Project description:The Eurasian spruce bark beetle Ips typographus is known for its devasting attack on the host tree Picea abies, a common conifer in Europe. The beetle uses various pheromone components (2-methyl-3-buten-2-ol and cis-verbenol) for mass aggregation to overcome the tree defence compounds such as terpenes. Though this aggregation pheromone biosynthesis and respective precursors via terpenes detoxification mechanism was investigated for a few decades, gene-level understanding behind these biosynthesis pathways are uncertain yet in I. typographus. Though, applying Juvenile hormone (JH III) on the beetles have induced specific pheromone biosynthesis in many bark beetle species, irrespective of their life stage, it is not uniform found in all Ips species. While investigating pheromone biosynthesis among various life stages of I. typographus, we have also reported recently about the JHIII induction of aggregation pheromone biosynthesis from the gut tissue of the beetle. Thus, in this study, we have applied the concept of JHIII induction on I. typographus and analyzed the respective pheromone and possible biosynthesis precursors from via pathway gene families from the gut tissue of the beetle. A comparative approach from transcriptome and proteome study has revealed the mevalonate pathway genes including isoprenyl-di-phosphate synthase (IPDS) gene (Ityp09271) was upregulated over 5-fold change after JHIII induction in I. typographus. The identified IPDS is suspected to directly involve in 2-methyl-3-buten-2-ol, a vital aggregation pheromone of I. typographus. Added to that, a hydrolase gene family was found upregulated over 2-fold change, specifically in the male gut tissue after JHIII treatment. Furthermore, another vital gene family, CytochromeP450 have shown the upregulated (transcript) in the male gut tissue after treatment. Especially Previously reported CyP450 candidates Ityp3140 and Ityp03153 for pheromone compounds cis/trans- verbenol and ipsdienol biosynthesis respectively. Along with CyP450 candidates, the hydrolase gene candidates could possibly involve in braking down the detox compounds such as diglycosylated terpenes and stored wax esters (verbenyl oleate) from the gut possibly provided from the of the beetle body as a reservoir. An added metabolomic analysis has confirmed these compounds abundance was in the gut tissue. Especially, the abundance of the related fatty acid ester (verbenyl oleate) has reduced half in male gut tissue after the treatment. Hence, we have shed light on three possible genes from different families for the respective pheromone and its precursors biosynthesis after JHIII application over I. typographus. This approach would lead us to elucidate the molecular basis of stored pheromone biosynthesis and the derived knowledge from this study would lead to eco-friendly pest management for this aggressive pest. Key words: Ips typographus, bark beetle, pheromone biosynthesis, de novo, Juvenile hormone treatment.

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