Mature dry seeds are highly resilient plant structures where the encapsulated embryo is kept protected and dormant to facilitate its ultimate dispersion. Seed viability is heavily dependent on the seed coat's capacity to shield living tissues from mechanical and oxidative stress. In Arabidopsis (), the seed coat, also called the testa, arises after the differentiation of maternal ovular integuments during seed development. We recently described a thick cuticle tightly embedded in the mature seed's endosperm cell wall. We show here that it is produced by the maternal inner integument 1 layer and, remarkably, transferred to the developing endosperm. Arabidopsis () mutations cause maternally derived seed coat pigmentation defects. gene products encode proteins involved in flavonoid metabolism and regulators of seed coat development. mutants have abnormally high seed coat permeability, resulting in lower seed viability and dormancy. However, the biochemical basis of this high permeability is not fully understood. We show that the cuticles of developing mutant integuments have profound structural defects, which are associated with enhanced cuticle permeability. Genetic analysis indicates that a functional proanthocyanidin synthesis pathway is required to limit cuticle permeability, and our results suggest that proanthocyanidins could be intrinsic components of the cuticle. Together, these results show that the formation of a maternal cuticle is an intrinsic part of the normal integumental differentiation program leading to testa formation and is essential for the seed's physiological properties.

The seed coat is specialized dead tissue protecting the plant embryo from mechanical and oxidative damage. Tannins, a type of flavonoids, are antioxidants known to accumulate in the Arabidopsis seed coat and transparent testa mutant seeds, deficient in flavonoid synthesis, exhibit low viability. However, their precise contribution to seed coat architecture and biophysics remains evasive. A seed coat cuticle, covering the endosperm outer surface and arising from the seed coat inner integument 1 cell layer was, intriguingly, previously shown to be more permeable in transparent testa mutants deficient not in cuticular component synthesis, but rather in flavonoid synthesis. Investigating the role of flavonoids in cuticle permeability led us to identify periclinal inner integument 1 tannic cell walls being attached, together with the cuticle, to the endosperm surface upon seed coat rupture. Hence, inner integument 1 tannic cell walls and the cuticle form two fused layers present at the surface of the exposed endosperm upon seed coat rupture, regulating its permeability. Their potential physiological role during seed germination is discussed.

Seeds are a late land plant evolution innovation that promoted the striking spread and diversity of angiosperms. The seed coat is a specialized dead tissue protecting the plant embryo from mechanical damage. In many species, including Arabidopsis thaliana, the seed coat also achieves a remarkable balancing act: it limits oxygen uptake, avoiding premature embryo oxidative damage, but not entirely so as to enable seed dormancy release. The seed coat biophysical features implementing the striking physiological properties of the seed remain poorly understood. Tannins, a type of flavonoids, are antioxidants known to accumulate in the Arabidopsis seed coat and transparent testa (tt) mutant seeds, deficient in flavonoid synthesis, exhibit low dormancy and viability. However, their precise contribution to seed coat architecture and biophysics remains evasive. A seed coat cuticle, covering the endosperm outer surface was, intriguingly, previously shown to be more permeable in tt mutants deficient not in cuticular component synthesis, but rather in flavonoid synthesis. Investigating the role of flavonoids in cuticle permeability led us to identify cell walls, originating from the seed coat inner integument 1 cells, impregnated with tannins. We found that tannic cell walls are tightly associated with the cuticle, forming two fused layers that regulate endosperm permeability. In addition, we show that tannic cell walls are prominent building blocks of the seed coat, constituting a continuous barrier around the seed living tissues. Altogether our findings reveal the existence of tannic cell walls as a previously unrecognized biological barrier sustaining the seed’s key physiological properties.One sentence summaryThe seed coat is largely composed of plant cell walls impregnated with tannins, forming a thick and continuous protective barrier surrounding the embryo promoting seed viability and dormancy.