Induction of potato steroidal glycoalkaloid biosynthetic pathway by overexpression of cDNA encoding primary metabolism HMG-CoA reductase and squalene synthase.
Potato steroidal glycoalkaloids (SGAs) are toxic secondary metabolites whose total content in tubers must be regulated. SGAs are biosynthesized by the sterol branch of the mevalonic acid/isoprenoid pathway. In a previous study, we showed a correlation between SGA levels and the abundance of transcript coding for HMG-CoA reductase 1 (HMG1) and squalene synthase 1 (SQS1) in potato tissues and potato genotypes varying in SGA content. Here, Solanum tuberosum cv. Desirée (low SGA producer) was transformed with a gene construct containing the coding region of either HMG1 or SQS1 of Solanum chacoense Bitt. clone 8380-1, a high SGA producer. SGA levels in transgenic HMG-plants were either greater than (in eight of 14 plants) or no different from untransformed controls, whereas only four of 12 SQS-transgenics had greater SGA levels than control, as determined by HPLC. Quantitative real-time PCR was used to estimate relative steady-state transcript levels of isoprenoid-, steroid-, and SGA-related genes in leaves of the transgenic plants compared to nontransgenic controls. HMG-transgenic plants exhibited increased transcript accumulation of SQS1, sterol C24-methyltransferase type1 (SMT1), and solanidine glycosyltransferase 2 (SGT2), whereas SQS-transgenic plants, had consistently lower transcript levels of HMG1 and variable SMT1 and SGT2 transcript abundance among different transgenics. HMG-transgenic plants exhibited changes in transcript accumulation for some sterol biosynthetic genes as well. Taken together, the data suggest coordinated regulation of isoprenoid metabolism and SGA secondary metabolism.
Institute of Plant Sciences, ARO, the Volcani Center, 50250 Bet Dagan, Israel. iditgin@volcani.agri.gov.il
Idit
I
Ginzberg
Muddarangappa
M
Thippeswamy
Edna
E
Fogelman
Ufuk
U
Demirel
Alice M
AM
Mweetwa
James
J
Tokuhisa
Richard E
RE
Veilleux
D000470:Alkaloids / Q000096:biosynthesis*; D053898:Biosynthetic Pathways / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid; D018076:DNA, Complementary / Q000235:genetics*; D013186:Farnesyl-Diphosphate Farnesyltransferase / Q000235:genetics* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics; D006903:Hydroxymethylglutaryl CoA Reductases / Q000235:genetics* / Q000378:metabolism; D010840:Phytosterols / Q000096:biosynthesis; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D012992:Solanine / Q000031:analogs & derivatives* / Q000378:metabolism; D011198:Solanum tuberosum / Q000201:enzymology* / Q000235:genetics*
D000470:Alkaloids; D018076:DNA, Complementary; D010840:Phytosterols; D012333:RNA, Messenger; C514919:alpha-solanine; D012992:Solanine; C011860:alpha-chaconine; D006903:Hydroxymethylglutaryl CoA Reductases; D013186:Farnesyl-Diphosphate Farnesyltransferase