Staff View: Assessment of co-transformation technique for Malaysian orchid variety (Dendrobium Savin White) with cymbidium mosaic virus coat protein gene

Assessment of co-transformation technique for Malaysian orchid variety (Dendrobium Savin White) with cymbidium mosaic virus coat protein gene

Enzyme linked immunosorbent assay (ELISA) was conducted to test 300 samples of orchid plants, collected from commercial orchid nurseries around Malaysia. The orchid samples were either apparently healthy or exhibited virus-like symptoms. ELISA results showed that 88% of the samples were infected wit...

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Main Authors:	Rogayah Sekeli, Khairun Hisam Nasir, Wee, C. Y., Vilasini Pillai
Format:	Journal Contribution
Language:	English
Published:	2020
Subjects:	Dendrobium Mosaic viruses Orchidaceae ELISA Enzyme linked immunosorbent assay Plant nurseries Hygromycin Transgenic plants Polymerase chain reaction PCR Vectors Greenhouses morphology
Online Access:	http://agris.upm.edu.my:8080/dspace/handle/0/17337
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spelling	oai:http:--agris.upm.edu.my:0-17337Assessment of co-transformation technique for Malaysian orchid variety (Dendrobium Savin White) with cymbidium mosaic virus coat protein geneRogayah SekeliKhairun Hisam NasirWee, C. Y.Vilasini PillaiDendrobiumMosaic virusesOrchidaceaeELISAEnzyme linked immunosorbent assayPlant nurseriesHygromycinTransgenic plantsPolymerase chain reactionPCRVectorsGreenhousesmorphologyEnzyme linked immunosorbent assay (ELISA) was conducted to test 300 samples of orchid plants, collected from commercial orchid nurseries around Malaysia. The orchid samples were either apparently healthy or exhibited virus-like symptoms. ELISA results showed that 88% of the samples were infected with cymbidium mosaic virus (CyMV) including the orchid samples that did not exhibit any viral symptoms and looked healthy in appearance. Shoot tips from virus free plants were used to induce protocorm-like bodies (PLBs). The generated PLBs were bombarded with gold particle coated with plasmid pRQ6 and pJIT.CyMVcp containing hygromycin phosphotransferase (hptII) selectable marker gene and CyMVcp gene respectively. A total of 500 PLBs were bombarded and after 5 months selection in hygromycin media, 63 putative transgenic lines were recovered. Based on polymerase chain reaction analysis (PCR) on putative transgenic lines, 53 showed positive for the presence of hptII and CyMVcp genes. This study revealed that cotransformation of hptII and CyMVcp genes in two different vectors manage to achieve 10.6% transformation efficiency. The transgenic orchid plants were established in pots and acclimatised in the greenhouse. The transgenic plants were found morphologically similar compared to untransformed plants.2020-06-03T06:57:28Z2020-06-03T06:57:28Z2016Journal ContributionArticleRefereedJournal of Tropical Agriculture and Food Science, 44 (2), p. 265-2761394-9829http://agris.upm.edu.my:8080/dspace/handle/0/17337MY2020050643enhttp://jtafs.mardi.gov.my/jtafs/44-2/dendrobium.pdfMalaysiahttp://www.oceandocs.org/license
institution	AGRIS
collection	AGRIS
language	English
topic	Dendrobium Mosaic viruses Orchidaceae ELISA Enzyme linked immunosorbent assay Plant nurseries Hygromycin Transgenic plants Polymerase chain reaction PCR Vectors Greenhouses morphology
spellingShingle	Dendrobium Mosaic viruses Orchidaceae ELISA Enzyme linked immunosorbent assay Plant nurseries Hygromycin Transgenic plants Polymerase chain reaction PCR Vectors Greenhouses morphology Rogayah Sekeli Khairun Hisam Nasir Wee, C. Y. Vilasini Pillai Assessment of co-transformation technique for Malaysian orchid variety (Dendrobium Savin White) with cymbidium mosaic virus coat protein gene
description	Enzyme linked immunosorbent assay (ELISA) was conducted to test 300 samples of orchid plants, collected from commercial orchid nurseries around Malaysia. The orchid samples were either apparently healthy or exhibited virus-like symptoms. ELISA results showed that 88% of the samples were infected with cymbidium mosaic virus (CyMV) including the orchid samples that did not exhibit any viral symptoms and looked healthy in appearance. Shoot tips from virus free plants were used to induce protocorm-like bodies (PLBs). The generated PLBs were bombarded with gold particle coated with plasmid pRQ6 and pJIT.CyMVcp containing hygromycin phosphotransferase (hptII) selectable marker gene and CyMVcp gene respectively. A total of 500 PLBs were bombarded and after 5 months selection in hygromycin media, 63 putative transgenic lines were recovered. Based on polymerase chain reaction analysis (PCR) on putative transgenic lines, 53 showed positive for the presence of hptII and CyMVcp genes. This study revealed that cotransformation of hptII and CyMVcp genes in two different vectors manage to achieve 10.6% transformation efficiency. The transgenic orchid plants were established in pots and acclimatised in the greenhouse. The transgenic plants were found morphologically similar compared to untransformed plants.
format	Journal Contribution
author	Rogayah Sekeli Khairun Hisam Nasir Wee, C. Y. Vilasini Pillai
author_facet	Rogayah Sekeli Khairun Hisam Nasir Wee, C. Y. Vilasini Pillai
author_sort	Rogayah Sekeli
title	Assessment of co-transformation technique for Malaysian orchid variety (Dendrobium Savin White) with cymbidium mosaic virus coat protein gene
title_short	Assessment of co-transformation technique for Malaysian orchid variety (Dendrobium Savin White) with cymbidium mosaic virus coat protein gene
title_full	Assessment of co-transformation technique for Malaysian orchid variety (Dendrobium Savin White) with cymbidium mosaic virus coat protein gene
title_fullStr	Assessment of co-transformation technique for Malaysian orchid variety (Dendrobium Savin White) with cymbidium mosaic virus coat protein gene
title_full_unstemmed	Assessment of co-transformation technique for Malaysian orchid variety (Dendrobium Savin White) with cymbidium mosaic virus coat protein gene
title_sort	assessment of co-transformation technique for malaysian orchid variety (dendrobium savin white) with cymbidium mosaic virus coat protein gene
publishDate	2020
url	http://agris.upm.edu.my:8080/dspace/handle/0/17337
_version_	1782772377064570880
score	12.935284

Assessment of co-transformation technique for Malaysian orchid variety (Dendrobium Savin White) with cymbidium mosaic virus coat protein gene

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