Global Advanced Research Journal of Biotechnology (GARJB)

August  2015 Vol. 4(1), pp 030-040

Copyright © 2015 Global Advanced Research Journals


 

Full Length Research Paper 

Development of Regeneration and Transformation Protocol for Local Sudan’s wheat (Triticum aestivum L) Cultivars

Badr Eldin AE Saeed1, Marmar A. El Siddig2*, Aisha Ohag Osman3 and Adil A. El Hussein2 

1Department of Biotechnology and Biology, Faculty of Science and Biotechnology, AL Neelain University, Sudan.

2Department of Botany, Faculty of Science, University of Khartoum, Sudan.

3Department of Biology, Faculty of Education, Kassala University, Sudan.

Corresponding author Email: marmarelsiddig@hotmail.com

Accepted 18 July, 2014

Abstract

The objective of this study was to optimize a simple and efficient Agrobacterium-mediated genetic transformation protocol for two locally grown wheat cultivars (Bohain and Elnielain) using mature embryos. Transformation was carried out with disarmed A. tumefaciens strain EHA105 harbouring a binary vector pCAMBIA1305.2.  Conditions for in vitro regeneration of Agrobacterium-infected explants were optimized using Murashige and Skoog (MS) medium supplemented with various concentrations and/or combinations of plant growth regulators (PGRs) and Hygromycin. Results indicated that 70 and 80% of the embryogenic explants  of Bohain and Elnielain, respectively, have survived when exposed to Agrobacterium inoculums of 0.1 O.D600 and selection on medium containing 15 (Elnielain) or 25 (Bohain) mg/l of Hygromycin. The putative transformed explants were regenerated on MS medium containing 4.0 mg/l 2,4-D for shoot initiation, transferred to MS medium with 4.0 mg/l 2,4-D plus 1.0 mg/l Zeatin for Shoot elongation, then maintained on MS medium supplemented with a combination of 1.0 mg/l of each of 2,4-D, Zeatin and gibrellic acid (GA3). This protocol resulted in transformation efficiency of 20% for Bohain and 23% for Elnielain.  GUS expression was observed in transformed shoots but never in the control plants. PCR amplification of DNA extracted from the transformed tissues demonstrated the generation of the expected amplicon, corresponding to neomycin phosphotransferase II (nptII), hygromycin phosphotransferase II (hptII), cauliflower mosaic virus-35S (CaMV-35S) promoter and nopaline synthase (NOS) terminator genes. This result strongly verifies the successful transformation of the two locally wheat cultivars and paves the way for problem solving-applications encompassing these or other Sudanese wheat cultivars of economic importance.

Keywords: wheat, regeneration, Agrobacterium, transformation, pCAMBIA.

Related Articles


Reviews
Artak Ayunts
The importance of conflict transformation between the parties in the Nagorno-Karabakh conflict
Glo. Adv. Res. J. Biotechnol. October 2012 Vol: 1(7): - [Abstract] [Full Text - PDF] (144 KB)
Made Antara and Made Sri Sumarniasih
Role of Bioscience and Biotechnology through Green Revolution to Increase the Rice Production in Indonesia
Glo. Adv. Res. J. Biotechnol. July 2017 Vol: 6(7): - [Abstract] [Full Text - PDF] (250 KB)
Original Research Articles
Mohammad Mehdi Fateh and Alireza Izadbakhsh
Task space control of flexible joint electrically driven robots
Glo. Adv. Res. J. Biotechnol. June 2013 Vol: 2(5): - [Abstract] [Full Text - PDF] (198 KB)
Sajedeh Nazari, Mehrdokht Mazdeh, Mohammad Taheri, Arezou Sayad, Abolfazl Movafagh and Romina Dastmalchi
A double-blind, placebo-controlled and randomized trial of Cerebrolysin in patients with acute ischemic stroke in Iran
Glo. Adv. Res. J. Biotechnol. May 2015 Vol: 4(5): - [Abstract] [Full Text - PDF] (104 KB)
Sumaira Hafeez, Liu Zhangyong, Jin Tao*, Shahid Iqbal Awan, Irshad Ahmad, Toheed Anwar, Rahat Sharif, Ntambo Mbuya Sylvain and Consolatha Chambi
Assessment of Drought Tolerance in Spring Wheat (Triticum Aestivum L.) At Different Growth Stages under the Rainfed Conditions of Rawalakot Azad Kashmir Pakistan
Glo. Adv. Res. J. Biotechnol. April 2016 Vol: 5(4): - [Abstract] [Full Text - PDF] (259 KB)
Mudavanhu C, Zinyandu T, Mudavanhu N, Mazorodze S, Chinyanganya TP, Manyani A, Maponga R, Pedzisai E, Phiri S
Smallholder gardening as a sustainable livelihood strategy in Chikwanda communal lands, Gutu, Zimbabwe
Glo. Adv. Res. J. Biotechnol. January 2013 Vol: 2(1): - [Abstract] [Full Text - PDF] (338 KB)

Current Issue

Viewing Options

View Full Article - PDF
Download Full Article - PDF

Search for Articles

Badr Eldin AE Saeed1 on Google Scholar
Badr Eldin AE Saeed1 on Pubmed
Marmar A. El Siddig2* on Google Scholar
Marmar A. El Siddig2* on Pubmed
Aisha Ohag Osman3 on Google Scholar
Aisha Ohag Osman3 on Pubmed
Adil A. El Hussein2 on Google Scholar
Adil A. El Hussein2 on Pubmed

Statistics

Viewed 1262
Printed 293
Downloaded 1263
Powered By iPortal Works