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# In vitro Shoot Regeneration from Cotyledons of Immature Ovules of Impatiens platypetala Lindl.

Kyungchul Han
In Vitro Cellular & Developmental Biology. Plant
Vol. 30P, No. 2 (Apr., 1994), pp. 108-112
Stable URL: http://www.jstor.org/stable/4293030
Page Count: 5
An efficient and reproducible protocol has been developed for in vitro shoot regeneration from cotyledonary explants derived by germinating immature ovules of Impatiens platypetala Lindl. 'TR6-27-2'. Cotyledonary explants were cultured on a modified Murashige and Skoog (MS) agar-solidified medium containing $7.5 g\cdot liter^{-1}$ sucrose, $22.2 \mu M N^6-benzyladenine (BA)$, and $0.54 \mu M \alpha-naphthaleneacetic acid (NAA)$. The induction of organogenic tissues occurred after 6 to 8 wk in culture. Exogenous auxin and cytokinin were essential for the induction of organogenic tissues and survival of explants, and BA was most effective for the induction of organogenic tissues, compared with other cytokinins tested. The addition of glutamine ($500 mg \cdot liter^{-1}$) was also important for growth of organogenic tissues after induction and for reducing explant death during culture. The induction of organogenic tissue was also influenced by the type of cotyledon cultured and the age of the donor seedlings. On average, eight shoots per explant were induced from organogenic tissues larger than 0.5 cm in diameter 6 to 8 wk after transfer to a modified MS agar-solidified medium without NAA and BA reduced to $4.44 \mu M$. Shoots longer than 0.5 cm in length were successfully rooted 2 to 4 wk after transfer to a basal MS medium containing $30 g \cdot liter^{-1} sucrose$.