Intra‐population variations of several characteristics related to fertility were assessed for a nature‐reserve population of Primula sieboldii in the flood plain of the Arakawa River. Although there were large intra‐ and inter‐clonal variations of fruit and seed sets as well as other female fertility components in the 13 clones examined, female fertility was generally low, except in a homomorph clone. Pollinator availability was assessed by continuous monitoring of insect visits with 8 mm film. No insect visits were recorded during the monitoring of total 68 flowers over 16 hours in the nature‐reserve population. In contrast, a high pollinator visitation rate was recorded in a reference population of P. sieboldii on the flank of Mt. Yatugatake. Thus, the low rates of seed production in the nature reserve was due to a shortage of compatible pollens as a result of the limitation of pollinator availability. Stigmas of flowers in various clones occasionally were loaded with pollen grains whose sizes were not significantly different from those of anther pollens of the same flowers. However, pollen germination on stigmas was observed only in the flowers of a homo‐morph clone whose flowers have pin‐type stigmas and anthers with compatible thrum‐type pollens at the same height. Exceptionally high fertility of the homo‐morph clone would be associated with its inbreeding habit.
Hybrid lethality expressed in the interspecific hybrid of Nicotiana suaveolens Lehm. x N. tabacum L. cv. Hicks-2 is one of the mechanisms for reproductive isolation and it is temperature-sensitive. Apoptotic changes were detected in the cells of hybrid seedlings and calli expressing lethality at 28 degrees C but not under high-temperature conditions (36 degrees C), when the lethality is suppressed. Condensation of chromatin, fragmentation of nuclei and cytoplasmic reduction are the cytological changes associated with apoptosis leading to hybrid lethality. Fragmentation of nuclei was correlated with the lethal symptoms in both hybrid seedlings and calli, as confirmed by fluorimetry of the nuclear DNA using laser scanning cytometry. Agarose gel analysis of DNA extracted from hybrid seedlings and calli showing lethal symptoms revealed a specific ladder pattern suggesting nucleosomal fragmentation which is one of the biochemical changes of apoptosis. In-situ detection using terminal deoxyribonucleotidyl transferase-mediated dUTP-fluorescein nick end labeling (TUNEL) showed that this process occurred in distinct stages on each organ of hybrid seedlings and centripetally in hybrid calli. From these results, we confirmed that cell death inducing hybrid lethality was indeed apoptosis.
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