A phylogenetic and taxonomic re-evaluation of the Bipolaris - Cochliobolus - Curvularia Complex

Three genera, Cochliobolus, Bipolaris and Curvularia form a complex that contains many plant pathogens, mostly on grasses (Poaceae) with a worldwide distribution. The taxonomy of this complex is confusing as frequent nomenclatural changes and refinements have occurred. There is no clear morphological boundary between the asexual genera Bipolaris and Curvularia, and some species show intermediate morphology. We investigated this complex based on a set of ex-type cultures and collections from northern Thailand. Combined gene analysis of rDNA ITS (internal transcribed spacer), GPDH (glyceraldehyde 3-phosphate dehydrogenase), LSU (large subunit) and EF1-α (translation elongation factor 1-α) shows that this generic complex divides into two groups. Bipolaris and Cochliobolus species clustered in Group 1 along with their type species, whereas Curvularia species (including species named as Bipolaris, Cochliobolus and Curvularia) clustered in Group 2, with its generic type. The nomenclatural conflict in this complex is resolved giving priority to the more commonly used established generic names Bipolaris and Curvularia. Modern descriptions of the genera Bipolaris and Curvularia are provided and species resolved in this study are transferred to one of these genera based on their phylogeny. © 2012 Mushroom Research Foundation.

Polyphasic characterization of four new plant pathogenic Phyllosticta species from China, Japan, and the United States

The black rot disease of Vitis species and other host genera of Vitacease is caused by Phyllosticta ampelicida and allied taxa which is considered to be a species complex. In this paper, we introduce four new species of Phyllosticta, including two from the P. ampelicida complex, based on a polyphasic characterization including disease symptoms and host association, morphology, and molecular phylogeny. The phylogenetic analysis was conducted based on the ribosomal internal transcribed spacer (ITS) region and a combined multi-locus alignment of the ITS, actin (ACT), partial translation elongation factor 1-alpha (TEF-1), and glyceraldehydes 3-phosphate dehydrogenase (GPDH) gene regions. Our study confirms the phylogenetic distinctions of the four new species, as well as their phenotypic differences with known species in the genus.

Successful fertility experiments with cryopreserved spermatozoa of barramundi, Lates calcarifer (Bloch), using dimethylsulfoxide and glycerol as cryoprotectants

The fertility of cryopreserved Lates calcarifer sperm was studied to increase the availability of semen for routine fertilization of stripped eggs and to provide a tool for selective breeding. Semen diluted (1:4 v/v) and frozen (-196 degrees C) with 5% dimethylsulfoxide (DMSO) or 10% glycerol (final concentration) as cryoprotectants was used to inseminate freshly stripped ova. Frozen-thawed sperm were motile for about 4 min after being mixed with seawater. In the DMSO medium, post-thaw sperm activation was immediate after dilution with seawater, but in the glycerol medium maximum motility intensity was delayed for up to 1 min. When eggs and sperm were mixed before the addition of seawater, semen frozen with DMSO as cryoprotectant gave a mean hatch rate (84.1%) no different (P > 0.05) from that of unfrozen semen diluted with Ringer's solution (80.7%) or with DMSO (83.7%), but higher (P < 0.05) than that of semen frozen with glycerol (60.9%). Adding sperm to seawater 30 s before mixing with eggs did not improve the fertility of sperm cryopreserved with glycerol. Eggs inseminated with glycerol-cryoprotected sperm showed higher mortality during incubation than those inseminated with DMSO-cryoprotected sperm. Sperm held in liquid nitrogen for 90 days with DMSO as cryoprotectant yielded acceptable fertilization and hatching rates with semen-to-ova ratios of up to 1:100 (v/v) , and produced fish with no apparent abnormalities over a 29-day period after hatch. These results show that cryopreservation of L. calcarifer sperm is feasible and well suited to a variety of hatchery purposes.

Phosphine resistance in India is characterised by a dihydrolipoamide dehydrogenase variant that is otherwise unobserved in eukaryotes

Phosphine (PH3) fumigation is the primary method worldwide for controlling insect pests of stored commodities. Over-reliance on phosphine, however, has led to the emergence of strong resistance. Detailed genetic studies previously identified two loci, rph1 and rph2, that interact synergistically to create a strong resistance phenotype. We compared the genetics of phosphine resistance in strains of Rhyzopertha dominica and Tribolium castaneum from India and Australia, countries having similar pest species but widely differing in pest management practices. Sequencing analysis of the rph2 locus, dihydrolipoamide dehydrogenase (dld), identified two structurally equivalent variants, Proline49>Serine (P49S) in one R. dominica strain and P45S in three strains of T. castaneum from India. These variants of the DLD protein likely affect FAD cofactor interaction with the enzyme. A survey of insects from storage facilities across southern India revealed that the P45/49S variant is distributed throughout the region at very high frequencies, in up to 94% of R. dominica and 97% of T. castaneum in the state of Tamil Nadu. The abundance of the P45/49S variant in insect populations contrasted sharply with the evolutionary record in which the variant was absent from eukaryotic DLD sequences. This suggests that the variant is unlikely to provide a strong selective advantage in the absence of phosphine fumigation.