Metabolic compensation of steroidal glycoalkaloid biosynthesis in transgenic potato tubers: using reverse genetics to confirm the in vivo enzyme function of a steroidal alkaloid galactosyltransferase

被引:75
作者
McCue, KF
Shepherd, LVT
Allen, PV
Maccree, MM
Rockhold, DR
Corsini, DL
Davies, HV
Belknap, WR
机构
[1] USDA, Crop Improvement & Utilizat Res Unit, ARS, Western Reg Res Ctr, Albany, CA 92710 USA
[2] Scottish Crop Res Inst, Qual Hlth & Nutr Programme, Dundee DD2 5DA, Scotland
[3] USDA, Small Grains & Potato Germplasm Res Unit, ARS, Aberdeen, ID 83210 USA
关键词
antisense; chaconine; galactosyl transferase; secondary metabolism; solanine; transgene;
D O I
10.1016/j.plantsci.2004.08.006
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Steroidal glycoalkaloids (SGAs) are secondary metabolites of Solanaceous plants. Two predominant glycoalkaloids, alpha-chaconine and alpha-solanine are produced in potatoes. An antisense transgene was constructed to down-regulate glycoalkaloid biosynthesis using a potato cDNA encoding a sterol alkaloid glycosyltransferase (Sgt1). Introduction of this construct into potatoes resulted in some lines with an almost complete inhibition of alpha-solanine accumulation. This inhibition was compensated by elevated levels of alpha-chaconine and resulted in wild type total SGA levels in the transgenic lines. In vitro assays with the recombinant SGT1 isolated from yeast demonstrated that Sgt1 encodes an enzyme capable of both glucosyltransferase and galactosyltransferase activity with a preference for UDP-galactose as the sugar donor. Together this data confirms SGT1's role in vivo as the solanidine:UDP-galactose galactosyltransferase. (C) 2004 Elsevier Ireland Ltd. All rights reserved.
引用
收藏
页码:267 / 273
页数:7
相关论文
共 28 条
[11]   Chalcone synthase cosuppression phenotypes in petunia flowers: Comparison of sense vs antisense constructs and single-copy vs complex T-DNA sequences [J].
Jorgensen, RA ;
Cluster, PD ;
English, J ;
Que, QD ;
Napoli, CA .
PLANT MOLECULAR BIOLOGY, 1996, 31 (05) :957-973
[12]  
MCCUE KF, 2003, ACTA HORTIC, V619, P77
[13]   ISOLATION AND SEQUENCE-ANALYSIS OF CDNAS FOR THE MAJOR POTATO-TUBER PROTEIN, PATATIN [J].
MIGNERY, GA ;
PIKAARD, CS ;
HANNAPEL, DJ ;
PARK, WD .
NUCLEIC ACIDS RESEARCH, 1984, 12 (21) :7987-8000
[14]   Cloning and expression of solanidine UDP-glucose glucosyltransferase from potato [J].
Moehs, CP ;
Allen, PV ;
Friedman, M ;
Belknap, WR .
PLANT JOURNAL, 1997, 11 (02) :227-236
[15]  
RICKEY TM, 1991, PLANT MOL BIOL, P16
[16]   ANTISENSE RNA INHIBITION OF BETA-GLUCURONIDASE GENE-EXPRESSION IN TRANSGENIC TOBACCO PLANTS [J].
ROBERT, LS ;
DONALDSON, PA ;
LADAIQUE, C ;
ALTOSAAR, I ;
ARNISON, PG ;
FABIJANSKI, SF .
PLANT MOLECULAR BIOLOGY, 1989, 13 (04) :399-409
[17]  
SNYDER GW, 1993, PLANT CELL REP, V12, P324, DOI 10.1007/BF00237428
[18]   PURIFICATION AND CHARACTERIZATION OF SOLANIDINE GLUCOSYLTRANSFERASE FROM THE POTATO (SOLANUM-TUBEROSUM) [J].
STAPLETON, A ;
ALLEN, PV ;
FRIEDMAN, M ;
BELKNAP, WR .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 1991, 39 (06) :1187-1193
[19]   A potato α-glucosidase gene encodes a glycoprotein-processing α-glucosidase II-like activity.: Demonstration of enzyme activity and effects of down-regulation in transgenic plants [J].
Taylor, MA ;
Ross, HA ;
McRae, D ;
Stewart, D ;
Roberts, I ;
Duncan, G ;
Wright, F ;
Millam, S ;
Davies, HV .
PLANT JOURNAL, 2000, 24 (03) :305-316
[20]   GLYCOALKALOIDS AS PEST RESISTANCE FACTORS [J].
TINGEY, WM .
AMERICAN POTATO JOURNAL, 1984, 61 (03) :157-167