Endogenous GUS-like activity in Capsicum chinense Jacq
The gene uidA, codes for β-glucuronidase, which is one of the reporters more frequently utilized in transgenic plants. However, this can only be use if the selected organism does not present endogenous GUS-like activity. In tissues of C. chinense we found a GUS-like activity showing differe...
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Pontificia Universidad Católica de Valparaíso
2010
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oai:scielo:S0717-345820100004000022010-08-04Endogenous GUS-like activity in Capsicum chinense JacqSolís-Ramos,Laura YeseniaGonzález-Estrada,TomasAndrade-Torres,AntonioGodoy-Hernández,GregorioCastaño de la Serna,EnriqueThe gene uidA, codes for β-glucuronidase, which is one of the reporters more frequently utilized in transgenic plants. However, this can only be use if the selected organism does not present endogenous GUS-like activity. In tissues of C. chinense we found a GUS-like activity showing different levels of intensity. Histochemical screening showed that endogenous GUS-like activity decreased, or reduced significantly, in almost all tissues with exception of stament, when phosphate buffer was adjusted to pH 8. Subsequently, C. chinense zygotic embryo explants were transient transformed with Agrobacterium tumefaciens LBA4404 (pCAMBIA2301) and plantlets regenerated were histochemically stained in phosphate buffer pH 8. Observations of incubated tissues of C. chinense regenerants showed blue staining, suggesting expression of uidA. Incubated tissues of non-transformed regenerants did not show blue staining in phosphate buffer pH 8. The results show that for transformation experiments of C. chinense with uidA gene, pH 8 is recommended for histochemical staining.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.13 n.4 20102010-07-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000400002en |
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Scielo Chile |
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Scielo Chile |
| language |
English |
| description |
The gene uidA, codes for β-glucuronidase, which is one of the reporters more frequently utilized in transgenic plants. However, this can only be use if the selected organism does not present endogenous GUS-like activity. In tissues of C. chinense we found a GUS-like activity showing different levels of intensity. Histochemical screening showed that endogenous GUS-like activity decreased, or reduced significantly, in almost all tissues with exception of stament, when phosphate buffer was adjusted to pH 8. Subsequently, C. chinense zygotic embryo explants were transient transformed with Agrobacterium tumefaciens LBA4404 (pCAMBIA2301) and plantlets regenerated were histochemically stained in phosphate buffer pH 8. Observations of incubated tissues of C. chinense regenerants showed blue staining, suggesting expression of uidA. Incubated tissues of non-transformed regenerants did not show blue staining in phosphate buffer pH 8. The results show that for transformation experiments of C. chinense with uidA gene, pH 8 is recommended for histochemical staining. |
| author |
Solís-Ramos,Laura Yesenia González-Estrada,Tomas Andrade-Torres,Antonio Godoy-Hernández,Gregorio Castaño de la Serna,Enrique |
| spellingShingle |
Solís-Ramos,Laura Yesenia González-Estrada,Tomas Andrade-Torres,Antonio Godoy-Hernández,Gregorio Castaño de la Serna,Enrique Endogenous GUS-like activity in Capsicum chinense Jacq |
| author_facet |
Solís-Ramos,Laura Yesenia González-Estrada,Tomas Andrade-Torres,Antonio Godoy-Hernández,Gregorio Castaño de la Serna,Enrique |
| author_sort |
Solís-Ramos,Laura Yesenia |
| title |
Endogenous GUS-like activity in Capsicum chinense Jacq |
| title_short |
Endogenous GUS-like activity in Capsicum chinense Jacq |
| title_full |
Endogenous GUS-like activity in Capsicum chinense Jacq |
| title_fullStr |
Endogenous GUS-like activity in Capsicum chinense Jacq |
| title_full_unstemmed |
Endogenous GUS-like activity in Capsicum chinense Jacq |
| title_sort |
endogenous gus-like activity in capsicum chinense jacq |
| publisher |
Pontificia Universidad Católica de Valparaíso |
| publishDate |
2010 |
| url |
http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000400002 |
| work_keys_str_mv |
AT solisramoslaurayesenia endogenousguslikeactivityincapsicumchinensejacq AT gonzalezestradatomas endogenousguslikeactivityincapsicumchinensejacq AT andradetorresantonio endogenousguslikeactivityincapsicumchinensejacq AT godoyhernandezgregorio endogenousguslikeactivityincapsicumchinensejacq AT castanodelasernaenrique endogenousguslikeactivityincapsicumchinensejacq |
| _version_ |
1718441816496275456 |