Abstract
Tef (Eragrostis tef) provides a major source of human nutrition in the Horn of Africa, but biotechnology has had little impact on its improvement to date. Here, we report the elaboration of an in vitro regeneration protocol, based on the use of immature zygotic embryos as explant. Explant size was an important determinant of in vitro regeneration efficiency, as was the formulation of the culture medium. Optimal results were obtained by culturing 0.2–0.35 mm embryo explants on a medium containing KBP minerals, 9.2–13.8 μM 2,4-dichlorophenoxyacetic acid, 6 mM glutamine, and 0.5% Phytagel. Although this protocol was effective for both the improved cultivar “DZ-01-196” and the landrace “Fesho”, the former produced consistently more embryogenic tissue and a higher number of regenerants. An average of more than 2,800 shoots could be obtained from each “DZ-01-196” explant after 12 weeks of in vitro culture. These shoots readily formed roots, and plantlets transferred to soil were able to develop into morphologically normal, fertile plants. This regeneration and multiplication system should allow for the application of a range of biotechnological methods to tef.
Highlights
The small-grained cereal tef [Eragrostis tef (Zucc.) Trotter] is indigenous to Ethiopia, which is its centre of diversity and site of domestication [1]
The large (0.35–0.75 mm) “Fesho” and “DZ-01-196” explants cultivated on media containing N6 and L3 minerals displayed a low level of somatic embryogenesis and very little, if any, differentiation of shoots
Both MS and KBP minerals were permissive of callus formation, somatic embryogenesis, and de novo shoot regeneration from both genetic stocks, with the latter medium giving significantly the best response (Table 1)
Summary
The small-grained cereal tef [Eragrostis tef (Zucc.) Trotter] is indigenous to Ethiopia, which is its centre of diversity and site of domestication [1]. While the crop is well adapted to a range of climatic and soil conditions and enjoys a good level of tolerance to many pests and diseases, its mean grain yield in cultivation lies below 1t ha−1 [4]. The crop is cultivated over about one-third of the arable area in Ethiopia and represents a staple food for much of the population, in the form of the traditional pancake injera. In vitro plant regeneration from explants such as roots, young leaf bases, or seeds has been demonstrated [5,6,7], but the levels of efficiency achieved remain too low for use in tef improvement
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