Evaluation of selection strategies alternative to nptII in genetic transformation of citrus
The neomycin phosphotransferase (nptII) selection system has proved successful in citrus transformation; however, it may be recommendable to replace it given the pressure exerted against antibiotic-resistance selectable marker genes in transgenic plants. The present work investigates three different...
| Autores principales: | , , |
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| Formato: | article |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/4650 |
| _version_ | 1855032155533475840 |
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| author | Ballester, Alida Cervera, Magdalena Pena, Leandro |
| author_browse | Ballester, Alida Cervera, Magdalena Pena, Leandro |
| author_facet | Ballester, Alida Cervera, Magdalena Pena, Leandro |
| author_sort | Ballester, Alida |
| collection | ReDivia |
| description | The neomycin phosphotransferase (nptII) selection system has proved successful in citrus transformation; however, it may be recommendable to replace it given the pressure exerted against antibiotic-resistance selectable marker genes in transgenic plants. The present work investigates three different selection alternatives, comparing them to nptII selection in two citrus genotypes, Carrizo citrange and Pineapple sweet orange. The first method used the beta-glucuronidase (uidA) reporter marker gene for selection; the second attempted to generate marker-free plants by transforming explants with a multi-auto-transformation (MAT) vector, combining an inducible R/RS-specific recombination system with transgenic-shoot selection through expression of isopentenyl transferase (ipt) and indoleacetamide hydrolase/tryptophan monooxygenase (iaaM/H) marker genes; while the third exploited the phosphomannose isomerase (PMI)/mannose conditional positive selection system. Firstly, GUS screening of all regenerated shoots in kanamycin-free medium gave 4.3% transformation efficiency for both genotypes. Secondly, workable transformation efficiencies were also achieved with the MAT system, 7.2% for citrange and 6.7% for sweet orange. This system affords an additional advantage as it enables selectable marker genes to be used during the in vitro culture phase and later removed from the transgenic plants by inducible recombination and site-specific excision. Thirdly, the highest transformation rates were obtained with the PMI/mannose system, 30% for citrange and 13% for sweet orange, which indicates that this marker is also an excellent candidate for citrus transformation. |
| format | article |
| id | ReDivia4650 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia46502025-04-25T14:44:05Z Evaluation of selection strategies alternative to nptII in genetic transformation of citrus Ballester, Alida Cervera, Magdalena Pena, Leandro The neomycin phosphotransferase (nptII) selection system has proved successful in citrus transformation; however, it may be recommendable to replace it given the pressure exerted against antibiotic-resistance selectable marker genes in transgenic plants. The present work investigates three different selection alternatives, comparing them to nptII selection in two citrus genotypes, Carrizo citrange and Pineapple sweet orange. The first method used the beta-glucuronidase (uidA) reporter marker gene for selection; the second attempted to generate marker-free plants by transforming explants with a multi-auto-transformation (MAT) vector, combining an inducible R/RS-specific recombination system with transgenic-shoot selection through expression of isopentenyl transferase (ipt) and indoleacetamide hydrolase/tryptophan monooxygenase (iaaM/H) marker genes; while the third exploited the phosphomannose isomerase (PMI)/mannose conditional positive selection system. Firstly, GUS screening of all regenerated shoots in kanamycin-free medium gave 4.3% transformation efficiency for both genotypes. Secondly, workable transformation efficiencies were also achieved with the MAT system, 7.2% for citrange and 6.7% for sweet orange. This system affords an additional advantage as it enables selectable marker genes to be used during the in vitro culture phase and later removed from the transgenic plants by inducible recombination and site-specific excision. Thirdly, the highest transformation rates were obtained with the PMI/mannose system, 30% for citrange and 13% for sweet orange, which indicates that this marker is also an excellent candidate for citrus transformation. 2017-06-01T10:10:39Z 2017-06-01T10:10:39Z 2008 JUN 2008 article BallE., Alida, Cervera, M., Pena, L. (2008). Evaluation of selection strategies alternative to nptII in genetic transformation of citrus. Plant Cell Reports, 27(6), 1005-1015. 0721-7714 http://hdl.handle.net/20.500.11939/4650 10.1007/s00299-008-0523-z en openAccess Impreso |
| spellingShingle | Ballester, Alida Cervera, Magdalena Pena, Leandro Evaluation of selection strategies alternative to nptII in genetic transformation of citrus |
| title | Evaluation of selection strategies alternative to nptII in genetic transformation of citrus |
| title_full | Evaluation of selection strategies alternative to nptII in genetic transformation of citrus |
| title_fullStr | Evaluation of selection strategies alternative to nptII in genetic transformation of citrus |
| title_full_unstemmed | Evaluation of selection strategies alternative to nptII in genetic transformation of citrus |
| title_short | Evaluation of selection strategies alternative to nptII in genetic transformation of citrus |
| title_sort | evaluation of selection strategies alternative to nptii in genetic transformation of citrus |
| url | http://hdl.handle.net/20.500.11939/4650 |
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