title: Transgenic increases in seed oil content are associated with the differential expression of novel Brassica-specific transcripts.
abstract: BACKGROUND
  Seed oil accumulates primarily as triacylglycerol (TAG). While the biochemical pathway for TAG biosynthesis is known, its regulation remains unclear. Previous research identified microsomal diacylglycerol acyltransferase 1 (DGAT1, EC 2.3.1.20) as controlling a rate-limiting step in the TAG biosynthesis pathway. Of note, overexpression of DGAT1 results in substantial increases in oil content and seed size. To further analyze the global consequences of manipulating DGAT1 levels during seed development, a concerted transcriptome and metabolome analysis of transgenic B. napus prototypes was performed.
  
  
  RESULTS
  Using a targeted Brassica cDNA microarray, about 200 genes were differentially expressed in two independent transgenic lines analyzed. Interestingly, 24-33% of the targets showing significant changes have no matching gene in Arabidopsis although these represent only 5% of the targets on the microarray. Further analysis of some of these novel transcripts indicated that several are inducible by ABA in microspore-derived embryos. Of the 200 Arabidopsis genes implicated in lipid biology present on the microarray, 36 were found to be differentially regulated in DGAT transgenic lines. Furthermore, kinetic reverse transcriptase Polymerase Chain Reaction (k-PCR) analysis revealed up-regulation of genes encoding enzymes of the Kennedy pathway involved in assembly of TAGs. Hormone profiling indicated that levels of auxins and cytokinins varied between transgenic lines and untransformed controls, while differences in the pool sizes of ABA and catabolites were only observed at later stages of development.
  
  
  CONCLUSIONS
  Our results indicate that the increased TAG accumulation observed in transgenic DGAT1 plants is associated with modest transcriptional and hormonal changes during seed development that are not limited to the TAG biosynthesis pathway. These might be associated with feedback or feed-forward effects due to altered levels of DGAT1 activity. The fact that a large fraction of significant amplicons have no matching genes in Arabidopsis compromised our ability to draw concrete inferences from the data at this stage, but has led to the identification of novel genes of potential interest.
authors.affiliation: National Research Council Canada, Plant Biotechnology Institute, NRC, 101 Gymnasium Place, Saskatoon SKS7N0W9, Canada. Nirmala.sharma@nrc-cnrc.gc.ca
authors.forename: Nirmala
authors.initials: N
authors.lastname: Sharma
authors.forename: Maureen
authors.initials: M
authors.lastname: Anderson
authors.forename: Arvind
authors.initials: A
authors.lastname: Kumar
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: E Michael
authors.initials: EM
authors.lastname: Giblin
authors.forename: Suzanne R
authors.initials: SR
authors.lastname: Abrams
authors.forename: L Irina
authors.initials: LI
authors.lastname: Zaharia
authors.forename: David C
authors.initials: DC
authors.lastname: Taylor
authors.forename: Pierre R
authors.initials: PR
authors.lastname: Fobert
mesh_terms: D000040:Abscisic Acid / Q000378:metabolism; D001937:Brassica / Q000201:enzymology / Q000235:genetics*; D003583:Cytokinins / Q000378:metabolism; D051048:Diacylglycerol O-Acyltransferase / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D007210:Indoleacetic Acids / Q000378:metabolism; D055442:Metabolome; D020411:Oligonucleotide Array Sequence Analysis; D010938:Plant Oils / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics; D012639:Seeds / Q000201:enzymology* / Q000235:genetics; D014280:Triglycerides / Q000096:biosynthesis*
chemical_list: D003583:Cytokinins; D007210:Indoleacetic Acids; D010938:Plant Oils; D010940:Plant Proteins; D018749:RNA, Plant; D014280:Triglycerides; D000040:Abscisic Acid; D051048:Diacylglycerol O-Acyltransferase