The stearoyl-acyl-carrier-protein desaturase promoter (Des) from oil palm confers fruit-specific GUS expression in transgenic tomato.
The stearoyl-acyl-carrier-protein (ACP) desaturase is a plastid-localized enzyme that catalyzes the conversion of stearoyl-ACP to oleoyl-ACP and plays an important role in the determination of the properties of the majority of cellular glycerolipids. Functional characterization of the fatty acid desaturase genes and their specific promoters is a prerequisite for altering the composition of unsaturated fatty acids of palm oil by genetic engineering. In this paper, the specificity and strength of the oil palm stearoyl-ACP desaturase gene promoter (Des) was evaluated in transgenic tomato plants. Transcriptional fusions between 5' deletions of the Des promoter (Des1-4) and the β-glucuronidase (GUS) reporter gene were generated and their expression analyzed in different tissues of stably transformed tomato plants. Histochemical analysis of the Des promoter deletion series revealed that GUS gene expression was confined to the tomato fruits. No expression was detected in vegetative tissues of the transgenic plants. The highest levels of GUS activity was observed in different tissues of ripe red fruits (vascular tissue, septa, endocarp, mesocarp and columella) and in seeds, which harbored the promoter region located between -590 and +10. A comparison of the promoter-deletion constructs showed that the Des4 promoter deletion (314bp) produced a markedly low level of GUS expression in fruits and seeds. Fluorometric analysis of the GUS activity revealed a 4-fold increase in the activity of the full-length Des promoter compared to the CaMV35S promoter. RNA-hybridization analyses provided additional evidence of increased GUS expression in fruits driven by a Des fragment. Taken together, these results demonstrate the potential of the Des promoter as a tool for the genetic engineering of oil palms and other species, including dicots, in improving the quality and nutritional value of the fruits.
School of Biosciences and Biotechnology, Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor, Malaysia.
Rima
R
Saed Taha
Ismanizan
I
Ismail
Zamri
Z
Zainal
Siti Nor Akmar
SN
Abdullah
D028023:Arecaceae / Q000201:enzymology*; D001483:Base Sequence; D005638:Fruit / Q000201:enzymology* / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter / Q000235:genetics; D005966:Glucuronidase / Q000737:chemistry / Q000235:genetics*; D018551:Solanum lycopersicum / Q000201:enzymology* / Q000235:genetics*; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D010937:Plant Growth Regulators / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000201:enzymology; D011401:Promoter Regions, Genetic; D012639:Seeds / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
D010937:Plant Growth Regulators; D006899:Mixed Function Oxygenases; C018747:acyl-(acyl-carrier-protein)desaturase; D005966:Glucuronidase