteiches mycelial and zoospore developmental stages under in vitro conditions and assess their potential as biocontrol agents against aphanomyces root rot in field pea under growth chamber conditions. In vitro screening of soil bacteria identified 184 antagonistic isolates, including 22 from an existing culture collection. Mean mycelial growth inhibition zones ranged from 1 to 12 mm, and mean zoospore germination inhibition ranged from 0% to 100%. Use of 16S rDNA sequence analysis placed isolates into 18 different bacterial genera. Screening of 47 bacteria that inhibited both infective stages identified 29 potential biocontrol strains, including Rhizobium spp. that significantly (α = 0.05) suppressed aphanomyces root rot in field pea grown in vermiculite, suggesting the intriguing possibility of using N-fixing Rhizobium inoculants as biocontrol agents for aphanomyces control. Further screening of 20 isolates as soil inoculants identified K-Hf-L9 (Pseudomonas fluorescens), PSV1-7 (Pantoea agglomerans), and K-Hf-H2 (Lysobacter capsici) isolates as having the highest biocontrol activity, significantly (α = 0.05) suppressing aphanomyces root rot in field pea in growth chamber trials. This study demonstrates the possibility of aphanomyces root rot management using biocontrol agents.

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