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

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 Root and Stem Rot Caused by Phytophthora nicotianae on Peperomia tetraphylla in China

Plant Disease ◽

10.1094/pdis-94-9-1171c ◽

2010 ◽

Vol 94 (9) ◽

pp. 1171-1171 ◽

Cited By ~ 1

Author(s):

D. X. Zeng ◽

X. L. Wu ◽

Y. H. Li

Keyword(s):

Southern China ◽

Aerial Mycelium ◽

Its Region ◽

Soil Extract ◽

Phytophthora Nicotianae ◽

Stem Rot ◽

Molecular Characteristics ◽

Its Sequence ◽

First Report ◽

Pure Cultures

Peperomia tetraphylla, an evergreen herb, is becoming increasingly popular as a potted ornamental plant in southern China. In the summer of 2008, in some commercial flower nurseries in Shenzhen, Guangdong Province, P. tetraphylla showed extensive black stem and root rot, with leaves dropping from the rotten stem. Small pieces (approximately 3 mm2) of stems and leaves were excised from the margins of the black lesions, surface disinfected for 30 s to 1 min in 0.1% HgCl2, plated onto potato dextrose agar (PDA), and incubated at 25°C in the dark. All the plated samples yielded Phytophthora, and microscopic examination of pure cultures grown on PDA plates showed arachnoid colonies with abundant aerial mycelium, chlamydospores, and a few sporangia. Numerous sporangia were formed in sterile soil extract. Sporangia were ovoid or obpyriform, noncaducous, with prominent solitary papillae, and measured 31 to 52 μm (average 38 μm) × 21 to 34 μm (average 27 μm). Chlamydospores were spherical and 21 to 34 μm in diameter (average 28 μm). The internal transcribed spacer (ITS) region of rDNA of a single isolate was amplified using primers ITS4/ITS5 and sequenced (2). The ITS sequence, when submitted for a BLAST search in the NCBI database, showed 100% homology with the sequences of two reference isolates of Phytophthora nicotianae (Accession Nos. AY833526 and EU433396) and the consensus ITS sequence was deposited in the NCBI as Accession No. GQ499373. The isolate was identified as Phytophthora nicotianae on the basis of morphological and molecular characteristics (1). Pathogenicity of the isolate was confirmed by inoculating 1-year-old plants of P. tetraphylla growing in pots. The isolate was grown for 7 days on PDA plates and mycelial plugs, 5 mm in diameter and taken from the advancing margins of the colonies, were buried approximately 1 cm deep near the base of the stem in such a way that the mycelium on the plugs was in contact with the surface of the stem, which had been wiped earlier with 70% ethanol and gently wounded with a needle. Plants treated the same way but inoculated with sterile PDA plugs served as control plants. Three plants in each pot were inoculated and there were five replications each for the treatment and the control. All plants were kept in a greenhouse at 22 to 32°C. After 6 to 7 days, the inoculated plants showed black lesions around the mycelial plugs; symptoms of root and stem rot developed rapidly thereafter and the plants collapsed within 2 weeks. All symptoms on the inoculated plants were identical to those observed in naturally diseased plants, whereas the control plants remained healthy. The same fungus was consistently reisolated from the inoculated plants. To our knowledge, this is the first report of Phytophthora nicotianae on P. tetraphylla in China. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (2) J. B. Ristaino et al. Appl. Environ. Microbiol. 64:948, 1998.

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First Report of Stem and Leaf Anthracnose on Blueberry Caused by Colletotrichum gloeosporioides in China

Plant Disease ◽

10.1094/pdis-11-12-1056-pdn ◽

2013 ◽

Vol 97 (6) ◽

pp. 845-845 ◽

Cited By ~ 1

Author(s):

C. N. Xu ◽

Z. S. Zhou ◽

Y. X. Wu ◽

F. M. Chi ◽

Z. R. Ji ◽

...

Keyword(s):

Colletotrichum Gloeosporioides ◽

Morphological Characteristics ◽

Its Region ◽

Pathogenic Fungi ◽

Molecular Characteristics ◽

First Report ◽

Potted Plants ◽

Leaf Spots ◽

Pathogenicity Tests ◽

Highbush Blueberries

Blueberry (Vaccinium spp.) is becoming increasingly popular in China as a nutritional berry crop. With the expansion of blueberry production, many diseases have become widespread in different regions of China. In August of 2012, stem and leaf spots symptomatic of anthracnose were sporadically observed on highbush blueberries in a field located in Liaoning, China, where approximately 15% of plants were diseased. Symptoms first appeared as yellow to reddish, irregularly-shaped lesions on leaves and stems. The lesions then expanded, becoming dark brown in the center and surrounded by a reddish halo. Leaf and stem tissues (5 × 5 mm) were cut from the lesion margins and surface-disinfected in 70% ethanol for 30 s, followed by three rinses with sterile water before placing on potato dextrose agar (PDA). Plates were incubated at 28°C. Colonies were initially white, becoming grayish-white to gray with yellow spore masses. Conidia were one-celled, hyaline, and cylindrical with rounded ends, measuring 15.0 to 25.0 × 4.0 to 7.5 μm. No teleomorph was observed. The fungus was tentatively identified as Colletotrichum gloeosporioides (PenZ.) PenZ & Sacc. (teleomorph Glomerella cingulata (Stoneman) Spauld. & H. Schrenk) based on morphological characteristics of the colony and conidia (1). Genomic DNA was extracted from isolate XCG1 and the internal transcribed spacer (ITS) region of the ribosomal DNA (ITS1–5.8S-ITS2) was amplified with primer pairs ITS1 and ITS4. BLAST searches showed 99% identity with C. gloeosporioides isolates in GenBank (Accession No. AF272779). The sequence of isolate XCG1 (C. gloeosporioides) was deposited into GenBank (JX878503). Pathogenicity tests were conducted on 2-year-old potted blueberries, cv. Berkeley. Stems and leaves of 10 potted blueberry plants were wounded with a sterilized needle and sprayed with a suspension of 105 conidia per ml of sterilized water. Five healthy potted plants were inoculated with sterilized water as control. Dark brown lesions surrounded by reddish halos developed on all inoculated leaves and stems after 7 days, and the pathogen was reisolated from lesions of 50% of inoculated plants as described above. The colony and conidial morphology were identical to the original isolate XCG1. No symptoms developed on the control plants. The causal agent of anthracnose on blueberry was identified as C. gloeosporioides on the basis of morphological and molecular characteristics, and its pathogenicity was confirmed with Koch's postulates. Worldwide, it has been reported that blueberry anthracnose might be caused by C. acutatum and C. gloeosporioides (2). However, we did not isolate C. acutatum during this study. To our knowledge, this is the first report of stem and leaf anthracnose of blueberry caused by C. gloeosporioides in China. References: (1) J. M. E. Mourde. No 315. CMI Descriptions of Pathogenic Fungi and Bacteria. Kew, Surrey, UK, 1971. (2) N. Verma, et al. Plant Pathol. 55:442, 2006.

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First Report of Stem Rot of Konjac Caused by Phytophthora nicotianae in China

Plant Disease ◽

10.1094/pdis-12-13-1204-pdn ◽

2015 ◽

Vol 99 (2) ◽

pp. 283-283

Author(s):

M. Shao ◽

W. F. Du ◽

D. C. Yu ◽

P. Du ◽

S. J. Ni ◽

...

Keyword(s):

Dna Analysis ◽

Morphological Characteristics ◽

Its Region ◽

Phytophthora Nicotianae ◽

Nuclear Ribosomal Dna ◽

Pathogenicity Test ◽

First Report ◽

Color Changes ◽

Health Products ◽

Light Green

Konjac (Amorphophallus) is an important economic crop widely used in health products and biomaterials in Asia (2). A serious foliage disease of Konjac was observed in Fuyuan County, Yunnan Province, China, in July 2012. The symptoms began with leaf color changes from light green to yellow, followed by discoloration on the stem base, plant wilting, bulb rotting, and ultimately plant death. Symptomatic tissues were cut into small pieces, surface-sterilized, and cultured on 20% V8 juice agar at 28°C. Five days after incubation, white fluffy colonies with the typical sporangium of Phytophthora sp. were observed from root and stem pieces. Isolates were identified as P. nicotianae based on morphological characteristics and DNA analysis. The growth rate of the colonies was 16 mm/d at 28°C. Sporangia were pyriform, ovoid to spherical, and papillate, and the dimensions of the 80 sporangia measured ranged from 23.7 to 60.4 × 19.4 to 45.7 μm (avg. 42.4 × 31.5 μm) with length-to-breadth ratios of 1.19 to 1.44 (avg. 1.34). The chlamydospores were spherical with a smooth surface, and their dimensions ranged from 20.3 to 47.3 × 18.9 to 45.9 μm (avg. 32.7 × 30.4 μm) (3). DNA was extracted from one colony containing spores and hyphae of the isolated P. nicotianae, and the nuclear ribosomal DNA internal transcribed spacer (ITS) region was amplified with primers ITS6 and ITS4 (4). The obtained 854-bp amplicon was purified and sequenced. NCBI BLAST retrieved a 100% identity with P. nicotianae (GenBank Accession No. KJ506732). A pathogenicity test of the isolated P. nicotianae was conducted in a greenhouse. After 7 days in a humidity-controlled greenhouse, all 10 inoculated plants showed similar symptoms as observed initially in the field, while control plants were symptomless. P. nicotianae was re-isolated from the inoculated stems, thus successfully completing Koch's postulates (1). To our knowledge, this is the first report of P. nicotianae as a pathogen of Konjac in China. References: (1) B. Alvarez-Rodriguez et al. Plant Dis. 97:1257, 2013. (2) H. Ban, et al. Plant Cell Rep. 28:1847, 2009. (3) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. APS Press, St. Paul, MN, 1996. (4) J. M. French et al. Plant Dis. 95:1028, 2011.

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First Report of Black Rot Caused by Phomopsis cucurbitae on Cantaloupe (Cucumis melo) in the Piedmont Region of Northern Italy

Plant Disease ◽

10.1094/pdis-06-11-0481 ◽

2011 ◽

Vol 95 (10) ◽

pp. 1317-1317

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

Cucumis Melo ◽

Morphological Characteristics ◽

Its Region ◽

Northern Italy ◽

Pathogenicity Test ◽

First Report ◽

Sterile Needle ◽

Pathogenicity Tests ◽

Cucumis Melo L ◽

Piedmont Region

Cucumis melo L., belonging to the Cucurbitaceae family, is cultivated on more than 23,000 ha in Italy. Cantaloupe (C. melo L. var. cantalupensis Naudin) is the most popular variety. In summer 2010, a previously unknown rot was observed on fruits produced in Italy and marketed in the Piedmont Region of northern Italy. Early symptoms on fruit consisted of irregular, brown, soft, sunken lesions up to 10 cm in diameter. No surface mold was visible and pycnidia were not present. Internally, the decay is adjacent to the sunken area of the fruit's surface and is soft, water soaked, spongy, with a nearly circular margin, and easily separated from healthy tissues. Fragments (approximately 3 mm3) were taken from the margin of the internal diseased tissues, cultured on potato dextrose agar (PDA), and incubated at 24 ± 1°C, (16 h of light and 8 h of darkness). Fungal colonies initially appeared coarse, at first whitish then buff brown, and produced dark pycnidia 0.5 mm in diameter, which exuded numerous conidia belonging to two types. Type A conidia were hyaline, unicellular, ellipsoidal to fusiform, sometimes slightly constricted in the middle, and measured 5.6 to 10.3 × 1.7 to 2.6 (average 8.0 × 2.1) μm. Type B conidia were hyaline, long, slender, curved, and measured 17.1 to 26.6 × 0.7 to 1.4 (average 22.0 × 1.0) μm. Sclerotia were not produced. The morphological characteristics of the fungus corresponded to those of the genus Phomopsis (1). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 and sequenced. BLAST analysis of the 543-bp segment showed a 99% similarity with the sequence of a Phomopsis sp. (GenBank Accession No. HM999947). The nucleotide sequence has been assigned the GenBank Accession No. JN032733. Both Phomopsis cucurbitae and P. sclerotioides are pathogenic to Cucurbitaceae, however P. cucurbitae is identifiable by the production of B conidia and the absence of sclerotia. Therefore, P. cucurbitae has been considered the causal agent of the disease. Pathogenicity tests were performed by inoculating three wounded cantaloupe fruits after surface disinfesting in 1% sodium hypochlorite. Six wounds per fruit, 1 cm deep, were made with a sterile needle. Mycelial disks (10 mm in diameter), obtained from PDA cultures of one strain, were placed on each wound. Three control fruits were inoculated with PDA. Fruits were incubated at 16 ± 1°C in the dark. The first symptoms developed 4 days after the artificial inoculation. Two days later, the rot developed at all inoculation points and the pathogen was consistently reisolated. Noninoculated fruit remained healthy. The pathogenicity test was performed twice with similar results. P. cucurbitae has been reported on melon in many countries (2,3). To our knowledge, this is the first report of the disease in Italy. Currently, the relevance of the disease in the country is not yet well known. However, attention must be paid considering that the pathogen can be transmitted through seeds. References: (1) H. L. Barnett and B. B. Hunter. Illustrated Genera of Imperfect Fungi. Burgess Publishing Company, Minneapolis, MN, 1972. (2) L. Beraha and M. J. O'Brien. Phytopathol. Z. 94:199, 1979. (3) E. Punithalingam and P. Holliday. Phomopsis cucurbitae. IMI Descriptions of Fungi and Bacteria. 47, Sheet 469, 1975.

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First Report of Powdery Mildew Caused by Golovinomyces sonchicola on Ixeris chinensis in Korea

Plant Disease ◽

10.1094/pdis-11-13-1132-pdn ◽

2014 ◽

Vol 98 (7) ◽

pp. 999-999 ◽

Cited By ~ 2

Author(s):

J. K. Choi ◽

B. S. Kim ◽

S. H. Hong ◽

S. E. Cho ◽

H. D. Shin

Keyword(s):

Powdery Mildew ◽

Sequence Similarity ◽

Folk Medicine ◽

Morphological Characteristics ◽

Its Region ◽

Width Ratio ◽

Lateral Part ◽

Potential Threat ◽

First Report ◽

Powdery Mildews

Ixeris chinensis (Thunb.) Nakai, known as Chinese ixeris, is distributed from Siberia to Japan, including Korea, Taiwan, and China. The whole plant has been used in folk medicine in Asia (4). In Korea, the plants of Chinese ixeris have been gathered and used as a wild root vegetable. During summer to autumn of 2011, Chinese ixeris leaves were found to be heavily infected with a powdery mildew in several locations of Korea. Symptoms first appeared as thin white colonies, which subsequently developed into abundant hyphal growth on both sides of the leaves, leading to drying of the leaves. The same symptoms on Chinese ixeris leaves were continuously observed in 2012 and 2013. Voucher specimens (n = 10) were deposited at Korea University Herbarium (KUS). Hyphal appressoria were moderately lobed or nipple-shaped. Conidiophores arose from the lateral part of the hyphae, measured 100 to 270 × 10 to 12.5 μm, and produced 2 to 6 immature conidia in chains with a sinuate outline. Basal parts of foot-cells in conidiophores were curved. Conidia were barrel-shaped to ellipsoid, measured 26 to 36 × 13 to 19 μm (length/width ratio = 1.7 to 2.4), lacked fibrosin bodies, and showed reticulate wrinkling of the outer walls. Primary conidia were ovate with conical-obtuse apex and subtruncate base. Germ tubes were produced on the perihilar position of conidia. Chasmothecia were not observed. The morphological characteristics were typical of the Euoidium type anamorph of the genus Golovinomyces, and the fungus measurements and structures were consistent with those of G. sonchicola U. Braun & R.T.A. Cook (1). To confirm the identification, internal transcribed spacer (ITS) region of rDNA sequences from a representative material (KUS-F26212) was amplified using primers ITS5/P3 and sequenced (3). The resulting 416-bp sequence was deposited in GenBank (Accession No. KF819857). A GenBank BLAST search revealed that the isolate showed >99% sequence similarity with those of G. cichoracearum from Sonchus spp. (e.g., AB453762, AF011296, JQ010848, etc.). G. sonchicola is currently confined to G. cichoracearum s. lat. on Sonchus spp., based on molecular sequence analyses (1). Pathogenicity was confirmed through inoculation by gently pressing a diseased leaf onto leaves of five healthy potted Chinese ixeris. Five non-inoculated plants served as controls. Inoculated plants developed symptoms after 6 days, whereas the controls remained symptomless. The fungus present on the inoculated plants was identical morphologically to that originally observed on diseased plants. Powdery mildew infections of I. chinensis associated with Golovinomyces have been known in China (2). To our knowledge, this is the first report of powdery mildew disease caused by G. sonchicola on I. chinensis in Korea. Farming of Chinese ixeris has recently started on a commercial scale in Korea. Though no statistical data are available, we postulate the cultivation area in Korea to be approximately 200 ha, mostly growing without chemical controls. Occurrence of powdery mildews poses a potential threat to safe production of this vegetable, especially in organic farming. References: (1) U. Braun and R. T. A. Cook. Taxonomic Manual of the Erysiphales (Powdery Mildews), CBS Biodiversity Series No.11. CBS, Utrecht, 2012. (2) F. L. Tai. Bull. Chinese Bot. Sci. 2:16, 1936. (3) S. Takamatsu et al. Mycol. Res. 113:117, 2009. (4) S. J. Zhang et al. J. Nat. Prod. 69:1425, 2006.

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First Confirmed Report of Cercospora Blight of Asparagus Caused by Cercospora asparagi in Korea

Plant Disease ◽

10.1094/pdis-10-12-0984-pdn ◽

2013 ◽

Vol 97 (3) ◽

pp. 428-428 ◽

Cited By ~ 4

Author(s):

K. S. Han ◽

B. S. Kim ◽

J. H. Park ◽

H. D. Shin

Keyword(s):

Morphological Characteristics ◽

Its Region ◽

Culture Collection ◽

Asparagus Officinalis ◽

Organic Farm ◽

Plastic Bags ◽

Japanese Workers ◽

Close Phylogenetic Relationship ◽

Voucher Specimens ◽

Pathogenicity Tests

In September 2011, hundreds of asparagus (Asparagus officinalis L.) showing symptoms of blight with nearly 100% incidence (percentage of plants affected) were found in polyethylene tunnels at an organic farm in Gangneung City of Korea. Lesions on needles and branches of the ferns were small, elliptic to subcircular, pale tan to brown with reddish brown borders, and became gray and cottony due to heavy sporulation under continuous high humidity. Infection caused premature defoliation and weakened plant vigor. The damage purportedly due to this disease has reappeared with confirmation of the causal agent made again in 2012. A cercosporoid fungus was consistently associated with disease symptoms. Stromata were well developed, consisting of brown cells, and were 10 to 30 μm wide. Conidiophores were fasciculate (n = 2 to 12), olivaceous brown, paler upwards, straight to mildly curved, not geniculate in shorter ones, or 1 to 2 times geniculate in longer ones, 40 to 260 μm long, 3.5 to 5.5 μm wide, and 1- to 6-septate. Conidia were hyaline, cylindric to acicular, straight in shorter ones, curved in longer ones, truncate to obconically truncate at the base with darkened hila, guttulate, 2- to 12-septate, and 40 to 220 × 3 to 5 μm. Morphological characteristics of the fungus were consistent with the previous reports of Cercospora asparagi Sacc. (1). Voucher specimens were housed at the Korea University herbarium (KUS). An isolate from KUS-F26046 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC46400). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequence of 497 bp was deposited in GenBank (Accession No. JX964995). This showed >99% similarity with sequences of many Cercospora species, indicating their close phylogenetic relationship. For pathogenicity tests, conidial suspensions (105 conidia/ml) were prepared by culturing the fungus on V8 juice agar (2) for 3 weeks. Five plants were inoculated with conidial suspensions and five plants were sprayed with sterile distilled water. The plants were covered with plastic bags to maintain 100% RH for 24 h and then transferred to a greenhouse. Typical symptoms of necrotic lesions appeared on the inoculated plants 6 days after treatment, and were identical to the ones observed in the field. C. asparagi was reisolated from symptomatic tissues, confirming Koch's postulates. No symptoms were observed on control plants. The disease has been reported through the regions of the world where asparagus is grown (3). In Korea, the disease was recorded in 1928 by Japanese workers under Cercosporina asparagicola Speg. (regarded as synonymous with Cercospora asparagi) with brief notes (4). Though one sample of asparagus was sent to the author (KSH) for diagnosis in 2009 summer and determined to be infected with C. asparagi (unpublished data), there has been no additional finding of the disease in Korea for the last 82 years. To our knowledge, this is the first confirmed report of Cercospora blight of asparagus caused by C. asparagi in Korea. References: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Ithaca, NY, 1953. (2) C. J. Cooperman and S. F. Jenkins. Phytopathology 76:617, 1986. (3) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, Retrieved October 20, 2012. (4) K. Nakata and S. Takimoto. Bull. Agric. Exp. Stat. Korea 15:1, 1928.

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First Report of Cherry Stem Rot and Leaf Necrosis Disease Caused by Phytophthora nicotianae in Yantai, China

Plant Disease ◽

10.1094/pdis-05-14-0489-pdn ◽

2015 ◽

Vol 99 (2) ◽

pp. 284-284

Author(s):

X. L. Yu ◽

X. Q. Liu ◽

P. S. Wang ◽

Y. Z. Wang

Keyword(s):

Average Length ◽

Morphological Characteristics ◽

Its Region ◽

Phytophthora Species ◽

Phytophthora Nicotianae ◽

Control Measures ◽

Stem Rot ◽

Economic Losses ◽

First Report ◽

Leaf Necrosis

Cherry (Cerasus avium (Linn.) Moench) is the third most economically important fruit in Yantai, Shandong Province, China. In August 2012, brown spots or necrosis on cherry seedling leaves, with an incidence of 8.2 to 34.3%, were observed in some fields of cherry seedlings in Yantai. Our survey indicated that the economic losses could reach up to 15.3% if disease conditions, such as a cool rainy summer season, were favorable. Conspicuous watery lesions on the stems turned to brown streaks; the leaves all wilted; and finally the plants collapsed. Diseased stem and leaf samples were surface-disinfected in 1% sodium hypochlorite for 1 min, rinsed three times in sterile water, which was absorbed with filter paper, and then transferred to 10% V8 juice agar medium containing 50 μg/ml ampicillin and 5 μg/ml carbendazim (1). The plates were incubated at 22°C in the dark for 5 days. The colonies consisted of white, loose, fluffy aerial mycelia. Eight isolates were obtained, and all were identified as Phytophthora nicotianae based on morphological characteristics and the sequence of the internal transcribed spacer (ITS) region of rDNA. The sporangia were ovoid/spherical, obturbinate with rounded bases and prominent papillae that were 37.5 to 62.5 × 30 to 50 μm (average 46.4 × 37.8 μm, n = 100) in size, with an average length-to-breadth ratio of 1.2. Chlamydospores were terminal, intercalary, and measured 19 to 42 μm (average 30.4 μm), which is typical of P. nicotianae (2). The genomic DNA of the eight isolates was extracted from mycelia. The ITS region of all eight isolates was amplified using primers ITS1 and ITS4, producing specific products that were directly sequenced. The sequence of a representative isolate P1401 (895 bp) was submitted to GenBank (Accession No. KJ754387). It was 100% similar to P. nicotianae strains NV-20T and TARI 22073 (KC768775 and GU111667). To confirm the pathogenicity, at least 10 cherry leaves and new stems were inoculated with mycelial plugs (5 × 5 mm) from each isolate. Necrosis of leaves and stems was observed 4 and 7 days after inoculation, respectively. No symptoms were observed on the control leaves and stems that were inoculated with blank agar plugs. P. nicotianae was re-isolated from the infected leaves, and the ITS sequence was analyzed to confirm its identity. Phytophthora species, such as P. cambivora, P. megasperma, and P. drechsleri, had been previously isolated from cherry (3), but to the best of our knowledge this is the first report of stem rot and leaf necrosis disease caused by P. nicotianae on cherry. Since the economic loss caused by this disease could reach 15% if an outbreak occurred in a rainy summer, control measures should be implemented. References: (1) Y. Balci et al. Mycol. Res. 112:906, 2008. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St Paul, MN, 1996. (2) S. M. Mircetich and M. E. Matheron. Phytopathology 66:549, 1976.

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First Report of Pineapple Heart Rot Caused by Phytophthora nicotianae in Hainan Province, China

Plant Disease ◽

10.1094/pdis-11-12-1017-pdn ◽

2013 ◽

Vol 97 (4) ◽

pp. 560-560 ◽

Cited By ~ 4

Author(s):

H. F. Shen ◽

B. R. Lin ◽

J. X. Zhan ◽

X. M. Pu

Keyword(s):

Morphological Characteristics ◽

Its Region ◽

Soft Rot ◽

Phytophthora Nicotianae ◽

Ananas Comosus ◽

Hainan Province ◽

Sterile Water ◽

Tropical Fruit ◽

First Report ◽

Research Technology

Pineapple (Ananas comosus) is an economically important tropical fruit in Hainan Province, China. During September to November 2011, heart rot disese of pineapple was found in Ledong and Wangning of Hainan Province. A survey of 150 ha producing areas of pineapple revealed that the fields were affected at an incidence ranging from 25% to 30%. Infected plants showed water-soaked lesions and soft rot on the base of heart leaves near the soil surface. Heart leaves of infected plants were easily pulled out. As the disease progressed, plants collapsed and died. Diseased tissue fragments (2 × 2 mm) were surface-disinfected for 10 min with 0.3% NaClO, then rinsed three times in sterile water, and plated to 10% V8 juice agar (4). Inoculated dishes were incubated at 26°C in the dark. After 5 days, Phytophthora (identified by the presence of coenocytic hyphae and papillate sporangia) were isolated from the tissue cultures, which has aseptate hyphae. Sporangia were papillate, noncaducous, oval or spherical, and 34.5 to 58.2 μm. Clamydospores, both terminal and intercalary, were also spherical, and were 23.4 to 34.0 μm (2). The ITS region of rDNA was amplified using primers ITS4/ITS5, and the 927-bp product of the ITS showed 99% sequence identity to Phytophthora nicotianae (GenBank Accession No. JF792540), and the sequence was accessed to NCBI (JX978446). Pathogenicity tests were confirmed by irrigating the wounded stem bases of 10 2-month-old pineapple plants with 50 ml of P. nicotionae zoospore solution (15,000 zoospores/ml), and another 10 plants of the same cultivar inoculated with sterile water served as controls. Plants were placed in pots in a greenhouse at 28°C and 90% relative humidity. After 9 days, soft rot was observed clearly on the base of heart leaves of all 10 inoculated plants, while the control plants appeared normal. P. nicotianae was reisolated from the infected pineapple plants, and confirmed to be the same as the inoculated pathogen by conducting a ITS rDNA sequence comparison and morphological characteristics. P. nicotianae was previously reported as the causal agent of heart rot of pineapple in Hawaii, U.S.A. (3) and Guangdong Province of China (1). To our knowledge, this is the first report of P. nicotianae on pineapple in Hainan Province, China. References: (1) J. Z. Chen et al. J. Yunnan Agric. Univ. 8:134, 2003. (2) H. H. Ho. Mycologia 73:705, 1981.(3) K. W. Howard et al. Plant Dis. Rep. 48:848, 1964. (4) X. B. Zheng. Page 81 in: Phytophthora and its Research Technology. China Agricultural Press, Beijing, 1997.

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Detection of a Phytophthora sp. Causing Asparagus Spear and Root Rot in Michigan

Plant Disease ◽

10.1094/pd-89-1011b ◽

2005 ◽

Vol 89 (9) ◽

pp. 1011-1011 ◽

Cited By ~ 1

Author(s):

C. Saude ◽

M. K. Hausbeck ◽

O. Hurtado-Gonzales ◽

K. H. Lamour

Keyword(s):

Tap Water ◽

Sequence Similarity ◽

Morphological Characteristics ◽

Its Region ◽

Asparagus Officinalis ◽

Storage Roots ◽

Phytophthora Spp ◽

Mycelial Plug ◽

Infected Material

In the spring of 2004, a Phytophthora sp. was isolated from asparagus (Asparagus officinalis) spears, roots, and dormant crowns from several fields in Oceana and Ingham counties in Michigan. Symptomatic spears were often curved, had water-soaked lesions slightly above or below the soil line or were shriveled at the site of infection or both. Infected storage roots had water-soaked lesions but were not soft at the lesion site. Infected crowns had fewer roots than healthy crowns. In the laboratory, plant tissues were rinsed in tap water and blotted dry. Sections from the edge of lesions were placed aseptically onto BARP (25 ppm of benomyl, 100 ppm of ampicillin, 30 ppm of rifampicin, and 100 ppm of pentachloroni-trobenzene) amended unclarified V8 juice agar and incubated at 25°C for up to 7 days. Phytophthora sp. isolates recovered from the infected material produced ovoid, nonpapillate, noncaducous sporangia and amphigy-nous oospores on isolation media. Single-sporangium cultures made for each isolate were stored long term in sterile 2-ml microcentrifuge tubes containing two 7-mm mycelial plugs, two sterile hemp seeds, and 1 ml of sterile distilled water. Sporangia produced on dilute V8 juice agar averaged 45 μm long × 26 μm wide and oospores were 25 to 30 μm in diameter. Chlamydospores were not observed. Five detached ‘Jersey Knight’ spears were inoculated with a 7-mm mycelial plug from the edge of actively growing 5-day-old cultures and incubated at 23 to 25°C for 5 to 7 days in a moist chamber. After 3 days, water-soaked lesions and shriveling and curving of the spears were visible on all inoculated spears. The pathogen was always reisolated from the lesion edge. No symptoms were observed when spears were inoculated with sterile V8 juice agar plugs. DNA was extracted from representative isolates, and the nuclear ribosomal internal transcribed spacer (ITS) region was amplified with ITS6 and ITS4 primers and sequenced. A BLAST search of the NCBI database with the ITS sequence revealed Phytophthora sp. UQ2141, Accession No. AF266795, as the closest match with 99% sequence similarity. These results, coupled with the morphological characteristics of the isolates, indicate that the Phytophthora sp. isolated from asparagus in Michigan is among the constituents of Phytophthora spp. included in the P. megasperma clade 6 (2), whose taxa are currently being reevaluated. Although a Phytophthora sp. has been described previously on asparagus (1,3), this is the first report, to our knowledge, of a Phytophthora sp. on asparagus in Michigan. The occurrence of excessive rainfall in the spring of 2004 is likely responsible for widespread disease and considerable yield losses in production fields. References: (1) P. A. Ark and J. T. Barrett. Phytopathology 28:754, 1938. (2) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (3) V. Vujanovic et al. Plant Dis. 87:447, 2003.

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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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