Putative resistance gene markers associated with quantitative trait loci for fire blight resistance in Malus 'Robusta 5' accessions

Background Breeding of fire blight resistant scions and rootstocks is a goal of several international apple breeding programs, as options are limited for management of this destructive disease caused by the bacterial pathogen Erwinia amylovora. A broad, large-effect quantitative trait locus (QTL) for fire blight resistance has been reported on linkage group 3 of Malus ‘Robusta 5’. In this study we identified candidate gene markers for fire blight resistance by integrating further genetic mapping studies with bioinformatics analysis of transcript profiling data and genome sequence databases. Results When several defined E.amylovora strains were used to inoculate three progenies from 62 international breeding programs, all with ‘Robusta 5’ as a common parent, two distinct QTLs were detected on linkage group 3, where only one had previously been mapped. In the New Zealand population, the proximal QTL co-located with SNP markers for a leucine-rich repeat, receptor-like protein (MxdRLP1) candidate resistance gene and a closely linked class 3 peroxidase gene. While the QTL detected in the German population was approximately 6 cM distal to this, directly below a SNP marker for a heat shock 90 family protein (HSP90). In the US population, the position of the LOD score peak on linkage group 3 was dependent upon the pathogen strains used for inoculation. One of the five MxdRLP1 alleles identified in fire blight resistant and susceptible cultivars was genetically associated with resistance and used to develop a high resolution melting PCR marker. A resistance QTL detected on linkage group 7 of the US population co-located with another HSP90 gene-family member and a WRKY transcription factor previously associated with fire blight resistance. However, this QTL was not observed in the New Zealand or German populations. Conclusions The results suggest that the upper region of ‘Robusta 5’ 76 linkage group 3 contains multiple genes contributing to fire blight resistance and that their contributions to resistance can vary depending upon pathogen virulence and other factors. Candidate gene mapping has proved a useful aid in defining these QTLs and developing markers for marker-assisted breeding of fire blight resistance

Towards a Practical Warning System for Fire Blight Blossom Infection

15 pages, 1 article*Towards a Practical Warning System for Fire Blight Blossom Infection* (Beer, S. V.; Schwager, S. J.; Norelli, J. L.; Aldwinckle, H. S.; Burr, T. J.) 15 page

RNAi mutants for determination of candidate gene function in resistance of apple to Erwinia amylovora (Fire blight)

To determine the function of candidate apple genes that confer resistance to fire blight, RNAi mutants of M.26 apple rootstock were produced using an efficient multiplex transformation system. Five RNAi EST-silencing vectors were used in each transformation experiment to allow selection of up to five types of mutants from a single experiment. RNAi-silencing constructs were created using ESTs associated with response of apple to Erwinia amylovora that were selected based upon transcript profiling data and bioinformatics. Candidate genes in six functional categories were evaluated by using the pHELLSGATE8 RNAi-mediated gene silencing vector to silence a specific EST derived sequence and then observing the resulting resistance phenotype. Silencing of candidate genes was confirmed by RT-qPCR. Transgenic lines were phenotyped following inoculation of young plants with E. amylovora by determining the area under the disease progress curve (AUDPC), the cumulative percentage of shoot length that was blighted and the population of E. amylovora in young plants as determined by qPCR. Some of transgenic lines with the p8L01 and pfbox silencing construct had a significantly lower population of E. amylovora than non-transformed M.26. The disease severity in the lines with the p4H09 silencing construct was significantly different from that of nontransformed M.26. Also some lines with the pfbox, p8L01, pPO53 and pSGT1 silencing constructs had significantly lower disease severity than non-transformed M.26. The genes associated with reduced disease-severity and/or lower E. amylovora population are potentially useful in maker assisted breeding and GE for cultivars and rootstocks with increased resistance to fire bligh