First report of Neofusicoccum parvum causing brown spots on gallnuts of Rhus potaninii in China

Plant Disease ◽

10.1094/pdis-07-21-1575-pdn ◽

2021 ◽

Author(s):

Tao Ma ◽

Zixiang Yang

Keyword(s):

Management Practices ◽

Sequence Similarity ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Neofusicoccum Parvum ◽

First Report ◽

Gene Coding ◽

Initial Symptoms ◽

Pathogenicity Tests ◽

Plastic Containers

Chinese gallnuts have been used as a source of tannic acids, which are widely used in medicinal and chemical products (Zhang et al. 1999). In summer 2020, a severe outbreak of brown spots was observed on Du-ensiform gall of Rhus potaninii, at the Dongyue village, Emei county, Sichuan province, China. Spots irregular brown to blackish brown mainly occurred on the gall surfaces and spread quickly on about 40% of Du-ensiform galls. Nine symptomatic galls were randomly collected in the field. Tissues from the infected galls (ca. 3×5 mm) were surface-disinfected by 75% ethyl alcohol and 10% sodium hypochlorite, then plated on potato dextrose agar PDA at 25°C in the dark for 6 days. Similar fungal colonies were isolated from 56% of the infected galls. Five isolates from different galls showed similar colony morphology. After sub-culturing of hyphal tips on PDA，the initially whitish colonies showed abundant gray to black aerial mycelium after 5-7 days, with no sporulation. A representative isolate RC82 was randomly selected to induce sporulation on sterilized pine needles placed on water agar at 25°C in the dark. Black pycnidia (up to 450 um diam.) developed after incubation for 28 days. Conidiogenous cells were holoblastic, hyaline and short subcylindrical. Conidia were hyaline, fusiform or elliptic, thin-walled and aseptated, and measured 15.2 ± 0.9 × 5.3 ± 0.3 µm (n=126). The morphological characteristics were consistent with Neofusicoccom parvum (Pennycook et al.1985; Crous et al. 2006). To classify this fungal pathogen genetically, sequence analyses were conducted using the ITS rDNA region and the gene coding for EF-1α with DNA samples from four isolates of infected samples. Genomic DNA was amplified with primers ITS1/ITS4 and EF1-728F (Carbone et al. 1999) /EF-2 (O’Donnell et al.1998). The sequences obtained were deposited in GenBank (accession Nos. ITS: MZ269214-MZ269217 and EF-1α: MZ269219-MZ269222, respectively). BLASTn searches revealed 100% similarity of each gene to related sequences of multiple reference isolates of N. parvum. Based on NJ phylogenetic tree analyses of the combined ITS and EF-1α datasets, the 4 isolates were clustered with N. parvum strains CMW9081 (ex-type), CMW27135, and CAA856 etc., with clear separation from other closely related Neofusicoccum spp. To perform pathogenicity tests, 3 isolates were randomly selected. The tests were conducted with mycelial plugs of a 7-day-old colony placed onto the surface of wounded and/or unwounded galls and host leaves. A total of 5 replicates were included/treatment. Sterile PDA agar plugs were inoculated as controls. The galls and leaves were incubated at 25°C in plastic containers lined with wet filter paper for high humidity. Initial symptoms appeared within 3 days on all wounded galls and leaves, while the unwounded galls, leaves and the controls remained asymptomatic. Some of the unwounded galls and leaves showed symptoms within 4-10 days after incubation. The fungus was re-isolated from the lesions of inoculated tissues. The re-isolated fungal colonies showed identical morphology and 100% sequence similarity of ITS and EF-1α with the initial isolate. No fungus was isolated from the controls. N. parvum is a common and cosmopolitan species on a wide variety of hosts (Phillips et al. 2013). To our knowledge, this is the first report of N. parvum causing gallnut brown spots on R. potaninii worldwide. It indicates that crop management practices need to be evaluated to control and or prevent the disease.

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First Report of Anthracnose of Papaya (Carica papaya L.) Caused by Colletotrichum siamense in China

Plant Disease ◽

10.1094/pdis-10-20-2154-pdn ◽

2021 ◽

Author(s):

Yujie Zhang ◽

Wenxiu Sun ◽

Ping Ning ◽

Tangxun Guo ◽

SuiPing Huang ◽

...

Keyword(s):

Carica Papaya ◽

Disease Incidence ◽

Aerial Mycelium ◽

Postharvest Disease ◽

Distilled Water ◽

First Report ◽

Surface Samples ◽

Initial Symptoms ◽

Colletotrichum Siamense ◽

Pathogenicity Tests

Papaya (Carica papaya L.) is a rosaceous plant widely grown in China, which is economically important. Anthracnose caused by Colletotrichum sp. is an important postharvest disease, which severely affects the quality of papaya fruits (Liu et al., 2019). During April 2020, some mature papaya fruits with typical anthracnose symptoms were observed in Fusui, Nanning, Guangxi, China with an average of 30% disease incidence (DI) and over 60% DI in some orchards. Initial symptoms of these papayas appeared as watery lesions, which turned dark brown, sunken, with a conidial mass appearing on the lesions under humid and warm conditions. The disease severity varied among fruits, with some showing tiny light brown spots, and some ripe fruits presenting brownish, rounded, necrotic and depressed lesions over part of their surface. Samples from two papaya plantations (107.54°E, 22.38°N) were collected, and brought to the laboratory. Symptomatic diseased tissues were cut into 5 × 5 mm pieces, surface sterilized with 2% (v/v) sodium hypochlorite for 1 minute, and rinsed three times with sterilized water. The pieces were then placed on potato dextrose agar (PDA). After incubation at 25°C in the dark for one week, colonies with uniform morphology were obtained. The aerial mycelium on PDA was white on top side, and concentric rings of salmon acervuli on the underside. A gelatinous layer of spores was observed on part of PDA plates after 7 days at 28°C. The conidia were elliptical, aseptate and hyaline (Zhang et al., 2020). The length and width of 60 conidia were measured for each of the two representative isolates, MG2-1 and MG3-1, and these averaged 13.10 × 5.11 μm and 14.45 × 5.95 μm. DNA was extracted from mycelia of these two isolates with the DNA secure Plant Kit (TIANGEN, Biotech, China). The internal transcribed spacer (ITS), partial actin (ACT), calmodulin (CAL), chitin synthase (CHS), β-tubulin 2 (TUB2) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) regions were amplified by PCR and sequenced. The sequences were deposited into GenBank with accessions MT904003, MT904004, and MT898650 to MT898659. BLASTN analyses against the GenBank database showed that they all had over 99% identity to the type strain of Colletotrichum siamense isolate ICMP 18642 (GenBank accession numbers JX010278, GQ856775, JX009709, GQ856730, JX010410, JX010019) (Weir et al., 2012). A phylogenetic tree based on the combined ITS, ACT, CAL, CHS, TUB2 and GAPDH sequences using the Neighbor-joining algorithm also showed that the isolates were C. siamense. Pathogenicity tests were conducted on 24 mature, healthy and surface-sterilized papaya fruits. On 12 papaya fruits, three well separated wounded sites were made for inoculation, and for each wounded site, six adjacent pinhole wounds were made in a 5-mm-diameter circular area using a sterilized needle. A 10 µl aliquot of 1 × 106 conidia/ml suspension of each of the isolates (MG2-1 and MG3-1) was inoculated into each wound. For each isolate, there were six replicate fruits. The control fruits were inoculated with sterile distilled water. The same inoculation was applied to 12 non-wound papaya fruits. Fruits were then placed in boxes which were first washed with 75% alcohol and lined with autoclaved filter paper moistened with sterilized distilled water to maintain high humidity. The boxes were then sealed and incubated at 28°C. After 10 days, all the inoculated fruits showed symptoms, while the fruits that were mock inoculated were without symptoms. Koch's postulates were fulfilled by re-isolation of C. siamense from diseased fruits. To our knowledge, this is the first report of C. siamense causing anthracnose of papaya in China. This finding will enable better control of anthracnose disease caused by C. siamense on papaya.

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First Report of Leaf Blight in Chinese Hickory (Carya cathayensis) Caused by Alternaria petroselini in China

Plant Disease ◽

10.1094/pdis-02-13-0176-pdn ◽

2013 ◽

Vol 97 (9) ◽

pp. 1253-1253 ◽

Cited By ~ 1

Author(s):

Y. H. Liu ◽

C. Q. Zhang ◽

B. C. Xu

Keyword(s):

Management Practices ◽

Morphological Characteristics ◽

Morphological Characterization ◽

Leaf Blight ◽

Internal Transcribed Spacers ◽

Infected Leaf ◽

First Report ◽

Plastic Bags ◽

Initial Symptoms ◽

Carya Cathayensis

Chinese hickory (Carya cathayensis) is one of the important economic forest crops in Zhejiang and Anhui Provinces, China. In 2012, nearly 40% of hickory orchards and 6.8% of hickory trees were affected by leaf blight in Zhejiang. Initial symptoms consisted of small, brown, water-soaked lesions, which subsequently enlarged and developed a black sporulating necrotic center surrounded by a chlorotic halo. Infected leaf samples collected from 25 different orchards in Lin'an and 13 different orchards in Chun'an were surface sterilized with 1.5% sodium hypochlorite for 1.5 min, rinsed in water, plated on 2% potato dextrose agar (PDA), and incubated at 25°C in the dark for 1 week. Single conidium cultures were consistently isolated and cultured on PDA and V8 agar for morphological characterization (1,3). On both agar media, colonies were dark olive brown with smooth margins and concentric rings of sporulation. Conidia were solitary, darkly pigmented, predominantly ovoid-subsphaeroid, and 23 to 52 × 13 to 23 μm with up to six or seven transepta and one to three longisepta. The ribosomal internal transcribed spacers ITS1 and ITS2 of 10 isolates were amplified using primers ITS1/ITS4 on DNA extracted from mycelium and nucleotide sequences showed 100% similarity to that of A. petroselini (GenBank Accession Nos. AY154685.1 and EU807868.1). To confirm pathogenicity, 10 uninfected leaves from each of 10 C. cathayensis trees were sprayed either with a conidia suspension (105 conidia per ml) or with distilled water only to serve as an un-inoculated control. Leaves were subsequently wrapped in plastic bags to retain moisture, and incubated for 48 h. After 1 week, 8 of 10 isolates caused lesions identical to those initially described whereas no symptoms developed on water inoculated leaves. Cultures reisolated from lesions and cultured on PDA exhibited morphological characteristics identical to A. petroselini (1,2,3), confirming Koch's postulates. To our knowledge, this is the first report of leaf blight in C. cathayensis, and this identification would allow producers to identify for appropriate management practices. References: (1) P. M. Kirk et al. The Dictionary of the Fungi, 10th edition, page 159. CABI Bioscience, UK, 2008. (2) B. M. Pryor et al. Mycologia 94:49, 2002. (3) E. G. Simmons. Alternaria: An Identification Manual. CBS Fungal Biodiversity Centre, Utrecht, The Netherlands, 2007.

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First Report of Colletotrichum clavatum Causing Quince Anthracnose in Serbia

Plant Disease ◽

10.1094/pdis-01-14-0052-pdn ◽

2014 ◽

Vol 98 (9) ◽

pp. 1272-1272

Author(s):

S. Živković ◽

V. Gavrilović ◽

T. Popović ◽

N. Dolovac ◽

N. Trkulja

Keyword(s):

Management Practices ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Fruit Trees ◽

Morphological Identification ◽

Distilled Water ◽

First Report ◽

Mediterranean Countries ◽

Wide Range ◽

Plant Pathol

Quince (Cydonia oblonga Mill.) tree is traditionally grown in Serbia. The fruits are used for compote, marmalade, and brandy production. In December 2012, quince fruits cv. Leskovacka with symptoms of postharvest anthracnose were collected in a storage facility in the area of Sabac, western Serbia. The symptoms were observed on fruits approximately 2 months after harvest. The incidence of the disease was about 3%, but the symptoms were severe. Affected fruits showed sunken, dark brown to black lesions with orange conidial masses produced in black acervuli. Small pieces (3 to 5 mm) of necrotic tissue were surface sterilized for 1 min in 1% NaOCl, washed twice with sterile distilled water, and placed on potato dextrose agar (PDA). Macroscopic and microscopic morphology characteristics of three isolates were observed after growth on PDA for 7 days at 25°C under a 12-h photoperiod. Fungal colonies developed white to gray dense aerial mycelium with orange conidial masses in the center of the colony. Conidia were hyaline, aseptate, clavate with rounded distal apices, 15.2 (12.8 to 16.8) × 4.5 (4.0 to 5.2) μm (mean L/W ratio = 3.3, n = 100). Morphological characteristics are consistent with the description of Colletotrichum clavatum (2). Fungal isolates were also characterized by sequencing of the internal transcribed spacer (ITS) rDNA region using ITS1/IT4 primers and β-tubuline 2 gene using T1/T2 primers. The nucleotide sequences were deposited in GenBank (ITS Accession Nos. KF908866, KF908867, and KF908868; β-tubuline 2 gene KF908869, KF908870, and KF908871). BLAST analyses of ITS and β-tubuline 2 gene sequences showed that isolates from quince were 100% identical to other C. clavatum in GenBank (ITS JN121126, JN121130, JN121132, and JN121180; β-tubuline 2 gene JN121213 to 17, JN121219, JN121228, JN121261 to 62, and JN121266 to 69), thus confirming the morphological identification. To fulfill Koch's postulates, asymptomatic fruits of quince cv. Leskovacka (five fruits per isolate) were surface sterilized with 70% ethanol, wounded with a sterile needle, and inoculated with 50 μl of a spore suspension (1 × 106 conidia/ml). Five control fruits were inoculated with 50 μl of sterile distilled water. The experiment was repeated twice. After 10 days of incubation in plastic containers, under high humidity (>90% RH) at 25°C, typical anthracnose symptoms developed on inoculated fruits, while control fruits remained symptomless. The isolates recovered from symptomatic fruits showed the same morphological features as original isolates. C. clavatum previously indicated as group B (3), or genetic group A4 within the C. acutatum sensu lato complex (4), is responsible for olive anthracnose in some Mediterranean countries (1,2), and has been reported as causal agent of anthracnose on a wide range of other hosts including woody and herbaceous plants, ornamentals, and fruit trees worldwide (4). To our knowledge, this is the first report of C. clavatum in Serbia, and the first report of quince anthracnose caused by this pathogen in Europe. Anthracnose caused by C. clavatum can endanger the production and storage of quince in the future, and may require investigation of new disease management practices to control this fungus. References: (1) S. O. Cacciola et al. J. Plant Pathol. 94:29, 2012. (2) R. Faedda et al. Phytopathol. Mediterr. 50:283, 2011. (3) R. Lardner et al. Mycol. Res. 103:275, 1999. (4) S. Sreenivasaprasad and P. Talhinhas. Mol. Plant Pathol. 6:361, 2005.

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First Report of Crown Rot of Banana Caused by Fusarium porliferatum in Georgia, USA

Plant Disease ◽

10.1094/pdis-08-21-1809-pdn ◽

2021 ◽

Author(s):

Sumyya Waliullah ◽

Greg E. Fonsah ◽

Jason Brock ◽

Yonggang Li ◽

Emran Ali

Keyword(s):

Sequence Similarity ◽

Morphological Characteristics ◽

Fusarium Proliferatum ◽

Crown Rot ◽

Post Inoculation ◽

Conidial Suspension ◽

Banana Fruit ◽

Single Spore ◽

First Report ◽

Initial Symptoms

Crown rot is one of the most damaging disease of banana fruit characterized by rot and necrosis of crown tissues. In severe cases, the disease can spread to the pedicel and banana pulp. Crown rot can be infected by several common fungi, including Lasiodiplodia theobromae, Musicillium theobromae, Colletotrichum musae, and a complex of Fusarium spp. and lead to softening and blackening of tissues (Lassois et al., 2010; Kamel et al., 2016; Triest et al., 2016; Snowdon, 1990). In November 2020, typical crown rot of banana fruits (cv. Pisang Awak, belonging to the tetraploid AABB genome) were observed from UGA Banana Research 12 Plots, Tifton, GA, with incidence rates of 15%. Initial symptoms appeared in the infected crown of green banana fruits. As the infection progressed, the crown tissues became blackened and softened, followed by an internal development of infection affecting the peduncle and the fruit, triggered early ripening of bananas. At last, the development of necrosis on the pedicels and fruits appeared and caused the fingers to fall off. To identify the pathogen, tissue pieces (~0.25 cm2) from the infected crown and pedicles were surface-sterilized in a 10% bleach solution for 1 min, followed by 30 s in 70% EtOH. The disinfected tissues were rinsed in sterile water 3 times and cultured on potato dextrose agar (PDA) amended with 50 µg/ml streptomycin at 25°C in the dark for 5–10 days. Isolates of the pathogen were purified using the single-spore isolation method (Leslie and Summerell 2006). Colonies on PDA produced fluffy aerial mycelium and developed an intense purple pigment when viewed from the underside. A range of colony pigmentation and growth rates were observed among the isolates. The microconidia were ovoid, hyaline, or ellipse in shape. The morphological features of the isolates were identified as Fusarium proliferatum (Leslie and Summerell, 2006). To further identify the isolates, genomic DNA was extracted from a representative isolate. And the internal transcribed spacer (ITS) region, the partial elongation factor (TEF1-α) gene and the β-tubulin gene (TUB2)were amplified and sequenced using the primers ITS1/ITS4 (Yin et al. 2012), EF-1 /EF-2 (O’Donnell et al. 1998) and B-tub1 /B-tub2 (O’Donnell and Cigelnik, 1997), respectively. The amplicons were sequenced and deposited in NCBI (accessions no. MZ292989, MZ293071 for ITS: MZ346602, MZ346603 for TEF1-α and MZ346600 and MZ346601 for B-tub). The ITS, TEF1-α, and B-tub sequences of the isolates showed 100% sequence similarity with Fusarium proliferatum isolates (accessions no. MT560212, LS42312, and LT575130, respectively) using BLASTn in Genbank. For pathogenicity testing, three whole bunched bananas sterilized with 10% bleach solutions and washed by sterilized water, were cut into 5 bananas per brunch. The cut surface of the banana crown was inoculated with conidial suspension (1.0 × 107 cfu/ml) of the pathogen with pipette tips. Equal number of bananas were treated with sterilized water in the same volume as a control. All bananas were sealed in a plastic bag and incubated at 25°C. After 7 days post inoculation, all inoculated bananas showed initial crown rot symptoms while no symptoms were observed on the control bananas. The fungus was re-isolated from the symptomatic tissues of infected bananas and confirmed to be genetically identical to F. proliferatum of the original inoculated strains according to morphological characteristics and molecular identification, fulfilling Koch’s postulates. To the best of our knowledge, this is the first report of F. proliferatum causing crown rot on bananas in Georgia, USA.

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First Report of Phytophthora nicotianae Causing Asparagus Spear and Root Rot in Peru

Plant Disease ◽

10.1094/pdis-92-6-0982b ◽

2008 ◽

Vol 92 (6) ◽

pp. 982-982 ◽

Cited By ~ 3

Author(s):

L. M. Aragon-Caballero ◽

O. P. Hurtado-Gonzales ◽

J. G. Flores-Torres ◽

W. Apaza-Tapia ◽

K. H. Lamour

Keyword(s):

Sequence Similarity ◽

Phytophthora Capsici ◽

Morphological Characteristics ◽

Its Region ◽

Infected Tissue ◽

Phytophthora Nicotianae ◽

Asparagus Officinalis ◽

Its Sequence ◽

First Report ◽

Pathogenicity Tests

During 2006, spears, roots, and crowns of asparagus (Asparagus officinalis) exhibiting brown necrotic lesions with water soaking were collected from several sites across Peru (Ica, Lima, and Trujillo). Small infected tissue sections were washed thoroughly with tap and sterile distilled water and transferred to corn meal agar plates (CMA) amended with PARP (100 ppm of pimaricin, 100 ppm of ampicillin, 30 ppm of rifampicin, and 100 ppm of pentachloronitrobenzene) and incubated for five days at 25°C. Hyphal tips were subcultured from actively expanding mycelium. Sporangia produced on CMA were papillate and averaged 38 μm long × 29 μm wide. Chlamydospores were terminal or intercalary and averaged 35 μm in diameter. Isolates incubated in the dark for more than 3 weeks did not produce oospores in single culture. Mating with Phytophthora capsici tester isolates CBS 121656 = A1 and CBS 121657 = A2 indicate that all five isolates were A2. For pathogenicity tests, inoculum was generated by incubating 300 g of autoclaved wheat seeds with four agar plugs (7 mm) of expanding mycelium in polyethylene bags for 1 month at 25°C. Nine-week-old asparagus plants (UC151 F1) were transferred into pots containing autoclaved substrate (1 part sand, 1 part potting soil, and 1 part peat). Inoculum was added as 1 g of inoculum per kilogram of substrate. Plants were maintained in the greenhouse at 23°C and watered daily. Decline symptoms as well as root and spear rot were observed after 7 days and a Phytophthora sp. was reisolated from infected tissue. No symptoms were observed on asparagus plants inoculated with sterile inoculum. DNA was isolated from two representative isolates, and the nuclear ribosomal internal transcribed spacer (ITS) region was amplified with ITS4 and ITS6 primers and sequenced. ITS sequence was submitted for a BLAST search in the NCBI database, showing Phytophthora nicotianae strain UQ848 Accession No AF266776 as the closest match with 99% sequence similarity (1). The consensus ITS sequence was deposited in NCBI (Accession No. EU433396). These results, together with the morphological characteristics, indicate that the Phytophthora sp. isolated from asparagus in Peru is P. nicotianae (Breda de Haan) (2). To our knowledge, this is the first report of P. nicotianae infecting asparagus and represents a new threat for asparagus growers in Peru. Control methods such as moderate watering and metalaxyl application are being applied to reduce Phytophthora outbreaks. References: (1) D. E. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society. St Paul, MN, 1996.

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First Report of Corynespora cassiicola Causing Leaf Spot on Akebia trifoliate

Plant Disease ◽

10.1094/pdis-04-13-0454-pdn ◽

2013 ◽

Vol 97 (12) ◽

pp. 1659-1659 ◽

Cited By ~ 2

Author(s):

Y. F. Ye ◽

N. Jiang ◽

G. Fu ◽

W. Liu ◽

F. Y. Hu ◽

...

Keyword(s):

Leaf Spot ◽

Sequence Similarity ◽

Yangtze River Basin ◽

Morphological Characteristics ◽

Colletotrichum Acutatum ◽

Leaf Spot Disease ◽

Corynespora Cassiicola ◽

High Sequence Similarity ◽

First Report ◽

Initial Symptoms

Akebia species have been used for centuries in medicinal practices in a few Asian countries such as China and Japan. The dried stems of Akebia trifoliata are known as mutong in the Chinese pharmacopoeia (4) and mokutsu in Kampo, the traditional Chinese medicine developed in Japan (2). In China, the plant is grown in the provinces of Shandong, Hebei, Shanxi, Henan, Gansu, and some provinces in the south of the Yangtze River basin. During the summer of 2012, a leaf spot disease was detected on A. trifoliata grown in Nanning, Guangxi, China. The disease occurred and spread rapidly in July under conditions of high temperature and high humidity. The symptoms appeared on three sites that we inspected; disease incidences were higher than 80%. Initial symptoms consisted of small (less than 5 mm in diameter), circular, purple-brown leaf spots. Spots later enlarged and became elliptical, circular, or irregular with gray-white centers and dark brown rims. The centers were slightly concave. The spots could coalesce with each other, resulting in leaf desiccation and wilting. A fungal isolate was obtained from symptomatic leaf tissue that taken from a field (22°50′N, 108°22′E) in Nanning, Guangxi, China. Single-spore culture of the isolate was incubated on potato dextrose agar (PDA) for 7 days in the dark at 28°C. Conidiophores were straight to slightly curved, unbranched, and pale brown. Conidia (19.0 to 140.5 μm long and 7.0 to 11.0 μm wide) were formed singly or in chains, obclavate to cylindrical, straight or curved, pale brown, with a rounded apex and truncate base, and 1 to 13 pseudosepta. Morphological characteristics of the isolate were similar to the descriptions of Corynespora cassiicola (Berk. & M.A. Curtis) C.T. Wei (1). Genomic DNA of the isolate was extracted and used for PCR amplification of rDNA-ITS (internal transcribed spacer) sequence with primers ITS1 and ITS4. The PCR products were purified and sequenced. The sequence (GenBank Accession No. KC977496) was used in BLAST searches to interrogate GenBank for sequence similarity. High sequence similarity of 100% was obtained with several C. cassiicola strains. Pathogenicity of the isolate was investigated to demonstrate Koch's postulate. Young, healthy, fully expanded green leaves of A. trifoliata were surface sterilized. Fifteen leaves were inoculated with 10-μl drops of conidia suspension (105 conidia per ml) and 10 leaves were inoculated with the same volume of sterile water to serve as controls. All the leaves were placed in a humid chamber for 5 days. Spots with similar symptoms to those observed in the field developed on all inoculated leaves. The pathogen was reisolated and identified as C. cassiicola. The controls remained symptomless. According to previous reports, A. trifoliata was infected by Alternaria tenuissima in China and by Colletotrichum acutatum in Japan (3). To our knowledge, this is the first report of C. cassiicola found on Akebia species worldwide. Furthermore, this new disease primarily affects plantations and reduces the quality and yield of the medicine. Some effective measures should be taken to control this disease. References: (1) M. B. Ellis and P. Holliday. CMI Description of Pathogenic Fungi and Bacteria, 303, 1971. (2) F. Kitaoka et al. J. Nat. Med. 63:374, 2009. (3) Y. Kobayshi et al. J. Gen. Plant Pathol. 70:295, 2004. (4) L. Li et al. HortScience 45:4, 2010.

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First Report of Neofusicoccum parvum Causing Panicle Blight and Leaf Spot on Vitis heyneana in China

Plant Disease ◽

10.1094/pdis-05-14-0515-pdn ◽

2015 ◽

Vol 99 (3) ◽

pp. 417-417 ◽

Cited By ~ 4

Author(s):

D. D. Wu ◽

G. Fu ◽

Y. F. Ye ◽

F. Y. Hu ◽

H. F. Mou ◽

...

Keyword(s):

Leaf Spot ◽

Severe Disease ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Its Region ◽

Average Diameter ◽

Wine Production ◽

Neofusicoccum Parvum ◽

First Report ◽

Panicle Blight

The climbing vine, Vitis heyneana Roem. & Schult, is a member of the grape family endemic to Asia. Its fruits are used in wine production, and its roots, stems, and leaves can be used in medicinal materials. This plant is grown in Southwest China, as well as in India, Bhutan, and Nepal. Mulao Autonomous County in Guangxi Province is the only artificial cultivation area in China. During the summer of 2013, a panicle blight and leaf spot were detected on V. heyneana on four farms in Mulao Autonomous County. The symptoms were observed from the onset of florescence through fruit harvest. Brown lesions initially appeared at the base of a panicle and then extended to the whole panicle, finally causing the panicle to die and fruit to drop. When the disease developed on leaves, the symptom initially appeared as small dark brown circular spots, later enlarging into irregular spots (average diameter 6 mm) with a light brown center and dark brown rim. With severe disease, some individual leaves were affected by numerous spots, leading to premature senescence. Small sections of diseased tissue excised from 10 panicle and 10 leaf samples were plated on potato dextrose agar (PDA) and incubated at 28°C. Fungal colonies developed, initially with abundant white aerial mycelium, which turned olivaceous gray after 5 days and formed black pycnidia after 25 days. The conidia were hyaline, ellipsoidal to fusiform, externally smooth, thin-walled, and nonseptate. Thirty conidia were measured; the dimensions were 12.0 to 17.5 × 4.0 to 6.0 μm. Morphological characteristics of the isolates were similar to the descriptions of Neofusicoccum parvum (3). The isolate MPT-1 was selected as a representative for molecular identification. Genomic DNA was extracted and used for PCR to amplify the internal transcribed spacer (ITS) region and partial translation elongation factor 1-alpha (EF1-α) gene, using primers ITS1/ITS4 and EF1-728F/EF1-986R, respectively. The obtained ITS sequence (GenBank Accession No. KJ599627) and EF1-α sequence (KM921768) showed >99% homology with several GenBank sequences of N. parvum. Morphological and molecular results confirmed the isolate as N. parvum. For pathogenicity tests, detached, young healthy panicles and leaves of V. heyneana were surface-sterilized, wounded by sterile needle, and inoculated with mycelial plugs (3 mm in diameter) of four N. parvum isolates. Ten panicles and 10 leaves were used for every isolate. Control panicles and leaves were treated with sterile PDA plugs. All the samples were placed in a humid chamber (RH 90%, 28°C, 12 h of light) for 3 days. Symptoms similar to those observed in the field developed on all panicles and leaves inoculated with N. parvum isolates. N. parvum was reisolated from all inoculated, symptomatic tissues. The controls remained symptomless. N. parvum has been reported to cause trunk canker on V. vinifera (2), dieback on Cupressus funebris (3), and a leaf spot on Myristica fragrans (1). To our knowledge, this is the first report of N. parvum causing panicle blight and leaf spot on V. heyneana in China. Panicle blight caused a large number of fruits to drop and reduced the yield seriously. Some effective measures should be taken to control this disease. References: (1) V. Jayakumar et al. New Dis. Rep. 23:19, 2011. (2) J. Kaliternam et al. Plant Dis. 97:1656, 2013. (3) S. B. Li et al. Plant Dis. 94:641, 2010.

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First Report of Damping-Off Caused by Rhizoctonia solani AG-4 on Pink Ipê (Tabebuia impetiginosa) in Italy

Plant Disease ◽

10.1094/pdis-10-10-0748 ◽

2011 ◽

Vol 95 (1) ◽

pp. 78-78 ◽

Cited By ~ 2

Author(s):

G. Polizzi ◽

D. Aiello ◽

A. Vitale ◽

V. Guarnaccia ◽

A. Panebianco ◽

...

Keyword(s):

Rhizoctonia Solani ◽

Morphological Characteristics ◽

Fluorescent Light ◽

Damping Off ◽

First Report ◽

Initial Symptoms ◽

Public Areas ◽

Pathogenicity Tests ◽

Stem Lesions ◽

Safranin O

Pink ipê or pink lapacho (Tabebuia impetiginosa Martius ex DC., family Bignoniaceae) is one of the most attractive blooming trees in the world. In Europe, pink ipê is widely used as an ornamental tree in landscaped gardens and public areas. In August 2010, a widespread damping-off was observed in a stock of approximately 100,000 potted 2-month-old seedlings in a nursery in eastern Sicily (Italy). The seedlings were being watered with overhead irrigation. More than 5% of the seedlings showed disease symptoms. Initial symptoms were black lesions at the seedling crown that expanded rapidly to girdle the stem. On infected seedlings, leaves turned black and gradually died. Black extended stem lesions were followed by death of the entire seedling in a few days. A fungus with mycelial and morphological characteristics of Rhizoctonia solani Kühn was consistently isolated from crown and stem lesions when plated on potato dextrose agar (PDA) amended with streptomycin sulfate at 100 μg/ml. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Mycelium was branched at right angles with a septum near the branch and a slight constriction at the branch base. Hyphal cells removed from cultures grown at 25°C on 2% water agar were determined to be multinucleate when stained with 1% safranin O and 3% KOH solution (1) and examined at ×400. Anastomosis groups were determined by pairing isolates with tester strains AG-1 IA, AG-2-2-1, AG-2-2IIIB, AG-2-2IV, AG-3, AG-4, AG-5, AG-6, and AG-11 on 2% water agar in petri plates (4). Anastomosis was observed only with tester isolates of AG-4, giving both C2 and C3 reactions (2). Pathogenicity tests were performed on container-grown, healthy, 3-month-old seedlings. Forty seedlings of T. impetiginosa were inoculated near the base of the stem with two 1-cm2 PDA plugs from 5-day-old mycelial cultures. The same number of plants only inoculated with PDA plugs served as controls. Plants were incubated in a growth chamber and maintained at 25°C and 95% relative humidity on a 12-h fluorescent light/dark regimen. Crown and stem lesions identical to those observed in the nursery appeared 5 days after inoculation and all plants died within 25 days. No disease was observed on control plants. R. solani AG-4 was reisolated from symptomatic tissues and identified as previously described. R. solani AG-4 was previously detected in the same nursery on Chamaerops humilis (3). To our knowledge, this is the first report of R. solani causing damping-off on T. impetiginosa. References: (1) R. J. Bandoni. Mycologia 71:873, 1979. (2) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (3) G. Polizzi et al. Plant Dis. 94:125, 2010. (4) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.

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First Report of Forsythia Shoot Blight Caused by Phytophthora nicotianae in Connecticut

Plant Disease ◽

10.1094/pdis-03-14-0297-pdn ◽

2014 ◽

Vol 98 (9) ◽

pp. 1278-1278

Author(s):

J. A. LaMondia ◽

D. W. Li ◽

A. M. Madeiras ◽

R. L. Wick

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Phytophthora Nicotianae ◽

First Report ◽

Sterile Needle ◽

Shoot Blight ◽

Pathogenicity Tests ◽

Half Strength ◽

Internal Necrosis

Blighting of Forsythia × intermedia ‘Showoff’ was first observed affecting several hundred plants in a commercial nursery in Connecticut in September 2012. Symptoms included wilting, leaf and stem blight, and dieback progressing to plant death. A Phytophthora sp. was isolated from symptomatic tissues on half-strength potato dextrose agar (½PDA). Colonies were white and cottony on ½PDA, reaching 9 mm in 15 days at 25°C, but colorless and inconspicuous on pimaricin, ampicillin, rifampicin, pentachloronitrobenzene agar (PARP) with sparse and limited aerial mycelium, reaching 60 mm in 15 days at 25°C. The characteristics of the pathogen were observed and measured from a 3-month-old colony on ½PDA. Sporangia were abundant, various in shape, ovoid, ellipsoid to pyriform or limoniform, occasionally gourd shaped or irregular; (17.9) 27.2 to 41.4 (47.3) × (12.6) 19.1 to 30.5 (36.7) μm (n = 30), length/breadth ratio 1.4 ± 0.2, papillate and noncaducous. Papillae measured 2.9 ± 0.8 × 3.4 ± 0.8 μm (n = 10). Chlamydospores were present, 23.4 ± 3.1 × 22 ± 3.3 μm (n = 10). Oogonia and oospores were not observed. Arachnoid mycelia were present. These morphological characteristics are consistent with Phytophthora nicotianae Breda de Haan (1). The identity of the pathogen was confirmed as P. nicotianae by BLAST analysis of ITS, Cox II, and beta tubulin gene sequences (99% match for the three sequences, E value = 0). Pathogenicity tests were conducted four times on healthy liners of Forsythia × intermedia ‘Showoff’ grown in 10-cm-diameter pots. Leaves and stems were wounded by pricking with a sterile needle and six plants were inoculated with 0.25 cm2 plugs of the pathogen growing on ½PDA placed on three leaves and in three stem nodes and covered with Parafilm. Controls consisted of an equal number of plants wounded and inoculated with ½PDA alone. All plants were placed inside high humidity chambers for 24 h and then transferred to a greenhouse for up to 1 month. Typical symptoms developed within 1 week of inoculation and the pathogen was re-isolated from diseased tissue. Disease incidence was nearly 100% of inoculated leaves and stems and not observed in control plants without the pathogen. Three replicate 6-week-old broadleaf tobacco ‘C9’ plants were each inoculated with tobacco or forsythia isolates of P. nicotianae or sterile media alone, by wounding stems and placing colonized 0.25 cm2 ½PDA plugs into wounds and covering with Parafilm. After 1 week, stems were split and the length of internal necrosis in the stem measured. Disease resulted from inoculation with both the tobacco and forsythia isolates and stem necrosis averaged 43 and 23 mm for tobacco or forsythia isolates, respectively. No necrosis was observed in the pathogen-free controls. P. nicotianae has been reported from the basal stem and roots of F. viridissima in Italy (2) and from shoots of Forsythia × intermedia in Virginia (3). To our knowledge, this is the first report of P. nicotianae causing shoot blight on Forsythia in the northeastern United States. References: (1) J. van. Breda de Haan. Mededeelingenuit's Lands Plantentuin Batavia. 15:57, 1896. (2) S. O. Cacciola et al. Plant Dis. 78:525, 1994. (3) C. X. Hong et al. Plant Dis. 89:430, 2005.

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First Report of Brown Rot on Crataegus pinnatifida var. major Caused by Monilia yunnanensis in China

Plant Disease ◽

10.1094/pdis-01-13-0085-pdn ◽

2013 ◽

Vol 97 (9) ◽

pp. 1249-1249 ◽

Cited By ~ 3

Author(s):

Y. Z. Zhao ◽

D. Wang ◽

Z. H. Liu

Keyword(s):

Morphological Characteristics ◽

Aerial Mycelium ◽

Brown Rot ◽

Its Region ◽

Northern China ◽

Its2 Sequence ◽

Liaoning Province ◽

First Report ◽

Crataegus Pinnatifida ◽

Initial Symptoms

Crataegus pinnatifida Bge. var. major N. is a time-honored herbal medicine and an important economic fruit that is processed into various foods. Cultivated fields are mostly situated in northern China. In August 2012, a fruit brown rot disease on mature plants was found in Fushun, Liaoning Province, China. This disease reduced the yield and the quality of fruits; the incidence of fruits affected in each tree was between 20% and 35%. The initial stage of infection was marked by regular or irregular light brown spots turning dark brown and gradually supporting many grey fluffy tufts of mycelium. At last, the lesions expanded, causing the fruits to become rotted or withered. The pathogen was isolated from infected fruits by a tissue isolation method (1) and cultured on potato dextrose agar (PDA) at 25°C under 12 h light/12 h dark for 7 days. The colonies reached 60 to 75 mm in diameter after 7 days and were pale green or pale yellow with a neat greyish white margin; the aerial mycelium sometimes formed annulations. The conidia were single, hyaline, lemon-shaped or oval, and 11.3 ± 1.4 (8.0 to 16.5) × 6.4 ± 0.8 (5.1 to 7.6) μm; sporulation was sparse and stromata that formed after 15 days were abundant, black, and spherical to elliptical in shape. Conidiophores were simple or branched, bearing a single conidium on the tip. The internal transcribed spacer (ITS) region of rDNA amplified with primers ITS1 and ITS4 was directly sequenced in both directions. The ITS1-5.8S-ITS2 sequence of rDNA was 100% identical to Monilinia yunnanensis (GenBank Accession No. HQ908788.1), which is distinguished from the closely related M. fructigena (AF150679.1) and M. polystroma (JX315717.1). The pathogen was identified as M. yunnanensis on the basis of morphological characteristics (2) and the ITS sequence of rDNA (2,3). Pathogenicity tests were performed on detached healthy fruits. Four fruits were wounded with a sterilized nail and inoculated by placing a PDA plug of mycelia on upper surfaces of the fruits. Another four fruits treated with sterile PDA plugs served as a control. Fruits were cultured in petri dishes with a 12-h photoperiod at 25°C and 90% relative humidity. The initial symptoms on inoculated fruits were observed after 5 days, while no symptoms showed on the controls. The pathogen was reisolated from the inoculated fruit and identified as M. yunnanensis by the above methods. While M. yunnanensis has been reported to cause brown rot on peach (2), to our knowledge, this is the first report of M. yunnanensis infecting C. pinnatifida Bge. var. major N. in China. References: (1) Z. D. Fang. Research Methods of Plant Disease, 1998. (2) M. J. Hu et al. Monilinia species causing brown rot of peach in China. PLoS ONE. Online publication. doi:10.1371/journal.pone.0024990, 2011. (3) G. C. M. van Leeuwen et al. Mycol. Res. 106:444, 2002.

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