Physiological aspects of seed recalcitrance: a case study on the tree Aesculus hippocastanum.

Recalcitrant seeds are typical of some tropical and subtropical trees. Their post-shedding life activity proceeds in humid air and wet litter. They are desiccation sensitive and, for this reason, have a short life span and need some special procedures for cryopreservation. This review is devoted to the post-shedding life strategy of recalcitrant seeds, which includes the maintenance of high hydration status, metabolic readiness and ability to rapidly germinate before desiccation-induced damage exerts a lethal effect. The main physiological aspects of recalcitrant seeds are considered starting from mature seeds, followed during dormancy if occurs and resulting in germination. The collected data embrace the metabolic processes in embryonic axes and whole seeds. The up-to-date results are integrated covering the main metabolic processes, namely water status and transport, protein and carbohydrate metabolism, antioxidant defense, axis-cotyledon relations, hormonal control and germination. Among the representatives of various taxa, the seeds of which exhibit recalcitrance, attention was given to horse chestnut seeds as one of most studied recalcitrants.

Vacuolar biogenesis and aquaporin expression at early germination of broad bean seeds.

A key event in seed germination is water uptake-mediated growth initiation in embryonic axes. Vicia faba var. minor (broad bean) seeds were used for studying cell growth, vacuolar biogenesis, expression and function of tonoplast water channel proteins (aquaporins) in embryonic axes during seed imbibition, radicle emergence and growth. Hypocotyl and radicle basal cells showed vacuole restoration from protein storage vacuoles, whereas de novo vacuole formation from provacuoles was observed in cells newly produced by root meristem. cDNA fragments of seven novel aquaporin isoforms including five Tonoplast Intrinsic Proteins (TIP) from three sub-types were amplified by PCR. The expression was probed using q-RT-PCR and when possible with isoform-specific antibodies. Decreased expression of TIP3s was associated to the transformation of protein storage vacuoles to vacuoles, whereas enhanced expression of a TIP2 homologue was closely linked to the fast cell elongation. Water channel functioning checked by inhibitory test with mercuric chloride showed closed water channels prior to growth initiation and active water transport into elongating cells. The data point to a crucial role of tonoplast aquaporins during germination, especially during growth of embryonic axes, due to accelerated water uptake and vacuole enlargement resulting in rapid cell elongation.