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Mapping disease resistance genes
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Identification of markers associated with race-specific resistance to Aphanomyces root rot in alfalfa

Poster presentation at the 2017 American Phytopathological Society meeting

Abstract
Aphanomyces root rot, caused by Aphanomyces euteiches, is one of the most important diseases of alfalfa in the United States. Two races of the pathogen are recognized and although most cultivars are resistant to race 1, fewer have resistance to race 2, the predominant race in North America. Molecular markers are needed to facilitate breeding for resistance and to clarify race/resistance gene structure. Resistant and susceptible seedlings were identified from three resistant cultivars, WAPH1, WAPH5 and 53V52, and used as parents to produce F1 populations. Severity of symptoms corresponded with amount of pathogen DNA and oospore density in roots. Race-specific resistance involves a hypersensitive response of individual epidermal or cortical cells upon pathogen attack followed by suberization of cells surrounding the stele and strong autofluorescence in cortical cells, indicating the presence of phenolic compounds. Segregation ratios of F1 populations suggested that resistance to race 1 in WAPH1 is conditioned by a single gene but resistance to race 1 is multigenic in WAPH5 and 53V52, and resistance to race 2 is multigenic in all three cultivars. Segregation for resistance to seven strains of A. euteiches in 70 F1 full-sib plants derived from 53V52 suggested the presence of clustered resistance genes and multiple race types. Identification of resistance gene loci is in progress using genotyping by sequencing and genetic mapping of F1 populations.


Characterization of the bacterial stem blight pathogen of alfalfa, Pseudomonas syringae pv. syringae ALF3

Poster presentation at the 2014 meeting of the American Phytopathological Society

Abstract
Bacterial stem blight of alfalfa occurs sporadically in the central and western U.S. Yield losses of up to 50% of the first harvest can occur with some cultivars. Developing resistant cultivars is hampered by lack of information on the pathogen and a standard test for evaluating plant germplasm. Bacteria producing a fluorescent pigment were isolated on King’s B agar from alfalfa with symptoms of bacterial stem blight from near Cheyenne, WY. The strain ALF3 was tentatively identified as Pseudomonas syringae pv. syringae based on 16S rDNA sequence and PCR amplification of syrB for lipodepsinonapeptide toxin production. Multilocus sequence analysis indicated that ALF3 falls within a clade containing strains of P. syringae pv. syringae with closest affinity to FF5 from pear. Comparison of a draft whole-genome sequence of ALF3 further confirmed that ALF3 most closely resembles FF5 (~96% sequence identity) and P. syringae pv. aptata DSM50252 from beet. Approximately 60 genes were unique to ALF3, including several predicted genes in the T3SS cluster such as a type III helper protein HrpZ (Pto) and phage-associated genes. ALF3 was highly pathogenic to snapbean pods but caused only mild symptoms on leaves of snapbean, pear, and sugarbeet. A standardized method for evaluating disease resistance in alfalfa was developed. Cultivars with fall dormancy ratings of 1 and 2 had higher percentages of resistant plants than cultivars with fall dormancy ratings of 8-11.