Abstract
SUMMARY Cotton is a high-value commercial crop in Australia. Although cotton is largely selfpollinating, previous researchers have reported that honeybees, Apis mellifera, can assist in cross-pollination and contribute to improved yield. Until recently, use of bees in cotton had, however, been greatly limited by excessive use of pesticides to control arthropod pests. With the widespread use of transgenic (Bt) cotton varieties and the associated reduction in pesticide use, I decided to investigate the role and importance of honeybees in Bt cotton, under Australian conditions. I conducted two major field trials at Narrabri, in the centre of one of Australia’s major cotton-growing areas, in the 2005-6 and 2006-7 seasons. In the first trial, I particularly assessed methods of manipulating honeybee colonies by feeding pollen supplements of pollen/soybean patties, and by restricting pollen influx by the fitting of 30% efficient pollen traps. I aimed to test whether either of these strategies increased honeybee flight activity and, thus, increased foraging on cotton flowers. My results showed that although supplementary feeding increased bee flight activity and brood production, it did not increase pollen collection on cotton. Pollen traps initially reduced flight activity. They also reduced the amount of pollen stored in colonies, slowed down brood rearing activity, and honey production. However, they did not contribute to increased pollen collection in cotton. In the second trial, I spent more time investigating honeybee behaviour in cotton as well as assessing the effect of providing flowering cotton plants with access to honeybees for different time periods (e.g. 25 d, 15 d, 0 d). In this year, I used double the hive stocking rate of (16 colonies / ha) than in the previous year, because in 2005-6 I observed few bees in cotton flowers. I also conducted a preliminary investigation to assess whether there was any gene flow over a 16 m distance from Bt cotton to conventional cotton, in the presence of a relatively high honeybee population. xvii Both of my field experiments showed that honeybees significantly increased cotton yield via increased boll set, mean weight of bolls, number of seeds / boll, and weight of lint / boll. It was obvious that cotton flowers, and particularly cotton pollen, were not attractive to honeybees, and this was also reflected in the low proportion (5.3% w/w) of pollen from cotton collected in the pollen traps. However, flower visitation rate was generally above the 0.5% level regarded as optimal for cross-pollination in cotton, and this was reflected in increased yield parameters. I recorded a gene flow of 1.7 % from Bollgard®II cotton to conventional cotton, over a distance of 16 m. This is much higher than had previously been reported for Australia, and may have been a result of high honeybee numbers in the vicinity, associated with my managed hives. In an attempt to attract more honeybees to cotton flowers, I conducted an investigation where I applied synthetic Queen Mandibular Pheromone (QMP) (Fruit Boost®) at two rates, 50 QEQ and 500 QEQ / ha, and for two applications, 2 d apart. Neither rate of QMP increased the level of bee visitation to flowers, either on the day of application or the subsequent day. There was also no increase in boll set or yield in plants treated with QMP. My observations of honeybee behaviour in cotton brought some interesting findings. First, honeybees totally ignored extrafloral nectaries. Second, most flower-visiting honeybees collected nectar, but the overwhelming majority of them (84%) collected floral nectar from outside flowers: this meant these bees did not contribute to pollination. Those nectar gatherers which entered flowers did contribute to pollination. However, they were observed to exhibit rejection of cotton pollen by scraping pollen grains from their body and discarding them, prior to returning to their hives. Pollen gatherers collected only small, loose pellets from cotton. xviii SEM studies showed that cotton pollen grains were the largest of all pollen commonly collected by bees in my investigations, and that they also had large spines. It is likely that these characteristics make cotton pollen unattractive to honeybees. Another possible reason for the unattractiveness of cotton flowers was the presence of pollen beetles, Carpophilus aterrimus, in them. I conducted a series of studies to determine the role of pollen beetles in pollination of cotton. I found that they did not contribute to pollination at low levels; at high populations they damaged flowers (with ≥ 10 beetles / flower, no flowers set bolls); and that honeybees, when given the choice, avoid flowers with pollen beetles. Because the insecticide fipronil was commonly used in Australian cotton at flowering time, and because I had some experience of its toxic effects against honeybees in my field investigations, I conducted a series of laboratory and potted plant bioassays, using young worker bees. The studies confirmed its highly toxic nature. I recorded an acute dermal LD50 of 1.9 ng / bee, and an acute oral LC50 of 0.62 ppm. Fipronil’s residual toxicity also remained high for an extended period in both laboratory and potted plant trials. For example, when applied to cotton leaves in weather-exposed potted cotton plants, it took 25 d and 20 d for full and half recommended rates of fipronil, respectively, to become non-toxic to honeybees. I had previously investigated whether a shorter period of exposure of cotton plants to honeybees would contribute adequately to increased yield, and concluded that a 10 d window within a 25 d flowering period would contribute 55% of the increase in total weight of bolls contributable to honeybee pollination, but only 36% of the increase in weight of lint. Given the highly residual activity of fipronil I recorded, the only opportunity for an insecticide-free period during flowering would be at its commencement. xix I concluded that, while there is evidence that honeybees can contribute to increased cotton yield in Bt cotton in Australia, this is unlikely with the continued use of fipronil at flowering