Production of transgenic rice (indica cv. MR219) overexpressing Abp57 Gene through Agrobacterium-mediated transformation

Agrobacterium-mediated transformation of indica rice is undoubtedly a challenging task due to the rice recalcitrant nature to transformation process. Therefore, optimization of the transformation protocol is important for specific indica rice cultivar to ensure effectiveness of the transformation. I...

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Main Authors:	Tan, Lay-Wen, Zuraida A. Rahman, Goh, Hoe-Han, Hwang, Duk-Ju, Ismanizan Ismail, Zamri Zainal
Format:	Article
Language:	English
Published:	Penerbit Universiti Kebangsaan Malaysia 2017
Online Access:	http://journalarticle.ukm.my/11057/ http://journalarticle.ukm.my/11057/ http://journalarticle.ukm.my/11057/1/04%20Lay-Wen%20Tan.pdf

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spelling	ukm-110572017-12-12T05:31:25Z http://journalarticle.ukm.my/11057/ Production of transgenic rice (indica cv. MR219) overexpressing Abp57 Gene through Agrobacterium-mediated transformation Tan, Lay-Wen Zuraida A. Rahman, Goh, Hoe-Han Hwang, Duk-Ju Ismanizan Ismail, Zamri Zainal, Agrobacterium-mediated transformation of indica rice is undoubtedly a challenging task due to the rice recalcitrant nature to transformation process. Therefore, optimization of the transformation protocol is important for specific indica rice cultivar to ensure effectiveness of the transformation. In this study, crucial parameters affecting Agrobacterium-mediated transformation were optimized to obtain transgenic rice of local rice cultivar (indica MR219). Embryogenic calli were chosen for inoculation with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pH2GW7-ABP57 containing gene of interest, Auxin binding protein 57 (Abp57). The parameters that have been optimized were the immersion time, co-cultivation period, acetosyringone concentration and co-cultivation temperature. A total of four days co-cultivation period and 30 min immersion of embryogenic callus are optimum for the transformation of MR219 with transformation efficiency of 26.4% and 16.0%, respectively. Acetosyringone at 200 μM and co-cultivation at 28°C also gave the highest transformation efficiency (14.4 and 18.4%, respectively). Meanwhile, inclusion of 20 g/L maltose+20 g/L sorbitol into the regeneration media has significantly improve the transformed somatic embryos growth and increase the regeneration efficiency up to 40.0%. The results of polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR) indicated that the transgene was successfully integrated and overexpressed in transgenic rice of MR219. In conclusion, significant improvement in transformation efficiency for rice cv. MR219 has been obtained by using the optimised protocol for transformation and regeneration developed in this study. Penerbit Universiti Kebangsaan Malaysia 2017-05 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/11057/1/04%20Lay-Wen%20Tan.pdf Tan, Lay-Wen and Zuraida A. Rahman, and Goh, Hoe-Han and Hwang, Duk-Ju and Ismanizan Ismail, and Zamri Zainal, (2017) Production of transgenic rice (indica cv. MR219) overexpressing Abp57 Gene through Agrobacterium-mediated transformation. Sains Malaysiana, 46 (5). pp. 703-711. ISSN 0126-6039 http://www.ukm.my/jsm/english_journals/vol46num5_2017/contentsVol46num5_2017.html
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Kebangasaan Malaysia
building	UKM Institutional Repository
collection	Online Access
language	English
description	Agrobacterium-mediated transformation of indica rice is undoubtedly a challenging task due to the rice recalcitrant nature to transformation process. Therefore, optimization of the transformation protocol is important for specific indica rice cultivar to ensure effectiveness of the transformation. In this study, crucial parameters affecting Agrobacterium-mediated transformation were optimized to obtain transgenic rice of local rice cultivar (indica MR219). Embryogenic calli were chosen for inoculation with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pH2GW7-ABP57 containing gene of interest, Auxin binding protein 57 (Abp57). The parameters that have been optimized were the immersion time, co-cultivation period, acetosyringone concentration and co-cultivation temperature. A total of four days co-cultivation period and 30 min immersion of embryogenic callus are optimum for the transformation of MR219 with transformation efficiency of 26.4% and 16.0%, respectively. Acetosyringone at 200 μM and co-cultivation at 28°C also gave the highest transformation efficiency (14.4 and 18.4%, respectively). Meanwhile, inclusion of 20 g/L maltose+20 g/L sorbitol into the regeneration media has significantly improve the transformed somatic embryos growth and increase the regeneration efficiency up to 40.0%. The results of polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR) indicated that the transgene was successfully integrated and overexpressed in transgenic rice of MR219. In conclusion, significant improvement in transformation efficiency for rice cv. MR219 has been obtained by using the optimised protocol for transformation and regeneration developed in this study.
format	Article
author	Tan, Lay-Wen Zuraida A. Rahman, Goh, Hoe-Han Hwang, Duk-Ju Ismanizan Ismail, Zamri Zainal,
spellingShingle	Tan, Lay-Wen Zuraida A. Rahman, Goh, Hoe-Han Hwang, Duk-Ju Ismanizan Ismail, Zamri Zainal, Production of transgenic rice (indica cv. MR219) overexpressing Abp57 Gene through Agrobacterium-mediated transformation
author_facet	Tan, Lay-Wen Zuraida A. Rahman, Goh, Hoe-Han Hwang, Duk-Ju Ismanizan Ismail, Zamri Zainal,
author_sort	Tan, Lay-Wen
title	Production of transgenic rice (indica cv. MR219) overexpressing Abp57 Gene through Agrobacterium-mediated transformation
title_short	Production of transgenic rice (indica cv. MR219) overexpressing Abp57 Gene through Agrobacterium-mediated transformation
title_full	Production of transgenic rice (indica cv. MR219) overexpressing Abp57 Gene through Agrobacterium-mediated transformation
title_fullStr	Production of transgenic rice (indica cv. MR219) overexpressing Abp57 Gene through Agrobacterium-mediated transformation
title_full_unstemmed	Production of transgenic rice (indica cv. MR219) overexpressing Abp57 Gene through Agrobacterium-mediated transformation
title_sort	production of transgenic rice (indica cv. mr219) overexpressing abp57 gene through agrobacterium-mediated transformation
publisher	Penerbit Universiti Kebangsaan Malaysia
publishDate	2017
url	http://journalarticle.ukm.my/11057/ http://journalarticle.ukm.my/11057/ http://journalarticle.ukm.my/11057/1/04%20Lay-Wen%20Tan.pdf
first_indexed	2023-09-18T19:59:12Z
last_indexed	2023-09-18T19:59:12Z
_version_	1777406734441119744

Production of transgenic rice (indica cv. MR219) overexpressing Abp57 Gene through Agrobacterium-mediated transformation

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