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The interaction of 2,4-D application and mannitol pretreatment in anther and microspore culture of Hordeum vulgare L. cv. igri

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Author: Hoekstra, S. · Bergen, S. van · Brouwershaven, I.R. van · Schilperoort, R.A. · Heidekamp, F.
Type:article
Date:1996
Institution: TNO Voeding
Source:Journal of Plant Physiology, 6, 148, 696-700
Identifier: 233343
Keywords: Nutrition · 2,4-D concentration and time · Anther and microspore culture · Growth regulator · Hordeum vulgare L. · Mannitol pretreatment · Pulse treatment

Abstract

The influence of 2,4-dichlorophenoxyacetic acid (2,4-D) on embryo-like structures (ELS) and plant development from barley microspores was determined. Microspores cultured on filters enabled simple modification of growth regulator concentrations. Regeneration frequencies obtained with 2,4- D as growth regulator were similar to the results achieved with the generally applied cytokinin 6-benzylaminopurine. If 2,4-D was applied after a regular mannitol pretreatment, maximal plant regeneration was achieved if 10-6 mol/L 2,4-D was present continuously or for 7 days. Alternatively, maximal plant formation was induced if 10-4 or 10-5 mol/L 2,4-D was present for 1 h or if present resp. 3 days or 1 day. Induction of plant regeneration by a l h treatment with 10-4 mol/L 2,4-D is a more generally observed phenomenon for single cells or small cell clusters of both dicotyledonous and monocotyledonous species. Without mannitol pretreatment, it was possible to induce plant production after 2,4-D treatment only in anther cultures. Without mannitol pretreatment no embryogenic type of microspores could be recognized at the moment of microspore isolation, and plating efficiency never reached 1%. In anther culture without mannitol pretreatment, a higher molarity and/or longer presence of 2,4-D was required and resulted only in about 1 green plant per anther. After application of a mannitol pretreatment, plant production increased at least 10 times. To our knowledge this is the first report on microspore-derived barley plants via androgenesis without any pretreatment. The combination of 2,4-D and anther pretreatment with mannitol as trigger for microspore differentiation is discussed.