scholarly journals First Report of Fusarium Wilt of Chicory (Cichorium intybus) Caused by Fusarium oxysporum in Italy

Plant Disease ◽  
2011 ◽  
Vol 95 (4) ◽  
pp. 496-496 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
A. Poli ◽  
M. L. Gullino

In the summer of 2009, a wilt of chicory was observed on 25 to 30% of 30-day-old Cichorium intybus L. cv. Clio plants grown outdoors on a commercial farm in Piedmont (northern Italy). Affected plants were chlorotic and stunted with poorly developed root systems compared with healthy plants. Black streaks were observed in the stem and proximal part of the leaf vascular system in wilted plants. Fusarium oxysporum Schltdl. was isolated from symptomatic vascular tissue on a Fusarium-selective medium (1) from 80% of samples. Grown on potato dextrose agar (PDA) for 4 days at 23°C, the colonies, initially white and later pale pink, produced hyaline microconidia that were oval-elliptical and cylindrical in shape measuring 5.6 to 14.9 (average 10.2) × 2.1 to 4.5 (3.0) μm, borne on short monophialides measuring 8.2 to 16.1 (average 13.2) × 2.1 to 4.2 (3.3) μm. Macroconidia were slightly curved, three-septate, with a slightly hooked apical cell and a foot-shaped basal cell measuring 24.9 to 41.6 (average 32.2) × 3.2 to 5.2 (4.3) μm. Chlamydospores were both terminally and intercalary, solitary but also in short chains (2 to 4 elements) measuring 21.1 to 41.0 (average 27.2) μm (2). The internal transcribed spacer (ITS) rDNA region was amplified using the primers ITS1/ITS4 and sequenced. BLASTn analysis of the 527-bp amplicon (GenBank Accession No. HQ644423) obtained had 98% sequence identity with F. oxysporum (GenBank Accession No. FJ605247). The translation elongation factor-1α (EF-1α) gene was amplified using primers EF-1/EF-2 and sequenced (GenBank Accession No. GU564259). The 663-bp fragment had 99% sequence identity with F. oxysporum (GenBank Accession Nos. EU313540, EU313539, and DQ837696). Pathogenicity tests were conducted on 15-day-old chicory plants from two cultivars (Clio and Katia). Thirty-five plants per cultivar were inoculated by dipping their roots in a 1 × 106 CFU/ml suspension of isolate FusCic45B recovered from wilted chicory. Inoculated and noninoculated plants were transplanted into five pots filled with 10 liters of steamed mix (peat/perlite/sand, 60:20:20 vol/vol) and were maintained in a glasshouse at 25 to 27°C. Wilt symptoms and vascular discoloration of the roots, crown, and veins developed 15 days after inoculation on all inoculated plants. Plants of cv. Clio were more susceptible. F. oxysporum was always reisolated from infected plants using the Fusarium-selective medium. All noninoculated plants remained healthy. The pathogenicity test was conducted twice. To our knowledge, this is the first report of wilt caused by F. oxysporum on chicory, C. intybus, in Italy as well as worldwide. References: (1) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (2) E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. The Pennsylvania State University Press, University Park, 1983.

Plant Disease ◽  
2009 ◽  
Vol 93 (10) ◽  
pp. 1078-1078 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. Troisi ◽  
M. L. Gullino

In the summer of 2007, a wilt of endive (Cichorium endivia) cv. Myrna plants was observed on several commercial farms near Alessandria in northern Italy. Approximately 40% of the plants on each farm were symptomatic. Affected plants were stunted and yellow and their root systems were poorly developed. Basal leaves eventually wilted. Black streaks were observed in the proximal part of the leaf vascular system. Fusarium oxysporum was consistently isolated from symptomatic vascular tissue on a Fusarium-selective medium (1). To confirm identification of the pathogen, the translation elongation factor 1α and the ribosomal DNA intergenic spacer (IGS) region were partially sequenced (2). Genomic DNA was extracted from mycelia growing on potato dextrose agar. Amplification of the EF-1α region generated a sequence of 648 bp; the IGS amplicon was 2,500 bp. The EF-1α sequence (GenBank Accession No GQ398152) was 99% similar to the sequence of a F. oxysporum strain isolated from soil and a strain pathogenic on cotton plants (GenBank Accession No. EU246574). The IGS sequence (GenBank Accession No GQ398153) was 97% similar to the sequence of a F. oxysporum strain (GenBank Accession No. EF661647). Pathogenicity tests were conducted on 15-day-old endive plants (cv. Myrna). Ten plants were inoculated by dipping their roots in a 1 × 106 CFU/ml suspension of one of the isolates recovered from a wilted endive plant. Inoculated and noninoculated plants were transplanted into pots filled with steamed soil and maintained in a glasshouse at 23 to 28°C. Wilt symptoms and vascular discoloration of the roots, crown, and veins developed 60 days after inoculation. F. oxysporum was consistently reisolated from infected plants. Noninoculated plants remained healthy. The pathogenicity test was conducted twice. To our knowledge, this is the first report of wilt caused by F. oxysporum on endive in Italy. References: (1) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (2) G. Mbofung et al. Phytopathology 97:87, 2007.


Plant Disease ◽  
2011 ◽  
Vol 95 (9) ◽  
pp. 1191-1191 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
P. Pensa ◽  
A. Poli ◽  
M. L. Gullino

During summer 2010, symptoms of a wilt disease were observed in a commercial farm in northern Italy on Crassula ovata (jade plant). First symptoms consisted of chlorosis and premature drop of still turgid leaves. As the disease progressed, leaves turned yellow and wilted before dropping off and the stem wilted, bent, and eventually rotted starting from the base. In some cases, the stem broke or the basal portion of the leaf rotted. Brown discolorations were observed in the vascular system. Of 10,000 plants, 65% (cv. Mini) and 5% of 600 plants (cv. Magical Tree) were affected. Premature dropping of leaves was more frequent on cv. Magical Tree. Using the Komada's Fusarium-selective medium, a fungus was consistently and readily isolated from symptomatic vascular tissues of plants belonging to both cultivars. Isolates obtained from both cultivars were purified, subcultured on potato dextrose agar (PDA), and single-spore cultures were obtained. On PDA, both isolates produced pale violet, abundant, aerial mycelium, felted in old cultures, with purple pigments in the agar. The isolates were grown on Spezieller Nährstoffarmer agar for characterization of macroconidia and microconidia (1). Both isolates produced sparse, 3 to 5 septate, nearly straight macroconidia measuring 30 to 56 × 3 to 5 (average 40 × 4) μm with a short apical cell and a foot-shaped basal cell. Sporodochia were not observed. Unicellular, oval-elliptical microconidia measuring 5 to 13 × 3 to 4 (average 8 × 3) μm were produced on short monophialides. Chlamydospores were abundant, single and sometime in pairs, terminal and intercalary, rough walled, and measured 6 to 9 μm. Such characteristics are typical of Fusarium oxysporum (3). The ITS region (internal transcribed spacer) of rDNA was amplified with primers ITS1/ITS4 (4) and sequenced. BLASTn analysis of an isolate from C. ovata cv. Mini (515 bp, Accession No. HQ682196) and C. ovata cv. Magical Tree (509 bp, Accession No. HQ682197) showed an E-value of 0.0 with F. oxysporum. For these sequences, pairwise alignment of EMBOSS (E.B.I. - The European Bioinformatics Institute) revealed identity and similarity of 99.0%. To confirm pathogenicity, tests were conducted on 5-month-old plants of cvs. Mini and Magical Tree. Plants (three per treatment) were inoculated by dipping roots in a 1 × 106 CFU/ml conidial suspension of the two isolates of F. oxysporum prepared from 10-day-old cultures grown on casein liquid medium (2), shaken (90 rpm) for 10 days at 24°C ± 1 (12-h fluorescent light, 12-h dark). Inoculated plants were transplanted into pots filled with steamed mix (sphagnum peat/perlite/pine bark/clay; 50:20:20:10) and maintained in a plant growth chamber at 25 ± 1°C under a regimen of 12 h per day of fluorescent light. Inoculated plants belonging to both cultivars showed typical first symptoms of Fusarium wilt after 13 days. In the following days, leaves dropped, stems wilted, and plants died. Noninoculated plants remained healthy. F. oxysporum was reisolated from inoculated plants. The pathogenicity test was conducted twice. This is, to our knowledge, the first report of F. oxysporum on C. ovata in Italy or worldwide. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Professional, Ames, IA, 2006. (2) A. Minuto et al. Phytoparasitica 36:294, 2008. (3) B. A. Summerell et al. Plant Dis. 87:117, 2003. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.


Plant Disease ◽  
2004 ◽  
Vol 88 (3) ◽  
pp. 309-309 ◽  
Author(s):  
A. Garibaldi ◽  
A. Minuto ◽  
M. L. Gullino

During the fall of 2002, African daisy (Osteospermum sp.) plants showing symptoms of a wilt disease were observed in a commercial, nonheated glasshouse in Albenga in northern Italy. Wilted plants were first observed when outside temperatures were between 15 and 28°C. Symptoms were first observed on seedlings 40 days after they had been transplanted into pots. The vascular tissues of affected plants appeared brown. These plants were stunted and developed yellowed leaves with brown or black streaks in the vascular system. The vascular streaks in the yellow leaves extended from the crown and were continuous with a brown discoloration in the vascular system of the crown and upper taproot. Fusarium oxysporum was consistently and readily isolated from symptomatic vascular tissue onto a Fusarium-selective medium (1). Healthy, rooted, 40-day-old plants were inoculated by root-dip with a conidial suspension (1 × 107 CFU/ml) of three isolates of F. oxysporum obtained from infected plants and transplanted into pots filled with steam-sterilized soil. Noninoculated plants served as control treatments. Plants (10 per treatment) were grown in a glasshouse at an average temperature of 25°C (minimum of 12°C and maximum of 39°C). Wilt symptoms and vascular discoloration in the roots, crown, and veins developed within 20 days on each inoculated plant, while noninoculated plants remained healthy. F. oxysporum was consistently reisolated from infected plants. The pathogenicity test was conducted twice. To our knowledge, this is the first report of F. oxysporum on Osteospermum sp. in Italy or elsewhere in the world. Reference: (1) H. Komada. Rev. Plant Prot. Res. 8:114, 1975.


Plant Disease ◽  
2002 ◽  
Vol 86 (9) ◽  
pp. 1052-1052 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

In spring 2001, plants of the lettuce cv. Salad Bowl showing symptoms of a wilt disease were observed in several commercial plastic greenhouses near Bergamo, in northern Italy. Wilted plants were first observed during the spring and summer of 2001 when temperatures were between 26 and 35°C. Symptoms were observed in the same area and the same farms in March 2002, in concomitance with a period of high temperatures. Although the distribution of the disease was generally uniform, symptoms were more severe in the central part of the greenhouses where temperatures were warmest. Symptoms were first observed at thinning, when seedlings (30 days old) appeared wilted. Vascular tissues of affected seedlings appeared red or brown. Affected plants were stunted and developed yellow leaves and brown or black streaks in the vascular system. The vascular streaks in the yellow leaves extended from the crown and were continuous with a red-brown discoloration in the vascular system of the crown and upper taproot. Symptoms were typically not visible on the outside of the crowns or roots. Fusarium oxysporum was consistently and readily isolated from symptomatic vascular tissues on a Fusarium-selective medium (2). Seeds of cv. Salad Bowl were planted in steam-sterilized soil artificially infested with 1 × 104 CFU/g soil of each of two isolates of F. oxysporum obtained from infected plants. Plants grown in noninfested soil served as noninoculated control treatments. Plants (25 per treatment) were grown at 25 to 28°C in growth chambers. Wilt symptoms and vascular discoloration of the roots, crown, and veins developed 20 days after seeding for all plants grown in the infested soil, and F. oxysporum was consistently reisolated from infected plants. The pathogenicity test was conducted twice. To our knowledge, this is the first report of F. oxysporum on lettuce in Europe. A wilt of lettuce attributed to F. oxysporum f. sp. lactucae was reported in Japan in 1967 (3) and in the United States in 1993 (1), in the latter case the incitant organism was described as F. oxysporum f. sp. lactucum. References: (1) J. C. Hubbard and J. S. Gerik. Plant Dis. 77:750, 1993. (2) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (3) T. Matuo and S. Motohashi. Trans. Mycol. Soc. Jpn. 8:13, 1967.


Plant Disease ◽  
2004 ◽  
Vol 88 (1) ◽  
pp. 83-83 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

Lamb's lettuce (Valerianella olitoria), also known as corn salad, is increasingly grown in Italy and used primarily in the preparation of mixed processed salad. In the summer of 2003, plants of lamb's lettuce cvs. Trophy and Palmares exhibiting wilt symptoms were observed in several commercial greenhouses near Bergamo in northern Italy. Wilted 30-day-old plants were observed first during the month of June, at the time of thinning when temperatures ranged between 28 and 35°C. Disease was generally uniform in the greenhouses and 30 to 50% of the plants were affected. Vascular tissue of affected seedlings appeared red or brown but later turned brown or black. Affected plants were stunted and developed yellowed leaves. Vascular discoloration was continuous from the upper taproot through the crown to the leaf. Fusarium oxysporum was consistently isolated from symptomatic vascular tissue onto a Fusarium-selective medium (1). Seeds of the same cultivars (Trophy and Palmares) affected by the wilt in the field were artificially inoculated by dipping them for 15 min into spore suspensions (1 × 106 conidia per ml) of three isolates of F. oxysporum obtained from infected plants. Noninoculated seeds served as control treatments. Forty seeds per treatment were sown in pots (1-liter volume) containing steam-sterilized soil and maintained at 25°C in a growth chamber programmed for 12 hours of light per day. Wilt symptoms developed on both cultivars 20 days after seeding, and F. oxysporum was consistently reisolated from infected plants. The plants obtained from noninoculated seeds remained healthy. The pathogenicity test was carried out twice with similar results. To our knowledge, this is the first report of F. oxysporum causing vascular wilt of lamb's lettuce and may warrant a new forma specialis designation. Reference: (1) H. Komada. Rev. Plant Prot. Res. 8:114, 1975.


Plant Disease ◽  
2003 ◽  
Vol 87 (2) ◽  
pp. 201-201 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

Two types of rocket are available on the market in Italy: (i) Eruca vesicaria (synonym E. sativa) known as ruchetta or cultivated garden rocket; and (ii) several species of Diplotaxis (Diplotaxis erucoides, D. muralis, and D. tenuifolia), which are wild plants now widely cultivated. Rocket is increasingly used in the mediterranean cuisine as salad and or to decorate dishes. In spring 2002, plants of the cultivated (E. vesicaria) and wild (Diplotaxis spp.) rocket showing symptoms of a wilt disease were observed in several commercial plastic greenhouses near Bergamo, in northern Italy. Wilted plants were first observed during the spring and summer of 2001 when temperatures were between 26 and 35°C. In May 2002, symptoms were again observed in the same area, on the same farm as well as other farms. Although the distribution of the disease was generally uniform, symptoms were more severe in the central part of the greenhouses where temperatures were warmest (30 to 60% of plants were affected). Diseased plants were stunted and chlorotic with brown or black streaks in the vascular system. Vascular tissues of affected plants appeared red or brown. Vascular streaks in the chlorotic leaves extended from the crown and were continuous with a red-brown discoloration in the vascular system of the crown and upper taproot. Fusarium oxysporum was consistently and readily isolated from symptomatic vascular tissues when plated on a Fusarium-selective medium (2). Microconidia measured 8.8 × 3.0 µm. E. vesicaria and Diplotaxis spp. were grown in steam-sterilized soil, and 10 days after emergence they were artificially inoculated by root dipping in a spore suspension (1 × 105 CFU/ml) of three F. oxysporum strains collected from infected plants. Uninoculated plants served as control. Plants (60 per treatment) were grown at 25 to 28°C in growth chambers. Wilt symptoms developed on all plants 20 days after inoculation, and F. oxysporum was consistently reisolated from infected plants. The pathogenicity test was conducted twice. To our knowledge, this is the first report of F. oxysporum on cultivated rocket in Europe and the first on wild rocket (Diplotaxis spp.) in the world. A wilt of E. sativa attributed to F. oxysporum f. sp. erucae was previously reported in India in 1973 (1). Studies are being carried out to determine if the Italian isolates of F. oxysporum belong to the same formae speciales. References: (1) C. Chatterjee and J. N. Rai. Indian Phytopathol 28:309, 1973. (2) H. Komada. Rev. Plant Prot. Res. 8:114, 1975.


Plant Disease ◽  
2005 ◽  
Vol 89 (6) ◽  
pp. 684-684 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

During the spring of 2004, bitterroot (Lewisia cotyledon (S. Wats.) B.L. Robins), a flowering perennial of interest for rock gardens, showed symptoms of a wilt disease in several commercial nurseries located in Piedmont (northern Italy). Ten to thirty percent of plants from several different nurseries were affected. Wilted plants were first observed approximately 20 days after being transplanted into containers when outside temperatures ranged from 15 to 22°C. Symptomatic plants were stunted with yellowed leaves and brown-to-black veins. The vascular browning extended into the crown and was continuous with a brown discoloration in the vascular system of the crown and upper taproot. Fusarium oxysporum, identified on the basis of colony and conidia morphology (1), was consistently and readily isolated from symptomatic vascular tissue onto a Fusarium-selective medium (2). Three isolates were transferred to casein hydrolysate and grown for 10 days to produce conidial inoculum. Healthy 30-day-old plants were inoculated by dipping roots into a (1 × 106 CFU/ml) conidial suspension. Plants were then transplanted into pots filled with steam-sterilized soil. Noninoculated plants served as a control. Plants (12 per treatment) were placed in a climatic chamber at 25°C. Wilt symptoms and vascular discoloration in the roots, crown, and veins developed within 20 days on each inoculated plant, while noninoculated plants remained healthy. F. oxysporum was consistently reisolated from symptomatic plants. The pathogenicity test was conducted twice with the same result. One isolate of the pathogen has been deposited in the ATCC culture collection. To our knowledge, this is the first report of F. oxysporum on L. cotyledon in Italy and elsewhere in the world. Currently, this disease is present in several nurseries in the Piedmont Region of Italy. References: (1) C. Booth. Fusarium. CMI, Kew, UK, 1977. (2) H. Komada. Rev. Plant Prot. Res. 8:114, 1975.


Plant Disease ◽  
2005 ◽  
Vol 89 (10) ◽  
pp. 1130-1130 ◽  
Author(s):  
S. T. Koike ◽  
T. R. Gordon

Cilantro, or coriander (Coriandrum sativum), is a leafy vegetable in the Apiaceae and is grown commercially in California primarily for use as a fresh herb. During 2002 and 2003 in coastal California (Santa Barbara County), commercial cilantro fields showed symptoms of a wilt disease. Affected plants grew poorly and were stunted. Lower foliage turned yellow with reddish tinges, and plants wilted during warmer times of the day. The main stem, crown, and taproot exhibited vascular discoloration that was reddish to light brown. As disease progressed, plants eventually died. For both years, the disease distribution was limited to isolated small patches (each patch measuring less than 1 m2 in area). A fungus was consistently isolated from symptomatic vascular tissue in crowns and taproots. On the basis of colony and conidial morphology, the isolates were identified as Fusarium oxysporum (2). No other fungi or bacteria were recovered from these plants. To test pathogenicity, suspensions containing 1 × 106 conidia/ml were prepared for five isolates. The roots of 30-day-old cilantro plants of four cultivars (30 plants each of Festival, Leisure, Santo, and LSO 14) were clipped and then soaked in the suspensions for 20 min. The roots of 30 plants of each cultivar were soaked in water as a control. Plants were repotted into new redwood bark + peat moss rooting medium and maintained in a greenhouse setting at 24 to 26°C. After 1 month, 95% or more of the inoculated plants showed yellowing and vascular discoloration symptoms similar to those seen in the field. F. oxysporum was reisolated from all inoculated plants. The four cilantro cultivars did not show differences in disease severity. Control plants showed no symptoms, and the fungus was not recovered from these plants. The experiment was repeated and the results were the same. Experiments also were conducted to determine if cilantro isolates could cause disease in celery (Apium graveolens var. dulce). Celery transplants and cilantro seedlings were prepared and inoculated as described above. However, after 2 months, celery plants did not show any disease symptoms, while the cilantro developed wilt symptoms and eventually died. A Fusarium wilt disease has been reported on coriander in Argentina and India where the pathogen was named F. oxysporum f. sp. coriandrii (1,3). To our knowledge, this is the first report of Fusarium wilt of cilantro in California. References: (1) M. Madia et al. Fitopatologia 34:155, 1999. (2) P. E. Nelson et al. Fusarium species: An Illustrated Manual for Identification. Pennsylvania State University Press, University Park, 1983. (3) U. S. Srivastava. Indian Phytopathol. 22:406, 1969.


Plant Disease ◽  
2010 ◽  
Vol 94 (9) ◽  
pp. 1163-1163 ◽  
Author(s):  
K. Perveen ◽  
N. Bokhari

In November 2008, a wilt of lavender (Lavandula pubescens) seedlings was observed in the greenhouse at King Saud University, Riyadh, Saudi Arabia. Affected seedlings were wilted and the root system was poorly developed. Diseased stems developed a dark coloration that extended down to the roots. Vascular tissue of the affected seedlings appeared red or brown. Isolations consistently yielded a fungus growing from the discolored stem tissue when placed on potato dextrose agar. The macroscopic characteristics of the colony, as well as microscopic structures, were used to identify the fungus as Fusarium oxysporum (2). Oval to elliptical microconidia without septa and originating from short phialides were used to distinguish the species from F. solani (1). The fungus was authenticated by the ITCC (Indian Type Collection Centre), Indian Agricultural Research Institute, New Delhi, India, and given I.D. No. 7532.09. For conducting further experiments, healthy seedlings of L. pubescens were obtained from the botanical garden of the King Saud University and grown in steam-sterilized soil. Healthy seedlings of lavender were inoculated using a root-dip method with a conidial suspension (1 × 107 CFU/ml) of one strain of F. oxysporum obtained from infected plants. Inoculated seedlings were then transplanted into steam-sterilized soil. Plants inoculated with sterilized water (1 ml per plant) served as control treatments. Wilt symptoms and vascular discoloration in the roots and crown developed within 20 days on all plants inoculated with the pathogen, while control plants remained asymptomatic. F. oxysporum was consistently reisolated from symptomatic plants. The pathogenicity test was conducted twice. To our knowledge, this is the first report of F. oxysporum on L. pubescens in Saudi Arabia or elsewhere in the world, and this newly identified disease may be a potential threat to commercial production of lavender. References: (1) J. F. Leslie and B. A. Summerell. Page 212 in: The Fusarium Laboratory Manual. Blackwell Publishing Professional, Hoboken, NJ, 2006. (2) P. C. Nelson et al. Clin. Microbiol. Rev. 7:479, 1994.


Plant Disease ◽  
2021 ◽  
Author(s):  
Laikun Xia ◽  
Yanyong Cao ◽  
Jie Wang ◽  
Jie Zhang ◽  
Shengbo Han ◽  
...  

Maize stalk rot has become one of the most important diseases in maize production in China. From 2017 to 2019, a survey was conducted to determine the population diversity of Fusarium species associated with maize diseases in 18 cities across Henan Province. Maize stalk rot with an incidence of more than 20% that caused yield losses up to 30% was observed on maize variety Zhengdan958, which was grown in two continuous maize fields in Zhumadian City, Henan Province. The stem tissues from the boundary between diseased and healthy pith were chopped into small pieces (3 × 8 mm), disinfected (70% ethanol for 1 min) and then placed onto potato dextrose agar (PDA) amended with L-(+)-Lactic-acid (1 g/L) and incubated at 25°C for 4 days. Colonies on PDA produced fluffy, light yellow aerial mycelium and purple to deep brick red pigment at 25°C (Fig 1A, 1B). On carnation leaf agar (CLA), macroconidia in orange sporodochia formed abundantly, but microconidia were absent. Macroconidia were short and thick-walled, had 3 to 5 septa, a poorly developed foot cell and rounded apical cell (Fig 1C). These characteristics matched the description of Fusarium culmorum (Leslie and Summerell 2006) and isolates DMA268-1-2 and HNZMD-12-7 were selected for further identity confirmation. Species identification was confirmed by partial sequences of three phylogenic loci (EF1-α, RPB1, and RPB2) using the primer pairs EF1/EF2, CULR1F/CULR1R, and CULR2F/CULR2R, respectively (O'Donnell et al., 1998). The consensus sequences from the two isolates were deposited in GenBank (MZ265416 and MZ265417 for TEF, respectively; MZ265412 and MZ265414 for RPB1, respectively; MZ265413 and MZ265415 for RPB2). BLASTn searches indicated that the nucleotide sequences of the three loci of the two isolates revealed 99% to 100% similarity to those of F. culmorum strains deposited in the GenBank, Fusarium-ID, and MLST databases (Supplementary Table 1~3). Pathogenicity test was conducted at the flowering-stage using Zhengdan958 and Xundan20 plants according to previously described method (Zhang et al., 2016; Cao et al., 2021; Zhang et al., 2021). The second or third internodes of thirty flowering plants were drilled to make a wound approximately 8 mm in diameter using an electric drill. Approximately 0.5 mL inoculum (125 mL colonized PDA homogenized with 75 mL sterilized distilled water) was injected into the wound and sealed with Vaseline and Parafilm to maintain moisture and avoid contamination. Sterile PDA slurry was used as a control. Thirty days after inoculation, the dark-brown, soft rot of pith tissues above and below the injection sites were observed, and some plants were severely rotten and lodged (Fig 1D, 1E). These symptoms were similar to those observed in the field. No symptoms were observed on control plants. The same pathogen was re-isolated from the inoculated stalk lesions but not from the control, thereby fulfilling Koch's postulates. To our knowledge, this is the first report of F. culmorum as the causal agent of stalk rot on maize plants in China. Also, this fungus has been reported to cause maize ear rot in China (Duan et al. 2016) and produce mycotoxins such as trichothecenes, nivalenol, and zearalenone that cause toxicosis in animals (Leslie and Summerell 2006). The occurrence of maize stalk rot and ear rot caused by F. culmorum should be monitored due to the potential risk for crop loss and mycotoxin contamination.


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