Late maturity alpha-amylase (LMA) in wheat is a genetic defect that may result in the accumulation of unacceptable levels of high pI alpha-amylase in grain in the absence of germination or weather damage. During germination, gibberellin produced in the embryo triggers expression of alpha-Amy genes, the synthesis of alpha-amylase and, subsequently, cell death in the aleurone. LMA also involves the aleurone and whilst LMA appears to be independent of the embryo there is nevertheless some evidence that gibberellin is involved. The aim of this investigation was to determine whether the increase in alpha-amylase activity in LMA-prone genotypes, like alpha-amylase synthesis by aleurone cells in germinating or GA-challenged grains, is followed by aleurone cell death. Programmed cell death was seen in aleurone layers from developing, ripe and germinated grains using confocal microscopy and fluorescent probes specific for dead or living cells. Small pockets of dying cells were observed distributed at random throughout the aleurone of ripening LMA-affected grains and by harvest-ripeness these cells were clearly dead. The first appearance of dying cells, 35 d post-anthesis, coincided with the later part of the 'window of sensitivity' in grain development in LMA-prone wheat cultivars. No dead or dying cells were present in ripening or fully ripe grains of control cultivars. In germinating grains, dying cells were observed in the aleurone adjacent to the scutellum and, as germination progressed, the number of dead cells increased and the affected area extended further towards the distal end of the grain. Aside from the obvious differences in spatial distribution, dying cells in 20-24 h germinated grains were similar to dying cells in developing LMA-affected grains, consistent with previous measurements of alpha-amylase activity. The increase in high pI alpha-amylase activity in developing grains of LMA-prone cultivars, like alpha-amylase synthesis in germinating grains, is associated with cell death, providing further evidence for the involvement of gibberellin in the LMA response.

Plants of malting barley variety Schooner were exposed to 5 days of high temperatures (up to 35°C) during mid grain filling under controlled environment conditions. Grains from heat treated plants accumulated c. 30% less starch than grains from control plants (21/16°C; 14 h day). Reduced starch deposition was not limited by assimilate levels in heat treated grains, but high temperature reduced the conversion of sucrose to starch. The reduction in starch synthesis appeared to result from the effects of diminished catalytic activity of a number of enzymes in the committed pathway of starch synthesis, and/or delayed recovery of enzyme activity in the cooler recovery conditions. Reductions of 11–75% in the activity of the enzymes under investigation followed high temperature exposure. In addition, ADPglucose pyrophosphorylase, branching enzyme and granule bound starch synthase showed increased activity during exposure to moderate temperatures (28–32°C), but reduced activity at high temperatures, while soluble starch synthase showed an immediate loss of activity, even at moderate temperatures. Sucrose synthase and UDPglucose pyrophosphorylase showed the greatest reduction in catalytic activity after plants were returned to cooler conditions. Individual enzymes showed variation in the level of recovery under the cooler temperature conditions which followed the heating period.

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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.