Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses
Acremonium implicatum is a seed-transmitted endophytic fungus that forms symbiotic associations with the economically significant tropical forage grasses, Brachiaria species. To take advantage of the endophyte's plant protective properties, we developed an efficient Agrobacterium-mediated transformation system for Acremonium implicatum, using green fluorescent protein (GFP) expression and vector pSK1019 (trpC promoter) or pCAMBIA1300 (CaMV35S promoter). We found that transformation efficiency doubled for both mycelial and conidial transformation as the co-cultivation period for Agrobacterium tumefaciens and Acremonium implicatum was increased from 48 to 72 h. Significantly, optimal results were obtained for either mycelial or conidial transformation with Agrobacterium tumefaciens strain AGL-1 and vector pSK1019 under the control of the trpC promoter. However, mycelial transformation consistently generated a significantly higher number of transformants than did conidial transformation. The mitotic stability of the transferred DNA was confirmed by growing ten transformants in liquid and agar media for six generations. In all cases, resistance to the selection pressure (hygromycin B) was maintained. Fluorescence emission was retained by the transformants and also expressed in Brachiaria tissues from plants inoculated with GFP-transformed A. implicatum. This technology will help in the transfer and expression of agronomically important genes in host plants.
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Elsevier
2008-03
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Subjects: | research, grasses, |
Online Access: | https://hdl.handle.net/10568/21177 https://doi.org/10.1016/j.mycres.2007.10.008 |
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dig-cgspace-10568-211772023-12-08T19:36:04Z Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses Abello, J.F. Kelemu, Segenet García, C. research grasses Acremonium implicatum is a seed-transmitted endophytic fungus that forms symbiotic associations with the economically significant tropical forage grasses, Brachiaria species. To take advantage of the endophyte's plant protective properties, we developed an efficient Agrobacterium-mediated transformation system for Acremonium implicatum, using green fluorescent protein (GFP) expression and vector pSK1019 (trpC promoter) or pCAMBIA1300 (CaMV35S promoter). We found that transformation efficiency doubled for both mycelial and conidial transformation as the co-cultivation period for Agrobacterium tumefaciens and Acremonium implicatum was increased from 48 to 72 h. Significantly, optimal results were obtained for either mycelial or conidial transformation with Agrobacterium tumefaciens strain AGL-1 and vector pSK1019 under the control of the trpC promoter. However, mycelial transformation consistently generated a significantly higher number of transformants than did conidial transformation. The mitotic stability of the transferred DNA was confirmed by growing ten transformants in liquid and agar media for six generations. In all cases, resistance to the selection pressure (hygromycin B) was maintained. Fluorescence emission was retained by the transformants and also expressed in Brachiaria tissues from plants inoculated with GFP-transformed A. implicatum. This technology will help in the transfer and expression of agronomically important genes in host plants. 2008-03 2012-07-09T05:54:53Z 2012-07-09T05:54:53Z Journal Article Abello, J.F., Kelemu, S. and Garcia, C. 2008. Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses. Mycological Research 112(3):407-413. 0953-7562 https://hdl.handle.net/10568/21177 https://doi.org/10.1016/j.mycres.2007.10.008 en Copyrighted; all rights reserved Limited Access p. 407-413 Elsevier Mycological Research |
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research grasses research grasses Abello, J.F. Kelemu, Segenet García, C. Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses |
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Acremonium implicatum is a seed-transmitted endophytic fungus that forms symbiotic associations with the economically significant tropical forage grasses, Brachiaria species. To take advantage of the endophyte's plant protective properties, we developed an efficient Agrobacterium-mediated transformation system for Acremonium implicatum, using green fluorescent protein (GFP) expression and vector pSK1019 (trpC promoter) or pCAMBIA1300 (CaMV35S promoter). We found that transformation efficiency doubled for both mycelial and conidial transformation as the co-cultivation period for Agrobacterium tumefaciens and Acremonium implicatum was increased from 48 to 72 h. Significantly, optimal results were obtained for either mycelial or conidial transformation with Agrobacterium tumefaciens strain AGL-1 and vector pSK1019 under the control of the trpC promoter. However, mycelial transformation consistently generated a significantly higher number of transformants than did conidial transformation. The mitotic stability of the transferred DNA was confirmed by growing ten transformants in liquid and agar media for six generations. In all cases, resistance to the selection pressure (hygromycin B) was maintained. Fluorescence emission was retained by the transformants and also expressed in Brachiaria tissues from plants inoculated with GFP-transformed A. implicatum. This technology will help in the transfer and expression of agronomically important genes in host plants. |
format |
Journal Article |
topic_facet |
research grasses |
author |
Abello, J.F. Kelemu, Segenet García, C. |
author_facet |
Abello, J.F. Kelemu, Segenet García, C. |
author_sort |
Abello, J.F. |
title |
Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses |
title_short |
Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses |
title_full |
Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses |
title_fullStr |
Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses |
title_full_unstemmed |
Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses |
title_sort |
agrobacterium-mediated transformation of the endophytic fungus acremonium implicatum associated with brachiaria grasses |
publisher |
Elsevier |
publishDate |
2008-03 |
url |
https://hdl.handle.net/10568/21177 https://doi.org/10.1016/j.mycres.2007.10.008 |
work_keys_str_mv |
AT abellojf agrobacteriummediatedtransformationoftheendophyticfungusacremoniumimplicatumassociatedwithbrachiariagrasses AT kelemusegenet agrobacteriummediatedtransformationoftheendophyticfungusacremoniumimplicatumassociatedwithbrachiariagrasses AT garciac agrobacteriummediatedtransformationoftheendophyticfungusacremoniumimplicatumassociatedwithbrachiariagrasses |
_version_ |
1787230995331678208 |