Abstract

Natural triploids of Populus tomentosa (2n = 3x = 57) are presumably the result of sexual polyploidization through the union of normal n female gametes and numerically unreduced (2n) male gametes. In our microscopic study of microspore mother cells (MMCs) of diploid P. tomentosa (2n = 2x = 38), we observed that the first meiotic division was normal but that the second division was characterized by frequent abnormal spindle orientation (parallel, tripolar, and fused spindles) and premature cytokinesis. The parallel, fused spindles and premature cytokinesis were considered to be leading dyad formation, and tripolar spindles seemed to be leading triad formation at the tetrad stage. There was a higher frequency of parallel spindles than other spindle forms, but there were no significant correlations between parallel spindles and dyads. An indirect immunofluorescence examination of meiosis II revealed that four tetragonally arranged nuclei were formed in MMCs with parallel spindles and that there were radial microtubules systems (RMSs) among these four nuclei, leading to the tetragonal tetrad. In some MMCs, however, the parallel spindles led to the gathering of one or two non-sister groups of chromosomes, causing an incorporation of RMSs from two daughter nuclei. Thus, the incorporated RMSs established three or two nuclear cytoplasmic domains for the control of division plane, resulting in either triad or dyad formation. These results provide new insights on the mechanism of parallel spindles leading to numerically unreduced pollen formation and on the selection and utilization of this type of pollen in polyploid breeding of P. tomentosa.

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