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T O T iP O T E N C Y A N D M O R P H O G E N E S IS OF P L A N T C E L L S IN V IT R O
HIGHEFFECTIVESOMATIC EMBRYOGENESISANDPLANT
REGENERATION OFDINITROANILINE-RESISTANT BIOTYPES OF
EI.EUSINEINDICA
A.I.Yemets,L.A .Klimkina,L.V.Tarasenko,Ya.B.Blume
Inst .Cell Biol. &Genet. Eng., Natl. Acad. Sci. ofUkraine, acad. Zabolotny str., 148, Kiev
GSP-22, 252650, Ukraine;
Natural mutant biotypes and mutant plants obtained by artificial ways
are valuable materials for various biotechnological approaches. Special
interest exists to plants with resistance to different types of herbicides. Last
decade there were found DNH-resistant biotypes of Eleusine indica weed as
a result of long use of dinitroaniline herbicides (DNH) on the fields of South
Carolina (USA). Recently it was established that the DNH-resistance of E.
indica connects with the mutation in one of a-tubulin gene (Baird et al.,
1996). Since genetically engineered transfer of this mutant tubulin gene
have not conducted up to now from technical difficulties it is actually to
obtain in v itrocell culture and regenerants of E. indica fo r somatic
hybridization.
In our experiments there were used the seeds of DNH-resistant biotypes
of E. indica kindly provided by Dr. W.V. Baird (Clemson Univ., SC, USA).
Calli were induced from mature embryos, hypocotyls, roots of young seedlings
and mesophyll tissue. For this purpose there were used different nutritional
media: on the basis of MS (Murasige and Skoog, 1962) salts with 1-2,5 mg/
1of 2,4-dichlorophenoxyacetic acid (2,4-D) and N6 (Chu et al., 1975) salts
with 1-6 mg/l of 2,4-D containing different organic supplements, it was
established that N6-media with 1-2 mg/l of 2,4-D, 2 mg/l glycine, 100 mg/l
asparagine, 100 mg/l casein hydrolisate were the most optimal for callus
induction. Calli formed only from mature embryos and hypocotyls. In order
to obtain embryogenic callus the concentration of 2,4-D were decreased in
2 times in media for callus induction. Nondifferentiated calli were cultivated
on such media for 1 week in the dark at 25 °C. Histological analysis
established the formation of embryolike structures in calli under these
conditions. For further regeneration embryogenic calli were placed for 2-5
days under the light and then replaced on MS-medium, containing 1 mg/l
kinetin and 0,1 mg/l indole-3-aceticacid. Rooted plants were cultivated on
hormonefree MS-medium. The experiments on protoplast isolation from
mesophyll and suspension culture of E. indica are continued now.
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