scholarly journals Pumpkin Fruit Rot in North Carolina Caused by Phytophthora nicotianae

Plant Disease ◽  
2000 ◽  
Vol 84 (8) ◽  
pp. 923-923
Author(s):  
G. J. Holmes

In 1999, during an evaluation of pumpkin (Cucurbita pepo) fruit for susceptibility to naturally occurring postharvest diseases, a soft rot of unknown etiology was noted. No fungal growth or sporulation was seen on the fruit surface and no root or crown rot was observed in the field. When fruit were cross-sectioned, masses of white, floccose mycelium covering large sections of the seed cavity were observed. Rot was observed in 21 fruit (6.4% of the total). The fungus was isolated from symptomatic fruit on a modified P10ARPH agar medium, semi-selective for Phytophthora spp. (2). Isolates from eight fruit formed papillate, ovoid sporangia, abundant chlamydospores, and colonies characteristic of P. nicotianae (1). No oospores were produced. Four sound pumpkin fruit (cv. Early Autumn) were inoculated with four isolates (one isolate per fruit). Each isolate was recovered from a different fruit. Pumpkins were surface sterilized at the point of inoculation by wetting with 70% ethanol. Inoculation was done by removing a small amount of mycelium from pure culture using a sterile, wooden toothpick and inserting it 2 cm deep into opposite sides of the mid section of sound fruit (two inoculations per fruit). Control fruit were punctured with sterile toothpicks (once per fruit). First symptoms appeared 4 days after inoculation at room temperature (22 to 24°C). Symptoms consisted of circular, water-soaked areas originating from the point of inoculation. Average diameter (based on four measurements on two fruit) of the water-soaked lesions were 3 cm at first appearance (i.e., 4 days) and 11 cm 10 days after inoculation. No symptoms developed on controls. When symptomatic fruit were cross-sectioned, masses of white, floccose mycelium were noted. Reisolation of this mycelium onto selective medium yielded P. nicotianae, thus fulfilling Koch's postulates. This is the first report of P. nicotianae causing fruit rot of pumpkins. References: (1) D. C. Erwin and O. K. Ribeiro. 1996. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN. (2) H. D. Shew. Phytopathology 77:1090, 1987.

Plant Disease ◽  
1998 ◽  
Vol 82 (6) ◽  
pp. 712-712 ◽  
Author(s):  
P. D. Roberts ◽  
E. Trujillo

Papaya trees (Carica papaya) were observed in 1997 with symptoms of acute chlorosis and wilting of foliage, circular whitish lesions with necrotic centers on fruits, and root and trunk rots that killed the diseased trees. Identical symptoms were observed on many trees in fields interplanted with other crops and in home gardens on two of the five islands of American Samoa. A Phytophthora sp. was consistently isolated on water agar from symptomatic stems, fruits, and roots. The fungus was grown in pure culture on V8 juice medium at 25°C under continuous fluorescent illumination, and was identified, on the basis of spherical to broadly ovoid (34 to 67 × 40 to 50 µm), intercalary to terminal sporangia, chlamydospores (20 × 40 µm), and uniform to uneven hyphae (5 to 7 µm wide), as Phytophthora nicotianae Breda de Haan (= P. parasitica Dastur) (1,2). Ten 4-week old papaya seedlings grown on pasteurized soil in 15-cm pots were inoculated with a 2-ml suspension of 100 sporangia per ml in sterile water applied to the soil surface. Six uninoculated plants were used as controls. Two mature fruit that had reached the climacteric point were inoculated with 200 µl of sterile water containing approximately 20 sporangia. Inoculum was placed inside a 1-cm-diameter rubber circle that had been attached to the surface of the fruit to prevent run-off. Two uninoculated fruits served as controls. Inoculations of seedlings and fruit were conducted three times. Wilting symptoms developed on inoculated plants in less than 5 days and plant death accompanied by crown rot occurred in 14 days. Inoculated fruit developed lesions covered with a whitish fungal growth within 10 days after inoculation. Plants and fruits exposed to sterile water were healthy. The pathogen was reisolated from the diseased plants and fruits and it was morphologically identical to the original isolate, confirming its role as the causative agent of the disease. The wide distribution of diseased plants over an estimated one-quarter of the area of the island of Tutuila (approximately 24 sq. km) and on the adjacent island of Ofu indicates that the disease was not of recent introduction. References: (1) H. H. Hob et al. The genus Phytophthora in Taiwan. Inst. Bot., Ac. Sinica, Monogr. Ser. 15., 1995. (2) G. M. Waterhouse and J. M. Waterhouse. C.M.I. Descriptions of Pathogenic Fungi and Bacteria No. 34, 1964.


Author(s):  
G. M. Waterhouse

Abstract A description is provided for Phytophthora nicotianae var. parasitica. Information is included on the disease caused by the organism, its transmission, geographical distribution, and hosts. HOSTS: On a very wide range of host plants comprising 58 families including: avocado, castor, Cinchona spp., citrus, cotton, eggplant, guava, lucerne, papaw, parsley, pineapple, Piper betle, rhubarb, sesame, strawberry, tomato. DISEASES: Damping-off of seedlings (tomato, castor, citrus, cotton); root rot (citrus, avocado, strawberry, lucerne); crown rot (parsley, rhubarb, strawberry, lucerne); brown stem rot of tobacco; stem canker and tip blight of Cinchona spp. ; leaf blight (castor, sesame, pineapple, Piper betle) and fruit rot (citrus, tomato, guava, papaw, eggplant). GEOGRAPHICAL DISTRIBUTION: Africa (Ethiopia, Mali, Madagascar, Mauritius, Morocco, Nigeria, Sierra Leone, Southern Rhodesia, Tanganyika); Asia (Burma, Ceylon, China, Formosa, India, Israel, Japan, Java, Malaya, Philippines); Australia & Oceania (Australia, Hawaii, Tasmania); Europe (Cyprus, France, Germany, Great Britain, Holland, Ireland, Italy, Poland, Portugal, U.S.S.R.); North America (Bermuda, Canada, Mexico, U.S.A.); Central America & West Indies (Costa Rica, Cuba, El Salvador, Guatemala, Jamaica, Montserrat, Puerto Rico, Trinidad);. South America (Argentina, Brazil, British Guiana, Colombia, Paraguay, Peru, Venezuela). TRANSMISSION: Soil-borne, spreading rapidly after heavy rain or where soil remains moist or water-logged (40: 470). Also recorded in drainage water in India and in reservoirs and canals supplying citrus groves in U.S.A. (23: 45; 39: 24). A method for determining a disease potential index in soil using lemon fruit has been described (38: 4). Also present in testas of seeds from diseased citrus fruit which may infect nursery seedbeds (37: 165).


Plant Disease ◽  
2014 ◽  
Vol 98 (7) ◽  
pp. 1011-1011 ◽  
Author(s):  
Y. Li ◽  
L. G. Mao ◽  
D. D. Yan ◽  
X. M. Liu ◽  
T. T. Ma ◽  
...  

Ginger (Zingiber officinale Roscoe) is an important commercial crop planted on more than 13,000 ha annually in Anqiu city, Shandong Province, China. From 2010 to 2011, the incidence of Pythium soft rot disease on cv. Laiwu Big Ginger reached 40 to 75% in Anqiu and yield losses of up to 60% were observed. The disease symptoms included brown spots on ginger rhizomes followed by soft rot, stems and leaves above ground becoming withered and yellow, and water soaking on the collar region. The soft rot did not produce offensive odors, which is different from bacterial rots (2). Forty symptomatic rhizomes were sampled from eight farms. Martin's method (1) was used to isolate the pathogen. Ten pieces from each rhizome were washed with sterile distilled water for 30 s and plated on Martin's selective medium at 26°C in a chamber without light. Colonies grew with cottony aerial mycelium. Main hyphae were 5.7 to 9.6 μm wide. Globose sporangia consisting of terminal complexes of swollen hyphal branches were 11.4 to 18.3 μm wide. The average diameter of zoospores was 9.2 μm. The oogonia were globose and smooth, with a diameter of 21 to 33 μm. The sequences of the rRNA gene internal transcribed spacer (ITS) regions 1 and 2 and the 5.8S gene of five isolates were amplified using primers ITS1 and ITS4 (4), and the nucleotide sequence was the same as isolate No. 2, which was deposited in GenBank (Accession No. KC594034). A BLAST search showed 99% identity with Pythium aphanidermatum strain 11-R-8 (Accession No. JQ898455.1). Pathogenicity tests of five isolates were carried out in a greenhouse. Sixty plants (cv. Laiwu Big Ginger) were grown for 30 days in plastic pots (diameter 20 cm) in sandy soil (pH 5.48) and inoculated. Ten plants were used as untreated controls. Five isolates were grown on Martin's liquid medium for 72 h and the spores were harvested in sterile distilled water. Aqueous spore suspensions of the five isolates were adjusted with deionized water to 1 × 108 CFU/ml and injected with a syringe into the soil around the rhizome of the plants. Plants were then placed in the greenhouse at 24 to 26°C and assessed for rhizome rot on the 14th day after inoculation. The inoculated isolates were recovered from the diseased rhizomes, confirming their pathogenicity. To our knowledge, this is the first report of ginger Pythium soft rot caused by P. aphanidermatum in China. Ginger Pythium soft rot caused by P. myriotylum is reported in Taiwan (3). References: (1) F. N. Martin. Page 39 in: The Genus Pythium. American Phytopathological Society, St. Paul, MN, 1992. (2) E. E. Trujillo. Diseases of Ginger (Zingiber officinale) in Hawaii, Circular 62, Hawaii Agricultural Experiment Station, University of Hawaii, December 1964. (3) P. H. Wang. Lett. Appl. Microbiol. 36:116, 2003. (4) T. J. White. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.


Plant Disease ◽  
2010 ◽  
Vol 94 (10) ◽  
pp. 1261-1261 ◽  
Author(s):  
İ. Kurbetli ◽  
K. Değirmenci

Almond (Prunus dulcis) production is currently increasing in Turkey. Losses of approximately 1% associated with root and crown rot of almond seedlings were observed in two commercial nurseries in Ankara and Düzce provinces in 2009. Aboveground symptoms were leaf chlorosis and wilt. Feeder roots were decayed, necrosis occurred on taproots and basal stems, and plants collapsed within several weeks. Roots were washed in tap water and 9 to 10 pieces (3 to 5 mm long) of root tissue taken from the margins of canker lesions, without surface disinfection, were placed on selective medium P5ARPH-CMA (2). Plates were incubated for 3 to 5 days at 20°C in darkness and a number of Phytophthora spp. were recovered. Actively growing mycelium was transferred to carrot piece agar containing β-sitosterol (per liter: carrot piece, 40 g; agar, 20 g; β-sitosterol, 20 mg). Isolates were identified as Phytophthora cactorum and P. citrophthora on the basis of morphological characteristics (1). P. cactorum produced abundant sporangia, oogonia, and paragynous antheridia on carrot piece agar plus β-sitosterol. It had conspicuously papillate and caducous sporangia with short pedicel. Sporangia were usually ovoid but sometimes nearly spherical. P. citrophthora did not produce sexual structures in single culture. It produced papillate, noncaducous sporangia, which were usually ovoid and obpyriform, often asymmetrically shaped and rarely possessed more than one apex. P. citrophthora did not grow at 35°C but it grew well at 30°C. Isolate identities were confirmed by sequence analysis of the ribosomal DNA internal transcribed spacers 1 and 2 (GenBank Accession Nos. HM357622, HM357623, HM357624, HM357625) using primers ITS1 and ITS2 (3). One representative isolate of each species was used to inoculate eight 2-year-old almond plants with an agar plug with actively growing mycelium that was attached to exposed cambium of basal stems. Agar plugs without mycelium were used for eight control plants. All plants inoculated with Phytophthora spp. collapsed within 3 to 4 weeks. Control plants remained healthy. Phytophthora spp. were reisolated from necrotic basal stems. To our knowledge, this is the first report of P. cactorum and P. citrophthora of almond in Turkey. References: (1) M. E. Gallegly and C. Hong. Phytophthora, Identifying Species by Morphology and DNA Fingerprints. The American Phytopathological Society, St. Paul, MN, 2008. (2) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986. (3) S. G. Roy et al. J. Phytopathol. 157:666, 2009.


Plant Disease ◽  
2009 ◽  
Vol 93 (8) ◽  
pp. 844-844 ◽  
Author(s):  
A. Pane ◽  
S. O. Cacciola ◽  
S. Scibetta ◽  
G. Bentivenga ◽  
G. Magnano di San Lio

In the summer of 2006, 1-year-old apricot (Prunus armeniaca L.) trees with leaf chlorosis, wilting, and defoliation associated with root and crown rot were observed in a nursery in Sicily (Italy). Of 3,000 plants, ~2% was affected. Four Phytophthora spp. (45, 25, 20, and 10% of the isolations of the first, second, third, and fourth species, respectively) were isolated from decayed roots and trunk bark on BNPRAH (3). Axenic cultures were obtained by single-hypha transfers. Isolates of the first species formed petaloid colonies on potato dextrose agar (PDA) and had an optimum growth temperature of 25°C. On V8 agar (VA), they produced persistent, papillate (often bipapillate), ovoid to limoniform sporangia (length/breadth ratio 1.4:1). They did not produce gametangia when paired with A1 and A2 isolates of Phytophthora nicotianae. The second species formed arachnoid colonies, had an optimum growth of 30°C, and produced uni- and bipapillate, ellipsoid, ovoid or pyriform sporangia (length/breadth ratio 1.3:1). All isolates were A2. The third species formed rosaceous colonies on PDA, had an optimum temperature of 28 to 30°C, and produced papillate (sometime bipapillate), ellipsoid or limoniform (length/breadth ratio 2:1), caducous sporangia with a tapered base and a long pedicel (as much as 150 μm). All isolates were A1 type. The fourth species formed petaloid-like colonies on PDA and had an optimum growth of 26 to 28°C. On VA, it produced papillate (sometimes bipapillate), ovoid (length/breadth ratio 1.3:1), and decidous sporangia with a short pedicel (<4 μm). The isolates were homothallic and produced oogonia (25 to 31 μm in diameter) with paragynous antheridia and aplerotic oospores. On the basis of morphological and cultural characters, the species were identified as P. citrophthora, P. nicotianae, P. tropicalis and P. cactorum. Identification was confirmed by the electrophoretic analysis of total mycelial proteins and four isozymes (acid and alkaline phosphatases, esterase, and malate dehydrogenase) on polyacrylamide gel (1). Analysis of internal transcribed spacer (ITS) regions of rDNA using the ITS 4 and ITS 6 primers for DNA amplification (2) revealed 99 to 100% similarity between apricot isolates of each species and reference isolates from GenBank (Nos. AF266785, AB367355, DQ118649, and AF266772). The ITS sequence of a P. citrophthora isolate from apricot (IMI 396200) was deposited in GenBank (No. FJ943417). In the summer of 2008, pathogenicity of apricot isolates IMI 396200 (P. citrophthora), IMI 396203 (P. nicotianae), IMI 396201 (P. tropicalis), and IMI 396202 (P. cactorum) was tested on 3-month-old apricot seedlings (10 plants for each isolate) that were transplanted into pots filled with soil prepared by mixing steam-sterilized sandy loam soil (4% vol/vol) with inoculum produced on autoclaved kernel seeds. Ten control seedlings were grown in autoclaved soil. Seedlings were maintained in a screenhouse and watered daily to field capacity. Within 40 days of the transplant, all inoculated seedlings showed leaf chlorosis, wilting, and root rot. Control seedlings remained healthy. All four Phytophthora spp. were reisolated solely from inoculated plants. To our knowledge, this is the first report of Phytophthora root and crown rot of apricot in Italy and of P. tropicalis on this host. References: (1) S. O. Cacciola et al. Plant Dis. 90:680, 2006. (2) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (3) H. Masago et al. Phytopathology 67:425, 1977.


2014 ◽  
Vol 15 (4) ◽  
pp. 159-165
Author(s):  
Daniel T. Drechsler ◽  
Steven N. Jeffers ◽  
William C. Bridges

Phytophthora nicotianae is a common pathogen of many herbaceous perennial plants, and this pathogen has been found causing disease on garden phlox (Phlox paniculata) in wholesale nurseries in South Carolina for a number of years. However, the relationship between P. nicotianae and garden phlox has not been studied or reported previously. Using Koch's postulates and standard inoculation methods for Phytophthora spp., P. nicotianae was found to cause crown rot on P. paniculata when potting medium was infested with colonized vermiculite and to cause foliage blight when aerial parts of the plant were inoculated with an aqueous suspension of zoospores. Foliage blight was more similar to symptoms we observed on garden phlox plants in wholesale nurseries, but crown rot also has been observed previously on plants in these nurseries. The cause of these two diseases was confirmed, but reproduction of Phytophthora foliage blight under experimental conditions was inconsistent. Thus, other factors not yet identified may play a role in the development of Phytophthora foliage blight on garden phlox in nurseries in South Carolina. Accepted for publication 1 September 2014. Published 1 November 2014.


Plant Disease ◽  
2008 ◽  
Vol 92 (2) ◽  
pp. 313-313 ◽  
Author(s):  
A. Pane ◽  
S. O. Cacciola ◽  
A. Chimento ◽  
C. Allatta ◽  
S. Scibetta ◽  
...  

In the summer of 2005, approximately 5% of a nursery stock of 12-month-old potted plants of bower vine (Pandorea jasminoides (Lindl.) K. Schum.) in Sicily (Italy) showed wilt, leaf chlorosis, defoliation, root rot, and collapse of the entire plant. Three Phytophthora spp. (20, 50, and 30% of the isolations of the first, second, and third species, respectively) were isolated from rotted roots on BNPRAH selective medium (2). Single-hypha isolates of the first species formed petaloid colonies on potato dextrose agar (PDA) and had an optimum growth temperature of 25°C (9.3 mm/day); on V8 juice agar, they produced uni- and bipapillate, ovoid to limoniform sporangia with mean dimensions of 45 × 30 μm and a mean length/width (l/w) ratio of 1.4:1. They did not produce gametangia when paired with A1 and A2 isolates of Phytophthora nicotianae. The second species formed arachnoides colonies on PDA, had an optimum growth temperature of 30°C (6.9 mm/day) and produced sporangia that were uni- and bipapillate, ellipsoid, ovoid, or pyriform to spherical (dimensions 44 × 34 μm; l/w ratio 1.3:1). All isolates were A2 mating type and produced amphyginous antheridia and spherical oogonia with smooth walls. The third species formed rosaceous colonies on PDA, had an optimum growth temperature of 28 to 30°C (11.9 mm/day), and produced uni- and bipapillate, ellipsoid or limoniform, caducous sporangia (dimensions 52 × 26 μm; l/w ratio 2.1:1) with a tapered base and a long pedicel (as much as 150 μm). All isolates were A1 type and produced amphigynous antheridia and spherical oogonia with smooth walls. The three species were identified as P. citrophthora, P. nicotianae, and P. tropicalis, respectively. The electrophoretic analysis of the mycelial proteins and four isozymes (1) confirmed the identification. Blast analysis of the sequence of the internal transcribed spacer region of the rDNA of a P. tropicalis isolate from bower vine (GenBank Accession No. EU076731) showed 99% similarity with the sequence of a P. tropicalis isolate from Cuphea ignea (GenBank Accession No. DQ118649). The pathogenicity of three isolates from bower vine, IMI 395552 (P. citrophthora), IMI 395553 (P. nicotianae), and IMI 395346 (P. tropicalis), was tested on 3-month-old potted bower vine plants (10 plants for each isolate) by applying 10 ml of a suspension (2 × 104 zoospores/ml) to the root crown. The plants were maintained at 24°C and 95 to 100% relative humidity. All inoculated plants wilted after 4 weeks. Noninoculated control plants remained healthy. The three Phytophthora spp. were reisolated from symptomatic plants. To our knowledge, this is the first report of Phytophthora root rot of bower vine in Italy. References: (1) S. O. Cacciola et al. Plant Dis. 90:680, 2006. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996.


Plant Disease ◽  
2010 ◽  
Vol 94 (6) ◽  
pp. 782-782 ◽  
Author(s):  
X. M. Yang ◽  
J. H. Wang ◽  
S. P. Qu ◽  
L. H. Wang ◽  
L. C. Peng

Lily (Lilium spp.) is an economically important cut flower in China. In August 2009, 30 to 40% of plants of lily cv. Siberia in a greenhouse for cut flower production in Yunnan, China were severely diseased. Infected plants developed water-soaked lesions and soft rot on the base of stems and leaves near the soil surface. As the disease progressed, stems bent and plants collapsed. Soft rot symptoms were observed on some bulbs and roots of severely diseased plants. Small, diseased tissue fragments (approximately 3 mm) were surface disinfected with 0.5% NaOCl and then plated to Phytophthora selective medium (10% V8 juice agar) (4). Inoculated dishes were incubated at 25°C in the dark. After 5 days, white colonies with abundant aerial mycelia developed from all plated tissue samples. The fungus had aseptate hyphae. Sporangia were papillate, both caducous and noncaducous, and the shape ranged from ovoid to spherical. The dimensions of sporangia were 30 to 62 × 21 to 46 μm. On the basis of morphological features, isolates were identified as Phytophthora nicotianae Breda de Haan. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS1/ITS4 and sequenced. BLAST analysis of the 835-bp fragment showed a 99% homology with the sequence of P. nicotianae AY833527. The nucleotide sequence has been assigned GenBank No. GU299778. PCR amplification of genomic DNAs using the P. nicotianae-specific primer pair ITS3-PNIC1 generated a 455-bp sequence (3). The result further confirmed the identity of P. nicotianae. Pathogenicity tests were conducted in the greenhouse on lily cv. Siberia grown in pots. Ten 3-month-old plantlets were inoculated by watering the wounded stem bases and soil surface with 30 ml of zoospore suspensions (105 spores per ml). Five uninoculated plantlets were used as controls. All plantlets were covered with plastic bags and incubated at room temperature (22 to 26°C) for 48 h. Inoculated plants developed initial symptoms of slight chlorosis and wilting of lower leaves. Within a 3-week period, all plants died due to soft rot of stem bases and leaves. The pathogen was reisolated from inoculated plants but not from control plants that were symptomless. P. nicotianae has been reported as the causal agent of Phytophthora blight on lily in Korea, Japan, and Hungary (1,2). To our knowledge, this is the first report of Phytophthora blight of lily in China. References: (1) J. Bakonyi et al. Plant Pathol. 50:795, 2001. (2) H. J. Jee and W. G. Kim. Plant Pathol. J. 14:452, 1998. (3) P. W. Tooley et al. Appl. Environ. Microbiol. 63:1467, 1997. (4) X. B. Zheng. Phytophthora and Its Research Technology. Beijing. China Agriculture Press, Beijing, 1997.


Plant Disease ◽  
2021 ◽  
Author(s):  
Jieying Xu ◽  
Xiao Yang ◽  
Cuiping Wu ◽  
Ziwei Zhou ◽  
Zhenpeng Chen ◽  
...  

Dianthus chinensis is a popular ornamental plant that is widely cultivated in China. From May 2020 to 2021, root rot and foliage blight were observed on approximately 50% groundcover plants at several landscape sites of Xuanwuhu Park and Nanjing Railway Station, China. Symptoms of wilting and chlorosis appeared in the initial stage, and severe infection caused the whole plant to die . To recover the causal pathogen, infected root and leaf samples were cut into 5×5 mm2 squares, surface-disinfected in 70% ethanol for 30 sec, placed onto 10% clarified V8 PARP agar at 25°C . After three days, Phytophthora-like hyphae were visibly emerged from both root and leaf tissues and growing into cV8A. Individual hyphal tips were transferred to new cV8A plates to obtain a total of 10 pure isolates. Colony morphology of all isolates on cV8A had slightly radiate to stellate patterns with cottony aerial mycelia. After four or five days all isolates had identical morphological traits including papillate and noncaducous sporangia on cV8, hyphal swellings, and intercalary and terminal chlamydospores. A representative isolate Pni-dc7 was examined for morphological measurements. Sporangia were mostly ovoid and sometimes obpyriform, averaging 28.9±5.6 µm in length and 24.9±5.8 µm in width (n=30). Chlamydospores were abundant and spherical with an average diameter at 29.2 ± 0.3 µm (n=30). Oogonia were not observed. For sequence analysis, the internal transcribed spacer (ITS) regions and large subunit (LSU) of the nuclear ribosomal RNA gene complex were amplified using the primer pairs ITS1/ITS4 and NL1/NL4 , respectively, while the mitochondrial cytochrome c oxidase subunit II (coxII) gene was amplified using FM58/FM66 (Martin et al. 2003). The ITS sequence of isolate Pni-dc7 (GenBank Acc. No. MZ519893) had a 100% identity to those of P. nicotianae (MH219914, KU172524, MT065839). The LSU sequence (MZ573547) had a 100% identity to those of P. nicotianae (KX250514, MZ348950, HQ665198).The cox2 sequence (MZ519893) had a 100% identity to those of P. nicotianae (MH221078, KJ506439, JF707072). Based on morphological and molecular evidence, Pni-dc7 was identified as P. nicotianae. Pathogenicity tests  were conducted using both detached leaves and whole plants. Asymptomatic leaves were collected from healthy plants.A 5×5 mm2 Pni-dc7-colonized cV8A plug was placed on each wound of five leaves. Sterile agar plugs were used for a non-inoculated control leaf. All six leaves were placed on a wet filter paper in a closed container at 25°C. All inoculated leaves had necrotic tissues around the wounds, the symptoms progressed from spots to the entire leaves after two days . The control leaves remained asymptomatic. In the whole-plant assay, a D. chinensis  plant (approx. 0.3 m in height) was inoculated with 5 mL of zoospore suspension that was mixed into the potting soil(500g). Three plants were inoculated and control plants were treated with sterile distilled water. After two weeks all three inoculated plants in three repeats of the assay had root and crown rot and foliage blight, whereas all control plants remained asymptomatic. P. nicotianae was reisolated from all inoculated plants. This is the first report of P. nicotianae causing root rot and foliage blight on D. chinensis in China. Considering the importance of D. chinensis to both ornamental nursery and landscaping industries in China, diseased plants at the landscape sites were removed to prevent the spread of P. nicotianae to production sites and other landscape locations.


2007 ◽  
Vol 97 (8) ◽  
pp. 958-963 ◽  
Author(s):  
G. C. Colburn ◽  
J. H. Graham

Phytophthora root rot of citrus in Florida is caused by Phytophthora nicotianae and P. palmivora. A naturally occurring isolate of P. nicotianae (Pn117) was characterized as hypovirulent on citrus roots. Pn117 infected and colonized fibrous roots, but caused significantly less disease than the virulent isolates P. nicotianae Pn198 and P. palmivora Pp99. Coincident inoculation of rootstock seedlings of Cleopatra mandarin (Citrus reticulata) or Swingle citrumelo (C. paradisi × Poncirus trifoliata) with the hypovirulent Pn117 and the virulent isolates Pn198 and Pp99 did not reduce the severity of disease caused by the virulent Phytophthora spp. When either rootstock was inoculated with the hypovirulent Pn117 for 3 days prior to inoculation with virulent isolates, preinoculated seedlings had significantly less disease and greater root weight compared with seedlings inoculated with the virulent isolates alone. Recovery of the different colony types of Phytophthora spp. from roots of sweet orange (C. sinensis) or Swingle citrumelo was evaluated on semiselective medium after sequential inoculations with the hypovirulent Pn117 and virulent Pp99. Pn117 was isolated from roots at the same level as the Pp99 at 3 days post inoculation. Preinoculation of Pn117 for 3 days followed by inoculation with Pp99 resulted in greater recovery of the hypovirulent isolate and lower recovery of the virulent compared with coincident inoculation.


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