bluuebunny/crossref_metadata_2025 huggingface dataset:
First Report of Lethal Yellowing Disease of Coconut Palms Caused by Phytoplasma on Nevis Island
Authors: Myrie W. A. , Paulraj L. , Dollet M. , Wray D. , Been B. O. , McLaughlin W.
Published: May 2007

Coconuts (Cocos nucifera) are an important small-holder's crop in many tropical countries and are used to enhance esthetics of coastal areas. Lethal yellowing (LY) is the single most important plant disease affecting the coconut industry in Jamaica. It affects many palm species in Jamaica, Florida, and Guatemala. This coconut disease was first recorded in Grand Cayman Island in 1834 and Jamaica in 1884. Symptoms of LY disease include premature nut fall, necrosis of the inflorescence, yellowing of the leaves, and defoliation. Thirty-eight coconut palms displaying symptoms indicative of LY disease were sampled in April, 2005 at several locations in Nevis. Immature leaf tissues (leaf bases adjacent to the apical meristem) and nondestructive (boring with a bit and braces) samples were collected from disease and healthy control coconut trees. DNA was extracted (2). The first round of polymerase chain reaction (PCR) with phytoplasma universal primer pair P1/P7 (1,3) resulted in an rDNA fragment of 1.8 kb, and a subsequent nested PCR using LY16-23Sr/LY16Sf primers yielded an amplicon of 1.74 kb (4). Purified product was cloned for sequencing. Sequences obtained were analyzed with Vector NTI Software Suite. The sequence of LYN 18-3 was entered in Genbank and Accession No. DQ378279 was assigned. LYN 18-3 has approximately 99% homology with LY Phytoplasma U18747 from Florida (Manila palm [Veitchia merrillii]). The disease-associated phytoplasma was reliably detected in immature tissues and trunk phloem at the onset of foliar symptoms in palms by PCR. On the basis of the results obtained from this study, it is clear that LY phytoplasma (16SrIV group) was found in the samples collected from Nevis. To our knowledge, this is the first report on lethal yellowing disease in Nevis.

References: (1) S. Deng and C. Hiruki. J. Microbiol. Methods 14:53 1991. (2) J. J. Doyle and J. L. Doyle. Focus 12:13, 1990. (3) N. A. Harrison et al. Ann. Appl. Biol. 141:183, 2002. (4) C. D. Smart et al. Appl. Environ. Microbiol. 62:2988, 1996.
Ground Vegetation Survey forXylella fastidiosain California Almond Orchards
Authors: Shapland Elaine B. , Daane Kent M. , Yokota Glenn Y. , Wistrom Christina , Connell Joseph H. , Duncan Roger A. , Viveros Mario A.
Published: May 2007

Xylella fastidiosa is a xylem-limited bacterium that causes almond leaf scorch (ALS), Pierce's disease of grapevines, and other plant diseases. We surveyed ground vegetation in ALS-infected almond orchards in California's Central Valley for the presence of this bacterium. Plant tissue samples were collected throughout a 2-year period and processed for the presence of X. fastidiosa using restriction enzyme digestion of RST31 and RST33 polymerase chain reaction (PCR) products and bacterial culture on selective media. Overall disease incidence was low in the ground vegetation species; only 63 of 1,369 samples tested positive. Of the 38 species of common ground vegetation tested, 11 tested positive for X. fastidiosa, including such common species as shepherd's purse (Capsella bursa-pastoris), filaree (Erodium spp.), cheeseweed (Malva parvifolia), burclover (Medicago polymorpha), annual bluegrass (Poa annua) London rocket (Sisymbrium irio), and chickweed (Stellaria media). There was a seasonal component to bacterial presence, with positive samples found only between November and March. Both ground vegetation and almond trees were most commonly infected with the almond strain of X. fastidiosa (six of seven surveyed sites). ALS-infected almond samples had an X. fastidiosa concentration within previously reported ranges (1.84 × 106to 2.15 × 107CFU/g); however, we were unable to accurately measure X. fastidiosa titer in sampled ground vegetation for comparison. These results are discussed with respect to ground vegetation management for ALS control.
First Report of Thielaviopsis Trunk Rot of Date Palm in Italy
Authors: Polizzi G. , Castello I. , Vitale A. , Catara V. , Tamburino V.
Published: May 2007

As an alternative to Canary Island date palm (Phoenix canariensis Chabaud), date palm (P. dactylifera L.) is being planted in Sicilian landscapes. In January 2006, severe symptoms of trunk rot were noticed on 10 7- to 8-m high mature date palms in the central square of Aci Bonaccorsi (Catania) in eastern Sicily. In June 2004, these palms were transplanted directly from Egypt. In 4 of 13 planted date palms, the canopy suddenly fell off the trunk. The canopy of all palms appeared normal and healthy with no stem bleeding observed before trunk collapse. Cross sections of affected date palms revealed a brown rot of nonlignified or lightly lignified tissues along with a strong odor of fermented fruit (amyl acetate) associated with the presence of Geotrichum candidum Link ex Pers. Brown rot was not detected in three of the date palms examined. Symptoms were not detected on lignified and external fibers. Internal tissue adjacent to the rotted areas was placed on carrot agar amended with 500 μl of streptomycin sulfate and acidified (lactic acid; pH = 3.6) potato dextrose agar. Large sections (10 to 18 cm) of affected palm tissues were maintained in a moist chamber for 8 days. Microscopic examinations of five single-conidia isolates on media and sporulation from affected tissues yielded Thielaviopsis paradoxa De Seyn. (Höhn) (1). Endoconidia measuring 3 to 5.5 × 7 to 11 μm were cylindrical to somewhat oval when mature, hyaline to mid brown and smooth walled. Endoconiodophores were usually straight, colorless to pale brown, as much as 150 μm long, with a terminal spore-bearing cell through which spores are born. Chlamydospores were smooth, thick walled, brown, in chains, and were 7.5 to 13 μm width × 10 to 18 μm length (values referred to 50 examined chlamydospores). T. paradoxa is a pathogen that can infect any part of a palm and its pathogenicity to date palm is well documented (3). T. paradoxa is endemic in northern Italy (Ligurian Riviera) on P. canariensis where it causes a disease known as bud rot (2). To our knowledge, this is the first record of trunk rot caused by T. paradoxa on date palm in Italy and is the first report of the fungus in Sicily. It is recommended to avoid the foreign trade of mature date palms from known infected areas because of the symptomless infections by this pathogen.

References: (1) A. R. Chase and T. K. Broschat, eds. Diseases and Disorders of Ornamental Palms. The American Phytopathological Society, St. Paul, MN, 1991. (2) A. Garibaldi et al., eds. Malattie Delle Piante Ornamentali. Calderini Edagricole, Bologna, Italy, 2000. (3) P. Suleman et al. Plant Dis. 85:80, 2001.
First Report of Papaya ringspot virus W in Sponge Gourd from India
Authors: Verma Raj , Baranwal V. K. , Prakash Satya , Tomer S. P. S. , Pant R. P. , Ahlawat Y. S.
Published: May 2007

During August 2004, symptoms resembling a virus disease were observed in commercial cultivation of sponge gourd (Luffa cylindrica (L.) M. Roem. [cv. Chikni]) in Pune, India. Affected plants exhibited mosaic mottling, vein banding, and downward marginal curling on leaves. The incidence of disease was 10 to 30% based on the symptoms and confirmed using enzyme-linked immunosorbent assay (ELISA). Six fields of sponge gourd were visited, and 30 samples were collected randomly. Samples from affected fields were analyzed for the presence of virus by mechanical inoculation. Symptoms typical of those observed in the original field plants appeared 10 days after inoculation on sponge gourd. The virus isolate produced local lesions on Chenopodium amaranticolor and systemic symptoms on Cucumis melo, Cucumis sativus, Cucurbita maxima, Cucurbita pepo, L. acutangula, and L. cylindrica, but did not produce symptoms on Carica papaya (cvs. CO2 and Red Lady), Nicotiana glutinosa, N. tabacum (cv. White Burley), or Vigna unguiculata. Field-infected sponge gourd and all indicator plants were tested using double-antibody sandwich (DAS)-ELISA. The virus was identified as Papaya ringspot virus-W (PRSV-W) using DAS-ELISA (Agdia, Elkhart, IN). A strong, positive reaction was obtained with antiserum to PRSV-W but not with antisera to PRSV-P, Cucumber mosaic virus, Squash mosaic virus, Zucchini yellow mosaic virus, and Groundnut bud necrosis virus. To ensure virus purity, the isolate was passed through three successive single-lesion transfers on C. amaranticolor. Flexuous filamentous particles 775 nm long were observed with electron microscopy of leaf-dip preparation from symptomatic sponge gourd leaves. The virus particles were then decorated with a second PRSV-W antiserum obtained from the Plant Virology Unit, IARI, New Delhi. PRSV-W on sponge gourd has previously been reported from Taiwan (1). To our knowledge, this is the first report of natural occurrence of PRSV-W on sponge gourd in India.

Reference: (1.) C. H. Huang et al. J. Agri. Res. China 36:413, 1987.
Carrot Purple Leaf: A New Spiroplasmal Disease Associated with Carrots in Washington State
Authors: Lee Ing-Ming , Bottner Kristi D. , Munyaneza Joseph E. , Davis Robert E. , Crosslin James M. , du Toit Lindsey J. , Crosby Todd
Published: May 2007

During the growing seasons of 2003 and 2004, a disease occurred in several carrot crops in south central Washington with symptoms suggestive of infection by phytopathogenic mollicutes (phytoplasmas and spiroplasmas). In the fall, many affected carrot plants exhibited extensive purple or yellow-purple leaf discoloration, general stunting of shoots and taproots, and formation of bunchy, fibrous secondary roots. For detection of the putative causal agents, polymerase chain reaction (PCR) assays were performed using primers specific to phytoplasmas as well as primers specific to plant-pathogenic spiroplasmas. Restriction fragment length polymorphism (RFLP) analyses of PCR-amplified 16S rDNA sequences revealed that about 81% of affected plants showing dark purple or yellow-purple leaf symptoms tested positive for Spiroplasma citri. Of affected plants showing mild purple discoloration of leaf margins, 18% tested positive for a phytoplasma strain belonging to the clover proliferation group (16SrVI), subgroup 16SrVI-A, and 11% for another phytoplasma strain belonging to the aster yellows group (16SrI), subgroup 16SrI-A. Nucleotide sequence analysis of cloned 16S rDNA confirmed the phytoplasma group affiliations. Some symptomatic plants were co-infected with S. citri and either aster yellows phytoplasma or clover proliferation group phytoplasma. To our knowledge, this is the first documentation of spiroplasma infection of carrot in the United States.
Holdover Inoculum of Pseudomonas syringae pv. alisalensis from Broccoli Raab Causes Disease in Subsequent Plantings
Authors: Cintas N. A. , Koike S. T. , Bunch R. A. , Bull C. T.
Published: May 2007

Uniform plots of broccoli raab (Brassica rapa subsp. rapa) seedlings were inoculated with a rifampicin-resistant strain of Pseudomonas syringae pv. alisalensis, the causal agent of bacterial blight on crucifers, resulting in 100% disease incidence in mature plants. Diseased plants were incorporated into the soil at maturity and smaller replicated plots were replanted at various times after incorporation. Rifampicin-resistant fluorescent pseudomonads with rep-PCR profiles identical to P. syringae pv. alisalensis were isolated from lesions on plants grown in soil into which the first diseased crop was incorporated. Disease incidence declined in mature plants as the length of time between incorporation of the first planting and seeding of the replanted plots increased. Bacterial population levels in soil decreased over time and bacteria were no longer detectable 3 weeks after incorporation of the diseased crop. In laboratory tests, population levels of P. syringae pv. alisalensis decreased in untreated soil but not in autoclaved soil. Greenhouse studies demonstrated a direct correlation between population levels of P. syringae pv. alisalensis applied to soil and disease incidence in seedlings. However, the decline in bacterial populations in field soils did not wholly account for the decline in disease incidence with subsequent plantings.
Resistance of Strawberry Cultivars to Crown Rot Caused by Colletotrichum gloeosporioides Isolates from Florida Is Nonspecific
Authors: MacKenzie S. J. , Legard D. E. , Timmer L. W. , Chandler C. K. , Peres N. A.
Published: May 2007

Isolates of Colletotrichum gloeosporioides from strawberry (Fragaria × ananassa) and native grape were tested for virulence on strawberry cultivars in field experiments for three seasons. Isolate aggressiveness and cultivar resistance were determined by the proportion of plants killed at a defined time. Each year, four to six isolates were inoculated on four to seven different cultivars, with a subset of isolates and cultivars evaluated again the next season. On the dates that disease was evaluated, incidence ranged from 10 to 84% for individual cultivars. Cultivar and isolate effects were significant in all three seasons, but there was no significant cultivar by isolate interaction in any season. Thus, resistance to C. gloeosporioides appears to be nonspecific. In the third season, one isolate of Colletotrichum fragariae from strawberry and one from oak were included. There was no significant cultivar by isolate interaction detected for this species, although there were significant differences among cultivars and isolates. When the resistance of cultivars to both species was compared, the rankings of cultivars were similar, but a modest cultivar by species interaction was evident. The cultivar Treasure was more resistant to crown rot caused by either species than any other cultivar tested.
Foliar and Cane Rot of Arundo donax Caused by Nigrospora oryzae in Europe
Authors: Widmer T. , Kirk A. , Kirk G. , Guermache F.
Published: May 2007

A fungus was isolated consistently from dead shoot tips and flag leaves of Arundo donax L. (Poaceae) in France, Crete, Cyprus, Italy, Morocco, and Spain from April of 2003 through September of 2005. The fungus was identified as Nigrospora oryzae (Berk. & Br.) Petch (teleomorph Khuskia oryzae) on the basis of morphological characteristics (1). The mean diameter of 80 conidia obtained from sporulating plant specimens collected in France, Crete, and Cyprus were 14, 15, and 15 μm, respectively. The mean diameters of 25 conidiogenous cells and conidiophores were 7 and 4 μm, respectively. Identification was confirmed by comparing the sequence of the ribosomal DNA internal transcribed spacer 1 and 4 regions from the French isolate (GenBank Accession No. DQ219433) with the sequence of a voucher specimen from the Museum National d'Histoire Naturelle, Paris, France. The isolate of N. oryzae from France was deposited at the CBS collection in Utrecht, the Netherlands (CBS 113884). N. oryzae is known to be a weak pathogen on a wide range of plants but has not been reported on A. donax, which is now a well-established weed in the United States and North America, probably originating from the Mediterranean Region. Herein, the possible use of N. oryzae as a biological control agent was investigated. Twenty young A. donax shoots growing in the greenhouse and 20 emerging canes in the field were selected on the basis of uniformity in size. A spore suspension in distilled water adjusted to 5 × 105 conidia/ml of the French isolate was prepared and 0.5 ml was injected with a syringe just below the growing point of the flag leaf in onehalf of the greenhouse and field plants. The remaining plants were injected with 0.5 ml of distilled water as controls. Infection and death of the flag leaf occurred in 30% of the shoots in the greenhouse and 50% of the canes in the field 21 days from inoculation. No disease developed on the control plants. Greenhouse inoculation tests were repeated once. N. oryzae was reisolated from dead leaves and the terminal node of inoculated shoots, satisfying Koch's postulate. Attempts made to induce disease symptoms by applying spore suspensions on the whorl of leaves surrounding the apical tip failed. This is an indication that an insect vector may be needed to carry and deposit N. oryzae spores into the tight, whorled flag leaf for infection and disease development to occur. To our knowledge, this is the first report of foliar and cane rot of A. donax caused by N. oryzae.

References: (1) K. H. Domsch et al. Page 515 in: Compendium of Soil Fungi. IHW-Verlag, Eching, Germany, 1993.
First Record of Verticillium dahliae on Potato in Malta
Authors: Pace-Lupi T. G. , Porta-Puglia A. , Ippolito A. , Nigro F.
Published: May 2007

Potato is an important and highly valued crop throughout the Maltese Archipelago. Much of the production is exported to Holland. In January 2005, minor wilts and chlorosis of potato plants were observed in a field at Hal-Farrug, Luqa (Malta). Verticillium dahliae Kleb (1) was isolated on potato dextrose agar (PDA) from vascular tissue excised from the base of the plants. Three different isolates were obtained, all of which were typically distinguished by verticillately shaped conidiophores and the abundant production of microsclerotia on PDA. In May 2005, colonies of these three isolates were cultured in potato dextrose broth (PDB), from which conidial suspensions of each isolate were prepared with sterile distilled water to a concentration of 107 ml-1. For each isolate, 10 7-day-old potato seedlings were inoculated via root immersion in the inoculum suspension and transplanted to 20-cm diameter plastic pots containing a soil/peat mixture (1:1 [v/v]). Seedlings treated in the same way with sterile distilled water were used as a control. All plants were kept under controlled glasshouse conditions (20 ± 3°C) and watered to field capacity as required. Minor chlorosis and wilt of the pair of lower-most leaves was noted 7 days after inoculation. During subsequent weeks, wilt began to appear in the typical half-leaf form, while chlorosis was noted on all organs of the plants, including the principal stem (3). Symptoms were absent on the control plants. Measuring the weight of the new tubers produced by each plant revealed no apparent difference between inoculated and healthy plants; nevertheless, inoculated plants resulted in more tubers with a smaller diameter in respect to those of the uninoculated plants. V. dahliae was never isolated from tubers. Little to no variation in symptom severity was noted among plants inoculated with the three individual isolates. At the end of June, V. dahliae was reisolated on PDA from all inoculated plants, in particular, from vascular tissues originating from principal and lateral stems, crowns, and roots. All attempts to isolate the pathogen from control plants were unsuccessful. Molecular detection of the pathogen by using species-specific primers and real-time Scorpion PCR (2) confirmed the results obtained by the classical isolation method. The low symptom severity observed by the growers in the field, usually mistaken for normal dieback of aged plants, might explain why V. dahliae was never reported before on potatoes in the Maltese Archipelago.

References: (1) D. L. Hawksworth and P. W. Talboys. No. 256. Descriptions of Pathogenic Fungi and Bacteria. Commonwealth Mycological Institute (CMI), Kew, Surrey, UK, 1970. (2) F. Nigro et al. Pages 454–461 in: Proc. Convegno Internazionale di Olivicoltura. VI Giornate Scientifiche SOI, Spoleto, 2002. (3) W. R. Stevenson et al., eds. Compendium of Potato Diseases. 2nd ed. The American Phytopathological Society, St Paul, MN, 2001.
Plum Pox Virus Mixed Infection Detected on Apricot in Pakistan
Authors: Kollerová E. , Nováková S. , Šubr Z. , Glasa M.
Published: May 2007

Sharka, caused by Plum pox virus (PPV), is the most detrimental viral disease of stone fruit trees. First reported from Bulgaria in 1917, the virus is now widespread in Europe, the Mediterranean Basin, and Asia Minor and is sporadically present in North and South America. On the basis of molecular and serological properties, six PPV subgroups are recognized, from which PPV-D, PPV-M, and PPV-Rec are the most common (1,2). Several apricot trees (Prunus armeniaca) showing mild, pale green rings and diffuse chlorotic spots on leaves were found in a small orchard in the Baltistan District in northern Pakistan at approximately 2,400 m above sea level. Dried leaf samples from one symptomatic tree randomly selected from the orchard were positive for PPV using double-antibody sandwich enzyme-linked immunosorbent assay with antisera prepared in the laboratory, immunoblot analysis, and reverse transcription-polymerase chain reaction (RT-PCR) targeting the capsid protein (CP) gene using standard procedures (1). To check the subgroup affiliation and evaluate the molecular variability, the 562-bp variable region spanning the C-terminus of NIb and the N-terminus of the CP was amplified, the RT-PCR product was cloned into the pGEM-T Easy vector (Promega, Madison, WI), and positive clones were analyzed by restriction and sequence analyses. Interestingly, sequence analysis of four clones revealed mixed infection, i.e., the presence of two different PPV isolates in the apricot sample. One isolate belonged to PPV-D (GenBank Accession No. DQ422147) and the other belonged to the PPV-Rec subgroup (GenBank Accession No. DQ422148). Multiple alignment of the sequenced genome portion of the Pakistan PPV-D isolate indicated 96 to 99% nt identity with various PPV-D isolates without unique, clear-cut differences. Similarly, the PPV-Rec isolate had 98 to 99% identity with European PPV-Rec isolates and retained the cross-over at nucleotide position 8450 in the 3′ terminus of NIb. This sequence had the amino acid signature at the N-terminus of the CP typical of the PPV-Rec subgroup (2). Moreover, no particular clustering of the Pakistan isolates within PPV-D and PPV-Rec could be observed after phylogenetic analysis. The DAG motif, essential for aphid transmission, was present in both sequences. To our knowledge, this is the first indication of PPV occurrence in Pakistan and first identification of the PPV-Rec isolate outside Europe. Together with previous reports on the PPV presence in China and Kazakhstan (3,4), this report indicates the need for more detailed epidemiological studies focusing the PPV spread and its molecular diversity in Asia.

References: (1) T. Candresse et al. Phytopathology 88:198, 1998. (2) M. Glasa et al. J. Gen. Virol. 85:2671, 2004. (3) M. Navrátil et al. Plant Dis. 89:338, 2005. (4) S. Spiegel et al. Plant Dis. 88:973, 2004.
First Report of Ravenelia spegazziniana on Acacia farnesiana from Brazil
Authors: Avanzato M. V. , da Silveira R. M. Borges , Hernández J. R.
Published: May 2007

Leaves, young branches, and fruits of Acacia farnesiana (L.) Willd. (Fabaceae) that were distorted because of hypertrophy causing witches' brooms and other abnormal growth were collected in the area surrounding the Panteón da Liberdade e da Democracia, Brasilia, Brazil, during August 2005. Uredinia and telia were observed in the symptomatic areas. The uredinia were small on leaflets and rachis and on pods in confluent groups as much as 1 cm in diameter, subcuticular becoming erumpent, and cinnamon-brown; paraphyses were mostly clavate and colorless. Urediniospores were 17 to 28 × 12 to 16 μm, ellipsoid to obovoid, with a wall 1.5 to 2 μm thick at the sides and 2 to 3 μm at the apex, cinnamon-brown, with hub and spoke pattern of ornamentation, and pores in two bands of four each above and below the equator. Telia were small, erumpent, on rachis and small branches, and dark cinnamon-brown. Teliospores were 84 to 100 × 65 to 90 μm in diameter, chestnut-brown, (4) 5 to 6 probasidial cells across, and smooth; cysts were globoid, same number as peripheral probasidial cells; pedicels were colorless, multihyphal, and deciduous. The rust was identified as Ravenelia spegazziniana J.C. Lindq, on the basis of the comparison with specimens deposited at the Instituto Spegazzini (LPS) and U.S. National Fungus Collection (BPI), as well as with published descriptions (2). R. spegazziniana has been reported on species of Acacia from Central, North, and South America, and Hawaii (1). A. farnesiana is probably native to tropical America, but it is naturalized and cultivated all over the world where it has become invasive in disturbed areas in some Pacific islands. This species is economically important for fuel wood, flowers used in the perfume industry, and bark and fruits used for tannin production. In Federal District (DF), Brasilia, Brazil, A. farnesiana is commonly used in urban forestation as an ornamental and shade tree. To our knowledge, this is the first report of R. spegazziniana in Brazil. The voucher specimen has been deposited in the herbarium of the Faculty of Biology of Porto Alegre from the Universidade Federal do Rio Grande do Sul as ICN 139085.

References: (1) D. F. Farr et al. Fungal Databases. Systematic Botany and Mycology Laboratory. On-line publication. ARS, USDA, 2006. (2) J. R. Hernandez and J. F. Hennen. Mycol. Res. 106:972. 2002.
A New Natural Host of Lisianthus necrosis virus in Taiwan
Authors: Chen Y. K. , Jan F. J. , Chen C. C. , Hsu H. T.
Published: May 2007

Lisianthus necrosis virus (LNV) was first identified as a fungus-borne virus that induced systemic necrosis in lisianthus (Eustoma russellianum) in Japan (2). In Taiwan, LNV causes systemic bright yellow chlorosis followed by necrosis in lisianthus (1). The disease was able to spread through the infested soil. Isolation of a fungus vector was attempted but was not successful (1). Calla lilies (Zantedeschia spp.) showing symptoms of systemic necrosis were observed in the fields of central Taiwan. A virus culture was established through single-lesion isolation from a local lesion host, Chenopodium quinoa, and maintained in Nicotiana benthamiana. Mechanical inoculation of the virus resulted in systemic infection in E. russellianum and Datura stramonium and local infection in Celosia argentea, Gomphrena globosa, Chenopodium amaranticolor, Zinnia elegans, Cucumis melo, Cucumis sativus, Cucurbita pepo, Vigna angularis, and Petunia hybrida. Electron microscopic examination of ultrathin sections of infected plant tissues revealed the presence of spherical viral particles approximately 33 nm in diameter. Scattered and aggregated virion particles were frequently observed in the cytoplasm of infected cells. Results of enzyme-linked immunosorbent assay, western blotting, and immunoelectron microscopy indicate that the virus is serologically related to LNV (1). Reverse transcription-polymerase chain reaction (RT-PCR) with degenerate primers (forward primer 5′-ATGGAAATCGTTAGG and reverse primer 5′-CTATAGCAATGTTGC) for LNV coat protein gene produced a cDNA of approximately 1.1 kb. The RT-PCR product was cloned into pGEM-T vector (Promega, Madison, WI) and sequenced. Sequence analysis showed that the cloned fragment (GenBank Accession No. DQ523229) was 1,167 bp long and shared 99% identity at the nucleotide and deduced amino acid levels with that of the LNV isolated from lisianthus (GenBank Accession No. DQ011234). To our knowledge, this is the first report of the natural occurrence of LNV infection in calla lily.

References: (1) C. C. Chen et al. Plant Dis. 84:506, 2000. (2) M. Iwaki et al. Phytopathology 77:867, 1987.
Natural Infection of Verbena and Phlox by a Recently Described Member of the Carmovirus Genus
Authors: Assis Filho F. M. , Harness A. , Tiffany M. , Gera A. , Spiegel S. , Adkins S.
Published: May 2007

A novel carmovirus infecting angelonia (Angelonia angustifolia) was recently described independently by researchers in the United States, Israel, and Germany (1,2,4). Angelonia flower break virus (AnFBV) and Angelonia flower mottle virus were proposed as appropriate names for this carmovirus. The virus, causing stunting, mild leaf mottle, flower mottling, and flower breaking symptoms has been detected in naturally infected angelonia in the United States, Israel, and Germany (2,4). Here we report the first detection of natural infection of verbena (in the United States and Israel) and phlox (in the United States) by using a recently developed double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA; Agdia, Elkhart, IN). Prior to this report, verbena was considered insusceptible to carmovirus infection (3) and phlox was known as an experimental host for AnFBV (2). A comparative serological study including 27 virus species, demonstrated that DAS-ELISA did not cross-react with any viruses that commonly infect ornamentals or are related to carmoviruses, showing that the polyclonal antibodies are specific to AnFBV. Antibody specificity was confirmed by the carmovirus group PCR test (Agdia). Furthermore, reverse transcription-polymerase chain reaction with AnFBV specific primers (2) produced the expected 1172-bp band from all ELISA-positive samples tested. Between November 2005 and March 2006, AnFBV was detected in 181 of 567 verbena, 26 of 143 phlox, and 193 of 267 angelonia samples submitted to Agdia Testing Services by commercial ornamental propagators for virus testing. Most samples were asymptomatic, although a few exhibited mild leaf mottle. It should be noted that the number of AnFBV-infected samples might not accurately reflect the actual number of commercially produced plants infected with AnFBV because most of the samples analyzed originated from virus elimination programs. The detection of natural AnFBV infection of verbena, phlox, and angelonia suggests that AnFBV may be more widespread in the ornamental industry than previously thought.

References: (1) S. Adkins et al. Phytopathology (Abstr.) 95(suppl.):S2, 2005. (2) S. Adkins et al. Phytopathology 96:460, 2006. (3) G. P. Martelli and M. Russo. Online publication. ICTVdB-The Universal Virus Database. 00.074.0.02, 2004. (4) S. Winter et al. New Disease Reports. Vol 12. Brit. Soc. Plant Pathol. Online publication, 2005.
Diversity of Phytophthora capsici in Northwest Spain: Analysis of Virulence, Metalaxyl Response, and Molecular Characterization
Authors: Silvar C. , Merino F. , Díaz J.
Published: May 2007

Phytophthora crown rot, caused by Phytophthora capsici, is potentially the most destructive disease of pepper in Spain. Phenotypic and genetic diversity of 16 P. capsici isolates collected from 11 farms in northwest Spain was characterized based on virulence, mating type, sensitivity to metalaxyl, and genetic analysis using random amplified polymorphic DNA (RAPD) methods. Low variability was observed among the isolates in their metalaxyl response, with 87.5% being highly sensitive. No isolates of the A2 mating type were detected. More variability was found in the virulence assay, and isolates were classified into two groups according to their pathogenicity on a set of four pepper cultivar differentials. Genetic variation examined with eight RAPD primers generated 92 polymorphic bands and revealed the existence of different patterns among isolates. Cluster analysis using the unweighted pair-group method with arithmetic averages (UPGMA) separated the Spanish isolates into three RAPD groups and established a relationship between the Spanish population and a representative worldwide group of isolates. No correlation was found between groups obtained by RAPD analysis and groups defined by virulence or metalaxyl response.
Effect of Crop Rotation on Take-all of Wheat in Double-Cropping Systems
Authors: Cunfer Barry M. , Buntin G. David , Phillips Daniel V.
Published: May 2007

Take-all of wheat (Triticum aestivum), caused by Gaeumannomyces graminis var. tritici, became a serious problem with the widespread adoption of wheat:soybean double-cropping and minimum tillage farming systems in the southeastern United States during the past 30 years. A long-term crop rotation study was initiated in 1994 with 12 double-cropping sequences incorporating wheat, rye, or canola as the fall-planted crop and soybean or grain pearl millet as the summer crop. Cotton and fallow were included in some summer rotations during the last 2 years of the study. The purpose was to identify sustainable alternatives to the continuous wheat:soybean system that would provide acceptable management of take-all. G. graminis var. tritici cultured on autoclaved oats was incorporated into soil prior to planting the first season's crop. Take-all was severe in rotations with continuous wheat each year. Pearl millet was compatible with the cropping system but did not affect incidence or severity of take-all in a following wheat crop. Soybean or pearl millet had little effect on yield loss due to take-all in a subsequent wheat crop. A 1-year rotation with canola significantly reduced take-all incidence and severity. At the end of the second and third seasons, in those rotations where wheat followed 1 year of canola, wheat grain yield was the same as that in control plots that had little or no take-all. Two consecutive years of canola did not suppress take-all or improve wheat yields any more than a single year of canola between wheat crops. Seedling assays for take-all incidence and severity in growth chambers were conducted using soil collected twice each year near the end of each crop's growing season. Results were similar to those observed in the field. However, canola in the rotation had a greater effect in suppressing disease severity than disease incidence. Canola can be a valuable rotational crop for management of take-all in wheat in the southeastern United States.
First Report of the Purple Variant of Curtobacterium flaccumfaciens pv. flaccumfaciens, Causal Agent of Bacterial Wilt of Bean, in Canada
Authors: Huang H. C. , Erickson R. S. , Yanke L. J. , Chelle C. D. , Mündel H.-H.
Published: May 2007

Bacterial wilt of bean (Phaseolus vulgaris L.) caused by the yellow and orange variants of Curtobacterium flaccumfaciens pv. Flaccumfaciens (Hedges) Collins & Jones was found in western Canada in 2002 (1). A purple variant was found in a pooled sample of discolored cull seeds of great northern bean (cv. US1140) from a crop grown near Bow Island, Alberta, Canada in 2005. Bacterial colonies isolated from purple seed using modified Burkholder's agar (MBA) (3) were convex, glistening, and smooth edged with blue pigment diffusing into the medium. Three isolates (V154, V155, and V254) were identified with conventional tests (2), carbohydrate oxidation (GP Microplates, Biolog Inc., Hayward, CA), and cellular fatty acids (CFA) (MIDI, Inc., Newark, DE). All were grampositive, motile, aerobic rods with yellow colonies producing extracellular blue pigment on MBA when grown at 20 ± 2°C. Bacterial isolates grew at 27°C but grew weakly at 37°C. They were positive for catalase and hydrolysis of hippurate and indoxyl acetate and negative for urease, gelatin liquification, and oxidase. CFA profiles were approximately 48% 15:0 anteiso, 40% 17:0 anteiso, 7% 16:0 iso, and 3% 15:0 iso; with 17:1 anteiso A variable but <1%. Many carbohydrates were oxidized in the Biolog microplates with little acid production. The results match C. flaccumfaciens (2) and the MIDI and Biolog databases, as well as the purple variant of C. flaccumfaciens found in Nebraska, the only previous report of this variant (4). The pathogenicity of the three isolates was tested. Seeds of great northern (cv. US1140) and navy (cv. Morden003) beans were soaked in a bacterial suspension (1 × 108 CFU/ml) or distilled water (control) for 1 h, planted in Cornell mix in root trainers, incubated at 28/22°C (16-h day/8-h night) in a growth cabinet for 14 days, and examined for seedling wilt. The test had three replicates per treatment and 20 seeds per replicate in a completely randomized design. All three isolates were pathogenic to both bean cultivars. The wilt incidences were 51, 57, and 56% on US1140 and 64, 76, and 69% on Morden003 for isolates V154, V155, and V254, respectively. The purple variant of C. flaccumfaciens was reisolated from hypocotyls of wilted seedlings but not from healthy controls. The experiment was repeated using the reisolated bacteria and the results were similar to the first experiment, thus fulfilling Koch's postulates. To our knowledge, this is the first report of the purple variant of C. flaccumfaciens pv. flaccumfaciens in Canada.

References: (1) T. F. Hsieh et al. Plant Dis. 86:1275, 2002. (2) K. Komagata et al. Page 1313 in: Bergey's Manual of Systematic Bacteriology. Vol. 2. Williams and Wilkens, Baltimore, MD, 1986. (3) G. A. Nelson and G. Semeniuk. Phytopathology 54:330, 1964. (4) M. L. Schuster et al. Can. J. Microbiol. 14:423, 1968.
Phoma Basal Rot of Romaine Lettuce in California Caused byPhoma exigua: Occurrence, Characterization, and Control
Authors: Koike Steven T. , Subbarao Krishna V. , Verkley Gerard J. M. , Fogle Diana , O'Neill Timothy M.
Published: May 2007

Beginning in 2000 and continuing into 2004, a previously undescribed disease caused significant damage to romaine lettuce (Lactuca sativa) in the coastal Salinas Valley of California. Symptoms were brownish black, sunken cavities on the crown and upper taproot. Cavities were firm, lacked signs of fungal growth, and resulted in cracking and weakening of the crown. Affected plants were stunted, uneven in growth, and unmarketable. Crisphead lettuce also developed these symptoms, although disease incidence was always lower than that for romaine. A fungus was consistently isolated from cavities. Using morphological features and molecular methods, the fungus was identified as Phoma exigua. Pathogenicity of isolates from romaine and crisphead lettuce was demonstrated on both of these lettuce types. Replicated field studies showed that azoxystrobin, boscalid, and cyprodinil + fludioxonil fungicides effectively controlled the disease. This is the first report of a crown disease caused by P. exigua in the United States. A similar disease, Phoma basal rot on greenhouse grown lettuce in the United Kingdom, had been previously observed, but no report has been published.
Influence of Temperature and Time of Year on Colonization of Bermudagrass Roots by Ophiosphaerella herpotricha
Authors: Walker N. R. , Mitchell T. K. , Morton A. N. , Marek S. M.
Published: May 2007

The influence of temperature on the infection of bermudagrass seedlings by Ophiosphaerella herpotricha and colonization of plants in the field was investigated. Bermudagrass seedlings (cv. Jackpot) inoculated with O. herpotricha exhibited dark lesions after 8 days. Root lesion length was greatest at 17°C and was similar for all temperatures examined below 21°C. Seedlings grown at 25 or 30°C had small lesions that remained similar in size when evaluated at 8 and 10 days post inoculation. Colonization of bermudagrass roots from field plots were examined in July, October, and November of 2003 and 2004. In 2003, no differences between sampling dates were observed for plants sampled from the edge of the spring patch in 5.4-cm increments to a total distance of 21.6 cm. In 2004, July and October samples were similar; however, an increase in root colonization was found between the October and November samplings. These studies suggest that infection and colonization of bermudagrass roots by O. herpotricha occurs over a wide range of cool soil temperatures, occurs in the spring, and can be variable in the autumn.
Genetic Diversity of Gibberella zeae Isolates from Manitoba
Authors: Fernando W. G. D. , Zhang J. X. , Dusabenyagasani M. , Guo X. W. , Ahmed H. , McCallum B.
Published: May 2007

Gibberella zeae (anamorph Fusarium graminearum) causes Fusarium head blight, one of the most important diseases of cereals in the Canadian prairies for the last decade. In 2002, 60 isolates of G. zeae were collected and single spored from naturally infected spikes of wheat from Carman and Winnipeg in Manitoba. These isolates were compared using vegetative compatibility analysis and polymerase chain reaction (PCR)-based sequence related amplified polymorphisms (SRAP). Sixteen vegetative compatibility groups (VCG) were found among the 50 isolates tested. Five VCGs were found in the two locations, five in Carman and six in Winnipeg. Eight SRAP primer pairs amplified 90 polymorphic DNA fragments from 60 isolates and identified 59 distinct haplotypes. Among seven pairs of isolates, each pair from a distinct spike, four had isolates with different VCGs and six comprised different SRAP haplotypes. Principal component analysis and UPGMA separated the dataset into two main groups, each with isolates from both locations. The analysis of molecular variance also revealed that 75 and 20% of the variance was associated with differences among individual isolates and varieties sampled, respectively. Geographic location was not a significant source of variation at P = 0.05 and accounted for only 4% of total variance. A low correlation between VCG and SRAP marker data was detected. This study showed that, although genetic diversity is high among G. zeae isolates, Carman and Winnipeg collections have a similar genetic makeup and are likely part of the same population. The significant proportion of variance accounted by the variety compared with the geographic origin of isolates suggests that seedborne inoculum might have contributed to the genetic diversity within the G. zeae collection under study.
First Report of Potato virus M and Chrysanthemum stunt viroid in Solanum jasminoides
Authors: Verhoeven J. Th. J. , Jansen C. C. C. , Roenhorst J. W.
Published: May 2007

In 2005, a plant of the ornamental crop Solanum jasminoides from the Netherlands was submitted for testing on viruses and viroids because of its intended use for propagation. Sap from this plant was mechanically inoculated to the test plant species Chenopodium quinoa, Nicotiana benthamiana, N. hesperis-67A, and N. occidentalis-P1 (3). N. hesperis-67A showed chlorotic local lesions and rugosity followed by vein necrosis, N. occidentalis-P1 showed necrotic local lesions and systemic leaf distortion, and the two other test plant species remained symptomless. Potato virus M (PVM) was identified by double antibody sandwich enzyme-linked immunosorbent assay using leaves from S. jasminoides and N. hesperis-67A. The plant of S. jasminoides was also tested for the presence of viroids by reverse transcriptase-polymerase chain reaction (RT-PCR) with universal pospiviroid primers Pospi1-RE/FW (2). This reaction yielded an amplicon of the expected size of 198 bp. The sequence showed 100% identity to an isolate of Chrysanthemum stunt viroid (CSVd; NCBI GenBank Accession No. AF394453). Subsequently, the complete sequence of our viroid isolate (GenBank Accession No. DQ406591) was determined from the amplicon obtained after RT-PCR using specific primers for the detection of CSVd (1). The viroid isolate from S. jasminoides consisted of 354 nucleotides and showed the highest identity (98.6%) to a chrysanthemum isolate of CSVd (GenBank Accession No. AB055974). Therefore, the viroid was identified as CSVd. To our knowledge, this is the first report of PVM and CSVd in S. jasminoides.

Reference: (1) R. Hooftman et al. Acta Hortic. 432:120, 1996. (2) J. Th. J. Verhoeven et al. Eur. J. Plant Pathol. 110:823, 2004. (3) J. Th. J. Verhoeven and J. W. Roenhorst, EPPO Bull. 33:305, 2003.
Introduction of the Exotic Monopartite Tomato yellow leaf curl virus into West Coast Mexico
Authors: Brown J. K. , Idris A. M.
Published: May 2007

Leaf curl symptoms that are reminiscent of begomovirus (genus Begomovirus, family Geminiviridae) infection were observed widespread in the tomato crop during the early fall 2005 through the spring 2006 growing seasons in Sinaloa State, Mexico. Symptoms were widespread in three major valleys (Culiacan, Guasave, and Los Mochis) that are largely dedicated to fresh-market tomato production for the U.S. market from October to June. Symptoms included stunting of leaves, shortened internodes, distortion of leaf margins, and green vein banding. Fruit set was reduced significantly (as much as 90%) on the portion of the plant that developed above the point of symptom expression. Tomato fields were heavily infested with the B biotype of the whitefly Bemisia tabaci (Genn.) vector and no other insect vectors were noted in the fields. Total DNA was extracted from six symptomatic tomato plants (two from each valley) and used as template to amplify, clone, and sequence the core region of the begomovirus CP. BLAST analysis of begomovirus sequences available in the NCBI GenBank database indicated the closest match was the Old World monopartite begomovirus Tomato yellow leaf curl virus (TYLCV) from Israel (Accession No. X15656) at 97.8% shared nucleotide (nt) identity. The full-length genome was amplified for each of six isolates using TempliPhi (Amersham Biosciences, Piscataway, NJ) and cloned into the pGEM7 vector (Promega, Madison, WI). The complete DNA genome sequence was determined for eight clones by primer walking. Cloned TempliPhi products sequenced represented two to three isolates from each valley. Results indicated that the isolates (n = 8) were 98.9 to 100% identical (Accession No. DQ631892) to each other, and they shared 98% identity with TYLCV isolates reported from the Caribbean Region and Florida. This highly virulent begomovirus of tomato, originating in Israel, was first reported in Mexico from 1996 to 1997 when it was identified in tomato plants in the Yucatan Peninsula (1) (>1,500 miles from Sinaloa). The latter report followed the introduction of TYLCV in tomato seedlings from Florida into several eastern U.S. states (3,4) and then into Puerto Rico (2). The introduction of TYLCV into Sinaloa where tomato production is highly concentrated is significant because the region supplies the majority (as much as 93%) of fresh-market tomatoes to the western United States from October to June (>$750 million dollars). Of equal importance is the immediate proximity of the pandemic to California where more than 90% of the processing tomatoes in the United States are grown.

References: (1) J. T. Ascencio-Ibáñez et al. Plant Dis. 83:1178, 1999. (2) J. Bird et al. Plant Dis. 85:1028, 2001. (3) M. T. Momol et al. Plant Dis 83:487, 1999. (4) J. E. Polston and P. K. Anderson, Plant Dis. 81:1358, 1997.
Differentiation of Strains of Xylella fastidiosa Infecting Grape, Almonds, and Oleander Using a Multiprimer PCR Assay
Authors: Hernandez-Martinez R. , Costa H. S. , Dumenyo C. K. , Cooksey D. A.
Published: May 2007

Xylella fastidiosa is a xylem-limited, nutritionally fastidious bacterium that causes several plant diseases including Pierce's disease (PD) in grape and leaf scorch in almond (ALS) and oleander (OLS). OLS strains belong to X. fastidiosa subsp. sandyi, PD strains belong to X. fastidiosa subsp. fastidiosa, and strains from almond designated as ALS strains are of two general types belonging either to X. fastidiosa subsp. multiplex or X. fastidiosa subsp. fastidiosa. The ALS strains assigned to X. fastidiosa subsp. multiplex belong to two different genotypes (ALSI and ALSII) below the subspecies level. The OLS strains do not infect grape or almond. PD strains produce diseases in grape, alfalfa, almond, and some weeds, but they do not infect oleander, oak, peach, or citrus. ALS strains that belong to X. fastidiosa subsp. multiplex do not produce disease on grape. In this study, a relatively simple polymerase chain reaction (PCR) based method was developed to distinguish among PD, OLS, and ALS strains. PCR performed with primers XF1968-L and XF1968-R amplified a 638-bp fragment from OLS strains but not from PD strains or ALS strains that belong to X. fastidiosa subsp. fastidiosa. PCR with primers XF2542-L and XF2542-R amplified a 412-bp fragment from PD strains, but not from OLS strains. PCR with primers ALM1 and ALM2 produced a fragment of 521 bp from strains isolated from almond that belong to X. fastidiosa subsp. multiplex. The combination of the three primer sets allowed the distinction of the two ALS genotypes of X. fastidiosa subsp. multiplex. These results are in agreement with those obtained from analysis of sequences of 16S-23S rDNA intergenic spacer regions sequence analysis and with previous results based on randomly amplified polymorphic DNA analysis.