Trichome specific promoters for the manipulation of cannabinoids and other compounds in glandular trichomes

Abstract

The present technology provides trichome specific promoters of cannabinoid biosynthesis enzyme genes from Cannabis, nucleotide sequences of the trichome specific promoters, and uses of the promoters for modulating the production of cannabinoids and other compounds in organisms. The present technology also provides chimeric genes, vectors, and transgenic cells and organisms, including plant cells and plants, comprising the trichome specific promoters. Also provided are methods for expressing nucleic acid sequences in cells and organisms using the trichome specific promoters.

1 - 35 . (canceled) 
     
     
         36 . A method for increasing a cannabinoid in a host plant trichome, comprising:
 (a) introducing into a host cell an expression vector comprising a nucleotide sequence selected from the group consisting of:
 (i) a nucleotide sequence set forth in any one of SEQ ID NOs: 1-3, 5, 6, 8, 9, 11-14, 16-19, 21-26, 28, 29, or 31-33; and 
 (ii) a nucleotide sequence that is at least about 90% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-3, 5, 6, 8, 9, 11-14, 16-19, 21-26, 28, 29, or 31-33, and which encodes a promoter that has plant trichome gland specific transcriptional activity; 
   wherein the nucleic acid sequence of (i) or (ii) is operably linked to one or more nucleic acid sequences encoding an enzyme of the cannabinoid biosynthetic pathway; and   (b) growing the plant under conditions which allow for the expression of the cannabinoid biosynthetic pathway enzyme;   wherein expression of the cannabinoid biosynthetic pathway enzyme results in the plant having an increased cannabinoid content relative to a control plant grown under similar conditions.   
     
     
         37 . The method of  claim 36 , wherein the cannabinoid biosynthetic pathway enzyme is cannabidiolic acid (CBDA) synthase, cannabichromenic acid (CBCA) synthase, or Δ9tetrahydrocannabinolic acid (THCA) synthase. 
     
     
         38 . The method of  claim 37 , further comprising providing the plant with cannabigerolic acid (CBGA). 
     
     
         39 . A genetically-engineered plant produced by the method of  claim 36 , wherein the plant has increased Δ9tetrahydrocannabinol (THC), cannabichromene (CBC), and/or cannabidiol (CBD) content relative to a control plant. 
     
     
         40 . A synthetic DNA molecule comprising a nucleotide sequence selected from the group consisting of:
 (a) a nucleotide sequence set forth in any one of SEQ ID NOs: 31, 32, or 33; and   (b) a nucleotide sequence that is at least about 90% identical to the nucleotide sequence of any one of SEQ ID NOs: 31, 32, or 33, and which encodes a promoter having plant trichome gland specific transcriptional activity,   wherein the nucleotide sequence is operably linked to a heterologous nucleic acid.   
     
     
         41 . An expression vector comprising the DNA molecule of  claim 40 , operably linked to one or more nucleic acid sequences encoding a polypeptide. 
     
     
         42 . A genetically engineered host cell comprising the expression vector of  claim 41 . 
     
     
         43 . The genetically engineered host cell of  claim 42 , wherein the cell is a  Cannabis sativa  cell. 
     
     
         44 . The genetically engineered host cell of  claim 42 , wherein the cell is a  Nicotiana tabacum  cell. 
     
     
         45 . A genetically engineered plant comprising a cell comprising a chimeric nucleic acid construct comprising the synthetic DNA molecule of  claim 40 . 
     
     
         46 . The engineered plant of  claim 45 , wherein the plant is an  N. tabacum  plant. 
     
     
         47 . The engineered plant of  claim 45 , wherein the plant is a  C. sativa  plant. 
     
     
         48 . Seeds from the engineered plant of  claim 45 , wherein the seeds comprise the chimeric nucleic acid construct.