Plant Genome II Conference
Town & Country Conference Center, San Diego, CA, January, 1994.
PG-II: GENETIC ENGINEERING OF WHEAT
GENETIC ENGINEERING OF WHEAT.
Kutty K. Kartha, Ravindra N. Chibbar and Narender S. Nehra, Plant
Biotechnology Institute, National Research Council of Canada, 110
Gymnasium Road, Saskatoon, Saskatchewan, Canada, S7N 0W9.
We have developed an enhanced regeneration system (ERS) that
enhances the induction of somatic embryos and regeneration of
plants from isolated scutellar tissue of wheat. This system is
genotype independent and is also applicable to other cereals such
as barley. We have successfully used ERS to produce several
self-fertile transgenic wheat plants incorporating bar, gus or
nptII gene into their genome [Nehra et al., The Plant Journal, in
press (1994)]. The procedure is rapid resulting in the
production of transgenic plantlets within 12 weeks from
initiation of cultures and it avoids the need for establishing
long-term callus, cell suspension or protoplast cultures.
Somatic embryos regenerated from scutella bombarded with plasmid
pBARGUS and pRC62 (Act-21D-gus::nptII-nos) were selected on
L-phosphinothricin (L-PPT) and geneticin respectively, to recover
self-fertile transgenic wheat plants. Phosphinothricin
acetyltransferase (PAT) activity was observed at varying levels
in 50% of the plants selected on L-PPT whereas none of the plants
showed B-glucuronidase (GUS) activity. However, three quarters
of plants selected on the media containing geneticin exhibited
both GUS and neomycin phosphotransferase (NPTII) activity, thus
demonstrating the presence of functional fusion gene in the
putative transformants. Southern blot hybridization of
PAT-positive plants confirmed stable integration of both bar and
gus genes in R0 and R1 progeny plants. Segregation of the PAT
activity and herbicide resistance in R1 and R2 progeny plants
confirmed the Mendelian inheritance of the bar gene. Similarly,
GUS- and NPTII- positive plants also showed the stable
integration of fusion gene in R0 and R1 progeny plants of wheat
transformants. This protocol has the potential for becoming a
practical system for gene transfer in wheat.
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