INDUSTRIAL HEMP AND MARIJUANAARE BOTHONE AND THE SAME PLANT SPECIESCANNABIS SATIVA,HOWEVER, REPRESENT DIFFERENT VARIETIES SHOWING HIGH GENETIC, MORPHOLOGICAL, AND CHEMICAL DIVERSITY. WHILE MARIJUANA CONTAINS HIGH LEVELS (UP TO 20% OF DRY WEIGHT) OF THE PSYCHOACTIVE AND INTOXICATING CANNABINOID DELTA 9-TETRAHYDROCANNABINOL (THC), IN CONTRAST TO BE LEGALLY CONSIDERED AS INDUSTRIAL HEMP IN THE US THE CROP MAY NOT CONTAIN MORE THAN 0.3% THC. THROUGHOUT HUMAN HISTORY HEMP HAS BEEN CULTIVATED FOR FOOD, FIBER AND MEDICINE, AND IS PRESENTLY USED IN OVER 25,000 PRODUCTS. HEMP SEEDS ARE PRESSED TO OBTAIN AN OILWELL SUITED FOR HUMAN DIET AND THE RESIDUAL SEED CAKE CAN BE USED AS ANIMAL FEED. WHILE HEMP FIBERS ARE PRIMARILY USED TO PRODUCE PAPERS AND TEXTILES, THEIR USE HAS RECENTLY BEEN EXPANDED TOWARDS THE PRODUCTION OF CARBON NANOSHEETS, PLASTICS, ABSORBENT MATERIALS AND CONSTRUCTION CONCRETE. HOWEVER, IN ADDITION TO SEEDS AND FIBERS SIGNIFICANT REVENUES CAN ALSO BE ACHIEVED FOR SOME OF THE METABOLITES HIGHLY ABUNDANT IN INDUSTRIAL HEMP INCLUDING NON-INTOXICATING CANNABINOIDS SUCH AS CANNABIDIOL (CBD) AND TERPENES (UP TO 5% OF DRY WEIGHT) THAT ARE VALUED AS PHARMACEUTICALS, FRAGRANCES AND CHEMICAL FEEDSTOCKS.DESPITE THE EXISTENCE OF LARGE GENETIC DIVERSITY AND IN CONSEQUENCE A WIDE RANGE OF TERPENE AND CANNABINOID LEVELSIN CANNABIS SATIVA, THE RESEARCH AND KNOWLEDGE ABOUT THE BIOSYNTHESIS OF THESE COMPOUNDSAND THE UNDERLYING GENETICSIS STILL QUITE LIMITED DUE TO PREVIOUS LEGAL RESTRICTIONS. THE RECENT LIFTING OF THESE RESTRICTIONS HAS NOT ONLY LEAD TO A REVITALIZED INTEREST IN INDUSTRIAL HEMP AS A CROP, BUT ALSO TO THE URGENT NEED TO UNDERSTAND HOW THE BIOSYNTHESIS OF SPECIFIC TERPENES AND CANNABINOIDS ARE REGULATED SO THAT THEY CAN BE PREFERENTIALLY OVERPRODUCED OR ELIMINATED IN HEMP VARIETIES GROWN FOR PARTICULAR USES.REMARKABLY LITTLE IS KNOWN ABOUT THE REGULATION OF THESE BIOSYNTHETICPATHWAYS BY ENDOGENOUS AND ENVIRONMENTAL FACTORS.THIS IS RELEVANT FOR THE CULTIVATION OF INDUSTRIAL HEMP CONSIDERING ON THE ONE SIDETHE ECONOMIC POTENTIAL FOR THE PRODUCTION OF TERPENES AND CBD, AND ON THE OTHER SIDELEGAL RESTRICTIONS ON THC CONTENT. IN PARTICULAR THE FREQUENT OCCURRENCE OF ENVIRONMENTAL STRESS INDUCED SPIKES IN THC CONTENTSURPASSING LEGAL LIMITS REPRESENTS A SERIOUS ECONOMIC RISK FOR GROWERS. THUS, TO ADDRESS THIS CRITICAL NEED TO ESTABLISH A BETTER UNDERSTANDING OF THE TERPENE-CANNABINOID BIOSYNTHETICNETWORK IN INDUSTRIAL HEMP AND ITS REGULATION, IN THE FIRST PART OF THIS PROJECT WE WILL ANALYZE THE ACCUMULATION OF TERPENES AND CANNABINOIDS IN INDUSTRIAL HEMP UNDER DIFFERENT ENVIRONMENTAL STRESSES. IN THE SECOND PART OF THE PROJECT WE WILL ANALYZE CHANGES IN THE ACTIVITY LEVELS OF KEY TERPENE AND CANNABINOID BIOSYNTHETIC GENES UNDER THESE STRESSES, AND IDENTIFY NOVEL REGULATORY FACTORS. IN THE THIRD PART OF THE PROJECT WE WILL ENGINEER HEMP LINES TO DETERMINE HOW INACTIVATION OR UPREGULATION OF TWO KEY BIOSYNTHETIC GENES IN INDUSTRIAL HEMP VIA A NANOPARTICLE-BASED APPROACH AFFECTS ACCUMULATION OF TERPENES AND CANNABINOIDS.THE LONG-TERMGOAL OF THIS PROPOSAL IS TO ESTABLISH A BETTER UNDERSTANDING OF THE TERPENE-CANNABINOID BIOSYNTHETICNETWORK IN INDUSTRIAL HEMP AND ITS GENETIC REGULATION WHICH WILL ULTIMATELY RESULT IN THE ENGINEERING AND DEVELOPMENT OF HEMP VARIETIES WITH IMPROVEDOR ELIMINATED PRODUCTIONOF SPECIFIC TERPENE AND CANNABINOID COMPOUNDS. THIS PROJECT NOW PROVIDES THE OPPORTUNITY TO OBTAIN BASE-LINE DATA FOR SEEKING FURTHERRESEARCH OPPORTUNITIES TO INVESTIGATE THE HEMP TERPENE-CANNABINOID BIOSYNTHETICNETWORK, ITS REGULATION AND POTENTIAL FORENGINEERING VARIETIES WITH IMPROVED PRODUCTION OF SPECIFIC COMPOUNDS.IN ADDITION, THE PLANT LINES GENERATED IN THIS PROJECT WILL ALSO PROVIDE FUTURE OPPORTUNITIESTO TEST THE PERFORMANCE OF HEMP VARIETIES WITH MODIFIED TERPENE AND CANNABINOID PROFILES UNDER FIELD CONDITIONS.