To determine female fitness, the flowers of the four transgenic plant lines were emasculated by removing the anthers. This enabled the researchers to measure only animal-mediated fertilization success rates, because self-pollination was prevented a method utilized by plant breeders. It could be shown that only the control plants were normally cross-pollinated by pollen of the surrounding wild-grown tobacco plants, whereas the transgenic nicotine- and benzyl acetone-deficient lines could only produce less than half of the seeds.
The scientists measured the male fitness of the four transgenic lines by emasculating flowers of plants and subsequently determining the origin of pollen which had fertilized their seeds with DNA probes. This paternity test allowed scientists to identify which of the transgenic plant lines were most successful at passing their pollen along to neighboring plants. Here it could also be shown that the control plants producing natural amounts of nicotine and benzyl acetone were the most potent ones; the big losers (almost five times less of cross-fertilized seeds) were the plants that produced neither nicotine nor benzyl acetone.
Interestingly, during the growing season, the male fertilization success switched from the attractant (benzyl acetone)-deficient to the nicotine-deficient plants. In other words: The influence of nicotine in the nectar on successful pollinator-mediated fertilization of tobacco plants decreased continuously, whereas the attractant became more and more important. These measurements were confirmed by video recordings which showed that early in the year, when hummingbirds visit tobacco, nicotine in the nectar causes them to drink less of the bitter nectar, and in turn visit other flowers, thus increasing pollen transfer. Later in the year, moths visit frequently, attracted by the odor of benzyl acetone. The bitter
|Contact: Ian T. Baldwin|
Max Planck Institute for Chemical Ecology