Comparative ovule and megagametophyte development in Hydatellaceae and water lilies reveal a mosaic of features among the earliest angiosperms

Background and Aims: The embryo sac, nucellus and integuments of the early-divergentangiosperms Hydatellaceae and other Nymphaeales are comparedwith those of other seed plants, in order to evaluate the evolutionaryorigin of these characters in the angiosperms. Methods: Using light microscopy, ovule and embryo sac development aredescribed in five (of 12) species of Trithuria, the sole genusof Hydatellaceae, and compared with those of Cabombaceae andNymphaeaceae. Key Results: The ovule of Trithuria is bitegmic and tenuinucellate, ratherthan bitegmic and crassinucellate as in most other Nymphaeales.The seed is operculate and possesses a perisperm that developsprecociously, which are both key features of Nymphaeales. However,in the Indian species T. konkanensis, perisperm is relativelypoorly developed by the time of fertilization. Perisperm cellsin Trithuria become multinucleate during development, a featureobserved also in other Nymphaeales. The outer integument issemi-annular (‘hood-shaped’), as in Cabombaceaeand some Nymphaeaceae, in contrast to the annular (‘cap-shaped’)outer integument of some other Nymphaeaceae (e.g. Barclaya)and Amborella. The megagametophyte in Trithuria is monosporicand four-nucleate; at the two-nucleate stage both nuclei occurin the micropylar domain. Double megagametophytes were frequentlyobserved, probably developed from different megaspores of thesame tetrad. Indirect, but strong evidence is presented forapomictic embryo development in T. filamentosa. Conclusions: Most features of the ovule and embryo sac of Trithuria are consistentwith a close relationship with other Nymphaeales, especiallyCabombaceae. The frequent occurrence of double megagametophytesin the same ovule indicates a high degree of developmental flexibility,and could provide a clue to the evolutionary origin of the Polygonum-typeof angiosperm embryo sac.