Bees play a vital role as pollinators, contributing to approximately one-third of our food supply. 80 percent of flowering plants are relying on bee pollination. (Dolezal, A.G., et al., 2018)The economic benefits of bee pollination are estimated to be around 212 billion US dollars annually, accounting for nearly 9.5% of the global crop output value. (Khalifa, S.A.M. et al., 2021)

Unfortunately, bees are adversely affected by a range of environmental pressures, resulting in significant impairments to their foraging behavior. (Le Conte, Y et al., 2008)Take honey bees in Taiwan for example, due to the climate change, it have reduced 40 percent of bees this year. (Chao-Xin Zhang, 2010)The environmental pressure results in significant impairments to their flight endurance and cognitive abilities. This phenomenon has given rise to the "pollination crisis," an urgent issue demanding immediate attention.

The situation is concerning, but our project is dedicated to addressing it. Pyrroloquinoline quinone, or PQQ, is a redox cofactor and antioxidant well-known for its potential to enhance mitochondrial regeneration and boost cognitive abilities, as demonstrated in mice studies (Mohamad Ishak, N.S., et al, 2023)(Itoh, Y, et al, 2016). To harness the benefits of PQQ for bees, we've constructed a plasmid and utilized it to transform Bacillus subtilis, a bacterium, with the PQQ biosynthesis pathway. We are then feeding the bees with the bacterial supernatant as a supplement to assess its effects.

For safety, we employed the Golden Gate method to construct the plasmid, ensuring the absence of additional restriction sites. Furthermore, we conducted codon usage optimization for Bacillus subtilis and devised a Blue-Light-inducible Kill Switch in tandem with the LacI/O Regulatory System.

Encouragingly, our preliminary data strongly support the positive effects of PQQ on bees, particularly in terms of cognitive ability and physical activity. With these benefits, bees may be better able to find suitable honey sources and return to the hive more easily. In other words, our project proposes a promising supplement to improve bees' resistance to environmental stress and alleviate the pollination crisis in the future.