Maclean et al. - 2002 - Rice almanac source book for the most important e
Maclean et al. - 2002 - Rice almanac source book for the most important e
Maclean et al. - 2002 - Rice almanac source book for the most important e
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An<strong>the</strong>r (mid-uninucleate)<br />
Explants<br />
Mature seeds<br />
Immature embryo<br />
(8—12 days)<br />
Transgenic rice<br />
H<strong>et</strong>erogeneous<br />
embryogenic c<strong>al</strong>li (EC)<br />
Agrobacterium<br />
trans<strong>for</strong>mation<br />
EC (selected)<br />
Sou<strong>the</strong>rn Western Enzymes<br />
Confirmation<br />
Embryogenic<br />
cell<br />
suspension<br />
Protoplasts + DNA<br />
(PEG/electroporation)<br />
Particle<br />
bombardment<br />
Selection, 50 µg/mL<br />
(2—3 wk) Hg/PPT<br />
Plant regeneration<br />
(with or without selection)<br />
Selection<br />
(8 wk) Hg/PPT<br />
Trans<strong>for</strong>med c<strong>al</strong>li<br />
Scutellum<br />
Selection, 50 µg/mL<br />
(4—6 wk) Hg/PPT<br />
Putative<br />
trans<strong>for</strong>med EC<br />
Fig. 3. A. schematic protocol <strong>for</strong> production of fertile transgenic rice plants using biolistic, protoplast, and<br />
Agrobacterium systems.<br />
But biotechnology’s <strong>most</strong> novel contribution<br />
will probably be in adding <strong>al</strong>ien genes to <strong>the</strong> rice<br />
gene pool through gen<strong>et</strong>ic engineering. Gen<strong>et</strong>ic<br />
engineering <strong>al</strong>so <strong>al</strong>lows <strong>the</strong> reintroduction of rice<br />
genes that have been extracted and modified to<br />
give <strong>al</strong>tered properties. Such gene transfers are<br />
impossible with convention<strong>al</strong> breeding m<strong>et</strong>hods.<br />
Gen<strong>et</strong>ic engineering <strong>al</strong>so <strong>al</strong>lows introducing one<br />
or two well-characterized genes at a time. There<br />
is no need <strong>for</strong> <strong>the</strong> extensive backcrossing done in<br />
convention<strong>al</strong> hybridization to remove<br />
undesirable genes.<br />
Three m<strong>et</strong>hods have been used successfully<br />
to transfer genes into rice: protoplast trans<strong>for</strong>mation,<br />
biolistics or particle bombardment, and<br />
Agrobacterium-mediated gene transfer (see Fig.<br />
3).<br />
Marker genes and promoters<br />
The first <strong>for</strong>eign genes expressed in rice were<br />
bacteri<strong>al</strong> genes conferring antibiotic (HPT,<br />
hygromycin phosphotransferase) or herbicide<br />
resistance (ppt, phosphinotricin ac<strong>et</strong>yl<br />
transferase). When <strong>the</strong> appropriate antibiotic or<br />
Internation<strong>al</strong> issues 41