Details
| Original language | English |
|---|---|
| Title of host publication | Proceedings of the Fifth nternational Symposium on In Vitro Culture and Horticultural Breeding |
| Publisher | International Society for Horticultural Science |
| Pages | 795-800 |
| Number of pages | 6 |
| ISBN (print) | 906605719X, 9789066057197 |
| Publication status | Published - Nov 2006 |
Publication series
| Name | Acta Horticulturae |
|---|---|
| Volume | 725 II |
| ISSN (Print) | 0567-7572 |
Abstract
Hippeastrum species (Amaryllidaceae) have significant floricultural importance and are one of the major bulb crops in the commercial market. Amaryllidales are monocotyledonous plants that have been shown to be generally recalcitrant to molecular genetic manipulation. Successful plant transformation requires efficient regeneration and selection system and these were developed for a new hybrid Hippeastrum x chmielii Chm. This hybrid shows very vigorous growth and flowers without undergoing a quiescent phase. In preparation of transformation experiments the aim of this study was to establish appropriate media for selecting transgenic cells/regenerants. Therefore, the effect of different concentrations of glufosinate (syn. phosphinothricin) on plant regeneration and propagation were analyzed. Glufosinate will be used in transformation experiments to select the cells expressing the pat gene. In addition the influence of cefotaxim, an antibiotic used for suppression of agrobacteria after co-culture, on the selective efficiency of glufosinate was tested. Two different kinds of explants were used: first, young flower stalk segments cultured on MS medium with 2 mg L-1 2iP and 0.2 mg L -1 NAA, and second quarters of in vitro bulblets on MS medium with 0.5 mg L-1 BA and 0.1 mg L-1 NAA. Glufosinate totally inhibited bulblet formation for both types of explants even at low concentrations of 1.0 mg L-1 to 2.0 mg L-1. Therefore, this level was found to be suitable for being used in the selection medium. The addition of 500 mg L-1 cefotaxim did neither inhibit regeneration nor did it interfere with the selective efficiency in glufosinate containing medium. The results obtained in this study are the basis for establishing an Agrobacterium-mediated transformation system. By genetic transformation Hippeastrum x chmielii Chm. can be improved regarding flower longevity or disease resistance.
Keywords
- Breeding, Cefotaxim, Gene transfer, Glufosinate, In vitro, Phosphinothricin, Regeneration
ASJC Scopus subject areas
- Agricultural and Biological Sciences(all)
- Horticulture
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Proceedings of the Fifth nternational Symposium on In Vitro Culture and Horticultural Breeding. International Society for Horticultural Science, 2006. p. 795-800 (Acta Horticulturae; Vol. 725 II).
Research output: Chapter in book/report/conference proceeding › Conference contribution › Research › peer review
}
TY - GEN
T1 - Towards Agrobacterium-mediated transformation of Hippeastrum x chmielii Chm. - Identification of appropriate selection media
AU - Ilczuk, A.
AU - Łukaszewska, A.
AU - Winkelmann, Traud
AU - Serek, Margrethe
PY - 2006/11
Y1 - 2006/11
N2 - Hippeastrum species (Amaryllidaceae) have significant floricultural importance and are one of the major bulb crops in the commercial market. Amaryllidales are monocotyledonous plants that have been shown to be generally recalcitrant to molecular genetic manipulation. Successful plant transformation requires efficient regeneration and selection system and these were developed for a new hybrid Hippeastrum x chmielii Chm. This hybrid shows very vigorous growth and flowers without undergoing a quiescent phase. In preparation of transformation experiments the aim of this study was to establish appropriate media for selecting transgenic cells/regenerants. Therefore, the effect of different concentrations of glufosinate (syn. phosphinothricin) on plant regeneration and propagation were analyzed. Glufosinate will be used in transformation experiments to select the cells expressing the pat gene. In addition the influence of cefotaxim, an antibiotic used for suppression of agrobacteria after co-culture, on the selective efficiency of glufosinate was tested. Two different kinds of explants were used: first, young flower stalk segments cultured on MS medium with 2 mg L-1 2iP and 0.2 mg L -1 NAA, and second quarters of in vitro bulblets on MS medium with 0.5 mg L-1 BA and 0.1 mg L-1 NAA. Glufosinate totally inhibited bulblet formation for both types of explants even at low concentrations of 1.0 mg L-1 to 2.0 mg L-1. Therefore, this level was found to be suitable for being used in the selection medium. The addition of 500 mg L-1 cefotaxim did neither inhibit regeneration nor did it interfere with the selective efficiency in glufosinate containing medium. The results obtained in this study are the basis for establishing an Agrobacterium-mediated transformation system. By genetic transformation Hippeastrum x chmielii Chm. can be improved regarding flower longevity or disease resistance.
AB - Hippeastrum species (Amaryllidaceae) have significant floricultural importance and are one of the major bulb crops in the commercial market. Amaryllidales are monocotyledonous plants that have been shown to be generally recalcitrant to molecular genetic manipulation. Successful plant transformation requires efficient regeneration and selection system and these were developed for a new hybrid Hippeastrum x chmielii Chm. This hybrid shows very vigorous growth and flowers without undergoing a quiescent phase. In preparation of transformation experiments the aim of this study was to establish appropriate media for selecting transgenic cells/regenerants. Therefore, the effect of different concentrations of glufosinate (syn. phosphinothricin) on plant regeneration and propagation were analyzed. Glufosinate will be used in transformation experiments to select the cells expressing the pat gene. In addition the influence of cefotaxim, an antibiotic used for suppression of agrobacteria after co-culture, on the selective efficiency of glufosinate was tested. Two different kinds of explants were used: first, young flower stalk segments cultured on MS medium with 2 mg L-1 2iP and 0.2 mg L -1 NAA, and second quarters of in vitro bulblets on MS medium with 0.5 mg L-1 BA and 0.1 mg L-1 NAA. Glufosinate totally inhibited bulblet formation for both types of explants even at low concentrations of 1.0 mg L-1 to 2.0 mg L-1. Therefore, this level was found to be suitable for being used in the selection medium. The addition of 500 mg L-1 cefotaxim did neither inhibit regeneration nor did it interfere with the selective efficiency in glufosinate containing medium. The results obtained in this study are the basis for establishing an Agrobacterium-mediated transformation system. By genetic transformation Hippeastrum x chmielii Chm. can be improved regarding flower longevity or disease resistance.
KW - Breeding
KW - Cefotaxim
KW - Gene transfer
KW - Glufosinate
KW - In vitro
KW - Phosphinothricin
KW - Regeneration
UR - http://www.scopus.com/inward/record.url?scp=33846550646&partnerID=8YFLogxK
U2 - 10.17660/ActaHortic.2006.725.110
DO - 10.17660/ActaHortic.2006.725.110
M3 - Conference contribution
AN - SCOPUS:33846550646
SN - 906605719X
SN - 9789066057197
T3 - Acta Horticulturae
SP - 795
EP - 800
BT - Proceedings of the Fifth nternational Symposium on In Vitro Culture and Horticultural Breeding
PB - International Society for Horticultural Science
ER -