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In vitro manipulation of tef 141
Table 7 In vitro culture responses of isolated tef another as influenced by induction medium,
genotype and gamma irradiation of seeds grown to give the anther donor plants in two studies
In vitro culture variables tested % callusing Reference
Induction Tef varieties Gamma ray
Medium dose (GY)
N6 DZ-01-354 - 7 mulu (1995)*
DZ-Cr-37 - 3
Ms DZ-01-354 - 0
DZ-Cr-37 - 0
N6 DZ-01-354 0 6.66 Hailu et al.(1999)**
700 2.72
Alba 0 12.57
6.94
Fesho 0 3.62
2.20
DZ-01-354 4.44
Alba 9.69
Gea Lammie 3.05
0 7.56
700 3.91
*in both cases semi-semi-solid (solid) media with 2 mg/1 2,4-D + 30 g/1 malthose, and dark
incubation ** media with 100 mg/1 myoinositol, 1.5 mg/` biotin, 30 g/1 maltose, and dark
incubation
Preliminary tef ovary in vitro culture investigations on N6 basal medium as used for
anthers, generally demonstrated poor callus induction performances of this type of
explant (Hailu et al., 1999). The results of these workers revealed that of the total number
of cultured ovaries of the three tef genotypes tested (i.e.DZ-01-354, cv. Alba and cv.
Fesho) following gamma irradiation of seeds at 0 and 700 GY, callus formation occurred
only in 3 out of 158 ovaries (about 3%) of cv. Fesho receiving 700 gy gamma ray, and in
1 out of 127 ovaries of DZ-01-354 receiving no gamma ray treatment.
Suspension cell cultures
Embryogenic cell suspension cultures were initiated for eight tef genotypes from calli
induced from leaf and root segments of eight-day-old tef seedlings in the look for
genotypes with the greatest capacity for somatic embryogenesis (Endashaw et al., 1995.
the cell suspension manifested composition of two cell types: one long, tubular,
vacuolated and usually occurring freely; and the other consisting of compact and firmly
packed cells. Ms liquid media supplemented with 0.8 mg/1 of 3,6-D or 2,4-D and subculturing
at 10-days intervals were sufficient to establish and maintain suspension cultres
of tef derived from leaf and root calli. Callus initiated from seed explants produced
suspension largely composed of non embryonic, elongated cells (Endashaw et al., 1995).
From utilization point of view embryogenic cell suspension cultures are primarily useful
for getting protoplasts which can serve in various in vitro manipulations including
somatic hybridization as well as other forms of genetic transformation.
Protoplast cultures
In electrofusion of protoplasts of desiccation tolerant and sensitive grass species Gaff et
al. (1986) reported very low release and with certain genotypes no yield at all of tef
protoplasts with the methods they used. Later, however, Endashaw (1995) attempted an