First Report of Sea buckthorn Stem Wilt Caused by Fusarium proliferatum in Liaoning, China

Sea buckthorn（Hippophae rhamnoides L.） is a flowering shrub native to cold-temperate regions of Eurasia, which is also valuable for its berries and leaves containing various vitamins and flavonoids (Pundir et al. 2021). In late June 2020, high mortality (more than 70%) was observed in sea buckthorn in a 1.6-ha seedling nursery in Chaoyang City, Liaoning province, China, where 16 Chinese and Russian cultivars (cv.) had been planted since 2014 (cv. Shenqiuhong, eshi01 through eshi15). The mortality of two introduced sea buckthorn varieties (eshi02, eshi04) was 100% (125 trees died in total). The symptoms include massive drooping leaves and dried-up stems on 6-year-old infected trees. Pieces of tree roots and stems with brown discoloration in the xylem vessels were selected. Small tissue fragments (0.2-0.5 cm) were surface disinfested (3 min in 75% ethanol, rinsed with sterile distilled water), air-dried, and placed on potato dextrose agar (PDA) medium for 5 days at 25°C in the dark. A fungus was consistently isolated from both diseased roots and stems tissues, and a representative isolate (LC-1) was harvested. Genomic DNA was extracted for amplification and sequencing of the partial translation elongation factor-1α (EF1 and EF2 primers, accession Nos. MZ669853) (O’Donnell et al. 1998) and RNA polymerase II second largest subunit (RPB2) (7cf/11aR primers, accession Nos. MZ669854) (O’Donnell et al. 2007). The sequences were further analyzed at the Fusarium MLST (https://fusarium.mycobank.org/) for identity confirmation, and showed 99.8% (over 95.2% query coverage) and 96.4% (over 88.4% query coverage) similarity to Fusarium proliferatum (NRRL 13584, 13591). Isolates on Spezieller Nahrstoffarmer agar (SNA) produced abundant aerial white mycelia and yellow pigmentation. The 30 macroconidia measured ranged from 28.5 - 62.5 × 3.2 – 5.4 μm, were thin, slender, with 3-5 septa. The aseptate microconidia ranged from 4.7 – 13.6 × 2.2 – 4.3 μm (n = 30). Pathogenicity tests were performed on healthy, potted 1-year-old sea buckthorn seedlings (cv. eshi05) using two isolates in a greenhouse at 25 °C, 80% relative humidity, and 12-hour light/dark photoperiod. Ten potted seedlings were inoculated on the stems by placing a 5-mm-diameter mycelial plug (5-day-old PDA cultures for each isolate) into the surface of a wound created with a needle, and the inoculation sites were covered with Parafilm to maintain moisture. Ten seedlings were inoculated with PDA plugs as controls. Six to ten days after inoculation, color of the leaves in the middle of the stems was variegated, and then dark necrotic lesions on leaf margins were observed. Three weeks after inoculation, 80% of inoculated stems were wilted, while control plants remained asymptomatic. The pathogen was consistently re-isolated and the recovered isolates were identified as F. proliferatum by amplifying the EF-1α gene. The typical symptoms on inoculated plants were dark to brown necrotic lesions on chlorotic leaves initially, and black withered stems in the terminal stage, similar to those observed on sea buckthorn trees infected with Fusarium sporotrichioides in Gansu and Heilongjiang provinces (Song et al. 2010; Xia et al. 2021). To our knowledge, this is the first report of sea buckthorn stem wilt caused by F. proliferatum in Liaoning province, China, which will be beneficial for expanding knowledge of Fusarium disease in sea buckthorn and provide more information for sustainable disease management in sea buckthorn.

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First Report of Sea buckthorn Stem Wilt Caused by Fusarium sporotrichioides in Gansu, China

Plant Disease ◽

10.1094/pdis-03-21-0627-pdn ◽

2021 ◽

Author(s):

Bo Xia ◽

Yue Liang ◽

Jianzhong Hu ◽

Xiaoling Yan ◽

Liqiang Yin ◽

...

Keyword(s):

Elongation Factor ◽

Morphological Characteristics ◽

Gansu Province ◽

Sea Buckthorn ◽

Molecular Characteristics ◽

Ear Rot ◽

Fusarium Sporotrichioides ◽

Tree Roots ◽

Internal Transcribed Spacer Region ◽

First Report

Sea buckthorn (Hippophae rhamnoides) is an important deciduous shrub for fruit and ecological restoration in arid and semi-arid regions of China. Twelve Chinese and Russian cultivars (cv. Shenqiuhong, eshi01, ... eshi11) were planted about 1.6 acre area in a seedling nursery, located in Qingyang City of Gansu province in northwest China, where high mortality (more than 70%) of sea buckthorn was observed in late July 2019. Symptoms consisted of massive chlorosis, drooping leaves and dried-up stems on 5-year-old trees. Pieces of tree roots and stems with irregular light-brown discoloration in the xylem vessels were selected. Small pieces of discolored tissue were surface disinfested (1 min in 1% sodium hypochlorite, followed by three rinses with sterile distilled water), air-dried, and placed on potato dextrose agar (PDA) medium for 5 days at 25°C in the dark. A fungus was consistently isolated from both diseased roots and stems tissues. Colonies on PDA grew rapidly. Dense mycelia were pinky-white initially, and became carmine red color with age on the undersurface of the plate. Macroconidia were moderately curved, 3 to 5 marked septa, hyaline, thick walled, and measuring 27.8± 3.6 µm × 4.8 ± 0.5 µm (n = 30). Microconidia were abundant, pear-shaped, ellipsoid to fusoid, often with a papilla at the base, and 8.4 ± 2.2 µm ×3.1 ± 0.3 µm (n = 30). Genomic DNA was extracted for amplification and sequencing of the internal transcribed spacer region (ITS1 and ITS4 primers) (White et al. 1990) of the ribosomal DNA (Accession Nos. MN160235 to MN160238) and translation elongation factor-1 alpha (EF1 and EF2 primers, accession Nos. MN429075 to MN429078) (O’Donnell et al. 1998). The sequences revealed 99% similarity to the sequences of the ITS (AY188917), and 100% identity with EF1-α (JF740808) regions of Fusarium sporotrichioides. Based on morphological and molecular characteristics, the fungus was identified as F. sporotrichioides (Leslie and Summerell 2006). Koch’s postulates were fulfilled on healthy, potted 1-year-old sea buckthorn seedings using two isolates in a greenhouse at 25 °C, 90% relative humidity, and 12-hour light/dark photoperiod. Ten potted seedings were inoculated on the stems by placing a 5-mm-diameter mycelial plug (5-day-old PDA cultures for each isolate) into the surface of a wound created with a needle, and the inoculation sites were covered with Parafilm to maintain moisture. Ten seedings were inoculated with PDA plugs as controls. Seven to ten days after inoculation, typical symptoms of dark-brown necrotic lesions on chlorotic leaf margins were observed. About 2 weeks after inoculation, the inoculated stems were gradually dry up, accompanied by withering and fallen leaves. Control plants remained asymptomatic. Pathogens were successfully isolated from the inoculated stems again, exhibiting morphological characteristics identical to those of F. sporotrichioides. Previous papers reported F. sporotrichioides as a common pathogen caused lavender wilt (Cosic et al. 2012), foliar spots on forage corn (Moya-Elizondo et al. 2013) and maize ear rot (Wang et al. 2019). To our knowledge, this is the first report of sea buckthorn stem wilt caused by F. sporotrichioides on several Chinese and Russian cultivars in Gansu province of China. In Heilongjiang province, the same disease was reported in 2010 (Song et al. 2010), nearly 30 longitudes away from Gansu province. Therefore, this disease appears to be a serious risk for future sea buckthorn production.

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Phylogeny and Mycotoxin Profile of Pathogenic Fusarium Species Isolated from Sudden Decline Syndrome and Leaf Wilt Symptoms on Date Palms (Phoenix dactylifera) in Tunisia

Toxins ◽

10.3390/toxins13070463 ◽

2021 ◽

Vol 13 (7) ◽

pp. 463

Author(s):

Amal Rabaaoui ◽

Chiara Dall’Asta ◽

Laura Righetti ◽

Antonia Susca ◽

Antonio Logrieco ◽

...

Keyword(s):

Rna Polymerase Ii ◽

Fusarium Solani ◽

Fusarium Species ◽

Elongation Factor ◽

Phoenix Dactylifera ◽

Fusarium Proliferatum ◽

Fruit Rot ◽

Morphological Identification ◽

Single Strain ◽

Date Palms

In 2017–2018, extensive symptoms of sudden decline and fruit rot were observed on date palms in southern Tunisia. Samples of diseased plants were randomly collected in six localities. Based on morphological identification, Fusarium was the most frequent fungal genus detected. A sequencing of translation elongation factor, calmodulin, and second largest subunit of RNA polymerase II genes was used to identify 63 representative Fusarium strains at species level and investigate their phylogenetic relationships. The main species detected was Fusarium proliferatum, and at a much lesser extent, Fusarium brachygibbosum, Fusarium caatingaense, Fusarium clavum, Fusarium incarnatum, and Fusarium solani. Pathogenicity on the Deglet Nour variety plantlets and the capability to produce mycotoxins were also assessed. All Fusarium species were pathogenic complying Koch’s postulates. Fusarium proliferatum strains produced mainly fumonisins (FBs), beauvericin (BEA), and, to a lesser extent, enniatins (ENNs) and moniliformin (MON). All F. brachygibbosum strains produced low levels of BEA, diacetoxyscirpenol, and neosolaniol; two strains produced also T-2 toxin, and a single strain produced HT-2 toxin. Fusarium caatingaense, F. clavum, F. incarnatum produced only BEA. Fusarium solani strains produced MON, BEA, and ENNs. This work reports for the first time a comprehensive multidisciplinary study of Fusarium species on date palms, concerning both phytopathological and food safety issues.

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First Report of Fusarium stem and root rot of Gerbera jamesonii caused by Fusarium incarnatum in China

Plant Disease ◽

10.1094/pdis-02-21-0440-pdn ◽

2021 ◽

Author(s):

Shuning Chen ◽

Wei Sun ◽

Huizhu Yuan ◽

Xiaojing Yan

Keyword(s):

Root Rot ◽

Vascular System ◽

Elongation Factor ◽

Morphological Observation ◽

Conidial Suspension ◽

Gerbera Jamesonii ◽

Gene Sequences ◽

Plastic Container ◽

First Report ◽

Brown Discoloration

Gerbera (Gerbera jamesonii Bolus) is an important cut flower grown globally. In 2020, gerbera plants (Redaicaoyuan, Baimawangzi, and Hongditan cultivars) with roots, crowns, and stems rot were found in a greenhouse in Nanping, Fujian, China. Approximately 30% of the 60,000 plants showed symptoms. Diseased plants were stunted with chlorotic leaves. The leaves and flower heads were wilted and withered. Brown discoloration with red to black streaks occurred in the vascular system of the crown and stem. The stem pieces (3×3 mm) showing the symptom were surface-disinfected with 1% NaClO for 1 min and washed three times with sterilized water. The stem pieces were then dried and placed on potato dextrose agar (PDA) at 25℃ inside a dark chamber. Ten single-spored isolates were identified as Fusarium incarnatum based on morphological features. White to light brown mycelia were observed among the isolates on PDA medium. Falculate, multicelluar, straight to slightly curved macroconidia produced in monophialide sporodochia without distinctive foot shaped basal cell; and chlamydospores produced in some isolates (Leslie and Summerell). The size of macroconidia was 36.4 ± 5.20 × 4.6 ± 1.3 μm (n = 100) with 3 to 5 septates. Microconidia were mostly 0 to 1 septate measured 14.6 ± 1.9 × 2.6 ± 0.5 μm (n=100). Based on the morphological observation, isolates were further identified by molecular method. The ITS1/4 region combined with partial gene fragments of translation elongation factor (EF-1α, primer EF1/EF2, Geiser et al.) and calmodulin (CAM, primer CL1/CL2A, O’Donnell.) from the isolates were amplified and sequenced. All of the three tested isolates showed identical gene sequences. Sequences amplified from one represented isolate FIN-1 were submitted to Genbank. BLAST searches revealed that ITS1/4 (MW527088), EF-1α (MW556488), and CAM (MW556487) had 99.22%, 99.53%, 99.42% identity compared to F. incarnatum (MN480497, MN233577, and LN901596, respectively) in GenBank. FUSARIUM-ID (Geiser et al. 2004) analysis also showed 99 to 100% similarity with sequences of the F. incarnatum-equiseti species complex (FIESC) (FD_01636 for CAM, FD_01643 for EF-1α). The phylogenetic analysis was conducted using neighbor-joining algorithm based on the ITS, EF-1α, and CAM gene sequences. The isolate was clustered with F. incarnatum clade. Then, the pathogenicity of the fungus was confirmed by performing Koch’s postulates. Pure single-spored cultures were grown on carboxymethyl-cellulose (CMC) medium for sporulation. G. jamesonii plants used for pathogenicity tests were grown on sterilized potting soil in a plastic container to the ten-leaf stage prior to inoculation. Spores harvested from the CMC medium were adjusted to a concentration of 1×105 conidial/ml. Twelve healthy rooted gerbera seedlings were inoculated by drenching 10 ml of the conidial suspension onto roots. Twelve gerbera seedlings treated with 10 ml sterile water served as control treatments. Plants were grown in the glasshouse at temperatures of 23°C, relative humidity >70%, and 16 h light per day. After 10 days, blackening stems and withered leaf edges began to appear on inoculated seedlings, whereas control seedlings remained healthy. F. incarnatum was consistently re-isolated from the symptomatic stems, whereas no isolates were obtained from the control seedlings. The assay was conducted twice. To the best of our knowledge, this is the first report of F. incarnatum causing stem and root rot on G. jamesonii.

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First report of Trichoderma afroharzianum causing seed rot on maize in Italy

Plant Disease ◽

10.1094/pdis-12-21-2697-pdn ◽

2022 ◽

Author(s):

Martina Sanna ◽

Massimo Pugliese ◽

Maria Lodovica GULLINO ◽

Monica Mezzalama

Keyword(s):

Rna Polymerase Ii ◽

Elongation Factor ◽

Aerial Mycelium ◽

Light Cycle ◽

Conidial Suspension ◽

Ear Rot ◽

Trichoderma Spp ◽

Trichoderma Species ◽

First Report ◽

Pathogenicity Tests

Maize (Zea mays L.) is a cereal crop of great economic importance in Italy; production is currently of 60,602,320 t, covering 588,597 ha (ISTAT 2021). Trichoderma species are widespread filamentous fungi in soil, well known and studied as biological control agents (Vinale et al., 2008). Seeds of a yellow grain hybrid (class FAO 700, 132 days) were collected in September 2020 from an experimental field located in Carmagnola (TO, Italy: GPS: 44°53'11.0"N 7°40'60.0"E) and tested with blotter test (Warham et al., 1996) to assess their phytosanitary condition. Over the 400 seeds tested, more than 50% showed rotting and development of green mycelium typical of the genus Trichoderma. Due to the high and unexpected percentage of decaying kernels, ten colonies were identified by morphological and molecular methods. Single conidia colonies of one Trichoderma (T5.1) strain were cultured on Potato Dextrose Agar (PDA) for pathogenicity tests, and on PDA and Synthetic Nutrient-Poor Agar (SNA) for morphological and molecular identification. The colonies grown on PDA and SNA showed green, abundant, cottony, and radiating aerial mycelium, and yellow pigmentation on the reverse. Colony radius after 72 h at 30°C was of 60-65 mm on PDA and of 50-55 mm on SNA. The isolates produced one cell conidia 2.8 - 3.8 µm long and 2.1 - 3.6 µm wide (n=50) on SNA. Conidiophores and phialides were lageniform to ampulliform and measured 4.5 – 9.7 µm long and 1.6 – 3.6 µm wide (n=50); the base measure 1.5 – 2.9 µm wide and the supporting cell 1.4 – 2.8 µm wide (n=50). The identity of one single-conidia strain was confirmed by sequence comparison of the internal transcribed spacer (ITS), the translation elongation factor-1α (tef-1α), and RNA polymerase II subunit (rpb2) gene fragments (Oskiera et al., 2015). BLASTn searches of GenBank using ITS (OL691534) the partial tef-1α (OL743117) and rpb2 (OL743116) sequences of the representative isolate T5.1, revealed 100% identity for rpb2 to T. afroharzianum TRS835 (KP009149) and 100% identity for tef-1α to T. afroharzianum Z19 (KR911897). Pathogenicity tests were carried out by suspending conidia from a 14-days old culture on PDA in sterile H2O to 1×106 CFU/ml. Twenty-five seeds were sown in pots filled with a steamed mix of white peat and perlite, 80:20 v/v, and maintained at 23°C under a seasonal day/night light cycle. Twenty primary ears were inoculated, by injection into the silk channel, with 1 ml of a conidial suspension of strain T5.1 seven days after silk channel emergence (BBCH 65) (Pfordt et al., 2020). Ears were removed four weeks after inoculation and disease severity, reaching up to 75% of the kernels of the twenty cobs, was assessed visually according to the EPPO guidelines (EPPO, 2015). Five control cobs, inoculated with 1 ml of sterile distilled water were healthy. T. afroharzianum was reisolated from kernels showing a green mold developing on their surface and identified by resequencing of tef-1α gene. T. afroharzianum has been already reported on maize in Germany and France as causal agent of ear rot of maize (Pfordt et al. 2020). Although several species of Trichoderma are known to be beneficial microorganisms, our results support other findings that report Trichoderma spp. causing ear rot on maize in tropical and subtropical areas of the world (Munkvold and White, 2016). The potential production of mycotoxins and the losses that can be caused by the pathogen during post-harvest need to be explored. To our knowledge this is the first report of T. afroharzianum as a pathogen of maize in Italy.

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First Report of Fusarium equiseti Causing Fusarium Wilt on Potato (Solanum tuberosum) in China

Plant Disease ◽

10.1094/pdis-02-21-0281-pdn ◽

2021 ◽

Author(s):

Lingxiao Cui ◽

Chengde Yang ◽

Liping Yang ◽

Mengjun Jin ◽

Lijuan Wei

Keyword(s):

Solanum Tuberosum ◽

Rna Polymerase Ii ◽

Fusarium Wilt ◽

Elongation Factor ◽

Morphological Characteristics ◽

Gansu Province ◽

Vascular Bundles ◽

First Report ◽

Potato Plants ◽

Pathogenicity Tests

Potato (Solanum tuberosum) is one of the most economically important crops in China, containing carbohydrates, protein, fiber, numerous vitamins and minerals, and is a heart healthy food (Raidl, 2020). Potato infected by Fusarium spp. exhibits quality and yield decline, and even death. In infected plants, the upper leaves exhibit chlorosis, the lower leaves wither and the vascular bundles of stems and tubers turn yellow, and then tan to brown. In August 2018, symptomatic potato stems and roots were collected from Zhangye city, Gansu province, China. Diseased stem tissues were surface sterilized with 75% alcohol for 30 s, and then rinsed in sterile water. The tissue pieces were placed on potato dextrose agar (PDA) and incubated at 25°C in darkness. Fusarium-like colonies were consistently isolated and three monoconidial isolates were obtained. Isolate 3SMJ-2 was selected as a representative for morphological characterization, molecular analysis, and pathogenicity tests. 3SMJ-2 was inoculated in PDA liquid medium, grown on a shaker for 7 days at 25℃ to obtain a mix suspension of hypha fragments and spores (107 spores/mL). Healthy potato plants, named “Xin Daping” and were planted in pots (17 cm diameter by 12 cm) filled with 2L of sterile soil per pot. After 8 weeks, the plants were inoculated with the inoculum or distilled water. Then they were incubated in growth chambers at 25°C under a 12-h/12-h day/night potato period with 90% relative humidity for 24 h. For each treatment, 3 pots were inoculated. After 50 days, 100% of the inoculated potato plants exhibited wilt symptoms similar to those in the field but the control plants were symptomless. A Fusarium identical to strain 3SMJ-2 was re-isolated from symptomatic potato plants to fulfilling Koch’s postulates. Morphological characteristics of the re-isolated strain were identical to the original isolate, which confirmed pathogenicity of strain 3SMJ-2 originally isolated from the potatoes. Colonies of 3SMJ-2 were white with short conidiophores, a few microconidia and sickle-shaped macroconidia (25.2 to 42.9× 3.1 to 4.6 µm) (n = 60) with 4~7 septa, and mostly 5 septa, after cultivated on PDA in an incubator at 25℃ for 14 days. Spherical terminal or intercalary chlamydospores were observed on the mycelium. Strain 3SMJ-2 was identified preliminarily as Fusarium sp. based on morphological characteristics (Leslie et al., 2006). Genomic DNA was extracted from 3SMJ-2 using the OMEGA Fungal DNA kit according to the manufacturer’s protocol. The internal transcribed spacer (ITS), translation elongation factor 1-alpha (TEF) and RNA polymerase II second largest subunit (RPB2) were amplified using ITS1/ITS4 (White et al., 1990), Ef728M/Tef1R (Stępień et al., 2012) and 5F2 /7cR (O'Donnell et al., 2007), respectively. After sequencing by Beijing TSINGKE Biological Technology Co., Ltd., 3 fragments of approximately 519 bp, 587 bp and 1059 bp from the strain 3SMJ-2 were deposited in GenBank as MN420681, MW561963 and MW561964. The ITS, TEF and RPB2 sequences were 100%, 100% and 99.8% identical to those of F.equiseti (KY365589, KF499577, and MH582110). Based on the pathogenicity tests, morphological characteristics and molecular analyses, we identified the strain 3SMJ-2 as F. equiseti, the pathogen causing Fusarium wilt on potato in Zhangye City. Although, F. equiseti has been reported to cause root rot of cowpea (Li et al., 2017) and sugar beet (Cao et al., 2018) in China. To our knowledge, this is the first report confirming F. equiseti causing potato wilt in China. Potato is an economically important crop in Gansu and the occurrence of the new disease caused by F. equiseti on potato needs to be properly managed to reduce yield loss.

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First report of Fusarium solani causing stem rot of Dracaena in Iran

Journal of Plant Protection Research ◽

10.1515/jppr-2016-0013 ◽

2016 ◽

Vol 56 (1) ◽

pp. 100-103 ◽

Cited By ~ 1

Author(s):

Mostafa Abedi-Tizaki ◽

Doustmorad Zafari ◽

Jamal Sadeghi

Keyword(s):

Fusarium Solani ◽

Elongation Factor ◽

Stem Rot ◽

Pathogenicity Test ◽

Translation Elongation Factor ◽

Translation Elongation ◽

First Report ◽

Brown Discoloration ◽

Elongation Factor 1 Alpha ◽

Elongation Factor 1

Abstract In July 2013, symptoms of stem rot were observed in the Dracaena sanderiana cuttings in greenhouses of Mahallat County, Markazi Province, Iran. The symptoms first appeared as severe wilting. Later, leaves became brown and necrotic. Symptoms on the cuttings were observed as rotted areas on the middle of the stems. The cortical tissues of the plants showed a distinct brown discoloration. Eventually, the infected plants died. The pathogen was isolated from Dracaena stems and identified as F. solani by a fragment of the translation elongation factor 1-alpha (EF-1α) gene. Fusarium solani was confirmed by a pathogenicity test, and the causal agent was re-isolated from infected D. sanderiana plants. To the best of our knowledge, this is the first report of stem rot caused by F. solani on the cuttings of D. sanderiana.

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Identification and First Report of Fusarium andiyazi Causing Sheath Rot of Zizania latifolia in China

Plants ◽

10.3390/plants10091844 ◽

2021 ◽

Vol 10 (9) ◽

pp. 1844

Author(s):

Ya-Min Ma ◽

Jun-Zi Zhu ◽

Xiao-Gang Li ◽

Lai-Liang Wang ◽

Jie Zhong

Keyword(s):

Rna Polymerase Ii ◽

Phylogenetic Analyses ◽

Elongation Factor ◽

Leaf Sheath ◽

Pathogenicity Test ◽

Zizania Latifolia ◽

First Report ◽

Perennial Plant ◽

Fungal Isolates ◽

Sheath Rot

Zizania latifolia is a perennial plant native to East Asia. The swollen culm of Z. latifolia is a popular vegetable and traditional herbal medicine consumed in China and some other Asian countries. From 2019 to 2021, a sheath rot disease was found in Zhejiang Province of China. Symptoms mainly occurred in the leaf sheath showing as brown necrotic lesions surrounded by yellow halos. The pathogen fungal isolates were isolated from the affected sheaths. Ten representative isolates were selected for morphological and molecular identification by phylogenetic analyses of the translation elongation factor 1-α (TEF1) and the RNA polymerase II subunit beta (RPB2) gene regions. Based on the combined datasets, the fungal isolates were identified as Fusarium andiyazi. Koch’s postulates were confirmed by pathogenicity test, re-isolation and re-identification of the fungal isolates. To the best of our knowledge, this is the first report of sheath rot caused by F. andiyazi in Z. latifolia in China.

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First Report of Neofusicoccum parvum Associated with Dieback of Mango Trees in Peru

Plant Disease ◽

10.1094/pdis-93-4-0426b ◽

2009 ◽

Vol 93 (4) ◽

pp. 426-426 ◽

Cited By ~ 12

Author(s):

J. Javier-Alva ◽

D. Gramaje ◽

L. A. Alvarez ◽

J. Armengol

Keyword(s):

Dna Sequences ◽

Mangifera Indica ◽

Elongation Factor ◽

Tree Canopy ◽

Internal Transcribed Spacer Region ◽

Neofusicoccum Parvum ◽

First Report ◽

Early Maturation ◽

Mycelial Plug ◽

Stem Lesions

Mango (Mangifera indica L) is one of the most important cash crops of northern Peru. Since 2003, adult mango trees (cvs. Criollo and Kent) located in Piura Province developed symptoms of dieback characterized by the death of twigs and branches in the tree canopy. Additional disease symptoms involved darkened, elongated lesions on the peduncle, causing an early maturation of the fruit, and in advanced symptoms, stem-end rot of fruits. Symptoms were frequent in the spring months (September to November) when the lesions expand rapidly. Diseased tissues from branches and fruits were collected and small pieces of necrotic tissues were surface disinfected and plated onto potato dextrose agar (PDA) with 0.5 g L–1 streptomycin sulfate. Plates were incubated at 25°C in the dark. All affected tissues consistently developed colonies with a white mycelium, moderately dense, and becoming olivaceous gray after 5 to 6 days. Pycnidia were produced on sterile mango twigs placed on the surface of potato carrot agar (PCA) after 10 days. Conidia were hyaline, guttulate, aseptate, measuring (15-) 18.5 (-22.5) × (4-) 5.2 (-7.5) μm. Conidia became olivaceous and developed one or two septa before germination. Isolates were identified as Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers, & A.J.L. Phillips (1). DNA sequences of the rDNA internal transcribed spacer region (ITS) and part of the translation elongation factor 1-alpha (EF1-α) genes were used to confirm the identification through BLAST searches in GenBank (ITS: 99% identity to Accession No. EU080928; EF1-α: 98% identity to Accession No. AY343367). Representative sequences of the studied DNA regions were deposited at GenBank (ITS: Accession No. FJ528596; EF1-α: Accession No. FJ528597). Pathogenicity tests were conducted on 18-month-old potted mango plants cv. Kent with two N. parvum strains (A4 and A5). A mycelial plug (3 cm in diameter) taken from the margin of an actively growing colony of each isolate was put in a wound made with a cork borer of the same diameter on the stem of each plant. Inoculation wounds were wrapped with Parafilm. Controls were inoculated with sterile PDA plugs. Ten replicates for each isolate were used with an equal number of control plants. Plants were maintained in a greenhouse with a temperature range of 22 to 28°C. After 4 weeks, mango plants showed necrotic stem lesions originating from the inoculation point affecting also the branches of the inoculated plants. No differences in lesion area between strains were obtained. No lesions developed in the control plants. Reisolations from necrotic tissues were successful and both isolates were morphologically identical to those used for inoculations. N. parvum was isolated from all symptomatic trees in all surveyed areas. This pathogen has already been reported on mango (2) and currently represents a serious problem in the mango-producing areas of Peru. To our knowledge, this is the first report of N. parvum affecting mango in Peru. References: (1) P. W. Crous et al. Stud. Mycol. 55:235, 2006. (2) B. Slippers et al. Mycologia 97:99, 2005.

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First report of Leaf Wilt caused by Fusarium proliferatum and F. brachygibbosum on Date Palm (Phoenix dactylifera) in Tunisia

Plant Disease ◽

10.1094/pdis-08-20-1791-pdn ◽

2020 ◽

Author(s):

Ahmed Namsi ◽

Amal Rabaoui ◽

Mario Masiello ◽

Antonio Moretti ◽

Ahmed Othmani ◽

...

Keyword(s):

Date Palm ◽

Elongation Factor ◽

Aerial Mycelium ◽

Phoenix Dactylifera ◽

Fusarium Proliferatum ◽

Morphological Identification ◽

First Report ◽

Leaf Yellowing ◽

Colony Color

Since 2017, a new leaf wilt syndrome was observed in plantations of date palm in Tunisia. Its incidence increases sharply from year to year, especially in ‘Deglet Nour’ trees, aged between 5 and 15 years. In severe cases, the large number of dried leaves per tree can lead to complete cessation of date production. Symptoms appear on one or more leaves in the center of the crown. Whitening and drying start at the top of the leaflets and proceed to their base, while the midrib remains green. Then the whole leaf dies. Small white-creamy leaflet fragments and roots were collected from five different regions in the Djerid Oases. They were disinfected with diluted bleach (0,8 % NaOCl) and ethanol (80%) (each 2 min), rinsed with sterile distilled water, dried and finally plated in Petri dishes containing Potato Dextrose Agar (PDA) amended with 50mg/l neomycin. After incubation for 7 days at 25ºC±2, emerging fungal colonies were single-spored by serial dilution. They were transferred to PDA, Carnation Leaf Agar (CLA) and Spezieller Nahrstoffarmer Agar (SNA) for morphological identification. Based on the colony color on PDA, conidial morphology and phialide structures on CLA and/or SNA, of the 85 Fusarium isolates, around 90% were identified as F. proliferatum and around 10% as F. brachygibbosum (Leslie and Summerell, 2006). Fusarium proliferatum colonies rapidly developed white aerial mycelium that became purple in old cultures. Microconidia were abundant in the aerial mycelium and formed chains of variable length, on monophialides and polyphialids, a characteristic that distinguishes F. proliferatum from F. verticilloides. Less often, they were observed in false heads. Chlamydospores were absent. On CLA, microconidia were mostly 2 × 15 µm in size, a large number of sickle shaped macroconidia (2 × 25 µm) had one septum, some were larger (2 × 50 µm) with 3 septa and tips at both ends. Molecular identification was carried out based on elongation factor (EF-1α) gene sequencing. The region between the EF1 and EF2 primers (O’Donnell et al., 1998) was ampliﬁed and the sequences were compared to Fusarium reference sequences (GenBank). The sequences of the isolates Fus 1953 (539 bp), Fus 1962 (618 bp), and Fus 1965 (605 bp) shared respectively 100%, 99.51% and 99.51% homology with that of F. proliferatum JF740713.1 and were deposited in GenBank with the following accession numbers: MT630418, MT630419, and MT630420, respectively. The sequences of isolates 7F, 28F, Fus 1955 and Fus 1956 shared 100 % homology with that of F. brachygibbosum (GQ505418.1) while those of Fus 1955 and Fus 1956 showed 99.02 and 98.91 % identity, respectively, with F. brachygibbosum JX118981.1. The sequences of 7F (535 bp), 28F (535 bp), Fus 1955 (608 bp), and Fus 1956 (647 bp) were deposited in GenBank with the following accession numbers: MT630409, MT630410, MT630411, and MT630412, respectively. Two ml suspension of 106 conidia / ml of each isolate was sprayed separately or in combinations on in vitro cloned ‘Deglet Nour’ plants, placed in a greenhouse at 28°±2 °C and 70% R.H.. Isolates of F. proliferatum led to dryness and wilting leaflets after 3 weeks. Fusarium brachygibbosum only induced mild leaf yellowing, while in combination they were more virulent. Fungal isolates of both species were re-isolated and their identity confirmed to be the same of those isolated from leaflets infected in the open field, confirming Koch’s postulates. Control plants lacked symptoms. Fusarium proliferatum is known as date palm pathogen in many countries (Saleh et al. 2017), however, to our knowledge, this is the first report of F. proliferatum and also F. brachygibbosum causing Leaf Wilt symptoms on P. dactylifera in Tunisia.

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First Report of Fusarium Rot of Garlic Bulbs Caused by Fusarium proliferatum in North Carolina

Plant Disease ◽

10.1094/pdis-01-14-0040-pdn ◽

2014 ◽

Vol 98 (7) ◽

pp. 1009-1009 ◽

Cited By ~ 2

Author(s):

L. M. Quesada-Ocampo ◽

S. Butler ◽

S. Withers ◽

K. Ivors

Keyword(s):

North Carolina ◽

Apical Cell ◽

Elongation Factor ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

The United States ◽

Fusarium Proliferatum ◽

First Report ◽

Its Regions ◽

Dry Conditions

In August of 2013, garlic bulbs (Allium sativum) of the variety Chesnok Red grown and stored under dry conditions by a commercial producer in Buncombe County showed water-soaked, tan to salmon-pink lesions. Lesions on cloves became soft over time, slightly sunken, and had mycelium near the center of the bulb, which is characteristic of Fusarium rots on garlic (1,2). Approximately 10 to 20% of the bulbs inspected in the drying storage room were affected. Surface-sterilized tissue was excised from the margin of lesions on eight bulbs, plated onto acid potato dextrose agar (APDA), and incubated in the dark at room temperature (21°C). White to light pink colonies with abundant aerial mycelium and a purple pigment were obtained from all samples after 2 to 3 days of incubation. Inspection of colony morphology and reproductive structures under a microscope revealed that isolate characteristics were consistent with Fusarium proliferatum (Matsushima) Nirenberg. Microscopic morphological characteristics of the isolate included hyaline, septate hyphae; slender, slightly curved macroconidia with three to five septae produced in sporodochia; curved apical cell; and club-shaped, aseptate microconidia (measuring 3.3 to 8.3 × 1.1 to 1.3 μm) produced in chains by mono and polyphyalides. To further define the identity of the isolate, the beta-tubulin (Btub), elongation factor 1a (EF1a), and internal transcribed spacer (ITS) regions were amplified and sequenced (3). The resulting sequences were compared against the GenBank nucleotide database by using a BLAST alignment, which revealed that the isolate had 100% identity with F. proliferatum for the Btub, EF1a, and ITS regions (GenBank Accession Nos. AF291055.1, JX118976.1, and HF930594.1, respectively). Sequences for the isolate were deposited in GenBank under accessions KJ128963, KJ128964, and KJ128965. While there have been other reports of F. proliferatum causing bulb rot of garlic in the United States (1), to our knowledge, this is the first report in North Carolina. The finding is significant since F. proliferatum can produce a broad range of mycotoxins, including fumonisins, when infecting its host, which is a concern for food safety in Allium crops. References: (1) F. M. Dugan et al. Plant Pathol. 52:426, 2003. (2) L. J. du Toit and F. M. Dugan. Page 15 in: Compendium of Onion and Garlic Diseases and Pests. H. F. Schwartz and S. K. Mohan, eds. The American Phytopathological Society, St. Paul, MN, 2008. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

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