Megasporocyte Callose in Apomictic Buffelgrass, Kentucky Bluegrass, Pennisetum squamulatum Fresen, Tripsacum L., and Weeping Lovegrass

Abstract

Callose, a β‐l,3‐glucan, is synthesized and deposited in the walls of megaspore mother cells (MMCs) of angiosperms that produce either normal monosporic (>90% of all angiosperm species) or bisporic embryo sacs. Such deposits are absent in angiosperms that produce tetrasporic embryo sacs and were recently shown to be absent in diplosporous Elymus rectisetus (Nees in Lehm.) A. Löve & Connor, Tripsacum L. spp., and Poa nemoralis L. We determined the extent of callose deposits in MMCs of diplosporous weeping lovegrass (Eragrostis curvula Schrad.), several additional diplosporous Tripsacum accessions and aposporous buffelgrass (Pennisetum ciliare (L.) Link), Kentucky bluegrass (Poa pratensis L.), and Pennisetum squamulatum Fresen. Callose deposition was studied in pistils cleared in an aqueous aniline blue clearing medium and observed by means of UV microscopy. Pistils cleared with conventional clearing media were studied by means of interference contrast microscopy. Some pistils were cleared and analyzed in one clearing medium and recleared and reanalyzed in the other. A callosic MMC was observed in only one of 376 appropriately staged pistils taken from diplosporous species. This pistil, one of 89 weeping lovegrass pistils studied, was probably facultatively sexual. An absence of callose during megasporogenesis may be characteristic of most types of diplospory. Though present, the distributions and quantities of callose observed in MMC walls of aposporous apomicts were erratically reduced, possibly as a result of early MMC abortion or early activation of nucellar initials. Callose deficiencies and vacuolization patterns support the following hypotheses: (i) apospory is caused by a moderately precocious gametophytization of nucellar cells, and (ii) bispory, tetraspory, and diplospory are caused by a moderately to extremely precocious gametophytization of the MMC that is nearly complete in the latter case and variably incomplete in the former two cases. Understanding how these precocious gametophytizations evolved and are regulated may be crucial to the eventual transfer of apomixis to major crop species and to its successful manipulation by breeders.