Identification and Description of a New Pathogen Causing Flower Dry Rot on Passiflora edulis in China

Plant Disease ◽  
2020 ◽  
Author(s):  
Yue Lian Liu ◽  
Jian Rong Tang ◽  
Yu Han Zhou
Keyword(s):  
Disease Incidence ◽  
Phylogenetic Analyses ◽  
Apical Cell ◽  
Management Strategies ◽  
Morphological Description ◽  
Pathogenicity Test ◽  
New Disease ◽  
Single Spore ◽  
Dry Rot ◽  
Passiflora Edulis

Passiflora edulis Sims (passion fruit) is an economically important fruit crop. A new disease, flower dry rot with an incidence of 30%–40%, has occurred in orchards located in Zhanjiang, China, and led to serious production loss. Disease incidence was about 30%–-40%. A total of 221 isolates of Fusarium sp. were obtained from samples of three types of symptomatic flowers. Three representative single-spore isolates (PaB-1, PaB-2, and PaB-3) from each type were used for pathogenicity test, multi-locus phylogenetic analyses, and morphological description. Pathogenicity test on bud of 5- month-old P. edulis plants reproducedshowed similar symptoms as those observed in nature, and Koch’’s postulates were fulfilledachieved.Results showed that the symptoms were observed on inoculated buds but not on the control, and Koch’s postulates were satisfied in all test isolates. By comparing 36 typical species from the FUSARIUM-ID database, multi-locus phylogenetic analyses showed that the sequences of TEF1, RPB2, and ITS of thethese isolates belong to the Incarnatum clade of the F. incarnatum–-equiseti species complex (FIESC-17-a) with an independent branch. Conidia of the isolates were falcate, smooth walled, curved apical cell, and foot-shaped basal cell, 1-5-septate. The conidial anastomosis tubes were first observed in the FIESC. Therefore, the pathogenic isolates were identifieddescribed as F. pernambucanum (FIESC-17-a). Moreover, in this study, the conidial anastomosis tubes were first observed in the FIESCIt is a new pathogen causing flower dry rot on P. edulis. This study representeds the first report of flower dry rot on P. edulis caused by F. pernambucanum. Further studies should be performed for the selection of effective disease management strategies.This finding provided crucial information on the threats of F. pernambucanum affecting P. edulis in China and management strategies for the new disease, which should be developed.

scholarly journals First Report of Fusarium pernambucanum Causing Fruit Rot of Muskmelon in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Xianping Zhang ◽  
Jiwen Xia ◽  
Jiakui Liu ◽  
Dan Zhao ◽  
Lingguang Kong ◽  
...  
Keyword(s):  
Phylogenetic Analyses ◽  
Apical Cell ◽  
Morphological Characteristics ◽  
Fruit Rot ◽  
Likelihood Method ◽  
Correct Identification ◽  
Pathogenicity Test ◽  
First Report ◽  
The Core ◽  
Representative Isolate

Muskmelon (Cucumis melo L.) is one of the most widely cultivated and economically important fruit crops in the world. However, many pathogens can cause decay of muskmelons; among them, Fusarium spp. is the most important pathogen, affecting fruit yield and quality (Wang et al. 2011). In May 2017, fruit rot symptoms were observed on ripening muskmelons (cv. Jipin Zaoxue) in several fields in Liaocheng of Shandong Province, China. Symptoms appeared as brown, water-soaked lesions, irregularly circular in shape, with the lesion size ranging from a small spot (1 to 2 cm) to the decay of the entire fruit. The core and the surface of the infected fruit were covered with white to rose-reddish mycelium. Two infected muskmelons were collected from each of two fields, 10 km apart. Tissues from the inside of the infected fruit were surface disinfected with 75% ethanol for 30 s, and cultured on potato dextrose agar (PDA) at 25 °C in the dark for 5 days. Four purified cultures were obtained using the single spore method. On carnation leaf agar (CLA), macroconidia had a pronounced dorsiventral curvature, falcate, 3 to 5 septa, with tapered apical cell, and foot-shaped basal cell, measuring 19 to 36 × 4 to 6 μm. Chlamydospores were abundant, 5.5–7.5 μm wide, and 5.5–10.5 μm long, ellipsoidal or subglobose. No microconidia were observed. These morphological characteristics were consistent with the descriptions of F. pernambucanum (Santos et al. 2019). Because these isolates had similar morphology, one representative isolate was selected for multilocus phylogenetic analyses. DNA was extracted from the representative isolate using the CTAB method. The nucleotide sequences of the internal transcribed spacers (ITS) (White et al. 1990), translation elongation factor 1-α gene (TEF1), RNA polymerase II second largest subunit gene (RPB2), calmodulin (CAM) (Xia et al. 2019) were amplified using specific primers, sequenced, and deposited in GenBank (MN822926, MN856619, MN856620, and MN865126). Based on the combined dataset of ITS, TEF1, RPB2, CAM, alignments were made using MAFFT v. 7, and phylogenetic analyses were processed in MEGA v. 7.0 using the maximum likelihood method. The studied isolate (XP1) clustered together with F. pernambucanum reference strain URM 7559 (99% bootstrap). To perform pathogenicity test, 10 μl of spore suspensions (1 × 106 conidia/ml) were injected into each muskmelon fruit using a syringe, and the control fruit was inoculated with 10 μl of sterile distilled water. There were ten replicated fruits for each treatment. The test was repeated three times. After 7 days at 25 °C, the interior of the inoculated muskmelons begun to rot, and the rot lesion was expanded from the core towards the surface of the fruit, then white mycelium produced on the surface. The same fungus was re-isolated from the infected tissues and confirmed to fulfill the Koch’s postulates. No symptoms were observed on the control muskmelons. To our knowledge, this is the first report of F. pernambucanum causing of fruit rot of muskmelon in China. Considering the economic value of the muskmelon crop, correct identification can help farmers select appropriate field management measures for control of this disease.

scholarly journals First Report of Colletotrichum siamense Causing Anthracnose of Monstera deliciosa in Zhanjiang, China

Plant Disease ◽  
2020 ◽  
Author(s):  
Yue Lian Liu ◽  
Jian Rong Tang ◽  
Yu Han Zhou
Keyword(s):  
Disease Incidence ◽  
Morphological Characteristics ◽  
Pathogenicity Test ◽  
Sterile Water ◽  
Single Spore ◽  
First Report ◽  
Rdna Its ◽  
Colletotrichum Siamense ◽  
Monstera Deliciosa ◽  
Pure Cultures

Monstera deliciosa Liebm is an ornamental foliage plant (Zhen et al. 2020De Lojo and De Benedetto 2014). In July of 2019, anthracnose lesions were observed on leaves of M. deliciosa cv. Duokong with 20% disease incidence of 100 plants at Guangdong Ocean University campus (21.17N,110.18E), Guangdong Province, China. Initially affected leaves showed chlorotic spots, which coalesced into larger irregular or circular lesions. The centers of spots were gray with a brown border surrounded by a yellow halo (Supplementary figure 1). Twenty diseased leaves were collected for pathogen isolation. Margins of diseased tissue was cut into 2 × 2 mm pieces, surface-disinfected with 75% ethanol for 30 s and 2% sodium hypochlorite (NaOCl) for 60 s, rinsed three times with sterile water before isolation. Potato dextrose agar (PDA) was used to culture pathogens at 28℃ in dark. Successively, pure cultures were obtained by transferring hyphal tips to new PDA plates. Fourteen isolates were obtained from 20 leaves. Three single-spore isolates (PSC-1, PSC-2, and PSC-3) were obtained ,obtained, which were identical in morphology and molecular analysis (ITS). Therefore, the representative isolate PSC-1 was used for further study. The culture of isolate PSC-1 on PDA was initially white and later became cottony, light gray in 4 days, at 28 °C. Conidia were single celled, hyaline, cylindrical, clavate, and measured 13.2 to 18.3 µm × 3.3 to 6.5 µm (n = 30). Appressoria were elliptical or subglobose, dark brown, and ranged from 6.3 to 9.5 µm × 5.7 to 6.5 µm (n = 30). Morphological characteristics of isolate PSC-1 were consistent with the description of Colletotrichum siamense (Prihastuti et al. 2009; Sharma et al. 2013). DNA of the isolate PSC-1 was extracted for PCR sequencing using primers for the rDNA ITS (ITS1/ITS4), GAPDH (GDF1/GDR1), ACT (ACT-512F/ACT-783R), CAL (CL1C/CL2C), and TUB2 (βT2a/βT2b) (Weir et al. 2012). Analysis of the ITS (accession no. MN243535), GAPDH (MN243538), ACT (MN512640), CAL (MT163731), and TUB2 (MN512643) sequences revealed a 97-100% identity with the corresponding ITS (JX010161), GAPDH (JX010002), ACT (FJ907423), CAL (JX009714) and TUB2 (KP703502) sequences of C. siamense in GenBank. A phylogenetic tree was generated based on the concatenated sequences of ITS, GAPDH, ACT, CAL, and TUB2 which clustered the isolate PSC-1 with C. siamense the type strain ICMP 18578 (Supplementary figure 2). Based on morphological characteristics and phylogenetic analysis, the isolate PSC-1 associated with anthracnose of M. deliciosa was identified as C. siamense. Pathogenicity test was performed in a greenhouse at 24 to 30oC with 80% relative humidity. Ten healthy plants of cv. Duokong (3-month-old) were grown in pots with one plant in each pot. Five plants were inoculated by spraying a spore suspension (105 spores ml-1) of the isolate PSC-1 onto leaves until runoff, and five plants were sprayed with sterile water as controls. The test was conducted three times. Anthracnose lesions as earlier were observed on the leaves after two weeks, whereas control plants remained symptomless. The pathogen re-isolated from all inoculated leaves was identical to the isolate PSC-1 by morphology and ITS analysis, but not from control plants. C. gloeosporioides has been reported to cause anthracnose of M. deliciosa (Katakam, et al. 2017). To the best of our knowledge, this is the first report of C. siamense causing anthracnose on M. deliciosa in ChinaC. siamense causes anthracnose on a variety of plant hosts, but not including M. deliciosa (Yanan, et al. 2019). To the best of our knowledge, this is the first report of C. siamense causing anthracnose on M. deliciosa, which provides a basis for focusing on the management of the disease in future.

scholarly journals Botryosphaeria Dothidea and Neofusicoccum Yunnanense Causing Canker and Die-Back of Sequoiadendron Giganteum in Croatia

Forests ◽  
2021 ◽  
Vol 12 (6) ◽  
pp. 695
Author(s):  
Marta Kovač ◽  
Danko Diminić ◽  
Saša Orlović ◽  
Milica Zlatković
Keyword(s):  
Sierra Nevada ◽  
Phylogenetic Analyses ◽  
Management Strategies ◽  
Housekeeping Genes ◽  
First Record ◽  
Pathogenicity Test ◽  
Botryosphaeria Dothidea ◽  
Ornamental Tree ◽  
Sequoiadendron Giganteum ◽  
The Impact

Sequoiadendron giganteum Lindl. [Buchholz] is a long-lived tree species endemic to the Sierra Nevada Mountains in California. Due to its massive size and beauty, S. giganteum is a popular ornamental tree planted in many parts of the world, including Europe. Since 2017, scattered branch die-back has been observed on S. giganteum trees in Zagreb, Croatia. Other symptoms included resinous branch cankers, reddish-brown discoloration of the sapwood and, in severe cases, crown die-back. Branches showing symptoms of die-back and cankers were collected from six S. giganteum trees in Zagreb and the aim of this study was to identify the causal agent of the disease. The constantly isolated fungi were identified using morphology and phylogenetic analyses based on the internal transcribed spacer (ITS) of the ribosomal DNA (rDNA), and partial sequencing of two housekeeping genes, i.e., translation elongation factor 1-α (TEF 1-α), and β tubulin 2 (TUB2). The fungi were identified as Botryosphaeria dothidea (Moug.) Ces. and De Not. and Neofusicoccum yunnanense G.Q. Li & S.F. Chen. The pathogenicity test was conducted in a plant growth chamber on S. giganteum seedlings and revealed that N. yunnanense was more aggressive compared to B. dothidea. N. yunnanense was able to reproduce symptoms of canker and die-back and kill plants seven weeks after inoculation whereas B. dothidea produced cankers. To the best of our knowledge, this is the first report of B. dothidea and N. yunnanense causing canker and die-back disease of S. giganteum in Croatia. It is also the first record on the identity and pathogenicity of any fungal species associated with S. giganteum in this country. The study expended the known host range of N. yunnanense to include S. giganteum, which is a valuable ornamental tree in Croatian landscapes. Disease management strategies should be developed to mitigate or reduce the impact of the disease.

scholarly journals Brown Culm Rot of Dendrocalamus latiflorus Munro Caused by Diaporthe guangxiensis in Sichuan Province, China

Plant Disease ◽  
2021 ◽  
Author(s):  
Wenjian Wei ◽  
Han Zhang ◽  
Liling Xie ◽  
Han Liu ◽  
Fengying Luo ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Southern China ◽  
Disease Incidence ◽  
Its Region ◽  
Sichuan Province ◽  
Control Measures ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Single Spore ◽  
Dendrocalamus Latiflorus

Dendrocalamus latiflorus Munro, the most widely cultivated bamboo species in southern China, has high ornamental value used in gardens, while culms are also used for buildings and as fibers and edibles (Gao et al. 2011). In June 2020, brown culm rot of bamboo was observed in Yibin city, Sichuan Province, in an area of approximately 1000 hectares. Disease incidence was approximately 60%, of which 30% of the plants had died. At the end of June, the lesions expanded but did not surround the base of the culm. From the end of June to the beginning of September, the lesions expanded upward and formed a streak, of which the color gradually deepened to purple-brown and black-brown. At the same time, the disease spots at the base of the culm also expanded horizontally. After the spots surrounded the base of the culm, the diseased bamboo died. Ten culms showing typical symptoms were collected and cut into 5×5 mm pieces at the junction of infected and healthy tissues. The tissues were sterilized for 1 to 2 min in 3% sodium hypochlorite, decontaminated in 75% alcohol for 3 to 5 min, placed on modified potato glucose agar (PDA) with streptomycin sulfate (50 μg/ml), and incubated at 26°C. Two isolates were obtained by the single-spore method (Sivan et al. 1992). The isolates both produced white round colonies similar to Diaporthe guangxiensis and two types of conidia: one was α type (5.5 to 8.2×1.0 to 2.8 µm, n=30), colourless, single-celled, undivided, and oval, containing two oil droplets; and β type (21.1 to 30.2×0.8 to 1.4 µm, n=30), colourless, single celled and hook shaped. Genomic DNA was extracted from the two isolates by using a fungal genomic DNA extraction kit (Solarbio, Beijing). The products were amplified by polymerase chain reaction (PCR) with primers for the internal transcribed spacer 1 (ITS) region (White et al. 1990), calmodulin (CAL) gene (Carbone and Kohn 1999), translation elongation factor 1-alpha (TEF) gene (Glass and Donaldson 1995) and beta-tubulin (TUB) gene (Soares et al. 2018). The amplified products were sequenced and blasted in GenBank (accession numbers MW380383, MW431318, MW431317 and MW431316 for ITS, CAL, TEF, and TUB, respectively). The ITS, CAL, TEF, and TUB sequences showed 100%, 99.33%, 100%, and 99.80% identity to D. guangxiensis JZB320094 (accession numbers MK335772.1, MK736727.1, MK523566.1, MK500168.1 in GenBank), respectively. To evaluate the pathogenicity of the isolates, five plants were each inoculated with two isolates. The cortex of potted bamboo were injured locally with sterilized needle, and the bamboo culms were inoculated with 100 μl of conidial suspension (105 cfu/ml). The surface of the inoculation wound was covered with gauze soaked with sterilized water. Five plants inoculated with sterile water were used as controls. The treated plants were maintained in a greenhouse at a temperature of 22 to 29°C and relative humidity of 70 to 80%. One month later, of all inoculated plants showed similar symptoms as those observed in the field. D. guangxiensis was re-isolated from all inoculated plants. The pathogenicity test was repeated three times with similar results. This is the first report of D. guangxiensis causing brown culm rot of D. latiflorus in China. These results will facilitate an enhanced understanding of factors affecting bamboo and the design of effective management strategies of the pathogenic species on bamboo and thus to develop corresponding control measures.

scholarly journals First Report of Fruit Rot Caused by Fusarium luffae in Muskmelon in China

Plant Disease ◽  
2022 ◽  
Author(s):  
Xianping Zhang ◽  
Xuedong Cao ◽  
Qingqing Dang ◽  
Yongguang Liu ◽  
Xiaoping Zhu ◽  
...  
Keyword(s):  
Phylogenetic Analyses ◽  
Apical Cell ◽  
Morphological Characteristics ◽  
Fruit Rot ◽  
Likelihood Method ◽  
Correct Identification ◽  
Pathogenicity Test ◽  
Fusarium Spp ◽  
First Report ◽  
The Core

Muskmelon (Cucumis melo L.) is one of the most widely cultivated and economically important fruit crops in the world. However, many pathogens can cause decay of muskmelon fruit, including Fusarium spp.. Fusarium spp. are the most important pathogen, affecting muskmelon fruit yield and quality (Wang et al. 2011). In August 2020, fruit rot symptoms were observed on ripening muskmelons (cv. Tianbao) in several fields in Jiyang District, Jinan City of Shandong Province, China. The incidences of infected muskmelon ranged from 15% to 30% and caused an average 20% yield loss. Symptoms appeared as pale brown, water-soaked lesions that were irregular in shape, with the lesion sizes ranging from a small spot (1 to 2 cm) to decay of the entire fruit. The core and surface of infected fruit were colonized and covered with white mycelia. Two infected muskmelons were collected from two fields, 3.5 km apart. Tissues removed from inside the infected fruit were surface disinfected with 75% ethanol for 30 s, and cultured on potato dextrose agar (PDA) at 25°C in the dark for 5 days. Four purified cultures were obtained using the single spore method. On carnation leaf agar (CLA), 3 to 5 septate, falcate, with a pronounced dorsiventral curvature macroconidia with tapered apical cell, and foot-shaped basal cell, measuring 20 to 40 × 3.5 to 4.5 μm. Microconidia and chlamydospores were not observed. These morphological characteristics were consistent with the description of F. luffae (Wang et al., 2019). Because these isolates had similar morphology, two representative isolates (XP11 and XP12) were selected for multilocus phylogenetic analyses. DNA was extracted from the representative isolates using a CTAB method. Nucleotide sequences of the internal transcribed spacers (ITS) (White et al. 1990), calmodulin (CAM), RNA polymerase II second largest subunit (RPB2), translation elongation factor 1-α gene (TEF1) (Xia et al. 2019) were amplified using specific primers, sequenced, and deposited in GenBank (ITS: MW391509 and MW391510, CAM: MW392789 and MW392790, RPB2: MW392797 and MW392798, TEF1: MW392793 and MW392794). Alignments of a combined dataset of ITS, CAM, RPB2 and TEF1 were made using MAFFT v. 7, and phylogenetic analyses were conducted in MEGA v. 7.0 using the maximum likelihood method. The muskmelon isolates (XP11 and XP12) clustered together with the F. luffae reference strain LC12167 (99% bootstrap). To perform a pathogenicity test, 10 μl of conidial suspensions (1 × 106 conidia/ml) were injected into each muskmelon fruit using a syringe, and the control fruit was inoculated with 10 μl of sterile distilled water. There were ten replicated fruits for each treatment. The test was repeated three times. After 7 days at 25°C, the interior of the inoculated muskmelons begun to rot, and the rot lesion expanded from the core towards the surface of the fruit, then white mycelia were produced on the surface. Ten isolations were re-isolated from the infected tissues and confirmed to fulfill Koch’s postulates. No symptoms were observed on the control muskmelons. To our knowledge, this is the first report of fruit rot caused by F. luffae in muskmelon in China. Considering the economic value of the muskmelon crop, correct identification can help farmers select appropriate field management measures for control of this disease.

scholarly journals First Report of Passalora Leaf Spot Caused by Passalora bougainvilleae on Bougainvillea in Mexico

Plant Disease ◽  
2011 ◽  
Vol 95 (6) ◽  
pp. 775-775 ◽  
Author(s):  
V. Ayala-Escobar ◽  
V. Santiago-Santiago ◽  
A. Madariaga-Navarrete ◽  
A. Castañeda-Vildozola ◽  
C. Nava-Diaz
Keyword(s):  
Uv Light ◽  
Disease Incidence ◽  
Morphological Characteristics ◽  
The United States ◽  
Pathogenicity Test ◽  
Commonwealth Mycological Institute ◽  
Conidial Suspension ◽  
Single Spore ◽  
First Report ◽  
Bougainvillea Spectabilis

Bougainvillea (Bougainvillea spectabilis Willd) growing in 28 gardens during 2009 showed 100% disease incidence and 3 to 7% disease severity. Bougainvilleas with white flowers were the most affected. Symptoms consisted of light brown spots with dark brown margins visible on adaxial and abaxial sides of the leaves. Spots were circular, 2 to 7 mm in diameter, often surrounded by a chlorotic halo, and delimited by major leaf veins. Single-spore cultures were incubated at 24°C under near UV light for 7 days to obtain conidia. Pathogenicity was confirmed by spraying a conidial suspension (1 × 104 spores/ml) on leaves of potted bougainvillea plants (white, red, yellow, and purple flowers), incubating the plants in a dew chamber for 48 h and maintaining them in a greenhouse (20 to 24°C). Identical symptoms to those observed at the residential gardens appeared on inoculated plants after 45 to 60 days. The fungus was reisolated from inoculated plants that showed typical symptoms. No symptoms developed on control plants treated with sterile distilled water. The fungus produced distinct stromata that were dark brown, spherical to irregular, and 20 to 24 μm in diameter. Conidiophores were simple, born from the stromata, loose to dense fascicles, brown, straight to curved, not branched, zero to two septate, 14 × 2 μm, with two to four conspicuous and darkened scars. The conidia formed singly, were brown, broad, ellipsoid, obclavate, straight to curved with three to four septa, 40 × 4 μm, and finely verrucous with thick hilum at the end. Fungal DNA from the single-spore cultures was obtained using a commercial DNA Extraction Kit (Qiagen, Valencia, CA); ribosomal DNA was amplified with ITS5 and ITS4 primers and sequenced. The sequence was deposited at the National Center for Biotechnology Information Database (GenBank Accession Nos. HQ231216 and HQ231217). The symptoms (4), morphological characteristics (1,2,4), and pathogenicity test confirm the identity of the fungus as Passalora bougainvilleae (Muntañola) Castañeda & Braun (= Cercosporidium bougainvilleae Muntañola). This pathogen has been reported from Argentina, Brazil, Brunei, China, Cuba, El Salvador, India, Indonesia, Jamaica, Japan, Thailand, the United States, and Venezuela (3). To our knowledge, this is the first report of this disease on B. spectabilis Willd in Mexico. P. bougainvilleae may become an important disease of bougainvillea plants in tropical and subtropical areas of Mexico. References: (1) U. Braun and R. R. Castañeda. Cryptogam. Bot. 2/3:289, 1991. (2) M. B. Ellis. More Dematiaceous Hypomycetes. Commonwealth Mycological Institute, Kew, Surrey, UK, 1976. (3) C. Nakashima et al. Fungal Divers. 26:257, 2007. (4) K. L. Nechet and B. A. Halfeld-Vieira. Acta Amazonica 38:585, 2008.

scholarly journals First Report of Diaporthe phaseolorum Causing Stem Canker of Hemp (Cannabis sativa)

Plant Disease ◽  
2021 ◽  
Author(s):  
Marcus Vinicius Marin ◽  
Nan-Yi Wang ◽  
Jacqueline Coburn ◽  
Johan Desaeger ◽  
Natalia A. Peres
Keyword(s):  
Cannabis Sativa ◽  
Phylogenetic Analyses ◽  
Stem Canker ◽  
Tween 20 ◽  
Future Research ◽  
Herbaceous Plant ◽  
Pathogenicity Test ◽  
Single Spore ◽  
Isolation Medium ◽  
First Report

Hemp is an annual herbaceous plant that is used for its fiber and oil in a variety of commercial and industrial products. In Florida, it is currently being explored as a new specialty crop. During a field trial from October to January 2019 in Wimauma, FL, a stem canker was observed on up to 60% of three-month-old plants of 'Eletta Campana', 'Carmagnola Selezionata', and 'Tygra'. Symptoms started on the main stems with light-to-dark brown lesions of different sizes and shapes. Over time, the lesions coalesced into large necrotic areas and bore pycnidia. Isolations were made from diseased stem tissues on General Isolation medium (Amiri et al. 2018) after surface disinfestation (Marin et al. 2020). The plates were placed in a growth chamber at 25°C under a 12/12 photoperiod. A fungus with white, floccose, aerial mycelium and pycnidia producing alpha and beta conidia was consistently isolated. Three single spore isolates were chosen for identification and pathogenicity tests. Pycnidia on PDA were globose to irregular and ranged from 170 to 250 μm long (210 ± 2.5, n = 50) and 140 to 220 μm wide (180 ± 2.7, n = 50). The alpha conidia were unicellular, hyaline, ellipsoidal to fusiform and ranged from 5.3 to 7.7 μm long (6.5 ± 1.6, n = 50) and 1.5 to 4.6 μm wide (2.8 ± 1.8, n = 50). The beta conidia were hyaline, elongated, filiform, straight or curved and ranged from 10.2 to 17.7 μm long (16.1 ± 2.2, n = 50) and 0.5 to 1.8 μm wide (0.8 ± 0.2, n = 50). Perithecia were not observed. Based on morphological features, the fungus was similar to anamorphs of Diaporthe spp. (Santos et al. 2011; Udayanga et al. 2015). DNA from the same three isolates was extracted using the FastDNA kit, and the ribosomal internal transcribed spacer (ITS), β-tubulin (TUB), and calmodulin (CAL) regions were amplified following Udayanga et al. (2014), and Sanger sequenced by Genewiz. Sequences were deposited in GenBank (accession no. MT497039 to MT497047 for ITS, TUB, and CAL). BLASTn searches revealed isolates 20-58, 20-59, and 20-60 were 96.34% identical to the epitype isolate D. phaseolorum AR4203 for ITS (KJ590738.1, 527 bp out of 547 bp), 100% for TUB (KJ610893.1, 459 bp out of 459 bp), and 100% for CAL (KJ612135.1, 522 bp out of 522 bp) (Udayanga et al. 2015). Their identity was confirmed by phylogenetic analyses using maximum likelihood and Bayesian inference methods. To complete Koch’s postulates, pycnidia of the same three isolates were harvested and crushed in 2 mL Eppendorf tubes containing 0.01% Tween 20. Conidia suspensions were adjusted to 106 spores/mL. Three 5-week-old potted plants of 'Eletta Campana' and 'Carmagnola Selezionata' per isolate were inoculated using a 1 mL syringe with a needle by injecting 200 µL of the suspension into the stem. Plants were placed inside clear plastic bags for 48 h and maintained in the greenhouse. Control plants were injected with sterile deionized water and kept under the same conditions. The pathogenicity test was repeated once. Four weeks after inoculation, inoculated plants developed stem cankers from which the same pathogen was isolated, whereas controls remained healthy. To our knowledge, this is the first report of D. phaseolorum causing stem canker on hemp. This pathogen has been reported causing canker on sunflower and Phaseolus spp. (Gomzhina and Gannibal 2018; Udayanga et al. 2015; Vrandecic et al. 2009). This discovery may help shape future research into disease epidemiology and management for a crop in which very limited disease information is available at the moment.

scholarly journals First Report of Mango Malformation Disease Caused by Fusarium tupiense in Senegal

Plant Disease ◽  
2012 ◽  
Vol 96 (10) ◽  
pp. 1582-1582 ◽  
Author(s):  
A. Lamine Senghor ◽  
K. Sharma ◽  
P. Lava Kumar ◽  
R. Bandyopadhyay
Keyword(s):  
Mangifera Indica ◽  
Apical Cell ◽  
Morphological Characteristics ◽  
Fungal Growth ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Single Spore ◽  
Southeastern Part ◽  
Isolation Medium ◽  
Mango Malformation

Mango (Mangifera indica L.) is an economically important export crop for Senegal, producing about 100,000 tons of fruit annually. In April 2009, severe outbreaks of a new disorder occurred in mango orchards in the southeastern part of Casamance. Diseased plants showed abnormal growth of vegetative shoots with short thickened internodes and malformed inflorescence with short leaves interspersed among thickened sterile flowers that aborted early. These symptoms resembled those caused by mango malformation disease (4). To identify the causal agent, floral and vegetative samples from symptomatic mango plants were collected from Kolda district (12°53' N, 14°56' W). Malformed tissues were cut into 4-mm2 pieces, surface sterilized with 75% ethanol for 2 min, dried, and plated on the Fusarium isolation medium Peptone PCNB Agar (PPA) (2). Fungal growth with Fusarium morphology were transferred on PPA and further purified on water agar as single spore isolates. Cultures were identified on the basis of spore characters on carnation leaf agar and colony morphology on PDA (2). Two isolates (I4 and I17) were similar to F. mangiferae/F. sterilihyphosum/F. tupiense complex (3). Macroconidia were slender, slightly falcate, three- to five-septate, 18.5 to 27.7 × 1.1 to 2.3 μm with slightly curved apical cell produced on cream to orange sporodochia. Microconidia were single-celled, oval, 3.7 to 13.6 × 0.75 to 1.1 μm produced on mono- and polyphialides in false heads. Chlamydospores were absent. To confirm the identity, genomic DNA was isolated from pure cultures of I4 and I17, used for amplification of portion of translation elongation factor (TEF-1α). Amplified products (241 bp) were purified and sequenced in both directions (GenBank Accession Nos. JX272929 and JX272930). A BLASTn search revealed 100% sequence identity with F. tupiense (DQ452860), 99% identity with F. mangiferae (HM135531) and F. sterilihyphosum (DQ452858) from Brazil. Phylogenetic analysis inferred from the Clustalw alignment of TEF-1α sequences clustered I4 and I17 isolates with F. tupiense (3). To confirm Koch's postulates, 2-year-old healthy mango seedlings var. Keitt and Kent (12 plants each) were inoculated by placing 20 μl conidial suspension (5 × 107 conidia ml–1) on micro-wounds created in apical and lateral buds. Inoculated buds were covered with filter paper soaked in the same spore suspension (1). Seedlings inoculated similarly with sterile distilled water served as control. Seven months after the inoculation, typical malformation symptoms were observed on vegetative parts on all inoculated plants, but not on control plants. F. tupiense was reisolated from symptomatic shoots of inoculated plants. Based on the morphological characteristics, sequence analysis, and pathogenicity test, the pathogen of mango malformation in Senegal was identified as F. tupiense (3). To our knowledge, this is the first confirmed record in Senegal of mango malformation caused by F. tupiense. This disease is a serious threat to mango production and trade of Senegal. Urgent actions are necessary to stop this emerging epidemic that can spread to other countries in West Africa. References: (1) S. Freeman et al. Phytopathol. 6:456, 1999. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual, Blackwell Publishing, Ames, IA, 2006. (4) C. S. Lima et al. Mycologia. 104: in press (doi: 10.3852/12-052). (2) W. F. O. Marasas et al. Phytopathol. 96:667, 2006.

scholarly journals Identification and Characterization of Fungal Pathogens Causing Fruit Rot of Deciduous Holly

Plant Disease ◽  
2018 ◽  
Vol 102 (12) ◽  
pp. 2430-2445 ◽  
Author(s):  
Shan Lin ◽  
Nancy J. Taylor ◽  
Francesca Peduto Hand
Keyword(s):  
Fungal Pathogens ◽  
Disease Incidence ◽  
Phylogenetic Analyses ◽  
Management Strategies ◽  
Morphological Characterization ◽  
Fruit Rot ◽  
Economic Losses ◽  
Yield Reduction ◽  
Inoculum Concentration ◽  
Fruit Samples

Cut branches of deciduous holly (Ilex spp. L.) harboring colorful berries are traditionally used as ornaments in holiday decorations. Since 2012, a fruit rot of unspecified cause has resulted in significant yield reduction and economic losses across Midwestern and Eastern U.S. nurseries. In this study, symptomatic fruit samples collected from nine different locations over five years were analyzed, and several fungal species were isolated. A combination of morphological characterization, multilocus phylogenetic analyses, and pathogenicity assays revealed that Alternaria alternata and Diaporthe ilicicola sp. nov. were the primary pathogens associated with symptomatic fruit. Other fungi including A. arborescens, Colletotrichum fioriniae, C. nymphaeae, Epicoccum nigrum, and species in the D. eres species complex appeared to be minor pathogens in this disease complex. In detached fruit pathogenicity assays testing the role of wounding and inoculum concentration on disease development, disease incidence and severity increased when fruit was wounded and inoculated with a higher inoculum concentration. These findings indicate that management strategies that can protect fruit from injury or reduce inoculum may lower disease levels in the field. This research established the basis for further studies on this emerging disease and the design of research-based management strategies. To our knowledge, it also represents the first report of species of Alternaria, Colletotrichum, Diaporthe, and Epicoccum causing fruit rot of deciduous holly.

scholarly journals First Report of Dry and Soft Rot of Cereus marginatus var. cristata Caused by Fusarium oxysporum in Italy

Plant Disease ◽  
2014 ◽  
Vol 98 (10) ◽  
pp. 1441-1441 ◽  
Author(s):  
A. Garibaldi ◽  
P. Pensa ◽  
D. Bertetti ◽  
G. Ortu ◽  
M. L. Gullino
Keyword(s):  
Relative Humidity ◽  
Fusarium Oxysporum ◽  
Apical Cell ◽  
Morphological Characteristics ◽  
Soft Rot ◽  
The United States ◽  
Selective Medium ◽  
Economic Losses ◽  
Pathogenicity Test ◽  
Dry Rot

During the winter of 2013, 50% of 20,000 plants of Cereus marginatus var. cristata, Cactaceae family, grown in a commercial farm located in Liguria (northern Italy) showed symptoms of a dry or soft rot. In the case of dry rot, affected plants showed on the stem superficial necrosis and dry rot, irregularly shaped, 1 to 10 mm, while epidermal and cortical tissues were wounded. Affected plants survived but they lost ornamental value. In the case of soft rot, associated with conditions of higher relative humidity, rots on the stem extended as far as 4 cm in width. The internal part of bark, cambium, and xylem tissues as far as about 3 cm in depth was rotted. Vascular tissues were not discolored. Plants died in about 20 days. A Fusarium sp. was consistently isolated from symptomatic tissue on Komada selective medium (2) from plants showing soft rot. The isolates were purified and subcultured on potato dextrose agar (PDA). On PDA, the cultures produced a thick and soft growth of white to light pink mycelium and pale pink pigments in the agar. On Spezieller Nährstoffarmer agar (SNA), cultures produced short monophialides with unicellular, ovoid-elliptical microconidia measuring 3.7 to 8.2 × 1.7 to 3.5 (average 5.4 × 2.5) μm. On carnation leaf-piece agar (CLA), chlamydospores were abundant, terminal or intercalary, single or paired, but frequently also aggregated. On the same medium, at temperatures ranging from 20 to 24°C (14 h daylight, 10 h dark), cultures produced light orange sporodochia with macroconidia. These were 3 to 4 (sometimes 5) septate, nearly straight with a foot-shaped basal cell and a short apical cell, and measured 28.5 to 41.4 × 3.3 to 4.9 (average 35.0 × 4.0) μm. Such characteristics are typical of Fusarium oxysporum Schlechtendahl emend. Snyder & Hansen (3). Amplification of the internal transcribed spacer (ITS) of the rDNA using primers ITS1/ITS4 yielded a 504-bp amplicon (GenBank Accession No. KJ909935). Sequencing and BLASTn analysis of this amplicon showed a 100% homology with the sequence of F. oxysporum KC304802. To confirm pathogenicity, two Fusarium isolates were tested. For each isolate, three 2-year-old healthy plants of C. marginatus were inoculated by introducing into lesions (4 lesions/plant) artificially produced on the stem sterile needles contaminated with the pathogen (4). Inoculum was obtained from pure cultures grown on PDA. Control plants were punctured with sterile needles without inoculum. All the plants were placed in a greenhouse, at temperatures ranging between 16 and 24°C. For both tested strains, the first necrosis of stem tissues developed around the needles 7 days after the artificial inoculation, while non-inoculated plants remained healthy. Then, necrosis extended causing soft rot on plants maintained at relative humidity ranging from 55 to 65%. F. oxysporum identified by morphological characteristics was consistently isolated from symptomatic plants. The pathogenicity test was conducted twice. F. oxysporum has been reported on Cereus sp. in the United States and on C. peruvianus monstruosus in Italy (1). Currently, this disease is present in a few commercial nurseries in Liguria, although it could spread further and cause important economic losses. References: (1) A. Garibaldi et al. Plant Dis. 95:877, 2011. (2) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell, Ames, IA, 2006. (4) V. Talgø and A. Stensvand. OEPP/EPPO Bulletin 43:276, 2013.

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