Analysis of leaf appearance, leaf death and phoma leaf spot, caused by Leptosphaeria maculans, on oilseed rape (Brassica napus) cultivars

The definitive version can be found at: http://onlinelibrary.wiley.com/ Copyright Association of Applied BiologistsDevelopment of phoma leaf spot (caused by Leptosphaeria maculans) on winter oilseed rape (canola, Brassica napus) was assessed in two experiments at Rothamsted in successive years (2003-04 and 2004-05 growing seasons). Both experiments compared oilseed rape cultivars Eurol, Darmor, Canberra and Lipton, which differ in their resistance to L. maculans. Data were analysed to describe disease development in terms of increasing numbers of leaves affected over thermal time from sowing. The cultivars showed similar patterns of leaf spot development in the 2003-04 experiment when inoculum concentration was relatively low (up to 133 ascospores m-3 air), Darmor developing 5.3 diseased leaves per plant by 5 May 2004, Canberra 6.6, Eurol 6.8 and Lipton 7.5. Inoculum concentration was up to sevenfold greater in 2004-05, with Eurol and Darmor developing 2.4 diseased leaves per plant by 16 February 2005, whereas Lipton and Canberra developed 2.8 and 3.0 diseased leaves, respectively. Based on three defined periods of crop development, a piece-wise linear statistical model was applied to the progress of the leaf spot disease (cumulative diseased leaves) in relation to appearance ('birth') and death of leaves for individual plants of each cultivar. Estimates of the thermal time from sowing until appearance of the first leaf or death of the first leaf, the rate of increase in number of diseased leaves and the area under the disease progress line (AUDPL) for the first time period were made. In 2004-05, Canberra (1025 leaves x degrees C days) and Lipton (879) had greater AUDPL values than Eurol (427) and Darmor (598). For Darmor and Lipton, the severity of leaf spotting could be related to the severity of stem canker at harvest. Eurol had less leaf spotting but severe stem canker, whereas Canberra had more leaf spotting but less severe canker.Peer reviewe

GCN2-dependent phosphorylation of eukaryotic translation initiation factor-2α in Arabidopsis

The yeast regulatory protein kinase, general control non-derepressible-2 (GCN2) plays a key role in general amino acid control. GCN2 phosphorylates the α subunit of the trimeric eukaryotic translation initiation factor-2 (eIF2), bringing about a decrease in the general rate of protein synthesis but an increase in the synthesis of GCN4, a transcription factor that promotes the expression of genes encoding enzymes for amino acid biosynthesis. The present study concerned the phosphorylation of Arabidopsis eIF2α (AteIF2α) by the Arabidopsis homologue of GCN2, AtGCN2, and the role of AtGCN2 in regulating genes encoding enzymes of amino acid biosynthesis and responding to virus infection. A null mutant for AtGCN2 called GT8359 was obtained and western analysis confirmed that it lacked AtGCN2 protein. GT8359 was more sensitive than wild-type Arabidopsis to herbicides that affect amino acid biosynthesis. Phosphorylation of AteIF2α occurred in response to herbicide treatment but only in wild-type Arabidopsis, not GT8359, showing it to be AtGCN2-dependent. Expression analysis of genes encoding key enzymes for amino acid biosynthesis and nitrate assimilation revealed little effect of loss of AtGCN2 function in GT8359 except that expression of a nitrate reductase gene, NIA1, was decreased. Analysis of wild-type and GT8359 plants infected with Turnip yellow mosaic virus or Turnip crinkle virus showed that AteIF2α was not phosphorylated

Functional analysis and expression profiling of HcrVf1 and HcrVf2 for development of scab resistant cisgenic and intragenic apples

Apple scab resistance genes, HcrVf1 and HcrVf2, were isolated including their native promoter, coding and terminator sequences. Two fragment lengths (short and long) of the native gene promoters and the strong apple rubisco gene promoter (PMdRbc) were used for both HcrVf genes to test their effect on expression and phenotype. The scab susceptible cultivar ‘Gala’ was used for plant transformations and after selection of transformants, they were micrografted onto apple seedling rootstocks for scab disease tests. Apple transformants were also tested for HcrVf expression by quantitative RT-PCR (qRT-PCR). For HcrVf1 the long native promoter gave significantly higher expression that the short one; in case of HcrVf2 the difference between the two was not significant. The apple rubisco gene promoter proved to give the highest expression of both HcrVf1 and HcrVf2. The top four expanding leaves were used initially for inoculation with monoconidial isolate EU-B05 which belongs to race 1 of V. inaequalis. Later six other V. inaequalis isolates were used to study the resistance spectra of the individual HcrVf genes. The scab disease assays showed that HcrVf1 did not give resistance against any of the isolates tested regardless of the expression level. The HcrVf2 gene appeared to be the only functional gene for resistance against Vf avirulent isolates of V. inaequalis. HcrVf2 did not provide any resistance to Vf virulent strains, even not in case of overexpression. In conclusion, transformants carrying the apple-derived HcrVf2 gene in a cisgenic as well as in an intragenic configuration were able to reach scab resistance levels comparable to the Vf resistant control cultivar obtained by classical breeding, cv. ‘Santana’

Production of extracellular enzymes by different isolates of Pochonia chlamydosporia

For the first time, the specific activities of chitinases, esterases, lipases and a serine protease (VCP1) produced by different isolates of the nematophagous fungus Pochonia chlamydosporia were quantified and compared. The isolates were grown for different time periods in a minimal liquid medium or media supplemented with 1 % chitin, 0.2 % gelatin or 2 % olive oil. Enzyme-specific activities were quantified in filtered culture supernatants using chromogenic p-nitrophenyl substrates (for chitinases, lipases and esterases) and a p-nitroanilide substrate (to measure the activity of the proteinase VCP1). Additionally, information on parasitic growth (nematode egg parasitism) and saprotrophic growth (plant rhizosphere colonisation) was collected. Results showed that the production of extracellular enzymes was influenced by the type of medium (p < 0.05) in which P. chlamydosporia was grown. Enzyme activity differed with time (p < 0.05), and significant differences were found between isolates (p < 0.001) and the amounts of enzymes produced (p < 0.001). However, no significant relationships were found between enzyme activities and parasitic or saprotrophic growth using Kendall's coefficient of concordance or Spearman rank correlation coefficient. The results provided new information about enzyme production in P. chlamydosporia and suggested that the mechanisms which regulate the trophic switch in this fungus are complex and dependent on several factors