The role of fractures and lipids in the seed coat in the loss of hardseededness of six Mediterranean legume species

Changes in the seed coat morphology of 12 annual legumes were studied using environmental scanning electron microscopy (ESEM). The seeds of Biserrula pelecinus L. cv. Casbah, Ornithopus sativus cv. Cadiz, Trifolium clypeatum L., T. spumosum L., T. subterraneum L. cv. Bacchus Marsh, Trigonella balansae Boiss. & Reuter., Trigonella monspeliaca L. and Vicia sativa subsp. amphicarpa Dorthes (morthes.) were examined by ESEM after exposure to field conditions for 6 months, while those of Medicago polymorpha L. cv. Circle Valley, Trifolium clypeatum L., T. glanduliferum Boiss., T. lappaceum L., T. spumosum L., and T. subterraneum L. cv. Dalkeith, were examined after 2 years' exposure. The entry of water into seeds was followed by covering various parts of the seed coat with petroleum jelly and soaking the treated seeds in dyes. As the seeds softened over time, more and larger fractures appeared on the seed coat. Water entered the seed either through fractures, over the seed coat as a whole or through the lens. It is hypothesized that the formation of fractures occurs after physicochemical changes in the seed coat, probably associated with changes in the amount and nature of seed coat lipids. The newly matured whole seeds of M. polymorpha cv. Circle Valley, T. clypeatum, T. glanduliferum, T. lappaceum, T. spumosum, and T. subterraneum cv. Dalkeith were analysed for lipid content in 1997. The seed coats of T. subterraneum cv. Dalkeith and T. spumosum were separated from the cotyledons and examined in detail for lipid content. The lipid content of whole seeds ranged from 48 (T. lappaceum) to 167 mg/g (T. subterraneum cv. Dalkeith). Total lipid of the whole seeds of T. subterraneum cv. Dalkeith and T. glanduliferum declined by about 9 mg/g over 2 years, while in T. spumosum it declined by about 17 mg/g. In contrast, the major fatty acids in the seed coat declined by 0.67 mg/g over the 2 years. Change in seed coat lipids showed a marked similarity to changes in hardseededness for both T. subterraneum cv. Dalkeith and T. spumosum. The results strongly suggest that seed softening is associated with loss of lipids in the seed coat, because lipids have physical characteristics that are altered at temperatures experienced in the field.