Controlled pollinations with self- and cross-pollen were applied to individual flowers of five mature Eucalyptus globulus Labill. ssp. globulus trees to investigate the site of action of the self-incompatibility mechanism. Growth of pollen tubes in styles at 2 weeks after pollination and ovule penetration by pollen tubes at 2 and 4 weeks after pollination were investigated by fluorescence microscopy. Some pollinated flowers were left to develop to seed maturity, then harvested to quantify the level of self-incompatibility of each tree. Trees ranged from 76 to 100% self-incompatible. There was no significant difference in the number of pollen tubes in the style between treatments although variation was present between trees. The number of pollen tubes present was similar to the number of ovules present within flowers. Penetration of ovules by pollen tubes over all five trees combined revealed no difference between treatments at 2 weeks after pollination; however, there was slightly greater penetration by cross-pollen tubes at 4 weeks after pollination. This difference was not large enough to account for the near complete lack of selfed-seed production, suggesting late pre- or post-zygotic arrest of selfed ovules.
Controlled self-and cross-pollinations were conducted on flowers of five mature Eucalyptus nitens trees. Levels of self-sterility of the trees ranged from 25.8 to 93.6%. Pollen tube numbers in styles and ovule penetration by pollen tubes was investigated 2 weeks after pollination by fluorescence microscopy. There were no significant differences between treatments in the number of pollen tubes present in styles or in the percentage of ovules penetrated by pollen tubes. Embryology of material harvested 2 and 4 weeks after pollination was investigated by bright-field microscopy. Fertilisation had taken place by 2 weeks after pollination with nearly every ovule showing evidence of fertilisation. Cross-pollination resulted in a greater proportion of healthy, developing ovules, at both 2 and 4 weeks after pollination, compared with self-pollination. The proportion of degenerating ovules increased from 2 to 4 weeks after pollination. The reduced ability of E. nitens to set self-pollinated seed compared with cross-pollinated seed appears to be controlled by a post-zygotic mechanism. Differences in ovule size may potentially assist in the identification of trees incapable of setting self-pollinated seed.
The study was conducted to identify the self-incompatibility mechanism in Eucalyptus globulus ssp. globulus. Controlled self- and cross-pollinations were conducted on individual flowers from three mature trees that had self-incompatibility levels of 76, 99.6 and 100%. Flowers were harvested at 4, 6 and 8 weeks after pollination. Embryology was investigated by bright field microscopy on material harvested at 4 and 6 weeks after pollination. Fertilization had taken place at 4 weeks after pollination with zygotes and free nuclear endosperm visible. There was a greater proportion of healthy, fertilized ovules in the cross- compared with the self-pollination treatment, and approx. half the ovules examined from both pollen treatments were not fertilized or were degenerating. By 6 weeks after pollination a few zygotes were starting to divide. The number of healthy, fertilized ovules was still greater in the cross-pollination treatment, but the number of healthy fertilized ovules was lower in both treatments compared with 4 weeks after pollination, and many ovules were degenerating. Fertilized ovules were significantly larger than non-fertilized or degenerating ovules and this difference was detectable by eye at 6 and 8 weeks after pollination. The mechanism of self-incompatibility appears to have both late pre- and post-zygotic components.
Physical dormancy is common in seeds of arid-land legumes. Improved understanding of germination requirements of hard-seeded species will further our understanding of arid lands and aid restoration projects. We studied the germination responses of Acacia papyrocarpa (Benth.), A. oswaldii (F.Muell) and Senna artemisioides (Gaudich. ex DC.) Randell ssp. × coriacea (Benth.) Randell from a chenopod shrubland in South Australia. Imbibition testing indicated that all three species had physical dormancy, but the proportion of dormant seeds was lower in A. oswaldii. This corresponded to a thinner testa in this species. Mechanisms tested to scarify seeds included mechanical scarification and different durations of wet or dry heat. Mechanically scarified seeds germinated readily, reaching maximum numbers in 10–15 days, independently of incubation temperatures, with the exception of S. artemisioides seeds, which germinated at a slower rate in cooler temperatures. Overall, wet heat was more effective than dry heat to alleviate physical dormancy, whereas dry heat in some cases resulted in seed mortality. On the basis of these results, it is recommended that seeds of A. papyrocarpa and S. artemisoides be pretreated with wet heat in future restoration programs. No pre-treatment is required for dormancy loss in A. oswaldii seeds. The different responses of seeds of these species suggest that their populations have varying strategies for persistence in this unpredictable environment.
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