TY - JOUR
T1 - Multiple traits of agronomic importance in transgenic indica rice plants
T2 - Analysis of transgene integration patterns, expression levels and stability
AU - Maqbool, Shahina Bano
AU - Christou, Paul
N1 - Funding Information:
We are grateful to Dr Illimar Altosaar (Biochemistry Department, University of Ottawa, Ottawa, Ontario, Canada), Dr Luke Masson (Biotechnology Research Institute, Montreal, QC, Canada) and Dr John Gatehouse (Department of Biological Sciences, University of Durham, Durham, UK) for making specific genes available for our studies. We thank our colleagues at the John Innes Centre (JIC), UK, and National Centre of Excellence in Molecular Biology (CEMB), Lahore, Pakistan, for helpful discussions and advice. We thank Dr Richard Twyman for discussion and help in preparing the manuscript. We also acknowledge the Rockefeller Foundation for providing a grant to S.B.M. and support from the JIC and CEMB. The JIC is supported in part by a grant-in-aid by the BBSRC.
PY - 1999
Y1 - 1999
N2 - We cotransformed indica rice (Oryza sativa L. cvs. Basmati 370 and M7) with three plasmids, carrying a total of four genes, by particle bombardment. The Bacillus thuringiensis (Bt) δ-endotoxin genes cryIAc and cry2A were carried on separate vectors, while the gna (snowdrop lectin) and hpt (hygromycin phosphotransferase) genes were linked on the same, cointegrate vector. We evaluated the molecular and expression profiles of 29 independently derived transgenic lines over two generations. The gna and hpt genes cointegrated with a frequency of 100% as expected. Furthermore, 60% of the transgenic plants carried all four genes. Southern blot analysis showed that transgene copy number ranged from 1 to 15. We used western blots to determine protein expression levels in R0 and R1 plants. We observed wide variation in the expression levels of the nonselected transgenes among independently-derived lines, but expression levels within lines derived from the same clone were similar. Consistent with previous reports, we observed no correlation between transgene copy number and the level or stability of protein expression. We show that the introduction of multiple agronomically valuable genes into the rice genome by cotransformation is a practical strategy for engineering elite rice varieties.
AB - We cotransformed indica rice (Oryza sativa L. cvs. Basmati 370 and M7) with three plasmids, carrying a total of four genes, by particle bombardment. The Bacillus thuringiensis (Bt) δ-endotoxin genes cryIAc and cry2A were carried on separate vectors, while the gna (snowdrop lectin) and hpt (hygromycin phosphotransferase) genes were linked on the same, cointegrate vector. We evaluated the molecular and expression profiles of 29 independently derived transgenic lines over two generations. The gna and hpt genes cointegrated with a frequency of 100% as expected. Furthermore, 60% of the transgenic plants carried all four genes. Southern blot analysis showed that transgene copy number ranged from 1 to 15. We used western blots to determine protein expression levels in R0 and R1 plants. We observed wide variation in the expression levels of the nonselected transgenes among independently-derived lines, but expression levels within lines derived from the same clone were similar. Consistent with previous reports, we observed no correlation between transgene copy number and the level or stability of protein expression. We show that the introduction of multiple agronomically valuable genes into the rice genome by cotransformation is a practical strategy for engineering elite rice varieties.
KW - Basmati 370 and M7 varieties
KW - Particle bombardment
KW - Snowdrop lectin (GNA)
KW - Transgenic rice
KW - δ-endotoxins (Cry1Ac and Cry2A)
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U2 - 10.1023/A:1009634226797
DO - 10.1023/A:1009634226797
M3 - Article
AN - SCOPUS:0032862493
SN - 1380-3743
VL - 5
SP - 471
EP - 480
JO - Molecular Breeding
JF - Molecular Breeding
IS - 5
ER -