D. J. Vakalounakis, Laboratory of Phytopathological Mycology, Plant Protection Institute, Hellenic Agricultural Organization ‘Demeter’ (NAGREF), P.O. Box 2228, GR-71003 Heraklio, Greece; and
A. G. Doulis, Laboratoty of Plant Biotechnology, Institute of Viticulture, Floriculture and Vegetable Crops, NAGREF, P.O. Box 2228, GR-71003, Heraklio, Greece
In early December 2012 and February 2013, severe symptoms of white rust were observed on several commercial crops of the spinach (Spinacia oleracea L.) cvs. Tahiti and Rembrandt in the Aghia Pelaghia and Elia areas, respectively, of Heraklio, Crete, Greece. Initially, small, chlorotic lesions developed on the upper side of the leaves. As disease progressed, small, glassy white pustules developed on the underside of each leaf, frequently in concentric rings. The pustules were blister-like and oval, irregularly oval, or elongated, ranging from 0.5 to 2.0 mm in diameter and up to 3 mm in length. Lesions often coalesced, and the pustules could cover the entire lower leaf surface, rendering them unmarketable. The dome-like epidermis of host tissue covering the white rust pustules ruptured and exposed a white, chalky ‘dust’ of numerous powdery spores in small, circular to elongate sori. The sporangia were arranged in basipetal chains, were globose to oval, with a smooth wall that was uniformly thick and measured 19.1 ± 1.93 (14.1 to 23.5) μm × 15.3 ± 1.49 (12.9 to 20.0) μm when hydrated. Oospores were absent from the leaves. The morphological characteristics closely resembled those reported for the white rust pathogen, Albugo occidentalis Wilson (4), as well as those of A. occidentalis measured from dried leaves of Chenopodium spp. and Monolepis nuttalliana (Schult.) Greene (Herb. IMI96980, IMI351202, and IMI26345, respectively), kindly loaned by the Royal Botanic Gardens, Kew, United Kingdom. Pathogenicity was confirmed by spraying a suspension of 105 sporangia/ml on 10 healthy 20-day-old potted spinach plants of cv. Tahiti. Inoculated plants were covered with polyethylene bags for 3 days and incubated in a growth chamber at 16 to 20°C with a 10-h photoperiod. White rust symptoms were observed on the lower surface of the leaves 10 days after inoculation. Ten control plants sprayed similarly with distilled water and maintained under the same conditions as inoculated plants showed no symptoms. The fungus present on the inoculated plants was morphologically identical to that originally observed on diseased plants, fulfilling Koch's postulates. Genomic DNA from spinach leaf sori was extracted, and the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA), ITS1-5,8S-ITS2, as well as the cytochrome oxidase subunit II (COX2) mitochondrial gene, were amplified. PCR products were sequenced and deposited in GenBank (KC676794 and KC676795, respectively). In a BLAST search, the ITS1-5,8S-ITS2 and COX2 sequences showed 99% similarities to 684 bp (AJ553900.1) and 599 bp (AY286220.1) sequences of the corresponding A. occidentalis genes in GenBank, respectively. Based on morphological characteristics, pathogenicity tests, and molecular sequencing data, it was concluded that the pathogen on spinach in Crete is A. occidentalis. This is an economically important pathogen of spinach in the United States, that has also has been recorded in Iran (2) and India on a Chenopodium sp. (IMI351202), and in Canada on M. nuttalliana (IMI26345). To our knowledge, this is the first report of this pathogen in Greece, and the first record on spinach in Europe. A voucher specimen has been deposited at the Royal Botanic Gardens, Kew, United Kingdom (Accession No. K(M) 181610).
References: (1) J. C. Correll et al. Eur. J. Plant Pathol. 129:193, 2011. (2) A. G. Ebrahimi and H. Afzali. Rostaniha 1:73, 2000. (3) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. American Phytopathological Society, St. Paul, MN, 1989. (4) G. W. Wilson. Bull. Torrey Bot. Club 34:61, 1907.