A triglyceride lipase gene (LIP1) was cloned from Stagonospora nodorum, the causal agent of wheat glume blotch. LIP1 encodes a 561 amino acid preproprotein with a predicted N‐terminal signal peptide. Its expression was up‐regulated during plant infection and in culture media supplemented with saturated fatty glycerides. The recombinant Lip1 protein possessed lipolytic activity against a broad range of lipid substrates. When applied to wheat leaves, recombinant Lip1 decreased the hydrophobicity of the leaf surface, probably by liquefaction of epicuticular wax. Pretreatment of wheat leaves with Lip1 decreased the rate of conidial adhesion from 69·5% to 22·7% and from 58·9% to 28·4% in two independent assays based on different protocols. LIP1 replacement strains showed decreased lipolytic activity on culture media relative to the wild‐type strain, and adhesion of the conidia to the wheat leaf surface was impaired in the gene replacement strains. In two experiments, adhesion rates were 54·3% and 41·6% in the LIP1 replacement strains, as opposed to 77·7% and 66·6%, respectively, in the wild‐type. Collectively, the data demonstrate that the secreted lipase Lip1 is important for the adhesion of S. nodorum infection to wheat leaves.

Feng, J., Hwang, R., Chang, K. F., Conner, R. L., Hwang, S. F., Strelkov, S. E., Gossen, B. D., McLaren, D. L. and Xue, A. G. 2011. Identification of microsatellite markers linked to quantitative trait loci controlling resistance to Fusarium root rot in field pea. Can. J. Plant Sci. 91: 199–204. Fusarium root rot, caused by Fusarium solani (Mart.) Sacc. f. sp. pisi (F. R. Jones) W. C. Snyder & H. N. Hans, is the most common root disease of field pea (Pisum sativum L.) in western Canada. In this study, a recombinant inbred line (RIL) population (n=71) of field pea, derived from crosses between a resistant cultivar Carman, and a susceptible cultivar Reward, was evaluated to identify quantitative trait loci (QTL) controlling resistance to Fusarium root rot. The parental genotypes and RILs were evaluated for resistance to root rot following inoculation with F. solani in field experiments during 2007 and 2008. The frequency distribution of disease severities among the RILs was continuous. Transgressive segregation for resistance was observed among the RILs, with five lines more resistant than Carman, but no lines were more susceptible than Reward. To identify DNA markers linked with the resistance, 213 microsatellite markers were screened with genomic DNA from the two parental cultivars. Only 14 markers were polymorphic between the two parents and were used to genotype each of the RILs. Quantitative trait loci analysis based on the mean disease severity data from 2007 and 2008 identified a QTL that explained 39.0% of the phenotypic variance in the RIL population. This QTL is flanked by markers AA416 and AB60 on linkage group VII. The microsatellite markers that are closely linked to this QTL may be useful for marker assisted selection to develop cultivars with superior Fusarium root rot resistance.

A rapid and efficient protocol for the extraction of genomic DNA from plant pathogenic fungi was developed. Key features of the protocol include the SDS-assisted lysis of fungal mycelium with inclusion of a glass bead to help break hyphal walls, followed by isopropanol precipitation of the DNA. The protocol was used to extract genomic DNA from a collection of 26 fungal species, representing many important plant pathogens. Yield of DNA ranged from 2.1-4.9 g per 20 mg of mycelium or 0.4-0.6 g per 20 mg of spores. The DNA was of sufficient purity to be digested by restriction enzymes, to serve as a template in the PCR-amplification of genomic fragments as large as 4.9 kb, and to be used in dot-blot hybridization for the detection of multiple- and single-copy genes.

Isolates of Fusarium spp. were recovered from the roots of field pea (Pisum sativum) collected from 15 commercial fields in Alberta, Canada. Most of the isolates (75 out of 96) were identified as F. avenaceum, based on morphology, phylogeny and species-specific PCR amplification. Molecular differences in the F. avenaceum isolates were detected based on putative mating type, and on ITS and CPN60 sequences. MAT-1 and MAT-2 were equally distributed among the isolates. Phylogenetic analysis based on ITS and CPN60 sequences clustered most of the F. avenaceum isolates into a single group. In some cases, isolates with low aggressiveness clustered together in additional groups. There was no correlation between phylogenetic profile and either mating type or geographic origin. This population of F. avenaceum has a low level of genetic variation and consists of isolates derived from the two mating types. Isolates with low aggressiveness are also retained in the population.

Rhodiola plants exhibiting symptoms typical of phytoplasma infection were observed at an experimental farm near Edmonton, Alberta, Canada, in 2007. Phytoplasma infection was confirmed with a polymerase chain reaction (PCR)-assay using 16S ribosomal RNA primers developed specifically for phytoplasmas; the primers could amplify a band from tissue samples derived from infected, but not from asymptomatic, plants. Comparison between the sequence of the PCR product and the 16S ribosomal RNA gene sequences of other phytoplasmas available in GenBank indicated that the phytoplasma identified in rhodiola is a member of the subgroup 16SrI-B (aster yellows subgroup B).