REAL, an LTR retrotransposon from the plant pathogenic fungus Alternaria alternata

A retrotransposon was isolated and characterized from strain 15A of the Japanese pear pathotype of Alternaria alternata, which causes black spot disease in certain cultivars of Japanese pear by producing a host-specific toxin known as AK-toxin. The element, which we have named REAL (Retrotransposon of Alternaria alternata), is 6046 bp in size and contains direct long terminal repeats (LTRs) of 218 bp. Target-site duplication of 5 bp was found. REAL contains two long overlapping ORFs. The first ORF shows homology to retroviral gag genes. The second ORF has homology to protease, reverse transcriptase, RNase H and integrase domains of the retroelement pol genes, in that order. Phylogenetic comparison of reverse transcriptase domains from retrotransposons placed REAL in the Ty3/gypsy group of LTR retrotransposons, most closely related to grasshopper from Magnaporthe grisea. Northern analysis detected REAL transcripts of about 2.0 and 6.0 kb. The 6.0-kb species corresponds to a full-length transcript of the element. The element was found by Southern analysis in 12 out of 13 strains of the Japanese pear pathotype, and the banding patterns, copy numbers and signal intensities in these strains were variable. REAL-related elements were also found in some, but not all, of the other strains tested, including nonpathogenic A. alternata and other pathotypes, which cause diseases on different plant species by producing distinct hostspecific toxins. These results suggest that the distribution of REAL in A. alternata is not pathotype specific.