First Report of Root and Stem Rot Caused by Phytophthora tentaculata on Mimulus aurantiacus in North America

Sticky monkey flower plant, Mimulus aurantiacus (Phrymaceae), is a small, perennial shrub that is widely distributed throughout California, especially in coastal and disturbed habitats. It is also found in native plantings in parks and landscapes. In October 2012, nearly all the M. aurantiacus plants grown in a Monterey County, CA nursery for a restoration project were stunted and had dull, yellowish leaves. Roots and stem collars had necrotic, sunken lesions with few feeder roots. Thirty percent of the plants had died. Samples of diseased plants were sent to the CDFA-PPDC Lab and tested positive for Phytophthora sp. using the Agdia ELISA Phytophthora kit (Agdia, Elkhart, IN). A Phytophthora sp. was consistently isolated from the tissue on corn meal agar-PARP (CMA-PARP) (2). Sporangia were spherical to ovoid, papillate to bipapillate and 17 to 42.5 (avg. 27.5) × 12 to 35 (avg. 22.9) μm, with a length/breadth ratio of 1.2:1. Chlamydospores, which were spherical, terminal to intercalary, thin walled and 27.5 to 40 μm, and hyphal swellings formed on CMA-PARP. Spherical oospores, 25 to 36 μm, with primarily paragynous antheridia formed readily on V8 juice agar. rDNA sequences of the internal transcribed spacer (ITS) region of the isolates (GenBank KF667505), amplified using primers ITS1 and ITS4, were 100% identical to Phytophthora tentaculata (CBS 552.96, GenBank AF266775) by a BLAST query (1,3). To assess pathogenicity, exposed root crowns of three 3.78-liter potted M. aurantiacus plants were inoculated with 20 ml of zoospore suspension (2 × 104 ml−1). Plants were maintained in a 23°C growth chamber with a 12-h photoperiod and watered daily. Sterile water was applied to the exposed crowns of three control plants. At 2 weeks, all inoculated plants were wilted with chlorotic foliage. After 3 weeks, the cortical tissue of the crowns and roots was discolored and sloughing and P. tentaculata was recovered on CMA-PARP. P. tentaculata did not grow from the asymptomatic control plants. Inoculations were repeated with similar results. P. tentaculata is a homothallic species in Phytophthora clade 1 that causes crown, root, and stalk rot of nursery plants in Europe and China (1,4). A USDA PERAL analysis lists it as one of the top 5 Phytophthora species of concern to the United States (4). Genera infected with P. tentaculata include Apium, Aucklandia, Chicorium, Chrysanthemum, Delphinium, Gerbera, Lavandula, Santolina, Origanum, and Verbena (4). To our knowledge, this is the first report of P. tentaculata in North America. The source of inoculum of P. tentaculata in California remains unknown. The nursery used seed and cuttings of M. aurantiacus from nearby native areas for propagation, and P. tentaculata was not found in neighboring plant hosts or by baiting soil and water at the nursery. All infected M. aurantiacus material was destroyed. The presence of P. tentaculata in California nurseries could have serious economic impacts on the nursery industry and environmental impacts on susceptible native hosts, if spread into the wildlands. References: (1) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (2) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986. (3) H. Krober and R. Z. Marwitz. Pflanzenkrankh. Pflanzenschutz 100:250, 1993. (4) U.S. Department of Agriculture, Animal and Plant Health Inspection Services (APHIS). Phytophthora species in the Environment and Nursery Settings New Pest Response Guidelines, USDA-APHIS-PPQ-Emergency and Domestic Programs-Emergency Management, Riverdale, MD, 2010.

Ultrastructural features of Mimulus aurantiacus (Scrophulariaceae) pollen tubes in vivo

The aim of this study is to give information on ultrastructure of in vivo pollen tubes of Mimulus aurantiacus which were collected from the Botanical Garden of the University of California at Berkeley. Materials were prepared according to electron microscopy methods and examined under Zeiss electron microscope. Four zones were examined in the pollen tubes of Mimulus aurantiacus. APICAL ZONE: Mitochondria, smooth endoplasmic reticulum, rough endoplasmic reticulum, dictyosomes and secretory vesicles were observed. SUBAPICAL ZONE: This area contained abundant rough endoplasmic reticulum and occasionally some smooth endoplasmic reticulum. The polysomes, mitochondria, proplastids that contain starch, small vacuoles and a few lipid bodies were detected. NUCLEAR ZONE: Both generative and vegetative cell nuclei lie in this zone. The vegetative cell nucleus was large and long. Rough endoplasmic reticulum, mitochondria, ribosomes, dictyosomes, and amyloplasts that are rich of starch were observed. VACUOLATION AND PLUG FORMATION ZONE: Cytoplasm of the tubes was full of large vacuoles. Few organelles such as mitochondria, dictyosome and rough endoplasmic reticulum were detected along their periphery.