Genetic diversity in cornsalad (Valerianella locusta) and related species as determined by AFLP markers

Fifteen amplified fragment length polymorphism (AFLP) EcoRI/MseI‐based primer combinations with five selective bases (Eco RI‐ANN, MseI‐CN) were used to estimate genetic diversity among 45 line varieties of cultivated cornsalad and 19 genebank accessions classified into nine different species related to cornsalad. Polymorphic fragments were scored for calculation of Jaccard's coefficient of genetic similarity (GS). The average GS estimate in elite germplasm (GS = 0.90) was substantially higher than in exotic germplasm (GS = 0.47). UPGMA‐cluster analysis revealed genetic relationships among recently bred varieties, old varieties and genebank accessions. Analysis of molecular variance indicated almost threefold variability within sets compared with between sets due to a high level of polymorphism among wild species. Sources for increasing genetic diversity in elite germplasm of cornsalad were suggested and a duplicate among the genebank accessions was detected. AFLPs could be considered a powerful tool for genetic diversity estimation in cornsalad germplasm and are recommended for systematic fingerprinting of remaining cornsalad species.     

Detection of Acidovorax valerianellae in corn-salad seeds, seed transmission of the pathogen and disease development in the field

Acidovorax valerianellae , the causal agent of bacterial black spot of corn salad and responsible for severe economic losses to this vegetable in France, was successfully transmitted to corn-salad plants by artificially inoculated seeds in glasshouse and field experiments. In the field experiments, climatic data recorded under plastic tunnels indicated that increasing temperature and relative humidity increased symptom development. To investigate the possible contamination of commercial seedlots of corn salad, a seed test was developed consisting of soaking batches of seeds (five batches each of 5000, 1000, 500 and 100 seeds) overnight at 4°C in distilled sterile water, followed by dilution-plating of seed extracts on TSAV (tryptic soya agar for A. valerianellae ) semiselective medium. Suspected colonies were identified by biochemical and pathogenicity tests or, within 24 h, using antibodies specific to A. valerianellae . Acidovorax valerianellae was detected in three lots. Seed infection levels ranged from 0·10 to 0·89% of contaminated seeds and a single seed carried up to 1800 A. valerianellae colony-forming units.     

A semiselective medium for the isolation of Acidovorax valerianellae from soil and plant debris

Bacterial black spot of corn-salad ( Valerianella locusta ) caused by Acidovorax valerianellae appeared in France in the beginning of the 1990s. The disease is now widespread in the main area of corn-salad production and is responsible for high depreciation and significant economic losses every year. A semiselective medium is described for the detection of the bacterium in various environments and to identify primary sources of inoculum. The medium TSAV (tryptic soy broth, 3 g L⁻¹, agar 15 g L⁻¹, 5-fluorouracyl 5 mg L⁻¹, novobiocin 5 mg L⁻¹, propiconazole 5 mg L⁻¹) increased chances of obtaining cultures of A. valerianellae from naturally infested soil and from root-debris. Using TSAV, the pathogen was detected in root-debris up to 39 days after the harvest of a diseased crop. As only a few days usually separates harvest from the next sowing date, soil is likely to be an important source of inoculum for the next crop.