First Report of Soft Rot Caused by Aspergillus niger sensu lato on Mother-in-law’s Tongue in China

“Mother-in-law’s tongue” (MLT) [Dracaena trifasciata (Prain) Mabb. (syn. Sansevieria trifasciata Prain.)], also known as “Saint George's sword", “snake plant”, “tiger's tail orchid", etc., is an evergreen perennial ornamental plant grown worldwide. In September 2016, severe soft rot occurred on the leaves of MLT in a flower market in Nanyang city (32º56´N, 112º32´E), Henan province, China with 25% disease incidence (n=100). Water-soaked spots initially appeared on the leaf margin, enlarged rapidly, and became soft rot under excessively watered conditions. A blight zone was visualized at the margin of a developing lesion in backlit conditions. Severely affected leaves folded down from the lesions. Lesion expansion stopped under dry conditions. Grey or dark brown mycelia were frequently seen on the lesions. Tissue pieces (4×4 mm2) at the margin of lesions were cut out, treated with 75% ethanol for 10 s, followed by 70 s in 0.1% HgCl2, rinsed eight times with sterile water, and plated on potato dextrose agar (PDA) medium. Pure Aspergillus cultures were obtained from the surface-disinfected lesions after 4 days of incubation at 26°C. Two single-spore-derived isolates (An-1 and An-2) were randomly selected and used for morphological and molecular identifications as well as pathogenicity tests. The isolates formed round dark brown colonies with a large number of conidia after 5 days of incubation on PDA at 28°C. Conidia were subsphaeroidal or oblate, unicellular, dark brown, 2.9-4.2(3.5) × 1.9-3.4(2.7) μm in size (n=100), developed from a two-series of strigmata born on a conidial head, with ridge or stab-shaped prominences. For pathogenicity tests, the two isolates were separately grown on oatmeal agar and incubated at 30°C for 6 days. Mycelial plugs (5 mm diam.) were inoculated on the scalpel incision X-shaped wounds of surface-disinfected leaves of MLT. The inoculated leaves were kept on a two-layer of wet napkin in a steel basin covered with a plastic film. Soft rot symptoms developed from the wounds 6 days after incubation, similar to those observed on naturally affected leaves. The An-1- and An-2-inoculated unwounded leaves remained symptomless during the pathogenicity tests. Fungal cultures with the same phenotypes as the inocula were consistently reisolated from the lesions of the leaves inoculated by each of the two isolates, verifying the isolates as the causal agent of the disease based on Koch’s postulates. Both β-tubulin gene and rDNA-ITS (internal transcribed spacer) sequences of the two isolates were separately amplified and sequenced. Sequences were submitted to GenBank with accession numbers MN259522 and MN259523 for the β-tubulin gene sequences, and accession numbers MN227322 and MN227324 for the rDNA-ITS sequences of An-1 and An-2, respectively. Both An-1 and An-2 were clustered with members of Aspergillus niger van Tieghem in the phylogenetic tree of rDNA-ITS, clearly separated from other Aspergillus spp. In the phylogenetic tree of β-tublin gene, both An-1 and An-2 formed a subclade inside a large clade consisting of members of A. niger in strict sense. Based on the molecular and morphological results, both An-1 and An-2 clearly separated from other Aspergillus spp. and can be considered as A. niger sensu lato. Foliar diseases of MLT are known to be caused by a few fungal species such as Chaetomella spp. (Li et al. 2014) and Colletotrichum sansevieriae (Nakamura et al. 2006). This is the first report of A. niger sensu lato causing soft rot on MLT in China.

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First Report of Lasidiplodia theobromae and Cryptovalsa ampelina Associated with Grapevine Decline from Castilla y León, Spain

Plant Disease ◽

10.1094/pdis-93-5-0545c ◽

2009 ◽

Vol 93 (5) ◽

pp. 545-545 ◽

Cited By ~ 4

Author(s):

M. T. Martin ◽

L. Martin ◽

M. T. de-Francisco ◽

R. Cobos

Keyword(s):

Dna Sequences ◽

Morphological Characteristics ◽

Tubulin Gene ◽

Current Season ◽

First Report ◽

Grapevine Decline ◽

Eutypa Dieback ◽

Castilla Y Leon ◽

Pathogenicity Tests ◽

Β Tubulin

Symptoms of grapevine decline were surveyed. Samples from mature vines exhibiting external symptoms of Eutypa dieback and Esca were collected, as were young plants with and without external symptoms, and fungal isolations were performed. In 2007, 3-year-old grapevines (cv. Tempranillo grafted onto 110R rootstock) with low vigor, reduced foliage, and vascular streaking in the wood were observed. Small pieces of discolored wood were placed onto malt extract agar supplemented with 0.25 g/liter of chloramphenicol, incubated at 25°C, and resulting colonies were transferred to potato dextrose agar (PDA). Isolates were characterized by abundant aerial and fast-growing mycelium covering the plate surface after 3 days, mycelium became dark green. Pycnidia contained thick-walled, aseptate conidia 15 to 35 × 10 to 15 μm. Lasidiplodia theobromae was identified based on morphological characteristics (3) and confirmed by banding patterns obtained after the digestion of the 1,200-bp amplicon generated with ITS1 and NL4 primers (2) using restriction endonucleases (2). Single-spore cultures were generated and DNA sequences of the rDNA internal transcribed spacer region, partial sequence of the 5′ end of the β-tubulin gene, and a fragment of the elongation factor further confirmed the identification and revealed genetic similarity with other isolates of L. theobromae. A sequence of each fragment was deposited in GenBank with Accession Nos. EU600925, EU597297, and EU597298, respectively. Pathogenicity tests were conducted on four replicate rootstocks (110R) and 15 canes of current-season growth (cv. Tempranillo). Plants were inoculated with an agar plug containing L. theobromae; controls were treated with agar only. Grapevines were maintained in a greenhouse at 20 to 25°C. After 3 months, L. theobromae was reisolated from internal vascular lesions in 100 and 66% of inoculated rootstocks and canes, respectively. Control plants were asymptomatic and L. theobromae was not recovered. Using the same methodology, a fungus identified based on morphological characteristics in culture as Cryptovalsa ampelina (1) was isolated from grapevines (cv. Tempranillo) planted in 1987. Cultures in PDA were white to creamy white and cottony with diffuse margins. Colonies covered the 90-mm-diameter petri dish surface in 5 days. Conidia were 20 to 23 × 1 to 1.5 μm, unicellular, hyaline, and filiform. PCR amplifications of the DNA extracts of C. ampelina with Camp-1 and Camp-2R primers gave a characteristic DNA fragment of 300 bp (3) and DNA sequences of the ITS4-ITS5 amplicons (GenBank Accession No. EU597296) confirmed the identification. For the first time, the 5′ end of the β-tubulin gene was sequenced and deposited in GenBank (Accession No. EU600926). Pathogenicity tests were conducted as described above for L. theobromae. Both pathogens were examined in the same experiment. C. ampelina was reisolated from internal brown streaking lesions in 25% of the rootstocks and 33% of the canes. Control plants exhibited no symptoms. L. theobromae appeared to be a more aggressive pathogen than C. ampelina on grapevine with more internal brown streaking and greater recovery of pathogen from inoculated samples. To our knowledge, this is the first report of L. theobromae and C. ampelina causing grapevine decline in Castilla y León. References: (1) J. Luque et al. Phytopathol. Mediterr. 45:S101, 2006. (2) M. T. Martin and R. Cobos. Phytopathol. Mediterr. 46:18, 2007. (3) D. Pavlic et al. Stud. Mycol. 50:313, 2004.

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First Report of Pulp Rot in the Externally Asymptomatic Pomegranate Fruit Caused by Talaromyces albobiverticillius in Henan Province, China

Plant Disease ◽

10.1094/pdis-09-21-1997-pdn ◽

2021 ◽

Author(s):

Siliang Huang ◽

Ting Li ◽

Tiantian Yang ◽

Xue Ling Zheng ◽

Di Yang ◽

...

Keyword(s):

Phylogenetic Trees ◽

Henan Province ◽

Fruit Rot ◽

Gene Sequences ◽

Tubulin Gene ◽

First Report ◽

Rdna Its ◽

Phenotypic Features ◽

Pomegranate Fruit ◽

Β Tubulin

As a popular deciduous fruit tree, pomegranate (Punica granatum L.) is grown from tropical to temperate zones worldwide, therein China has at least 120000 hm2 cultivation area. In August 2020, severe pulp rot occurred in the externally asymptomatic pre-harvest pomegranate fruit on a 3-year-old soft-seeded variety (Tunisia) in the Zhanghe village (32º40´34˝N, 111º44´20˝E) of Jiuchong township, Xichuan county in Henan province, China with 6.4-20 (av. 12.6) % pulp rot incidence evaluated from 11 freshly sampled fruits (360 pulps per fruit investigated). The fruits showed no external symptoms, however, browning occurred on part of their pulps before harvest compared to the normal ones with white or pink color. The surface of the externally asymptomatic fruits was sterilized with 75% ethanol, and air-dried in a clean bench. The surface-disinfected fruits were dissected with a sterilized knife. Brown pulps from the fruits were picked up using flame-sterilized tweezers and placed on potato dextrose agar (PDA) plates. After five days of incubation at 28 °C, pure fungal cultures with similar phenotypic features developed from the affected pulps. Two randomly selected isolates Tp-2 and Tp-8 were used for the study. The colony surface of the isolates was greyish-green with claret-red exudates. Claret-red pigments were commonly secreted into the medium from the colonies. Conidia were unicellular, hyaline to greyish, mostly rugby ball-shaped with a dimension of 2.2-3.5 (2.7) µm × 1.6-2.0 (1.8) µm (n=50) for Tp-2, and 2.2-3.1 (2.6) µm × 1.6-2.2 (1.8) µm (n=50) for Tp-8. The rDNA internal transcribed spacer (ITS) and β-tubulin gene sequences of the isolates were amplified with primers ITS1/ITS4 and Bt2a/Bt2b, respectively. Sequences were submitted to GenBank with accession numbers MW132153 and MW132077 for the rDNA-ITS sequences, and MW507822 and MW507823 for the β-tubulin gene sequences of Tp-2 and Tp-8, respectively, with a maximal sequence identity greater than 99 % to multiple strains of Talaromyces albobiverticillius (TA) based on BLAST analyses. In the Neighbor-joining phylogenetic trees constructed using rDNA-ITS and β-tubulin gene sequences, both Tp-2 and Tp-8 formed a clade with mutiple strains of TA, clearly separated from other Talaromyces spp. Conidial suspensions (106 spores ml-1) of Tp-2 and Tp-8 were separately injected into five pomegranate fruits (Tunisia) sampled from an orchard free of the disease with a sterilized syringe. Five fruits inoculated with sterilized water were used as control (CK). The inoculated fruits were incubated at 25 °C for 10 days and cut out through the inoculated sites. Pulp rot symptoms occurred in the Tp-2/Tp-8-inoculated fruits, being similar to the naturally affected pulps. The CK pulps remained symptomless during the inoculation tests. Fungal cultures with the same phenotypic features as the inocula were constantly isolated from the brown pulps of the inoculated fruits, verifying both Tp-2 and Tp-8 as the causal agents of the disease based on Koch’s postulates. During a long-term (30-40 days) storage at ambient conditions, fruits sampled from affected orchards developed brown lesions on their peels from which TA cultures could be isolated. TA was reported as the pathogen causing postharvest fruit rot on pomegranate in Italy (Mincuzzi et al. 2017). This is the first report of TA causing pulp rot in the externally asymptomatic pomegranate fruit in China.

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First Report of Black Soft Rot of Indian Gooseberry Caused by Syncephalastrum racemosum

Plant Disease ◽

10.1094/pdis.2004.88.5.575c ◽

2004 ◽

Vol 88 (5) ◽

pp. 575-575 ◽

Cited By ~ 2

Author(s):

Neelima Garg ◽

Om Prakash ◽

B. K. Pandey ◽

B. P. Singh ◽

G. Pandey

Keyword(s):

Cellulase Activity ◽

Basal Medium ◽

Petri Dish ◽

Soft Rot ◽

Cellulolytic Activity ◽

Academic Press ◽

First Report ◽

Syncephalastrum Racemosum ◽

Indian Gooseberry ◽

Pathogenicity Tests

Indian gooseberry (Emblica officinalis Gaertn.) is a medicinal plant with high nutraceutical value. During November and December 2003, soft rot was noticed on harvested and stored (20 ± 5°C and 65 ± 5% relative humidity) fruits at the experimental farm in Rehmanhera, Lucknow, India (26°50′N, 80°54′E). These fruits had numerous, minute brown necrotic lesions showing white mycelial growth. A pronounced halo of water-soaked, faded tissue surrounded the lesion between the fringe of mycelium and healthy tissues. The rotted surface was covered with a black, powdery layer of spores. On Czapek yeast extract agar, fungal colonies were blackish grey, moderately dense, and covered the entire petri dish. The fungus produced aseptate mycelium. The sporangial heads were 30 to 50 μm in diameter with sporangiospores found linearly within cylindrical sacs (merosporangia) borne on spicules around the columella. Sporangiospores, spherical to cylindrical in shape and borne in chains, measured 3.0 to 5.0 μm long. The fungus was morphologically and physiologically identified as Syncephalastrum racemosum Schr. (2). For pathogenicity tests, healthy fruits (10 replicates) were surface sterilized and punctured inoculated aseptically with 1.0 × 106 conidia and incubated at 20 ± 5°C Typical symptoms of the disease appeared after 4 days. The fungus exhibited a strong level of cellulolytic activity as indicated by prolific growth on Indian gooseberry fiber waste under solid-state fermentation conditions. The level of cellulase activity (1) was 21 filter paper activity unit per ml at 72 hr in culture supernatant of basal medium having carboxymethyl cellulose as the carbon source. The fungus showed resistance to tannins (as much as 2%), since it could grow well in liquid growth medium (Czapek Dox broth) with 2% tannins and aonla juice with 1.8% tannins. Since Indian gooseberry is rich in fiber (2.5 to 3.4%) and tannins (1.5 to 2.0%), this may be an important pathogen. To our knowledge, this is the first report of the occurrence of Syncephalastrum racemosum on Indian gooseberry fruits. References: (1) T. K. Ghose. Pure Appl. Chem. 59(2):257, 1987. (2) J. I. Pitt and A. D. Hocking. Fungi and Food Spoilage. Academic Press. North Ryde, Australia, 1985.

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First Report of Fusarium Maize Ear Rot Caused by Fusarium kyushuense in China

Plant Disease ◽

10.1094/pdis-05-13-0558-pdn ◽

2014 ◽

Vol 98 (2) ◽

pp. 279-279 ◽

Cited By ~ 2

Author(s):

J.-H. Wang ◽

H.-P. Li ◽

J.-B. Zhang ◽

B.-T. Wang ◽

Y.-C. Liao

Keyword(s):

Fusarium Species ◽

Morphological Characteristics ◽

Distilled Water ◽

Ear Rot ◽

Tubulin Gene ◽

Average Growth ◽

First Report ◽

Maize Ear Rot ◽

Maize Ear ◽

Β Tubulin

From September 2009 to October 2012, surveys to determine population structure of Fusarium species on maize were conducted in 22 provinces in China, where the disease incidence ranged from 5 to 20% in individual fields. Maize ears with clear symptoms of Fusarium ear rot (with a white to pink- or salmon-colored mold at the ear tip) were collected from fields. Symptomatic kernels were surface-sterilized (1 min in 0.1% HgCl2, and 30 s in 70% ethanol, followed by three rinses with sterile distilled water), dried, and placed on PDA. After incubation for 3 to 5 days at 28°C in the dark, fungal colonies displaying morphological characteristics of Fusarium spp. (2) were purified by transferring single spores and identified to species level by morphological characteristics (2), and DNA sequence analysis of translation elongation factor-1α (TEF) and β-tubulin genes. A large number of Fusarium species (mainly F. graminearum species complex, F. verticillioides, and F. proliferatum) were identified. These Fusarium species are the main causal agents of maize ear rot (2). Morphological characteristics of six strains from Anhui, Hubei, and Yunnan provinces were found to be identical to those of F. kyushuense (1), which was mixed with other Fusarium species in the natural infection in the field. Colonies grew fast on PDA with reddish-white and floccose mycelia. The average growth rate was 7 to 9 mm per day at 25°C in the dark. Reverse pigmentation was deep red. Microconidia were obovate, ellipsoidal to clavate, and 5.4 to 13.6 (average 8.8) μm in length. Macroconidia were straight or slightly curved, 3- to 5-septate, with a curved and acute apical cell, and 26.0 to 50.3 (average 38.7) μm in length. No chlamydospores were observed. Identity of the fungus was further investigated by sequence comparison of the partial TEF gene (primers EF1/2) and β-tubulin gene (primers T1/22) of one isolate (3). BLASTn analysis of the TEF amplicon (KC964133) and β-tubulin gene (KC964152) obtained with cognate sequences available in GenBank database revealed 99.3 and 99.8% sequence identity, respectively, to F. kyushuense. Pathogenicity tests were conducted twice by injecting 2 ml of a prepared spore suspension (5 × 105 spores/ml) into maize ears (10 per isolate of cv. Zhengdan958) through silk channel 4 days post-silk emergence under field conditions in Wuhan, China. Control plants were inoculated with sterile distilled water. The ears were harvested and evaluated 30 days post-inoculation. Reddish-white mold was observed on inoculated ears and the infected kernels were brown. No symptoms were observed on water controls. Koch's postulates were fulfilled by re-isolating the pathogen from infected kernels. F. kyushuense, first described on wheat in Japan (1), has also been isolated from rice seeds in China (4). It was reported to produce both Type A and Type B trichothecene mycotoxins (1), which cause toxicosis in animals. To our knowledge, this is the first report of F. kyushuense causing maize ear rot in China and this disease could represent a serious risk of yield losses and mycotoxin contamination in maize and other crops. The disease must be considered in existing disease management practices. References: (1) T. Aoki and K. O'Donnell. Mycoscience 39:1, 1998. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (3) F. Van Hove et al. Mycologia 103:570, 2011. (4) Z. H. Zhao and G. Z. Lu. Mycotaxon 102:119, 2007.

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First Report of Eutypella spp. Associated with Branch Canker of Citrus in California

Plant Disease ◽

10.1094/pdis-01-11-0047 ◽

2011 ◽

Vol 95 (9) ◽

pp. 1187-1187 ◽

Cited By ~ 1

Author(s):

A. O. Adesemoye ◽

A. Eskalen

Keyword(s):

San Diego ◽

Its Sequences ◽

Control Treatment ◽

Molecular Characteristics ◽

Its2 Region ◽

Petroleum Jelly ◽

Tubulin Gene ◽

First Report ◽

Β Tubulin ◽

Branch Canker

Eutypella is one of the few genera in the Diatrypaceae considered plant pathogens (1). In California, E. vitis and other members of the Diatrypaceae cause branch and trunk canker on grapevine (3,4). Eutypella spp. have not previously been documented as pathogens of citrus. In a 2010 survey on citrus branch canker and dieback in six citrus-growing counties of California, four isolates of Eutypella species were detected in Riverside and San Diego counties. Canker symptoms included dieback and bark cracking, and cuts made through symptomatic trees showed that the cankers were expanding through the center of the tree. Branch samples were collected from 10 trees per orchard and 5 to 10 orchards per county (102 trees for two counties). Pieces of symptomatic tissue (1 to 2 mm2) were plated onto potato dextrose agar amended with 0.01% tetracycline (PDA-tet) and incubated at 25°C for 4 days. All isolates were identified by morphological and molecular characteristics. PCR of isolates was performed in a thermal cycler using two primer pairs, ITS4/5 and Bt2a/2b for amplifying the internal transcribed spacer (ITS1), 5.8S, and ITS2 region and the β-tubulin gene, respectively (2,3). PCR products were sequenced at the University of California, Riverside Genomics Core and the sequences compared in a BLAST search. Four isolates identified as Eutypella spp. included two (UCR1088 and UCR1101) from San Diego County and two (UCR1148 and UCR1149) from the Riverside County samples. The sequences were deposited in GenBank (HQ880579, JF758610, HQ880581, and HQ880582 and HQ880583, JF758611, HQ880585, and HQ880586 for the ITS regions and β-tubulin gene, respectively. ITS sequences for UCR1088 and UCR1101 had 98 and 100% match, respectively, to Eutypella spp. ITS sequences in GenBank (GQ293959 to GQ293961), while UCR1148 and UCR1149 matched 99% (GQ293956 to GQ293958). On the basis of morphological characteristics, UCR1088 and UCR1101 were similar to Eutypella spp. group 1, while UCR1148 and UCR1149 were similar to Eutypella spp. group 3 (4). Pathogenicity tests were conducted with all four isolates on detached shoots from healthy citrus trees of the same cultivar/rootstock from which each isolate was obtained. One wound per shoot was made on 1-year-old, green, detached shoots using a 3-mm-diameter cork borer and the wounded surfaces were inoculated with 3-mm-diameter mycelial plugs of 5-day-old cultures of each isolate growing on PDA-tet. Inoculated wounds and shoot ends were covered with petroleum jelly and wrapped with Parafilm (3). Control shoots were inoculated with sterile agar plugs. There were 10 inoculated shoots per isolate and noninoculated control treatment. Shoots were incubated at 25°C in moist chambers for 6 weeks. Lesions similar to those on the original infected shoots were observed on all inoculated shoots except the control treatment. Reisolation and identification of fungi from inoculated and control shoots were done using methods described above. Inoculated isolates were recovered from 100% of inoculated shoots but none was recovered from noninoculated shoots, indicating association of Eutypella spp. with citrus branch canker. To our knowledge, this is the first report of Eutypella spp. associated with cankers on citrus in California. References: (1) B. Piskur et al. Plant Dis. 91:1579, 2007. (2) B. Slippers et al. Mycologia 96:83, 2004. (3) F. P. Trouillas and W. D. Gubler. Plant Dis. 94:867, 2010. (4) F. P. Trouillas et al. Mycologia 102:319, 2010.

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First Report of Leaf Blight Caused by Nigrospora sphaerica on Curcuma in China

Plant Disease ◽

10.1094/pdis-04-11-0284 ◽

2011 ◽

Vol 95 (9) ◽

pp. 1190-1190

Author(s):

L. X. Zhang ◽

J. H. Song ◽

G. J. Tan ◽

S. S. Li

Keyword(s):

Disease Incidence ◽

Its Region ◽

Leaf Blight ◽

Morphological Data ◽

Future Research ◽

Leaf Margin ◽

Academic Press ◽

First Report ◽

Pathogenicity Tests ◽

Leaf Pathogen

Curcuma (family Zingiberaceae) is commonly cultivated for the use of rhizomes within traditional Chinese medicines. In October 2009 and 2010, severe leaf blight was observed on Curcuma wenyujin Y.H. Chen & C. Ling (4) in fields located in Ruian, China. The area of cultivation in Ruian encompasses 90% of the production in Zhejiang Province. Disease incidence was approximately 90% of plants observed in affected fields. Early symptoms were yellow-to-brown, irregular-shaped lesions on the leaf margin or tip. After several days, lesions expanded along the mid-vein until the entire leaf was destroyed. Blighted leaves turned grayish to dark brown and withered, and severely affected plants died. Eight fungal isolates were recovered from symptomatic C. wenyujin leaves, collected from eight different fields, on potato dextrose agar (PDA). These fungal colonies were initially white, becoming light to dark gray and produced black, spherical to subspherical, single-celled conidia (14 to 17 × 12 to 15 μm), which were borne on a hyaline vesicle at the tip of the conidiophores. On the basis of these morphological features, the isolates appeared to be similar to Nigrospora sphaerica (2). Strain ZJW-1 was selected as a representative for molecular identification. Genomic DNA was extracted from the isolate, and the internal transcribed spacer (ITS) region of the ribosomal DNA (ITS1-5.8S-ITS2) was amplified using ITS1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) primers (3). The ITS region was further cloned and sequenced (GenBank Accession No. JF738028) and was 99% identical to N. sphaerica (GenBank Accession No. FJ478134.1). On the basis of morphological data and the ITS rDNA sequence, the isolate was determined to be N. sphaerica. Pathogenicity tests were conducted on four leaves of four C. wenyujin plants by placing agar pieces (5 mm in diameter) from 8-day-old cultures on pushpin-wounded leaves. An equal number of control plants were wounded and inoculated with noncolonized PDA agar pieces. Plants were placed in moist chambers at 25°C with a 12-h photoperiod. Brown-to-black lesions were observed on wounded leaves after 3 days and expanded to an average of 56 × 40 mm 15 days after inoculation. No symptoms developed on the control leaves. The pathogen was reisolated from the margins of necrotic tissues but not from the controls. The pathogen has been reported as a leaf pathogen on several hosts worldwide (1). To our knowledge, this is the first report of N. sphaerica as a leaf pathogen of C. wenyujin in China. Future research will focus primarily on management of this disease. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory, USDA-ARS, Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , March 31, 2011. (2) E. W. Mason. Trans. Brit. Mycol. Soc. 12:152, 1927. (3) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990. (4) J. Zhao et al. Molecules 15:7547, 2010.

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First Report of Blueberry Leaf Spot Caused by Cylindrocladium colhounii in China

Plant Disease ◽

10.1094/pd-90-1553a ◽

2006 ◽

Vol 90 (12) ◽

pp. 1553-1553 ◽

Cited By ~ 2

Author(s):

Y. S. Luan ◽

L. Feng ◽

L. J. An

Keyword(s):

Leaf Spot ◽

Morphological Traits ◽

Internal Transcribed Spacers ◽

Fluorescent Light ◽

Adaxial Side ◽

Single Spore ◽

Tubulin Gene ◽

First Report ◽

Plastic Bags ◽

Β Tubulin

During late July and early August of 2005, leaf spot symptoms were observed in a blueberry nursery at a plantation in Dalian, which to our knowledge, lies within the largest blueberry-production area in China. Symptoms were observed primarily on lowbush species, for example Blomidon, as well as half-highbush cultivars. A slow-growing, white mycelium from the margin of necrotic leaf spots was recovered on potato dextrose agar (PDA). The following morphological traits were observed: erect conidiophores that branch twice and were terminated in a stiped, clavate phialide; hyaline, cylindrical, four-celled conidia; and globose, reddish brown, aggregated chlamydospores. Conidiophores (including stipes and terminal phialides) were 305 to 420 × 5 to 9 μm; primary branches were 9 to 45 × 5 to 6.3 μm; secondary branches were 9 to 17.3 × 3.1 to 4.5 μm; phialides were 7.8 to 17.5 × 2.5 to 6 μm; stipes (from the highest branch area to vesicle) were 150 to 270 μm long; and vesicles were 13 to 30 × 2 to 4.5 μm. Conidia were 50 to 72 × 4 to 5.5 μm. Chlamydospores were 15 to 20 μm in diameter. Koch's postulates were fulfilled by spray inoculating two healthy cultivars with conidiophores homogenized in axenic water. As a control, two healthy plants were sprayed with axenic water. Plants were placed inside plastic bags to maintain humidity and incubated in a growth chamber at 26°C under fluorescent light for 14 h and 20°C in darkness for 10 h. After 2 days, the plastic bags were removed and plants were maintained under the same conditions. After 4 days, small-to-medium brown spots with purplish margins were observed on the adaxial side of leaves from inoculated plants, but not from control plants. Fungi isolated from these lesions had the same morphological traits as the ones isolated previously from field plants. The morphological descriptions and measurements were similar to Cylindorocladium colhounii (2). The 5.8S subunit and flanking internal transcribed spacers (ITS1 and ITS2) of rDNA and the β-tubulin gene were amplified from DNA extracted from single-spore cultures using the ITS1/ITS4 primers and T1/Bt2b primers, respectively, and sequenced (1). The ITS and β-tubulin gene sequences were similar to C. colhounii STE-U 1237 (99%; GenBank Accession No. AF231953) and C. colhounii STE-U 705 (99%; GenBank Accession No. AF231954), respectively. The morphology, secondary conidiation, and sequences of ITS and β-tubulin gene identify the causal fungus as C. colhounii. To our knowledge, this is the first report of C. colhounii on blueberry in China or in the world. References: (1) P. W. Crous et al. Can. J. Bot. 77:1813, 1999. (2) T. Watanabe. Page 222 in: Dictorial Atlas of Soil and Seed Fungi. CRC Press, Inc., Boca Raton, Fl, 1994.

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First Report of a Pestalotiopsis sp. Causing Leaf Spot of Blueberry in China

Plant Disease ◽

10.1094/pdis-92-1-0171a ◽

2008 ◽

Vol 92 (1) ◽

pp. 171-171 ◽

Cited By ~ 10

Author(s):

Y. S. Luan ◽

Z. T. Shang ◽

Q. Su ◽

L. Feng ◽

L. J. An

Keyword(s):

Leaf Spot ◽

Apical Cell ◽

Vaccinium Corymbosum ◽

Internal Transcribed Spacers ◽

Fluorescent Light ◽

Tubulin Gene ◽

First Report ◽

Plastic Bags ◽

Plant Nursery ◽

Β Tubulin

In August 2006, leaf spots were observed on half-high blueberry (Vaccinium corymbosum) in a plant nursery in Dalian, China. The symptomatic potted 1-year-old blueberry plants were located in parts of a plant nursery with poor ventilation. The primary symptom was a leaf spot, 0.4 to 0.8 cm in diameter, with brown margins that enlarged and coalesced. Mycelium grew from symptomatic and green leaf tissue removed from the margin of a necrotic leaf spot. Plant tissues were surface disinfested with 0.1% mercuric chloride for 3 min and 70% ethyl alcohol for 30 s before plating onto potato dextrose agar. The resulting colonies were white with a regular margin and a rough surface. The cultures were covered with black and globular acervuli with a diameter of 100 to 200 μm. The base of each conidiophore was swollen and globose with phialides growing from the apical end. Mature conidia were straight to fusiform, measuring 19.0 to 27.5 × 6.3 to 9.2 μm, and five-celled with the three middle cells brown and darker than the end cells. The apical cell was triangular and hyaline with three simple setulae that were 17.2 to 29.7 μm long. The base cell terminated in a point 4.0 to 8.6 μm long. Koch's postulates were fulfilled for the fungus by spray inoculating two healthy young plants with 2 × 105 conidia per ml of sterile distilled water. As a control, two similar plants were sprayed with sterile water. Plants were placed inside plastic bags to maintain humidity and incubated in a growth chamber at 26°C under fluorescent light for 14 h and at 20°C in darkness for 10 h. After 3 days, the plastic bags were removed and plants were maintained under the same conditions. More than 20 days after inoculation, symptoms on inoculated plants were similar to those previously described in the nursery. Control plants did not show any symptoms. Cultures isolated from the lesions were similar to those isolated previously from plants in the nursery. The morphological descriptions and measurements were similar to Pestalotiopsis clavispora (1). The 5.8S subunit and flanking internal transcribed spacers (ITS1 and ITS2) of rDNA and partial β-tubulin gene were amplified from DNA extracted from single-spore cultures using the ITS1/ITS4 and T1/Bt2b primers (2) respectively, and sequenced (GenBank Accession Nos. EF119336 and EF152585). The ITS sequences were most similar to the ITS regions of P. clavispora TA-8 (98%; GenBank Accession No. AY924264), P. clavispora TA-6 (98%; GenBank Accession No. AY924263), and P. clavispora PSHI 2002 Endo 389 (96%; GenBank Accession No. AY682929). The partial β-tubulin gene sequence was identical to Pestalotiopsis sp. isolate PSHI 2004 Endo 86 (100%; GenBank Accession No. DQ657901). The morphology and sequence data support the identity of the causal fungus as P. clavispora. To our knowledge, this is the first report on the presence of a Pestalotiopsis sp. causing a disease of blueberry in China. References: (1) E. F. Guba. Monograph of Monochaetia and Pestalotia. Harvard University Press, Cambridge, MA, 1961. (2) W. Tao et al. Mol. Cell Biol. 27:689, 2007.

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Pectobacterium carotovorum subsp. brasiliense Causes Soft Rot and Death of Neobuxbaumia tetetzo in Zapotitlan Salinas Valley, Puebla, Mexico

Plant Disease ◽

10.1094/pdis-02-18-0370-re ◽

2019 ◽

Vol 103 (3) ◽

pp. 398-403 ◽

Cited By ~ 3

Author(s):

Dimas Mejía-Sánchez ◽

Sergio Aranda-Ocampo ◽

Cristian Nava-Díaz ◽

Daniel Teliz-Ortiz ◽

Manuel Livera-Muñoz ◽

...

Keyword(s):

Soft Rot ◽

Genetic Identification ◽

Pectobacterium Carotovorum ◽

Bacterial Isolates ◽

First Report ◽

Whole Plant ◽

Salinas Valley ◽

Pathogenicity Tests ◽

Carotovorum Subsp ◽

Rdna Igs

Neobuxbaumia tetetzo (Coulter) Backeberg (tetecho) is a columnar cactus endemic to Mexico. Tetecho plants, flowers, fruits, and seeds play an important role in the semiarid ecosystem, as they serve as a refuge and food for insects, bats, and birds, and are widely used by ethnic groups since pre-Hispanic times. Tetecho is affected by a soft rot that damages the whole plant and causes its fall and disintegration. Eight bacterial colonies of similar morphology were isolated from plants showing soft rot and inoculated in healthy tetecho plants, reproducing typical symptoms of soft rot 9 days after inoculation. Ten representative isolates were selected for phenotypic and genetic identification using 16s rDNA, IGS 16S-23S rDNA, and rpoS genes and for pathogenicity tests on several members of the cactus family and other plants. Based on the results, these bacterial isolates were identified as Pectobacterium carotovorum subsp. brasiliense. Inoculation of this bacteria caused soft rot in different cacti, fruits, leaves, and roots of other plants. This is the first report of the subspecies brasiliense of P. carotovorum causing soft rot and death in cacti in the world and the first report of this subspecies in Mexico.

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First Report of Soft Rot on Dendrobium officinale caused by Epicoccum sorghinum in China

Plant Disease ◽

10.1094/pdis-04-21-0866-pdn ◽

2021 ◽

Author(s):

Caiyun Xiao ◽

Rongyu Li ◽

Xingchen Song ◽

Xujun Tian ◽

Qijun Zhao

Keyword(s):

Internal Transcribed Spacer ◽

Elongation Factor ◽

Morphological Characteristics ◽

Reference Value ◽

Soft Rot ◽

Dendrobium Officinale ◽

First Report ◽

Rdna Its ◽

And Control ◽

Internal Transcribed Spacer Rdna

In recent years, soft rot is one of the most serious diseases in the production of Dendrobium officinale. In this study, we took the diseased plants of Dendrobium officinale in Guizhou as samples, through Koch's rule and sequence analysis of rDNA internal transcribed spacer (rDNA-ITS), calmodulin (cmdA), the second largest subunit of RNA polymerase Ⅱ (RPB2), elongation factor EF-1 α and β-tubulin (β-Tub), it was determined that the pathogen of Dendrobium officinale soft rot was sorghum accessory cocci. This is our first report on the soft rot of Dendrobium officinale caused by Epicoccum sorghinum in China. The morphological characteristics of the pathogen shown in the study will have a certain reference value for the prevention and control of the soft rot of Dendrobium officinale in the future.

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