First report of Didymella americanaon baby lima bean (Phaseolus lunatus)

A new foliar disease was observed on baby lima bean (Phaseolus lunatus) in fields across western New York State, USA. The disease occurred in 10 fields with variable incidence and severity. Symptoms were initially necrotic, tan spots on leaves with red to reddish brown irregular margins that coalesced to encompass the entire leaf and cause abscission. Pycnidia were observed within the lesions. Isolations from diseased leaves yielded several pycnidial forming fungi, including a Didymella species. These isolates were characterized by morphology and sequencing of multiple reference genes (internal transcribed spacer (ITS), partial actin, β- tubulin (tub2), translation elongation factor 1-α (TEF), 28S rDNA large subunit (LSU), rpb2, and calmodulin). A four gene phylogeny (ITS, tub2, LSU, and rpb2) showed that the isolates from baby lima bean belonged to a well-supported clade that contained the type culture of Didymella americana. Pathogenicity of the isolates on three commonly grown cultivars of baby lima bean was confirmed. Symptoms that developed on inoculated plants were similar to those observed on diseased plants in the field. This is the first report of D. americana on baby lima bean.

High-resolution electrophoretic procedures for the identification of five Eimeria species from chickens, and detection of population variation

To overcome limitations of conventional approaches for the identification of Eimeria species of chickens, we have established high resolution electrophoretic procedures using genetic markers in ribosomal DNA. The first and second internal transcribed spacer (ITS-1 and ITS-2) regions of ribosomal DNA were amplified by polymerase chain reaction (PCR) from genomic DNA samples representing five species of Eimeria (E. acervulina, E. brunetti, E. maxima, E. necatrix and E. tenella), denatured and then subjected to denaturing polyacrylamide gel electrophoresis (D-PAGE) or single-strand conformation polymorphism (SSCP) analysis. Differences in D-PAGE profiles for both the ITS-1 and ITS-2 fragments (combined with an apparent lack of variation within individual species) enabled the unequivocal identification of the five species, and SSCP allowed the detection of population variation between some isolates representing E. acervulina, which remained undetected by D-PAGE. The establishment of these approaches has important implications for controlling the purity of laboratory lines of Eimeria, for diagnosis and for studying the epidemiology of coccidiosis.

Dry flower disease of Macadamia in Australia caused by Neopestalotiopsis macadamiae sp. nov. and Pestalotiopsis macadamiae sp. nov

Incidence of dry flower disease of macadamia (Macadamia integrifolia), expressed as blight of the flowers, necrosis and dieback of the rachis, is increasing in Australia. In the 2012/13 production season, incidence of dry flower disease resulted in 10% to 30% yield loss in the affected orchards. Etiology of the disease has not been established. This study was established to characterise the disease and identify the causal pathogen. A survey of the major macadamia producing regions in Australia revealed dry flower disease symptoms, regardless of cultivar or location at all stages of raceme development. Based on colony and conidial morphology, the majority (41%) of fungal isolates obtained from tissue samples were identified as Pestalotiopsis and Neopestalotiopsis spp. The phylogeny of the combined partial sequence of the internal transcribed spacer, beta-tubulin and translation elongation factor 1-alpha gene loci, segregated the isolates into two well supported clades, independent of location or part of the inflorescence affected. Further morphological examination supported the establishment of two new species, which are formally described as Neopestalotiopsis macadamiae sp. nov. and Pestalotiopsis macadamiae sp. nov. Using spore suspensions of isolates of both species, Koch?s postulates were fulfilled on three macadamia cultivars at all stages of raceme development. To our knowledge, this is the first report of species of Neopestalotiopsis and Pestalotiopsis as causal agents of inflorescence disease in macadamia.