Detection of Ralstonia solanacearum in Asymptomatic Imported Seed Potato using a DNA-based Method

Potato is an economically important crop among vegetables grown in Sri Lanka that mainly relies on healthy seed potatoes. About 40% of the annual seed potato requirement is fulfilled by the import of seed potatoes from Netherland, USA, Germany and France. Import of seed potatoes makes possibilities to enter plant pathogenic pests and diseases to Sri Lanka. Bacterial wilt is one of the most destructive diseases of potato. Ralstonia solanacearum, which causes bacterial wilt of potato, is considered as an important quarantine significant plant pathogen in Sri Lanka. The currently available conventional methods such as culture methods, biochemical methods are time consuming, very laborious and not sensitive for the detection of R. solanacearum in imported seed potatoes. Although immunodiagnostic methods are rapid, the sensitivity is not enough to detect the bacterium in asymptomatic or latently infected seed potatoes. In this study, a DNA-based detection method was applied to screen seed potatoes imported into Sri Lanka and 5 out of 30 tested samples (17%) were positive for R. solanacearum. The seed potato samples detected as infected with R. solanacearum were further studied and it revealed that the Asian phylotype I and the American phylotype II were detected from seed potato samples imported to the country. Phylotype II (Race 3/biovar 2) was detected in seed potatoes imported from USA and France and both phylotype I and phylotype II (Race 3/biovar 2) were detected in seed potatoes imported from Netherland from where majority of seed potatoes are imported into the country. The quarantine measures should be strictly followed to avoid the spread and establishment of phylotype II, Race 3/biovar 2 strains within the country, as it is the extremely destructive potato pathogen which have a restricted distribution in higher elevations of Sri Lanka. Further, the DNA-based method can be used to identify the pathogen to avoid the introduction or entry of R. solanacearum into the country for the betterment of potato cultivation in Sri Lanka.KEYWORDS: Seed potato, Bacterial wilt, Ralstonia solanacearum, Quarantine pest, Rsol_fli

An eQTL Analysis of Partial Resistance to Puccinia hordei in Barley

Background - Genetic resistance to barley leaf rust caused by Puccinia hordei involves both R genes and quantitative trait loci. The R genes provide higher but less durable resistance than the quantitative trait loci. Consequently, exploring quantitative or partial resistance has become a favorable alternative for controlling disease. Four quantitative trait loci for partial resistance to leaf rust have been identified in the doubled haploid Steptoe (St)/Morex (Mx) mapping population. Further investigations are required to study the molecular mechanisms underpinning partial resistance and ultimately identify the causal genes.Methodology/Principal Findings - We explored partial resistance to barley leaf rust using a genetical genomics approach. We recorded RNA transcript abundance corresponding to each probe on a 15K Agilent custom barley microarray in seedlings from St and Mx and 144 doubled haploid lines of the St/Mx population. A total of 1154 and 1037 genes were, respectively, identified as being P. hordei-responsive among the St and Mx and differentially expressed between P. hordei-infected St and Mx. Normalized ratios from 72 distant-pair hybridisations were used to map the genetic determinants of variation in transcript abundance by expression quantitative trait locus (eQTL) mapping generating 15685 eQTL from 9557 genes. Correlation analysis identified 128 genes that were correlated with resistance, of which 89 had eQTL co-locating with the phenotypic quantitative trait loci (pQTL). Transcript abundance in the parents and conservation of synteny with rice allowed us to prioritise six genes as candidates for Rphq11, the pQTL of largest effect, and highlight one, a phospholipid hydroperoxide glutathione peroxidase (HvPHGPx) for detailed analysis.Conclusions/Significance - The eQTL approach yielded information that led to the identification of strong candidate genes underlying pQTL for resistance to leaf rust in barley and on the general pathogen response pathway. The dataset will facilitate a systems appraisal of this host-pathogen interaction and, potentially, for other traits measured in this populatio