Effects of the environment, cultivar, maturity, and preservation method on red clover isoflavone concentration

Red clover (Trifolium pratense L.) contains isoflavones that are of interest because of their benefits for human health as well as their adverse effects on the fertility of farm animals. A series of field experiments was conducted in Sainte-Anne-de-Bellevue, QC, Canada, to determine the effects of the environment, cultivar, plant maturity, plant part, and preservation method on the concentration of the two predominant isoflavones in red clover, formononetin and biochanin A. In a multi-year, multisite trial, the total isoflavone concentration in 10 cultivars ranged between 8923 and 12 753 mu g g(-1) of DM averaged across sites, harvests, and years. Despite strong environmental effects, the cultivar Start consistently had the lowest isoflavone concentrations, with few differences observed among other cultivars. Across stages of maturity, leaves were found to have the highest isoflavone concentration followed by stems and inflorescences (11 970, 4896, and 3297 mu g g(-1) of DM respectively). Changes in isoflavone concentrations with increasing maturity varied depending on the plant part. Overall, highest isoflavone concentrations were found in leaves and stems during the vegetative stages, with the formononetin concentration declining until plants initiated flowering, especially in stems, with concentrations then stabilized in both parts. Upon initiation, inflorescences contained similar isoflavone concentrations than leaves, but concentrations decreased rapidly during flower development to fall even below those observed in stems. Inflorescences then had isoflavone concentrations that were as much as 11 times lower than leaves. Fresh herbage contained higher formononetin and total isoflavone concentrations than did silage and hay (114 464, 12 200, and 11 604 mu g g(-1) of DM, respectively). The isoflavone concentration in field-grown red clover is thus high but can be affected by a range of agronomic factors.