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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

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