Phytophthora hedraiandra Detected from Irrigation Water at a Perennial Ornamental Plant Nursery in Virginia.

Abstract

Water survey for Phytophthora spp. by baiting with rhododendron leaves in April 2006 at a perennial ornamental plant nursery in Virginia detected five isolates showing a unique, previously unknown single-strand conformation polymorphism (SSCP) fingerprint (1). These cultures were isolated from two reservoirs at different depths of water column from surface to 2 m. They were homothallic and produced smooth-surfaced spherical oogonia with an average diameter of 27 μm on 10% V8 agar. Oospores were aplerotic. The paragynous antheridia were averaging 12 μm in diameter. Sporangia were papillate, spherical to ovoid, averaging 39 by 28 μm (length by width). They were caducous with short (<4 μm) pedicels. Chlamydospores and hyphal swellings were not observed. Two isolates were sequenced for rDNA internal transcribed spacer (ITS) 1 and 2 regions and cytochrome oxidase subunit 1 (Cox 1) gene. ITS sequences of both isolates (GenBank Accession Nos. JN376065 and JN376066) were identical to that of Phytophthora hedraiandra type culture (GenBank Accession No. AY707987). Also, the Cox 1 sequence of an isolate (Accession No. JN376067) had 99% homology with that of the type culture (GenBank Accession No. AY69115). Pathogenicity of both isolates was tested on Rhododendron catawbiense and Viburnum tinus, two known hosts of P. hedraiandra (2). For each isolate and host, five leaves and stems on potted plants were wounded by needles and then inoculated by placing over each wound a 5-mm2 mycelial plug from a 7-day-old culture and securing with Parafilm. V8 agar was used instead of mycelial plugs on control plants. After inoculation, each plant was enclosed in a plastic bag for 1 day and then incubated at 22°C with a 12-h photoperiod. Distilled water was sprayed daily for 5 days postinoculation (dpi) until disease symptoms were observed. At 15 dpi, 3 of the 10 inoculated rhododendron leaves and 6 of the 10 stems showed leaf lesions, wilting, dieback, and cankers, eventually leading to rhododendron death. Two of the 10 viburnum leaves and 4 of the 10 stems showed similar symptoms. Leaf lesions were approximately 3 to 5 cm in diameter. P. hedraiandra was recovered from diseased tissues and all resulting cultures showed an identical SSCP fingerprint to tested isolates as well as a P. hedraiandra isolate from Minnesota (3). No symptom developed on control plants. To our knowledge, this is the first report of P. hedraiandra in Virginia. Considering neither host plant has been grown or bought for resale by this nursery, this study indicates that P. hedraiandra may have a wider host range than is currently known. This possibility and the importance of water dispersal for P. hedraiandra in disease epidemiology warrant further investigation.