Triacylglycerol (TAG) production in the microspore derived (MD) cell
suspension culture ofBrassica napus L. cv Jet Neuf was enhanced when the sucrose
concentration in the growth medium was increased from 2 to 14 % (w/v). mRNA
differential display by polymerase chain reaction was used to examine gene expression in
cells grown at different sucrose concentrations in order to identify mRNAs which
could be associated with oil formation. The anchored primer, T12AA, was used to screen
one subset, representing approximately one twelfth of the transcript population, isolated
from cultures grown in media supplemented to 2, 6 and 14 % (w/v) sucrose. Analysis of
this mRNA subset revealed thirteen cDNAs which appeared to be upregulated as the
sucrose concentration was increased. Cloning and sequencing revealed multiple cDNA
fragments for each signal detected by differential display. RT-PCR analysis of sixteen
different cDNAs revealed that eight encoded mRNAs which were upregulated in parallel
to the increase in media sucrose. Comparison of the eight upregulated cDNAs to other
sequences in GenBank revealed the following: (1) BSS8A had a 100% identity with the
last 25 amino acids of an acyl carrier protein from Arabidopsis thaliana, (2) BSS1A
displayed homology to a number of sequences of unknown function, (3) BSS1 IB
displayed weak but significant homology to a number of sequences of unknown function,
(4) BSS13A displayed homology to four members of the thioredoxin family from ,4.
thaliana and (5) four Had no significant homology to previously reported sequences
which makes them potential candidates to encode lipogenic enzymes. These results
indicate that differential display of mRNA may be a simple and rapid method for the
identification of sucrose-modulated gene expression changes in this system and for the
characterization of novel sequences potentially encoding lipogenic proteins. / xxi, 256 leaves : ill. ; 28 cm.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:ALU.w.uleth.ca/dspace#10133/345 |
| Date | January 1997 |
| Creators | Davoren, Jonathan M., University of Lethbridge. Faculty of Arts and Science |
| Contributors | Weselake, Randall, Laroche, André |
| Publisher | Lethbridge, Alta. : University of Lethbridge, Faculty of Arts and Science, 1997, Arts and Science, Department of Chemistry and Biochemistry |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | en_US |
| Detected Language | English |
| Type | Thesis |
| Relation | Thesis (University of Lethbridge. Faculty of Arts and Science) |
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