First Report of Fusarium andiyazi Associated with Rice Bakanae in Italy

Abstract

Rice (Oryza sativa L.) is cultivated on approximately 230,000 ha in northern Italy. Since 2001, increasing economical losses presumably caused by Fusarium fujikuroi Nirenberg (Gibberella fujikuroi mating population C), an exotic fungus known as the etiological agent of Bakanae disease, have been reported in Italy. The spread of this disease is primarily seedborne. In 2009, during an annual survey of Italian rice seed, 69 samples were tested for the presence of strains belonging to the G. fujikuroi species complex. Four hundred seeds per sample were surface sterilized and then placed in 90-mm Petri dishes containing potato dextrose agar and incubated for 7 days at 21°C. Thirty two putative G. fujikuroi strains were single-spore purified and identified on the basis of their morphological features on Spezieller Nährstoffarmer agar plates with a piece of sterile filter paper. Strains were characterized at species level by morphological observations (1,2) and translation elongation factor 1-α (TEF) gene sequencing. Unexpectedly, 60% of the strains evaluated belonged to the species F. andiyazi Marasas, Rheeder, Lampr., K.A. Zeller & J.F. Leslie. This fungus, first described on sorghum (Sorghum bicolor L.) in Africa and the United States (1), has been reported to be one of the species associated with Bakanae in Asia and Africa (3). Two F. andiyazi strains, (E432 and E439), isolated in the district of Modena were chosen for pathogenicity testing and their TEF gene sequences were deposited in GenBank (Accession Nos. GU827420 and GU827419). A conidial suspension was produced on Mung-bean liquid media and adjusted to a concentration of 1 × 106 CFU/ml. Italian cv. Galileo was used in the test because of its high susceptibility to Bakanae (Ente Nazionale delle Sementi Elette, Verona, Italy, data unpublished). Rice seeds were heat sterilized for 20 min at 60°C, submerged for 30 min in the conidial suspensions, dried, and subjected to a blotter test. Uninoculated, sterilized seeds served as a control. Seeds were incubated for 15 days in a growth chamber (26°C, 80% relative humidity, and 12-h photoperiod). For each strain, the experiment was repeated three times on samples of 25 seedlings. Results were analyzed by analysis of variance and Tukey test. Symptoms consisted of a generic seedling wilt, a root length reduction ranging from 21 to 48%, and the presence of root discoloration. Seed germination was reduced by 9%. Shoot development was not significantly altered. Proof of pathogenicity was obtained through reisolation of F. andiyazi from symptomatic tissues. To our knowledge, this is the first report of F. andiyazi on rice in Europe. References: (1) W. F. O. Marasas et al. Mycologia 93:1203, 2001. (2) H. I. Niremberg and K. O'Donnell. Mycologia 90:434, 1998. (3) E. G. Wulff et al. Environ. Microbiol. 12:649, 2009.