First Report of Erwinia carotovora subsp. carotovora Causing Bacterial Root Rot of Sweetpotato (Ipomoea batatas) in Louisiana

Bacterial root and stem rot of sweetpotato (Ipomoea batatas (L.) Lam.) was first fully characterized in the U.S. in 1977 (2). It was thought to be caused exclusively by Erwinia chrysanthemi. Although a previous report described sweetpotato as a host for E. carotovora subsp. carotovora, based on artificial inoculations, others have reported that neither E. carotovora subsp. carotovora nor E. carotovora subsp. atroseptica decay sweetpotato storage roots (1). In October 1995, storage roots of sweetpotato cv. Beauregard were received from St. Landry Parish, LA, that displayed typical bacterial root rot. Isolations from these roots yielded bacteria that showed a similarity of 0.945 to E. carotovora subsp. carotovora with the Biolog GN Bacterial Identification System (version 3.50). This isolate (Ecc-LH) also differed from isolates of E. chrysanthemi (Ech) from sweetpotato and other hosts in that it was insensitive to erythromycin, did not produce phosphatase or lecithinase, and did not produce gas from glucose. Ecc-LH differed from known strains of E. carotovora subsp. atroseptica in that it did not produce reducing substances from sucrose or acid from palatinose. When Beauregard storage roots were inoculated by inserting micropipette tips containing 50 μl of 1.0 × 108 CFU/ml, both Ecc-LH and Ech-48 produced typical bacterial root rot symptoms. However, when they were compared by infectivity titrations at 28 to 32°C, Ecc-LH was less virulent than Ech-48. Ecc-LH had an ED50 of approximately 1.0 × 106 CFU/ml and did not cause appreciable disease below inoculum concentrations of 1.0 × 105, whereas Ech-48 had an ED50 of approximately 1.0 × 108 and caused soft rot at the lowest concentration tested, 1.0 × 103. Similar disease incidence was observed in infectivity titrations at 22 to 24°C, but Ech-48 caused less severe soft rot. E. carotovora subsp. carotovora was reisolated from inoculated storage roots and its identity was reconfirmed by Biolog. When terminal vine cuttings of Beauregard were dipped in 1.0 × 108 CFU/ml and planted in a greenhouse, bacterial stem rot symptoms developed on plants inoculated with Ech-48 at about 4 weeks postinoculation, or when new growth began. However, no symptoms developed on plants inoculated with Ecc-LH. This is the first report of natural occurrence of E. carotovora subsp. carotovora causing bacterial root rot of sweetpotato in Louisiana. E. chrysanthemi remains the most important pathogen causing bacterial soft rot in sweetpotato since it is widely associated with sweetpotato, is more virulent on storage roots and also causes a stem rot. E. carotovora subsp. carotovora can cause root rot, but has been isolated in only one location to date, is less virulent on storage roots, and apparently does not cause stem rot on the predominant cultivar in U.S. sweetpotato production, Beauregard. References: (1) C. A. Clark and J. W. Moyer. 1988. Compendium of Sweet Potato Diseases. American Phytopathological Society, St. Paul, MN. (2) N. W. Schaad and D. Brenner. Phytopathology 67:302, 1977.

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First Report of Stem Rot and Wilt of Sunflower Caused by Sclerotinia minor in Spain

Plant Disease ◽

10.1094/pdis.2002.86.6.697d ◽

2002 ◽

Vol 86 (6) ◽

pp. 697-697

Author(s):

M. L. Molinero-Ruiz ◽

J. M. Melero-Vara

Keyword(s):

Helianthus Annuus ◽

Root Rot ◽

Mycelial Growth ◽

Disease Incidence ◽

Potato Dextrose Agar ◽

Stem Rot ◽

Sclerotinia Minor ◽

Stem Base ◽

First Report ◽

White Mycelium

In 2001, sunflower (Helianthus annuus L.) plants with symptoms of stem and root rot and wilt were observed in Soria, Spain. Light brown, water-soaked lesions developed on the collar of infected plants and extended along the stem, affecting the pith and causing early and sudden wilt. White mycelium and sclerotia (0.5 to 2 mm long) formed in the pith of stems. The sclerotia were disinfested in NaClO (10% vol/vol) for 1 min, transferred to potato dextrose agar (PDA), and incubated at 20°C. The fungus consistently obtained was identified as Sclerotinia minor Jagger (1). Pathogenicity was confirmed in a greenhouse experiment (15 to 25°C, 13 h light). Seven-week-old plants of six genotypes of sunflower (‘Peredovik’, HA89, HA821, HA61, RHA274, and HA337) were inoculated by placing one PDA disk with active mycelial growth adjacent to each basal stem just below the soil line and covering it with peat/sand/silt (2:2:1, vol/vol). Six plants of each genotype were inoculated without wounding, and another six were inoculated immediately after stem base wounding with a scalpel; six wounded and uninoculated plants were used as controls. First symptoms (wilting) appeared 4 days after inoculation in all genotypes. Two weeks after inoculation, the percentage of dead plants ranged from 33 to 92% (depending on cultivar), white mycelium was observed at the base of affected plants, and sclerotia were present in the pith of diseased plants. There was no effect of plant wounding on disease incidence or severity, and the fungus was reisolated from inoculated plants. To our knowledge, this is the first report of S. minor in Spain. Reference: (1) L. M. Kohn. Mycotaxon IX 2:365, 1979.

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First Report of Pectobacterium versatile Causing Aerial Stem Rot of Potato in China

Plant Disease ◽

10.1094/pdis-06-21-1264-pdn ◽

2021 ◽

Author(s):

Wanxin Han ◽

Jinhui Wang ◽

Zheng Li ◽

Yang Pan ◽

Dai Zhang ◽

...

Keyword(s):

16S Rdna ◽

Disease Incidence ◽

New York State ◽

Housekeeping Genes ◽

Soft Rot ◽

Stem Rot ◽

Post Inoculation ◽

First Report ◽

Potato Plants ◽

Aerial Stem

Pectobacterium species cause blackleg, soft rot and stem rot in potato and many other vegetable crops (Charkowski 2015). In July 2020, potato plants showing characteristic symptoms of aerial stem rot were observed in a field (cv. Xisen 6) in Fengning Manchu Autonomous County, Chengde, Hebei Province (North China). The disease incidence in that field (5 ha in size) was more than 50%. Putative pectolytic bacteria were obtained from symptomatic stem tissues (light brown and water-soaked stem sections) by culturing on the crystal violet pectate (CVP) medium. Bacterial colonies producing pits, were restreaked and purified on Luria-Bertani (LB) agar. The isolates causing stem rot were gram negative and rod shaped, negative for oxidase, urease, indole production, gelatin liquefaction and acid production from maltose and D-sorbitol. All isolates were catalase positive, produced acid from lactose, rhamnose, saccharose, raffinose and D-arabinose, and were tolerant to 5% NaCl, and able to utilize citrate. The bacterial gDNA was extracted using the EasyPure Bacteria Genomic DNA Kit (TransGen Biotech). The 16S rDNA region was amplified by PCR using the universal primer pair 27F/1492R and sequenced. Result of the Blastn analysis of the 16S rDNA amplicons (MZ379788, MZ379789) suggested that the isolates FN20111 and FN20121 belonged to the genus Pectobacterium. To determine the species of the stem rot Pectobacterium isolates, multi-locus sequence analysis (MLSA) was performed with six housekeeping genes acnA, gapA, icdA, mdh, proA and rpoS (MZ403781-MZ403792), and phylogenetic tree was reconstructed using RAxML v8.2.12 (https://github.com/stamatak/standard-RAxML). The result of phylogenetic analysis showed that the stem rot Pectobacterium isolates FN20111 and FN20121 clustered with P. versatile (syn. ‘Candidatus Pectobacterium maceratum’) strains CFBP6051T (Portier et al. 2019), SCC1 (Niemi et al. 2017) and F131 (Shirshikov et al. 2018). And the isolates FN20111 and FN20121 were more closely related to the type strain CFBP6051T than to strains SCC1 and F131. Potato seedlings (cv. Xisen 6 and Favorita) were inoculated with the isolates FN20111 and FN20121 by injecting 100 µl of bacterial suspensions (108 CFU·mL-1) into the upper parts of the stems of potato plants, or injected with 100 µl of 0.9% saline solution as control. The seedlings were grown at 28°C and 50% relative humidity. Three days post-inoculation, only the bacteria-inoculated seedlings showed diseased symptoms resembling to those observed in the field. Bacterial colonies were obtained from the infected stems and were identified using the same PCR primers of housekeeping genes as described above, fulfill Koch’s postulates. P. versatile causing soft rot and blackleg on potato plants has been reported in Finland (Niemi et al. 2017), Russia (Shirshikov et al. 2018), Netherlands (Portier et al. 2019), Poland (Waleron et al. 2019) and in New York State (Ma et al. 2021). To our knowledge, this is the first report of P. versatile causing aerial stem rot of potato in China.

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An Outbreak of Bacterial Stem Rot of Dieffenbachia amoena Caused by Erwinia carotovora subsp. carotovora in the Eastern Mediterranean Region of Turkey

Plant Disease ◽

10.1094/pdis.2004.88.3.310c ◽

2004 ◽

Vol 88 (3) ◽

pp. 310-310 ◽

Cited By ~ 4

Author(s):

R. Cetinkaya-Yildiz ◽

M. Mirik ◽

Y. Aysan ◽

M. Kusek ◽

F. Sahin

Keyword(s):

Mediterranean Region ◽

Eastern Mediterranean ◽

Disease Incidence ◽

Eastern Mediterranean Region ◽

Acid Methyl Ester ◽

Erwinia Carotovora ◽

Soft Rot ◽

Stem Rot ◽

Tomato Plants ◽

Rot Disease

Severe outbreaks of bacterial stem rot disease occurred on dieffenbachia plants (Dieffenbachia amoena cv. Tropic Snow) during the autumn and spring seasons of 2002 and 2003 in two commercial glasshouses (3.5 ha) near Adana and Mersin in the Eastern Mediterranean Region of Turkey. Characteristic symptoms of the disease were wilting of the lower leaves, darkening and water soaking of the leaves and stem at or below the soil level, and browning in the vessel and pith of the diseased plants. Eventually, the stem and leaves completely rotted, and the plants collapsed. Nearly 30 and 40% (2002 and 2003, respectively) of the 20,000 potted plants in the glasshouses were destroyed because of the disease. Cuttings often developed a typical soft rot during propagation. Disease incidence was estimated at approximately 50% on propagating material during 2003. Isolations were made from rotted stems, leaves, and discolored vessels of the dieffenbachia plants on King's medium B. Bacteria consistently isolated from the diseased tissues formed white-to-cream colonies on the medium. Bacteria from purified colonies were gram, oxidase, and arginine dyhidrolase negative, catalase positive, and facultative anaerobic. Ten representative strains all fermented glucose and reduced nitrates to nitrites. The strains caused soft rot of potato slices within 24 h at 25°C. All strains were resistant to erythromycin in an antibiotic disk (15 μg) assay. Negative results were obtained from utilization of α-methyl glycoside, reducing substance from sucrose, and indole production from tryptophane and phosphathase activity. Positive results were obtained from pectate, aesculin, and gelatine liquefaction for all strains. Acid was produced from glucose, sucrose, mannitol, mannose, lactose, raffinose, melibiose, trehalose, and L(+)-arabinose but not Darabinose, sorbitol, inulin, and maltose. Pathogenicity was confirmed by needle-stab inoculation at the stem on three plants each of dieffenbachia and tomato plants (5-week-old cv. H-2274). Sterile distilled water was used as a negative control. All plants were covered with polyethylene bags for 48 h at 25°C. Within 72 h after inoculation, water-soaking and soft-rot symptoms were observed on dieffenbachia and tomato plants. All of the bacterial strains isolated in the present study were identified as Erwinia carotovora subsp. carotovora (Jones) based on fatty acid methyl ester analysis with similarity indices ranging from 80 to 94%. Furthermore, Biolog GN (Department of Plant Protection, Faculty of Agriculture, Ataturk University, Erzurum, Turkey) profiles identified them as the same pathovar with similarity values of 67 to 72%. All of the test results were similar to those of reference strain GSPB 435 (Gottinger Sammlung phytopathogener Bakterien, Georg-August University, Gottingen, Germany) of E. carotovora subsp. carotovora used in this study. To our knowledge, this is the first report of the occurrence and outbreak of a bacterial rot disease on dieffenbachia grown in the Eastern Mediterranean Region of Turkey. Contaminated cuttings may be the primary source of inoculum within and between glasshouses.

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Relationships of Preharvest Weather Conditions and Soil Factors to Susceptibility of Sweetpotato to Postharvest Decay Caused by Rhizopus stolonifer and Dickeya dadantii

Plant Disease ◽

10.1094/pdis-11-14-1143-re ◽

2015 ◽

Vol 99 (6) ◽

pp. 848-857 ◽

Cited By ~ 6

Author(s):

Brooke A. Edmunds ◽

Christopher A. Clark ◽

Arthur Q. Villordon ◽

Gerald J. Holmes

Keyword(s):

North Carolina ◽

Soil Moisture ◽

Soil Temperature ◽

Air Temperature ◽

Root Rot ◽

Disease Incidence ◽

Soft Rot ◽

Storage Roots ◽

Dickeya Dadantii ◽

High Soil Temperature

Postharvest soft rots of sweetpotato caused by Rhizopus stolonifer (Rhizopus soft rot) and Dickeya dadantii (bacterial root rot) occur sporadically and can result in significant losses. A 3-year field study related preharvest conditions, including soil texture, chemistry, and fertility; air temperature; soil temperature and moisture; and various cultural practices (153 total variables), to postharvest susceptibility to both diseases in 75 sweetpotato fields in North Carolina and 63 sweetpotato fields in Louisiana. Storage roots were sampled from each field, cured, stored, and inoculated with each pathogen after 100 to 120 days in storage. Disease susceptibility was measured as incidence of diseased storage roots 10 days following inoculation. There was wide variation from field to field in incidence of both diseases (0 to 100% for Rhizopus soft rot and 5 to 95% for bacterial root rot) in both states in each year. Correlations between disease incidence and each of the preharvest variables revealed numerous significant correlations but the variables that correlated with disease incidence were different between North Carolina and Louisiana. Models for both diseases were built by first using forward stepwise regression to identify variables of interest, followed by a mixed-model analysis to produce a final reduced model. For North Carolina fields, postharvest Rhizopus soft rot susceptibility was described by the percentage of the soil cation exchange capacity occupied by calcium, amount of plant-available soil phosphorus, percent soil humic matter, mean air temperature, mean volumetric soil moisture at 40 cm in depth, and mean soil temperature at 2 cm in depth. Postharvest bacterial soft rot susceptibility was described by soil pH and the number of days of high soil temperature late in the season. For Louisiana fields, Rhizopus soft rot susceptibility was described by a complex of variables, including late-season air and soil temperature and late-season days of extreme soil moisture. For bacterial root rot, days of low air temperature and days of high soil temperature late in the season as well as days of low soil moisture best described variation. Although the influence of preharvest variables on postharvest susceptibility was profound for each disease, the complexity of factors involved and differences between the data for the two states makes development of a predictive system extremely difficult.

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Modeling the impact of sweetpotato weevils on storage root yield

Open Agriculture ◽

10.1515/opag-2018-0035 ◽

2018 ◽

Vol 3 (1) ◽

pp. 319-325

Author(s):

Daniel A. Akansake ◽

Putri E. Abidin ◽

E. E. Carey

Keyword(s):

Ipomoea Batatas ◽

Soft Rot ◽

Northern Ghana ◽

Root Yield ◽

Storage Roots ◽

Yield Data ◽

Level Of Significance ◽

Attainable Yield ◽

The Impact ◽

Model Approach

Abstract This study estimated the amount of loss in storage roots caused by various levels of damage caused by sweetpotato weevils (Cylas spp). Seven varieties of sweetpotato (Ipomoea batatas L. (Lam)) were evaluated in three production sites in northern Ghana for two years (2014 and 2015). Yield data for each experimental plot were collected. A regression analysis was carried out using the generalized linear model approach. In the study, nonmarketable roots were classified as all undersized roots (<100g) and spoilt roots due to weevil, millipede, and soft rot. The results indicated weevil damage as the only significant predictor of nonmarketable yield at 5% level of significance. From the study, the average values for total root yield, marketable root yield, and nonmarketable root yield were 9.39, 6.71, and 2.67 ton/ha respectively. The minimum weevil damage (score 2) resulted in a yield loss of 2 ton/ha which represents 8.3% while severe damage at score 9 could cause a loss of 7.43 ton/ha of storage roots representing 31% of the attainable yield of sweetpotato. Weevil susceptibility needs to be treated as a serious trait when evaluating sweetpotato genotypes to be released as varieties.

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First Report of a Fruit Rot of Pumpkin Caused by Acidivorax avenae subsp. citrulli in Georgia

Plant Disease ◽

10.1094/pdis.1999.83.2.199b ◽

1999 ◽

Vol 83 (2) ◽

pp. 199-199 ◽

Cited By ~ 31

Author(s):

D. B. Langston ◽

R. D. Walcott ◽

R. D. Gitaitis ◽

F. H. Sanders

Keyword(s):

Polyclonal Antibodies ◽

Pcr Amplification ◽

Tween 80 ◽

Soft Rot ◽

Fruit Rot ◽

Identification System ◽

Banding Patterns ◽

First Report ◽

Microbial Identification ◽

Necrotic Spots

In September 1998, a fruit rot was reported affecting pumpkin (Cucurbita pepo) in a commercial field in Terrell Co., Georgia. Symptoms on the surface of fruit occurred as round, necrotic spots or cracks a few millimeters in diameter. With age, the tissue surrounding these lesions became soft and wrinkled. A soft rot expanded into the flesh of the pumpkin, originating from the lesions observed on the surface. In time, infected pumpkins totally collapsed. V-shaped, necrotic lesions occurred at the margin of the leaf and extended inward toward the mid-rib. Samples were collected from the field and bacteria were isolated from fruit and leaf lesions onto King's medium B (1). The bacterium isolated was rod shaped, gram negative, nonflourescent, oxidase positive, Tween 80 positive, carboxymethyl cellulose positive, β-OH butyrate positive, and malonate negative. The bacterium reacted positively with polyclonal antibodies specific for the watermelon fruit blotch pathogen Acidivorax avenae subsp. citrulli and was identified as A. avenae subsp. citrulli by MIDI (Microbial Identification System, Newark, DE) according to statistical analysis of fatty acid data. Results from polymerase chain reaction (PCR) amplification of the bacterium isolated from pumpkin yielded 360-bp fragments that, when digested with the restriction enzyme HaeIII, had DNA banding patterns identical to those of stock A. avenae subsp. citrulli DNA. Koch's postulates were completed successfully with 2-week-old watermelon seedlings. This is the first report of A. avenae subsp. citrulli causing fruit rot of pumpkin in Georgia. Reference: (1) E. O. King et al. J. Lab. Clin. Med. 44:301, 1954.

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First Report of Fusarium solani Causing Root Rot on Coptis chinensis in Southwestern China

Plant Disease ◽

10.1094/pdis-02-14-0164-pdn ◽

2014 ◽

Vol 98 (9) ◽

pp. 1273-1273 ◽

Cited By ~ 2

Author(s):

X.-M. Luo ◽

J.-L. Li ◽

J.-Y. Dong ◽

A.-P. Sui ◽

M.-L. Sheng ◽

...

Keyword(s):

Fusarium Solani ◽

Root Rot ◽

Southwestern China ◽

Molecular Characteristics ◽

Distilled Water ◽

Conidial Suspension ◽

Storage Roots ◽

Coptis Chinensis ◽

Causal Organism ◽

First Report

China is the world's largest producer country of coptis (Coptis chinensis), the rhizomes of which are used in traditional Chinese medicine. Since 2008, however, root rot symptoms, including severe necrosis and wilting, have been observed on coptis plants in Chongqing, southwestern China. Of the plants examined from March 2011 to May 2013 in 27 fields, 15 to 30% were covered with black necrotic lesions. The leaves of infected plants showed wilt, necrotic lesions, drying, and death. The fibrous roots, storage roots, and rhizomes exhibited brown discoloration and progressive necrosis that caused mortality of the infected plants. Infected plants were analyzed to identify the causal organism. Discoloration of the internal vascular and cortical tissues of the rhizomes and taproots was also evident. Symptomatic taproots of the diseased coptis were surface sterilized in 1% sodium hypochlorite for 2 min, rinsed in sterile distilled water for 2 min, and then air-dried in sterilized atmosphere/laminar flow. Small pieces of disinfested tissue (0.3 cm in length) were transferred to petri dishes containing potato dextrose agar (PDA) supplemented with 125 μg ml–1 streptomycin sulfate and 100 μg ml–1 ampicillin, and incubated for 5 days at 25°C with a 12-h photoperiod. Four distinct species of fungal isolates (HL1 to 4) derived from single spores were isolated from 30 plants with root rot symptoms collected from the study sites. To verify the pathogenicity of individual isolates, healthy coptis plants were inoculated by dipping roots into a conidial suspension (106 conidia/ml) for 30 min (15 plants per isolate), as described previously (1). Inoculated plants were potted in a mixture of sterilized quartz sand-vermiculite-perlite (4:2:1, v/v) and incubated at 25/18°C and 85 to 90% relative humidity (day/night) in a growth chamber with a daily 16-h photoperiod of fluorescent light. Plants dipped in sterile distilled water were used as controls. After 15 days, symptoms similar to those observed in the field were observed on all plants (n = 15) that were inoculated with HL1, but symptoms were not observed on plants inoculated with HL2, HL3, and HL4, nor on control plants. HL1 was re-isolated from symptomatic plants but not from any other plants. Morphological characterization of HL1 was performed by microscopic examination. The septate hyphae, blunt microconidia (2 to 3 septa) in the foot cell and slightly curved microconidia in the apical cell, and chlamydospores were consistent with descriptions of Fusarium solani (2). The pathogen was confirmed to be F. solani by amplification and sequencing of the ribosomal DNA internal transcribed spacer (rDNA-ITS) using the universal primer pair ITS4 and ITS5. Sequencing of the PCR product revealed a 99 to 100% similarity with the ITS sequences of F. solani in GenBank (JQ724444.1 and EU273504.1). Phylogenetic analysis (MEGA 5.1) using the neighbor-joining algorithm placed the HL1 isolate in a well-supported cluster (97% bootstrap value based on 1,000 replicates) with JQ724444.1 and EU273504.1. The pathogen was thus identified as F. solani based on its morphological and molecular characteristics. To our knowledge, this is the first report of root rot of coptis caused by F. solani in the world. References: (1) K. Dobinson et al. Can. J. Plant Pathol. 18:55, 1996. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, 2006.

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First Report of Mucor spp. Causing Root Rot of Radix pseudstellariae in Guizhou Province of China

Plant Disease ◽

10.1094/pdis-07-20-1634-pdn ◽

2020 ◽

Author(s):

Hongmiao Wu ◽

Jiachun Wu ◽

Feng Li ◽

Ling Zheng ◽

Jingkai Fan ◽

...

Keyword(s):

Root Rot ◽

Disease Incidence ◽

Acid Soils ◽

Its Region ◽

Significant Loss ◽

Day Length ◽

Guizhou Province ◽

First Report ◽

Chinese Medicinal Plants ◽

Branched Chains

Radix pseudostellariae L. is one of the most common and highly-prized Chinese medicinal plants with various pharmacological effects, and mainly produced in acid soils in the Guizhou and Fujian provinces of southwestern and southeastern China, respectively (Wu et al. 2020). However, consecutive monoculture of R. pseudostellariae results in severe root rot and decline in biomass and quality of underground tubers. Root tubers of R. pseudostellariae are typically planted in December and harvested in next June. Root rot commonly starts developing in May. The disease incidence of root rot was ranging from 37 to 46% in root portions and basal stem of R. pseudostellariae under the consecutive monoculture fields in Shibing County, Guizhou Province, China (108°12ʹE, 27°03ʹN) (Li et al. 2017). Severe root rot was observed in Shibing County in May 2018. Infected plants displayed curly, withered, and yellow leaves, blight, retarded growth, root rot, and yield losses. Abundant whitish mycelia were observed on roots and surrounding soil. Two fungal isolates, designated GZ20190123 and GZ20190124, were obtained from symptomatic roots cultured on potato dextrose agar (PDA). The optimum temperature range for growth of the two isolates was 25 to 30°C. The optimum pH range for the growth of GZ20190123 was 5 to 5.5, whereas GZ20190124 grew better between pH 5 to 8.5. The mean mycelial growth rates of GZ20190123 and GZ20190124 at 30°C were 2.1 and 1.5 cm/day, respectively. Conidia of the two isolates were ovoid or obclavate and were produced in single or branched chains. The internal transcribed spacer (ITS) region was amplified with primers ITS1 and ITS4 (White et al. 1990). The sequences were deposited in GenBank as accession No. MN726736 for GZ20190123 and MN726738 for GZ20190124. Sequence comparison revealed 99% (GZ20190123) and 97% (GZ20190124) identity with previously reported isolate xsd08071 of Mucor racemosus Bull. (accession No. FJ582639.1) and isolate BM3 of Mucor fragilis Bainier (accession No. MK910058.1), respectively, which was confirmed by phylogenetic analysis. The two isolates were tested for pathogenicity on R. pseudostellariae. Six roots of R. pseudostellariae were surface-sterilized with 75% ethanol and stab inoculated with mycelia using a sterile toothpick for each isolate. Sterile distilled water was stab inoculated to twelve roots to serve as the control. Treated roots were incubated in a greenhouse with 16 h day length [light intensity 146.5 μmol/(m2·s)] and day/night temperature 26°C/18°C. The inoculated roots showed the expected symptoms on roots and sprouts 7 days after inoculation, whereas the control roots with sprouts did not show any symptom. The fungi were re-isolated from the diseased roots and confirmed as expected M. racemosus or M. fragilis based on the ITS sequences, which satisfied Koch’s postulates. Thus, isolate GZ20190123 was identified as M. racemosus and GZ20190124 as M. fragilis. Previously, M. racemosus and M. fragilis have been reported as a pathogen on tomato (Kwon and Hong 2005) and grape (Ghuffar et al. 2018), respectively. To our knowledge, this is the first report of M. racemosus and M. fragilis causing root rot of R. pseudostellariae in southwestern China, where the disease could cause a significant loss to production of this important medicinal plant.

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First Report of Fusarium asiaticum Causing Stem Rot of Ligusticum chuanxiong in China

Plant Disease ◽

10.1094/pdis-05-21-1026-pdn ◽

2021 ◽

Author(s):

Tingting Zhu ◽

Linxuan Li ◽

Antonios Petridis ◽

George Xydis ◽

Maozhi Ren

Keyword(s):

Disease Incidence ◽

Elongation Factor ◽

Stem Rot ◽

Post Inoculation ◽

Pathogenicity Test ◽

First Report ◽

Ligusticum Chuanxiong ◽

Fusarium Asiaticum ◽

Sterile Needle ◽

Yellow Pigments

Ligusticum chuanxiong (known as Chuanxiong in China) is a traditional edible-medicinal herb, which has been playing important roles in fighting against COVID-19 (Ma et al. 2020). In March 2021, we investigated stem rot of Chuanxiong in six adjacent fields (~100 ha) in Chengdu, Sichuan Province, China. The disease incidence was above 5% in each field. Symptomatic plants showed stem rot, watersoaked lesions, and blackening with white hyphae present on the stems. Twelve symptomatic Chuanxiong plants (2 plants/field) were sampled. Diseased tissues from the margins of necrotic lesions were surface sterilized in 75% ethanol for 45 s, and 2% NaClO for 5 min. Samples were then rinsed three times in sterile distilled water and cultured on potato dextrose agar (PDA) at 25ºC for 72 h. Fourteen fungal cultures were isolated from 18 diseased tissues, of which eight monosporic isolates showed uniform characteristics. The eight fungal isolates showed fluffy white aerial mycelia and produced yellow pigments with age. Mung bean broth was used to induce sporulation. Macroconidia were sickle-shaped, slender, 3- to 5-septate, and averaged 50 to 70 μm in length. Based on morphological features of colonies and conidia, the isolates were tentatively identified as Fusarium spp. (Leslie and Summerell 2006). To identify the species, the partial translation elongation factor 1 alpha (TEF1-α) gene was amplified and sequenced (O’Donnell et al. 1998). TEF1-α sequences of LCSR01, LCSR02 and LCSR05 isolates (GenBank nos. MZ169386, MZ169388 and MZ169387) were 100%, 99.72% and 99.86% identical to that of F. asiaticum strain NRRL 26156, respectively. The phylogenetic tree based on TEF1-α sequences showed these isolates clustered with F. asiaticum using Neighbor-Joining algorithm. Furthermore, these isolates were identified using the specific primer pair Fg16 F/R (Nicholson et al. 1998). The results showed these isolates (GenBank nos. MZ164938, MZ164939 and MZ164940) were 100% identical to F. asiaticum NRRL 26156. Pathogenicity test of the isolate LCSR01 was conducted on Chuanxiong. After wounding Chuanxiong stalks and rhizomes with a sterile needle, the wounds were inoculated with mycelia PDA plugs. A total of 30 Chuanxiong rhizomes and stalks were inoculated with mycelia PDA plugs, and five mock-inoculated Chuanxiong rhizomes and stalks served as controls. After inoculation, the stalks and rhizomes were kept in a moist chamber at 25°C in the dark. At 8 days post inoculation (dpi), all inoculated stalks and rhizomes exhibited water-soaked and blackened lesions. At 10 dpi, the stalks turned soft and decayed, and abundant hyphae grew on the exterior of infected plants, similar to those observed in the field. No disease symptoms were observed on the control plants. The pathogen was re-isolated from the inoculated tissues and the identity was confirmed as described above. Ten fungal cultures were re-isolated from the 10 inoculated tissues, of which nine fungal cultures were F. asiaticum, fulfilling Koch’s postulates. To our knowledge, this is the first report of F. asiaticum causing stem rot of Chuanxiong in China. Chuanxiong has been cultivated in rotation with rice over multiple years. This rotation may have played a role in the increase in inoculum density in soil and stem rot epidemics in Chuanxiong. Diseased Chuanxiong may be contaminated with the mycotoxins produced by F. asciaticum, 3-acetyldeoxynivalenol or nivalenol, which may deleteriously affect human health. Therefore, crop rotations should be considered carefully to reduce disease impacts.

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First Report of Leaf Spot of Dieffenbachia picta and Aglaonema commutatum Caused by Burkholderia gladioli in Argentina

Plant Disease ◽

10.1094/pdis-93-5-0550c ◽

2009 ◽

Vol 93 (5) ◽

pp. 550-550 ◽

Cited By ~ 1

Author(s):

A. M. Alippi ◽

A. C. López

Keyword(s):

Burkholderia Cepacia ◽

Disease Incidence ◽

Soft Rot ◽

Distilled Water ◽

First Report ◽

Burkholderia Gladioli ◽

Sterile Needle ◽

Pcr Products ◽

Humidity Chamber ◽

Physiological Tests

During May of 2008 (austral autumn), an uncharacterized disease was observed on Dieffenbachia picta (Lodd.) Schott and Aglaonema commutatum Schott in commercial greenhouses in Pontevedra (34°45′6″S, 58°42′42″W), Argentina. Affected plants showed irregular, brown lesions on leaves, approximately 15 to 20 mm in diameter, surrounded by water-soaked haloes that progressed inward from the margins. Water-soaked rotting symptoms were also observed in petioles. Disease incidence approached 80%. Abundant bacterial streaming was observed from lesions when examined at ×100. Bacteria consistently isolated from lesions formed cream-colored, glistening, convex colonies on sucrose peptone agar and produced a yellowish green, diffusible, nonfluorescent pigment on King's medium B. Four isolates from different symptomatic plants were selected for further study. All were aerobic, gram-negative rods that accumulated poly-β-hydroxybutyrate inclusions. In LOPAT tests, all induced a hypersensitive response in tobacco plants, caused soft rot of potato tubers, and were positive for levan, negative for arginine dihydrolase, and variable for oxidase. All isolates oxidized glucose, did not hydrolyze starch and were able to rot onion slices. Colonies developed at 41°C but not at 4°C. With the API 20NE test strips and database (bioMerieux, Buenos Aires, Argentina), all isolates matched (99% identity) Burkholderia cepacia, but their inability to metabolize cellobiose and sucrose further identified them as B. gladioli. For molecular identification, 23S rDNA was amplified by PCR using B. gladioli-specific primers LP1 and LP4, which yielded a 700-bp product (3), and PCR-restriction fragment length polymorphism of 16S rDNA using AluI (2). PCR products were identical to those from the type strain for B. gladioli, ICMP 3950, isolated from Gladiolus spp. that had been included in all tests for comparison. Pathogenicity was verified on D. picta and A. commutatum by spraying the plants with bacterial suspensions in sterile distilled water at 108 CFU/ml with and without wounding the leaves with a sterile needle and also by injection-infiltration of bacterial suspensions at 105 CFU/ml. In addition, another host plant, Gladiolus communis L., was inoculated in the same manner. Controls were sprayed or infiltrated with sterile distilled water. After 48 h in a humidity chamber, plants were kept at 25 ± 3°C in a greenhouse. In all hosts, symptoms were first detected 3 days after inoculation and lesions expanded to resemble natural infections within 4 to 7 days. All strains caused necrosis around the inoculation sites and lesions were identical to those induced by the ICMP reference strain. Bacteria were reisolated from each host tested and then the original and reisolated strains were compared by enterobacterial repetitive intergeneric consensus-PCR (1); DNA fingerprints of the reisolated strains were identical to those of the original strains, thereby fulfilling Koch's postulates. No lesions were observed on controls or on plants inoculated by spraying without wounding, suggesting that bacteria gain entry through wounds. On the basis of PCR and physiological tests the pathogen was identified as B. gladioli (2–4). To our knowledge, this is the first report of B. gladioli on Dieffenbachia and Aglaonema spp. References: (1) F. J. Louws et al. Appl. Environ. Microbiol. 60:2286, 1994. (2) C. Van Pelt et al. J. Clin. Microbiol. 37:2158, 1999. (3) P. W. Whitby et al. J. Clin. Microbiol. 38:282, 2000. (4) E. Yabuuchi et al. Microbiol. Immunol. 36:1251, 1992.

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