First Report of Leaf Spot of Dracaena reflexa Caused by Burkholderia gladioli Worldwide.

In December 2008 (austral summer), a new disease of Dracaena reflexa Lam. cv. Anita was observed in a postentry quarantine greenhouse near Auckland, New Zealand on plants imported from Costa Rica. Symptoms included rust-colored, water-soaked lesions with chlorotic margins approximately 5 by 10 mm. When the disease was first noticed, incidence approached 80%, but subsequent reduction in greenhouse temperature dramatically reduced symptom expression and lesions were only visible on some leaf tips. Bacteria consistently isolated from the lesions on King's medium B (KB) were cream-colored, shiny, and produced a yellow, diffusible, nonfluorescent pigment. All isolates were able to rot onion slices. On the basis of BIOLOG (Hayward, CA) carbon utilization profiles, isolates were initially identified as Burkholderia gladioli (Severini 1913) Yabuuchi et al. 1993 with a probability index of 100% and a similarity index of 0.85. For molecular identification, a near full-length sequence of the 16S rDNA gene was amplified from all isolates using primers fD2 and rP1 (1), obtaining a PCR product of approximately 1,500 bp. The nucleotide sequences were 100% identical to a number of B. gladioli GenBank entries, including Accession Nos. EF193645 and EF088209. To confirm pathogenicity, three isolates (two isolated prior to greenhouse temperature reduction and one after) were used. Three D. reflexa plants were inoculated per bacterial isolate by wounding three young fully expanded leaves on each plant (four wounds per leaf) and spraying the leaves with a bacterial suspension in sterile distilled water at 108 CFU/ml. At the same time, Gladiolus nanus plants were inoculated in a similar manner. Control plants (D. reflexa and G. nanus) were wounded and sprayed with sterile distilled water. All inoculated plants were covered with plastic bags to maintain humidity and placed in a growth chamber at 25°C. At 3 days, all inoculated plants began to show water soaking and reddish coloration around the inoculation sites, and by 7 days, the lesions had expanded to resemble natural infection. Bacteria isolated on KB from the leading edge of each lesion were morphologically identical to the initial isolates. No bacteria were recovered from the wound sites on the control plants. The 16S rDNA sequences of selected isolates from inoculated plants showed 100% identity to the sequences of the initial isolates, thereby fulfilling Koch's postulates. To our knowledge, this is the first report of B. gladioli causing leaf spot of D. reflexa in the world. Reference: (1) W. G. Weisburg et al. J. Bacteriol. 173:697, 1991.

First Report of Potato spindle tuber viroid in Cape Gooseberry (Physalis peruviana) in New Zealand.

In February 2009, 10 cape gooseberry plants (Physalis peruviana) grown from seed on a domestic property in Christchurch, New Zealand, showed severe leaf distortion, fasciation and etiolation of growing tips, and weak flowering. Symptoms were first observed in the emerging seedlings. No virus particles were observed in sap from infected plants with the electron microscope. Total RNA was isolated from leaves of the 10 plants with a Qiagen RNeasy Plant Mini Kit (Valencia, CA). All 10 plants tested positive for Potato spindle tuber viroid (PSTVd) by real-time reverse transcription (RT)-PCR (1) and by RT-PCR with PSTVd-specific primers (3) and generic pospiviroid primers (4). For both conventional PCRs, the expected 359-bp amplicons were sequenced directly and sequences were aligned together to create a consensus sequence (GenBank Accession No. FJ797614). BLASTn analysis showed 98% nucleotide identity to PSTVd (EU862231, DQ308556, X17268, and AY532801-AY532804). Sap from one of the infected plants was mechanically inoculated onto healthy P. peruviana, Solanum lycopersicum 'Rutgers', Chenopodium amaranticolor, C. quinoa, Cucumis sativum 'Crystal Apple', Gomphrena globosa, Nicotiana benthamiana, N. clevelandii, N. occidentalis '37B', N. tabacum 'WB', N. sylvestris, and Phaseolus vulgaris 'Prince'. After 4 weeks, the leaves of the 'Rutgers' tomato plants were showing severe distortion, purpling, and necrosis of mid-veins and P. peruviana plants were showing distortion of newly emerging apical leaves. Healthy control P. peruviana were asymptomatic. Symptoms appeared milder than that observed in the original P. peruviana plants, but this may be related to different environmental conditions or age or growth stage of the plants when inoculated. All other indicator plants were symptomless, but along with P. peruviana, tested positive for PSTVd by real-time RT-PCR (1). The presence of PSTVd was further confirmed in one original symptomatic and the mechanically inoculated P. peruviana plants and in the indicator plants by dot-blot hybridization with a digoxygenin-labeled synthetic ssRNA probe specific to the full-length PSTVd genome. PSTVd has been reported in New Zealand previously in commercial glasshouse crops of tomatoes and peppers (2), but was eradicated and so remains a regulated pest. The plants were grown from seeds imported from Germany and it is possible that the infection was seedborne. PSTVd was reported in young cape gooseberry seedlings in Germany and Turkey but the infection was asymptomatic (5). Symptoms were associated with the PSTVd-infected cape gooseberry in New Zealand. To our knowledge, this is the first report of the viroid in domestically grown plants in New Zealand, and only the second report of PSTVd in cape gooseberry worldwide. Our findings suggest that this species is an emerging host for PSTVd and that dissemination of seed may provide a pathway for international movement of the viroid. References: (1) N. Boonham et al. J. Virol. Methods 116:139, 2004. (2) B. S. M. Lebas et al. Australas. Plant Pathol. 34:129, 2005. (3) A. M. Shamoul et al. Can. J. Plant Pathol. 19:89, 1997. (4) J. T. H. Verhoeven et al. Eur. J. Plant Pathol. 110:823, 2004. (5) J. T. H. Verhoeven et al. Plant Dis. 93:316, 2009.

First Report of Passiflora latent virus in Banana Passionfruit (Passiflora tarminiana) in New Zealand.

Passiflora latent virus (PLV) naturally infects cultivated and wild Passiflora species in Australia, Germany, Israel and the United States (1-3). In March 2004, chlorotic lesions were observed on leaves of three vines of Passiflora tarminiana on one site in Auckland, New Zealand. Chenopodium amaranticolor and C. quinoa inoculated with sap from symptomatic leaves developed chlorotic local spots, followed by systemic leaf chlorosis and necrosis. Local symptoms appeared more quickly on C. quinoa (12 days) than on C. amaranticolor (20 days). No symptoms were observed on inoculated plants of Nicotiana benthamiana, N. clevelandii, N. occidentalis, N. tabacum, or Phaseolus vulgaris. Electron microscopy of crude sap preparations from infected C. quinoa, C. amaranticolor, N. occidentalis, and P. tarminiana showed flexuous, filamentous virus particles approximately 650 nm long. Plants of P. tarminiana and the three inoculated indicator species containing virus particles tested positive by PLV polyclonal antiserum in double-antibody sandwich (DAS)-ELISA (DSMZ, Braunschweig, Germany) and immunosorbent electron microscopy (Stephan Winter, DSMZ, personal communication). Nucleic acid was extracted from leaves of plants of each of the four viruliferous species with an RNeasy Plant Mini Kit (Qiagen, Doncaster, Australia) and then used in reverse transcription (RT)-PCR tests with novel forward (5'-CGAGACACACGCAAACGAA-3') and reverse (5'-CAGCAAAGCAAAGACACGA-3') primers specific to a 523-bp fragment of the PLV polyprotein. PCR products of the expected size were obtained, and an amplicon from P. tarminiana was directly sequenced (GenBank Accession No. EU257510). A BLAST search in GenBank showed 94% nucleotide sequence identity with a PLV isolate from Israel (GenBank Accession No. DQ455582). To our knowledge, this is the first finding of PLV in P. tarminiana and the first report of the virus in New Zealand. Chenopodium spp. have been reported previously as experimental hosts (2,3), and this study revealed that N. occidentalis also can be infected latently with PLV. P. tarminiana is a weed in New Zealand and subject to active control measures to manage the species. Economically important species such as P. edulis and P. ligularis are potentially susceptible to the virus. These species are not grown commercially in the surrounding area but are common in domestic Auckland gardens. Infected vines were removed from the site and destroyed, and symptomatic vines have not been observed at other sites. References: (1) R. D. Pares et al. Plant Dis. 81:348, 1997. (2) S. Spiegel et al. Arch. Virol. 152:181, 2007. (3) A. A. Stihll et al. Plant Dis. 76:843, 1992.

Occurrence of Hibiscus chlorotic ringspot virus in Hibiscus spp. in New Zealand.

Hibiscus spp. are popular ornamental plants in New Zealand. The genus is susceptible to Hibiscus chlorotic ringspot virus (HCRSV), a member of the genus Carmovirus, which has been reported in Australia, El Salvador, Singapore, the South Pacific Islands, Taiwan, Thailand, and the United States (1-4). In May of 2004, chlorotic spotting and ringspots were observed on the leaves of two H. rosa-sinensis plants in a home garden in Auckland, New Zealand. When inoculated with sap from symptomatic leaves, Chenopodium quinoa and C. amaranticolor developed faint chlorotic local lesions 12 to 15 days later. Phaseolus vulgaris exhibited small necrotic local spots 10 days postinoculation. No symptoms were observed on inoculated plants of Cucumis sativus, Gomphrena globosa, Nicotiana Clevelandii, N. tabacum, or N. sylvestris. Plants of H. rosa-sinensis and the three symptomatic indicator species tested positive for HCRSV using polyclonal antiserum (Agdia Inc., Elkhart, IN) in a double antibody sandwich (DAS)-ELISA. Forward (5'-GGAACCCGTCCTGTTACTTC-3') and reverse (5'-ATCACATCCACATCCCCTTC-3') primers were designed on the basis of a conserved region in the coat protein gene (nt 2722-3278) of HCRSV isolates in GenBank (Accession Nos. X86448 and DQ392986). A product of the expected size (557 bp) was amplified by reverse transcription (RT)-PCR with total RNA extracted from the four infected species. Comparison of the sequence of the amplicon from H. rosa-sinensis (GenBank Accession No. EU554660) with HCRSV isolates from Singapore and Taiwan (GenBank Accession Nos. X86448 and DQ392986) showed 99 and 94% nucleotide identity, respectively. From 2006 to 2008, samples from a further 25 symptomatic hibiscus plants were collected from different locations in the Auckland region. Nineteen, including plants of H. diversifolius, H. rosa-sinensis, and H. syriacus, tested positive for HCRSV by RT-PCR. To our knowledge, this is the first report of HCRSV in New Zealand and of the virus in H. diversifolius and H. syriacus. HCRSV is considered to be widespread in New Zealand. References: (1) A. A. Brunt et al. Plant Pathol. 49:798, 2000. (2) S. C. Li et al. Plant Pathol. 51:803, 2002. (3) H. Waterworth. No.227 in: Descriptions of Plant Viruses. CMI/AAB, Surrey, UK, 1980. (4) S. M. Wong et al. Acta Hortic. 432:76, 1996.

Relationship of severe dysplasia to stage A (incidental) adenocarcinoma of the prostate.

Totally embedded radical prostatectomy specimens and their transurethral resections (TUR) from 32 Stage A (incidental) carcinomas were studied for severe dysplasia. These cases were selected to be representative of the majority of Stage A cancers and hence consisted of cases with predominantly low-grade centrally located tumor. In 43.75% of the cases, severe dysplasia was minimal in amount, 43.75% intermediate, and 12.5% extensive. Severe dysplasia was spatially associated with the main tumor in only 31.25% of cases. In 43.75% of the cases all of the severe dysplasia was isolated (i.e., not adjacent or intermingled with any carcinoma). Whereas this study confirms one general association between severe dysplasia and the presence of carcinoma within the gland, the data do not strongly support the concept that the dysplasia is a direct precursor to centrally located low-grade Stage A carcinomas. From a practical viewpoint, severe dysplasia in the TUR was seen in 15.6% of cases and was not predictive of the amount of tumor within the gland.

Relationship of severe dysplasia to stage B adenocarcinoma of the prostate.

Severe dysplasia was studied in totally embedded radical prostatectomy specimens from 40 Stage B, intermediate-grade prostate cancers. All 40 cases had severe dysplasia which varied in amount (extensive in 35% of cases; intermediate in 55%; minimal in 10%). Severe dysplasia was predominantly concentrated in peripheral and posterior regions of the gland. Those cases with extensive severe dysplasia had more multifocal small cancers (14.6/case) compared with cases with intermediate (7.5/case) or minimal severe dysplasia (5.5/case). Most dominant tumor nodules had some intermingled severe dysplasia, and in 30% of the dominant tumor nodules, intermingled severe dysplasia occupied between 10% and 25% of the tumor nodule. Severe dysplasia was located next to the dominant tumor nodule in all cases. Nineteen percent of dominant tumor nodules had extensive adjacent severe dysplasia ranging from 5 to 14 mm in maximum diameter, showing that the potential exists for needle biopsy of prostate cancers to obtain only severe dysplasia. These data show a close relationship between severe dysplasia and Stage B cancers. In contrast, centrally located low-grade multifocal cancers, as seen in Stage A (incidental) carcinoma, had a significantly weaker association with severe dysplasia as compared with central, intermediate-grade, multifocal tumors or peripheral multifocal tumors of any grade.

Intradermal nodular fasciitis presenting as an eyelid mass.

BACKGROUND: Nodular fasciitis is a fibroblastic proliferation in which nodules, most commonly appearing on the extremities, develop suddenly and rapidly. Although excisional biopsy is curative, the nodules will often resolve spontaneously; however, the histologic appearance of a pleomorphic spindle cell neoplasm with frequent mitotic figures may raise concern of a more malignant neoplasm and lead to unnecessary and overly aggressive therapy. METHODS: A case of nodular fasciitis presenting in an unusual location, intradermally at the lateral canthus, is discussed. RESULTS: Review of the recent literature from other disciplines reveals new insights into the etiology and diagnostic options (fine needle aspiration, tomography, other imaging techniques) for this puzzling disease. CONCLUSIONS: When nodular fasciitis occurs in an unusual location, such as on the head and neck or at an intradermal location, it will not present as the deep, mobile, nontender nodule described in the dermatologic literature. Fine-needle aspiration and modern imaging techniques may help in the diagnosis and prevent unnecessary surgery for a self-limited condition.