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TO T IPO T EN CY AND M O R PH O G E N E SIS O F PLANT C E L L S IN V IT R O
PLANTREGENERATIONFROMHYPOCOTYLPROTOPLASTSOF
SUNFLOWER (HELIANTHUSANNUUS L .)
LenutaRakosy-Tican*, Genevieve Jeannin**, GuntherHahne**
*Universiiatea "Babes-Bolyai", Facultatea deBiologie si Geologie, Cluj-Napoca. E-
Mail:
**CNRS, Institutde BiologieMoleculairedesPlantes, Strasbourg, France.
Sunflower (Helianthus annuus L.) is an important crop and therefore
one o f the targets for plant genetic engineering. Although, plant regeneration
from different multicellular explants i.e. cotyledons, hypocotyls, shoot tips,
and zygotic embryos (Alibert et al., 1994; Hahne, 1995) has been achieved
the culture of protoplasts was facing more difficulties (Fischer et al., 1996).
There are to date only three reports of successful plant regeneration from
hypocotyl and cotyledon protoplasts (Burrus et al., 1991; Fischer et al.,
1992; Krasnyanski and Menczel, 1993). In each case different culture
protocols for specific cultivars have been used. In this study a comparison of
regeneration ability of hypocotyl protoplasts isolated from different cultivars
and cultured in two sequences of media and at two different densities was
done. Four cultivars were used for protoplast isolation: line 47302bcd, Florom
328, Turbo, and Select. Hypocotyl and cotyledon protoplasts of line 47302bcd
were previously reported to regenerate fertile plants (Fischer et al., 1992) by
using one particular sequence of media. Florom 328 was the only cultivar
which gave hypocotyl protoplasts able to regenerate embryos and then fertile
plants in VKM media (Krasnyanski and Menczel, 1993). Turbo and Select
are two hybrids related through male parent with Florom 328. The results
obtained so far revealed that shoots can be regenerated from protoplasts of
Florom 328 cultured at 5x104 protoplasts/ml in the sequence of media
previously used for line 47302bcd, or cultured at 8x105 protoplasts/ml in
VKM media. The last protocol was also successful fo r the cultivar Turbo.
The experiments are in progress and more regenerated plants are to be
expected.
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