Effects of Light, Growth Media, and Seedling Orientation on Bioassays of Alfalfa Autotoxicity

Most assessments of allelopathy involve bioassays. Our objective was to improve the sensitivity of an alfalfa ( Medicago sativa L.) seedling bioassay for evaluating genetic tolerance to autotoxic leaf extracts. In a petri dish assay on imbibed seed, light inhibited hypocotyl elongation of controls and increased root elongation. Root growth was sensitive to the autotoxin in both light and darkness. An agar medium gave better root growth of controls and lower standard errors than did filter paper when petri dishes were placed on edge to encourage downward root growth or were placed flat where roots grew laterally. Hypocotyl growth was not very sensitive to the autotoxic chemical(s) on either agar or paper medium when the plate was flat, because the hypocotyl arched upward to escape contact with the extract. Hypocotyl growth was sensitive in a rolled paper towel treatment held vertically because the hypocotyl remained in continuous contact with the extract. On agar plates placed flat, 50% inhibition of root length occurred at an extract concentration that was about 8% of that needed for 50% inhibition of germination at 36 and 48 h. Root growth was stimulated up to 15% above controls at very low concentrations of leaf extract. Root length at 120 h was the best indicator of autotoxic effects of alfalfa leaf extracts. We evaluated 17 germplasms and three cultivars of alfalfa for root growth response to the autotoxic chemical and found a twofold range ( P < 0.05) in tolerance.