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 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.

Saved in:
Bibliographic Details
Main Authors: Solís-Ramos,Laura Yesenia, González-Estrada,Tomas, Andrade-Torres,Antonio, Godoy-Hernández,Gregorio, Castaño de la Serna,Enrique
Format: Digital revista
Language:English
Published: Pontificia Universidad Católica de Valparaíso 2010
Online Access:http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000400002
Tags: Add Tag
No Tags, Be the first to tag this record!
id oai:scielo:S0717-34582010000400002
record_format ojs
spelling 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
institution SCIELO
collection OJS
country Chile
countrycode CL
component Revista
access En linea
databasecode rev-scielo-cl
tag revista
region America del Sur
libraryname SciELO
language English
format Digital
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
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.
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_ 1755991083981996032