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The paper reviews female gametophyte development in vitro, the pathways of gynogenetic embryogeny, and autonomous endosperm development in vitro, and presents some implications for future investigation of apomictic processes. In most species studied so far, the pattern of embryo sac development was not altered by in vitro conditions, and some rare deviations could be additional sources of gynogenetic embryos. Meiosis in vitro resulted in megaspore formation followed by embryo sac development or direct embryogenesis. Gynogenesis in vitro occurs via various embryological processes. Embryos developed parthenogenetically from egg cells in twelve species. Apogamic development from the synergid or from antipodal cells was found in six species. Early development of gynogenetic embryos showed some similarities to zygotic embryogenesis, but then the embryos often remained undifferentiated or formed microcalli and protocorm-like structures. In nine species the central cell in ovary/ovule cultures was sporadically stimulated to divide into free-nuclear endosperm. Generally only one gynogenetic structure developed per embryo sac, and no embryos appeared in ovules containing autonomous endosperm. Cellular programs for the autonomous development of endosperm and embryo seem quite different or even competitive in vitro. Various approaches should be taken in future research on apomixis. Classical and novel cytological methods, and in vitro organ and cell cultures should be combined with molecular techniques and genetic studies.
Poor seed set is a limiting factor in alfalfa breeding, as it slows the selection response. One strategy used to overcome this problem is to search for mutations of inflorescence morphology. Long-peduncle (lp), branched-raceme (br) and top-flowering (tf) inflorescence mutations increase the number of flowers per inflorescence, but they do not improve seed set per flower. Here we assessed pollen tube growth in styles of those inflorescence mutants and we observed embryo and endosperm development in seeds 1 to 16 days after pollination (DAP). The number of pollen tubes penetrating the style and the ovary was similar in all tested mutants and in the reference cultivar Radius. At 2 DAP, fertilized ovules were 2.7–3.9 times less numerous in certain inflorescence mutants than in the short-raceme cv. Radius. Ovule degeneration progressed at 2–4 DAP in all analyzed plants. Most ovules were not properly developed in the control cultivar (62%), nor in the forms with mutated inflorescence morphology (69–86%). The number of seeds per pod was lowest in the tf form despite its having the highest number of ovules per ovary. It appears that the number of ovules per pistil is not a crucial factor in seed set in alfalfa when fertilization efficiency is very low. Both poor fertilization and gradual ovule degeneration were factors causing poor seed set in the investigated alfalfa genotypes.
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