Reduced levels of cadinane sesquiterpenoids in cotton plants expressing antisense (+)-delta-cadinene synthase.
Cotton plants were transformed with an antisense construct of cdn1-Cl, a member of a complex gene family of delta-(+)cadinene (CDN) synthase. This synthase catalyzes the cyclization of (E,E)-farnesyl diphosphate to form CDN, and in cotton, it occupies the committed step in the biosynthesis of cadinane sesquiterpenoids and heliocides (sesterterpenoids). Southern analyses of the digestion of leaf DNA from R(o), T(o), and T(1) plants with Hind III, Pst I and Kpn I restriction enzymes show the integration of antisense cdn1-C1 cDNA driven by the CaMV 35S promoter into the cotton genome. Northern blots demonstrate the appearance of cdn synthase mRNA preceding CDN synthase activity and the formation of gossypol in developing cottonseed. T(2) cottonseed show a reduced CDN synthase activity and up to a 70% reduction in gossypol. In T(1) leaves the accumulated amounts of gossypol, hemigossypolone and heliocides are reduced 92.4, 83.3 and 68.4%, respectively. These data demonstrate that the integration of antisense cdn1-C1 cDNA into the cotton genome leads to a reduction of CDN synthase activity and negatively impacts on the biosynthesis of cadinane sesquiterpenoids and heliocides in cotton plants.
Martin
Gail S
GS
Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX 77843-2128, USA.
Liu
Jinggao
J
Benedict
Chauncey R
CR
Stipanovic
Robert D
RD
Magill
Clint W
CW
D015139:Blotting, Southern; D003368:Gossypium / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D007535:Isomerases / Q000235:genetics* / Q000378:metabolism*; D015394:Molecular Structure; D016372:RNA, Antisense / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012717:Sesquiterpenes / Q000737:chemistry / Q000378:metabolism*
D016372:RNA, Antisense; D012333:RNA, Messenger; D012717:Sesquiterpenes; D007535:Isomerases; C097608:delta-cadinene synthase