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Materials and Methods for the Alteration of Enzyme and Acetyl Coa Levels in Plants Basil J

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Materials and Methods for the Alteration of Enzyme and Acetyl Coa Levels in Plants Basil J Iowa State University Patents Iowa State University Research Foundation, Inc. 4-28-2009 Materials and methods for the alteration of enzyme and acetyl CoA levels in plants Basil J. Nikolau Iowa State University, [email protected] Eve S. Wurtele Iowa State University, [email protected] David J. Oliver Iowa State University, [email protected] Patrick S. Schnable Iowa State University, [email protected] Tsui-Jung Wen Iowa State University Follow this and additional works at: http://lib.dr.iastate.edu/patents Part of the Agriculture Commons, Agronomy and Crop Sciences Commons, Biochemistry Commons, Botany Commons, Developmental Biology Commons, and the Molecular Biology Commons Recommended Citation Nikolau, Basil J.; Wurtele, Eve S.; Oliver, David J.; Schnable, Patrick S.; and Wen, Tsui-Jung, "Materials and methods for the alteration of enzyme and acetyl CoA levels in plants" (2009). Iowa State University Patents. 30. http://lib.dr.iastate.edu/patents/30 This Patent is brought to you for free and open access by the Iowa State University Research Foundation, Inc. at Iowa State University Digital Repository. It has been accepted for inclusion in Iowa State University Patents by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Materials and methods for the alteration of enzyme and acetyl CoA levels in plants Abstract The present invention provides nucleic acid and amino acid sequences of acetyl CoA synthetase (ACS), plastidic pyruvate dehydrogenase (pPDH), ATP citrate lyase (ACL),Arabidopsis pyruvate decarboxylase (PDC), and Arabidopsis aldehyde dehydrogenase (ALDH), specifically ALDH-2 and ALDH-4. The present invention also provides a recombinant vector comprising a nucleic acid sequence encoding one of the aforementioned enzymes, an antisense sequence thereto or a ribozyme therefor, a cell transformed with such a vector, antibodies to the enzymes, a plant cell, a plant tissue, a plant organ or a plant in which the level of an enzyme has been altered, and a method of producing such a plant cell, plant tissue, plant organ or plant. Desirably, alteration of the level of enzyme results in an alteration of the level of acetyl CoA in the plant cell, plant tissue, plant organ or plant. In addition, the present invention provides a recombinant vector comprising an antisense sequence of a nucleic acid sequence encoding pyruvate decarboxylase (PDC), the E1α subunit of pPDH, the E1β subunit of pPDH, the E2 subunit of pPDH, mitochondrial pyruvate dehydrogenase (mtPDH) or aldehyde dehydrogenase (ALDH) or a ribozyme that can cleave an RNA molecule encoding PDC, E1α pPDH, E1β pPDH, E2 pPDH, mtPDH or ALDH. Keywords Agronomy, Biochemistry Biophysics and Molecular Biology, Genetics Development and Cell Biology Disciplines Agriculture | Agronomy and Crop Sciences | Biochemistry | Botany | Developmental Biology | Molecular Biology This patent is available at Iowa State University Digital Repository: http://lib.dr.iastate.edu/patents/30 111111 1111111111111111111111111111111111111111111111111111111111111 US007524678B2 c12) United States Patent (10) Patent No.: US 7,524,678 B2 Nikolau et al. (45) Date of Patent: Apr. 28, 2009 (54) MATERIALS AND METHODS FOR THE Bourguignon et al, Eur. J Biochem, 204, 865-873 (1992). ALTERATION OF ENZYME AND ACETYL Bucher et al., EMBO Journal, 13 (12), 2755-2763 (1994). COA LEVELS IN PLANTS Camp eta!., Plant Physiol, 77, 571-577 (1985). Conner et a!., Database Biosis (Online), Database Accession No. (75) Inventors: Basil J. Nikolau, Ames, IA (US); Eve S. PREV199799296162, XP002134361, Abstract (1996) & Planta, Wurtele, Ames, IA (US); David J. 200(2), 195-202 (1996). Oliver, Ames, IA (US); Patrick S. Cui et al., Science, 272, 1334-1336 (1996). Schnable, Ames, IA (US); Tsui-Jung DBEST ID: 1276510, XP002134354 (Sep. 19, 1997) & EMBL Wen, Ames, IA (US) Accession No. Aa597828 (Sep. 24, 1997). DBEST ID: 1436389, XP002134345 (Jan. 5, 1998) & EMBLAcces­ (73) Assignee: Iowa State University Research sion No. N96514 (Apr. 19, 1996). Foundation, Inc., Ames, IA (US) DBEST ID: 1436723, XP002134344 (Jan. 5, 1998) & EMBLAcces­ sion No. N97183 (Apr. 19, 1996). ( *) Notice: Subject to any disclaimer, the term of this DBEST ID: 1437099, XP002134349 (Jan. 5, 1998) & EMBLAcces­ sion No. W43461 (May 27, 1996). patent is extended or adjusted under 35 DBEST ID: 14439, XP002134341 (Nov. 10, 1992) & EMBL Acces­ U.S.C. 154(b) by 393 days. sion No. Z18045 (Nov. 6, 1992). DBEST ID: 316729, XP002134342 (Apr. 14, 1993) &EMBLAcces­ (21) Appl. No.: 11/167,856 sion No. R30081 (Aug. 11, 1995). DBEST ID: 31724, XP002134350 (Sep. 9, 1993) & EMBL Acces­ (22) Filed: Jun.27,2005 sion No. Z26027 (Sep. 8, 1993). DBEST ID: 328510, XP002134352 (Jul. 25, 1995) & EMBL Acces­ (65) Prior Publication Data sion No. R90074 (Aug. 28, 1995). US 2005/0268352 AI Dec. 1, 2005 DBEST ID: 378529, XP002134343 (Nov. 6, 1995) & EMBL Acces­ sion No. H76244 (Nov. 10, 1995). Related U.S. Application Data DBEST ID: 551062, XP002134357 (Aug. 16, 1995) & EMBL Accession No. W43179 (May 27, 1996). (62) Division of application No. 10/293,865, filed on Nov. DBEST ID: 504075, XP002134353 (Aug. 16, 1995) & EMBL 13, 2002, now Pat. No. 6,942,994, which is a division Accession No. N97215 (Apr. 19, 1996). of application No. 09/344,882, filed on Jun. 25, 1999, Desprez eta!., EMBL Accession No. Z25661, XP002134347 (Aug. now Pat. No. 6,764,851. 24, 1993) & Trembl Accession No. Q42036 (Nov. 1, 1996). (60) Provisional application No. 60/090,717, filed on Jun. (Continued) 26, 1998. Primary Examiner-James S Ketter (51) Int. Cl. (74) Attorney, Agent, or Firm--Carol Larcher; Larcher & C12N 5110 (2006.01) Chao Law Group (52) U.S. Cl. ....................... 435/375; 536/23.6; 800/298 (58) Field of Classification Search ....................... None (57) ABSTRACT See application file for complete search history. (56) References Cited The present invention provides nucleic acid and amino acid sequences of acetyl CoA synthetase (ACS), plastidic pyru­ U.S. PATENT DOCUMENTS vate dehydrogenase (pPDH), ATP citrate lyase (ACL), Ara­ bidopsis pyruvate decarboxylase (PDC), and Arabidopsis 5,270,175 A 12/1993 Moll aldehyde dehydrogenase (ALDH), specifically ALDH-2 and 5,684,242 A 1111997 Schnable et al. 5,695,973 A 12/1997 Subbiah ALDH-4. The present invention also provides a recombinant vector comprising a nucleic acid sequence encoding one of FOREIGN PATENT DOCUMENTS the aforementioned enzymes, an antisense sequence thereto or a ribozyme therefor, a cell transformed with such a vector, wo WO 96/12029 A1 4/1996 antibodies to the enzymes, a plant cell, a plant tissue, a plant wo wo 97/27295 7/1997 organ or a plant in which the level of an enzyme has been wo wo 97/32986 9/1997 altered, and a method of producing such a plant cell, plant wo wo 98/00557 111998 tissue, plant organ or plant. Desirably, alterationofthe level of wo wo 98/06831 2/1998 wo wo 98/53086 1111998 enzyme results in an alteration of the level of acetyl CoA in wo wo 99/00505 111999 the plant cell, plant tissue, plant organ or plant. In addition, wo wo 99/13067 3/1999 the present invention provides a recombinant vector compris­ wo W000/11199 3/2000 ing an antisense sequence of a nucleic acid sequence encod­ ing pyruvate decarboxylase (PDC), theE 1a subunit ofpPD H, OTHER PUBLICATIONS the El ~subunit ofpPDH, the E2 subunit of pPDH, mitochon­ drial pyruvate dehydrogenase (mtPDH) or aldehyde dehydro­ Behal et al, EMBLAccession No. AF036618, XP002126950 (Jan. 5, genase (ALDH) or a ribozyme that can cleave an RNA mol­ 1998). ecule encoding PDC, Ela pPDH, El~ pPDH, E2 pPDH, Bevan eta!, EMBLAccession No. AL031804, XP002134360 (Oct. 1, mtPDH or ALDH. 1998) & Trembl Accession No. 082647 (Nov. 1, 1998). Bevan et al, EMBL Accession No. Z97340, XP002134339 (Jul. 4, 1997). 9 Claims, 15 Drawing Sheets US 7,524,678 B2 Page 2 OTHER PUBLICATIONS Nakamura, EMBLAccession No. AB023034, XP002134359 (Feb. 5, 1999). Desprez eta!., EMBL Accession No. Z33810, XP002134348 (May Nakamura, EMBL Accession No. AB026636, XP002134358 (May 23, 1994). 7, 1999). Dolferus eta!., EMBL Accession No. U71121, XP002134351 (Oct. Nelson eta!., Plant Physiol., 55, 69-72 (1975). 11, 1996) & Trembl Accession No. Q96535 (Feb. 1, 1997). Newman eta!. Accession No. N38599 Jan. 5, 1998. Dolferus eta!, EMBL Accession No. U71122, XP002134356 (Oct. Newman, DBEST ID: 1427622, XP002134337 (Dec. 30, 1997) & 11, 1996) & Trenbl Accession No. Q96536 (Feb. 1, 1997). EMBL Accession No. AA720359 (Jan. 7, 1998). Elshourbagy eta!., Eur. J Biochern, 204,491-499 (1992). Newman et a!., EMBL Accession No. AA585946, XP002134335 Elshourbagy eta!., J Bioi. Chern., 265 (3), 1430-1435 (1990). (Sep. 13, 1997). Engels eta!., Microbiology, 143, 3543-3553 (1997). Newman et al., EMBL Accession No. Tl3703, XP002134337 (Mar. Etchegaray eta!., Database Bios is (Online), Database Accession No. 6, 1994). 179045, XP002127203, Abstract (1998) & Biochemistry and Newman eta!., EMBL Accession No. T44006, XP002134338 (Feb. Molecular Biology International, 44(2), 235-243 (1998). 4, 1995). Federspiel et a!., EMBL Accession No. AC02292, XP002134340 Newman eta!., Plant Physiol., 106, 1241-1255 (1994). (Jun. 20, 1997) & Trembl Accession No. 022718 (Jan. 1, 1998). Nicholl, D.S.T. An introduction to genetic engineering pp. 77-85 Fritsch eta!., Plant Physiol, 63, 687-691 (1979). 1994. Guan eta!., J Bioi. Chern., 270(10), 5412-5417 (1995). Rangasamy et al., Database Bios is (Online), Database Accession No. Hofte et al., EMBL Accession No. Z26232, XP002134346 (Sep. 10, PREV199799687761, XP002134366, Abstract (1997) & Plant Cell 1993) & Trembl Accession No. Q42079 (Nov. 1, 1996). Reports, 16(10), 700-704 (1997). Huang eta!., Arch. Biochern. Biophys., 140, 158-173 (1970). Ratledge eta!., Lipids, 321(1), 7-12 (1997). Johnston et a!., Biochirnica et Biophysica Acta, 1321, 200-206 Reid eta!., Plant Physiol., 59, 842-848 (1977). (1997). Kaethner et al., Planta, 163, 290-294 (1985). Roughan et al., Analyt. Biochern., 216, 77-82 (1994). Lee et al., J Bioi.
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