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Direct Evidence for Rapid Degradation of
Bacillus
thuringiensis Toxin mRNA as a Cause of
Poor Expression in
Plants1
E. Jay De Rocher,
Tracy C. Vargo-Gogola2,
Scott H. Diehn3, and
Pamela J. Green*
Michigan State University-Department of Energy Plant Research
Laboratory (E.J.D.R., S.H.D., P.J.G.), Department of Physiology
(T.C.V.-G.), Department of Botany and Plant Pathology (S.H.D.), and
Department of Biochemistry (P.J.G.), Michigan State University,
East Lansing, Michigan 48824
It
is well established that the expression of Bacillus
thuringiensis (B.t.) toxin genes in higher
plants is severely limited at the mRNA level, but the cause remains
controversial. Elucidating whether mRNA accumulation is limited
transcriptionally or posttranscriptionally could contribute to
effective gene design as well as provide insights about endogenous
plant gene-expression mechanisms. To resolve this controversy, we
compared the expression of an A/U-rich wild-type cryIA(c) gene and a G/C-rich synthetic cryIA(c)
B.t.-toxin gene under the control of identical 5 and 3
flanking sequences. Transcriptional activities of the genes were equal
as determined by nuclear run-on transcription assays. In contrast, mRNA
half-life measurements demonstrated directly that the wild-type
transcript was markedly less stable than that encoded by the synthetic
gene. Sequences that limit mRNA accumulation were located at more than
one site within the coding region, and some appeared to be recognized
in Arabidopsis but not in tobacco (Nicotiana tabacum).
These results support previous observations that some A/U-rich
sequences can contribute to mRNA instability in plants. Our studies
further indicate that some of these sequences may be differentially
recognized in tobacco cells and Arabidopsis.
1
This work was supported by grants from the
Department of Energy, the U.S. Department of Agriculture, Project
Green, Michigan State University Research Excellence Funds, Midwest
Plant Biotechnology Consortium, Consortium for Plant Biotechnology
Research, and matching funds from Ciba-Geigy, Sandoz, Pioneer,
Anheuser-Busch, ICI, Agrigenetics, and DowElanco. S.H.D. and E.J.D.R.
were supported in part by a National Institutes of Health predoctoral
traineeship and a U.S. Department of Agriculture postdoctoral
fellowship, respectively.
2
Present address: Department of Cell Biology,
Vanderbilt University, 1161 21st Avenue South, C2310 Medical Center
North, Nashville, TN 37232-2175.
3
Present address: Pioneer Hi-Bred International,
Inc., Traits and Technology Development, 7300 NW 62nd Avenue, P.O. Box
1004, Johnston, IA 50131-1004.
*
Corresponding author; e-mail 22313pjg{at}msu.edu; fax
1-517-355-9298.
Plant Physiol. (1998) 117: 1445-1461
Copyright Clearance Center: 0032-0889/98/117//17
© 1998 American Society of Plant Physiologists
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