Simple Search:

Heat and hydrolytic enzymes treatment improved the Agrobacterium-mediated transformation of recalcitrant indica rice (Oryza sativa L.)


Citation

Wong, Clement Kiing Fook and Lai, Kok Song and Wong, Mui Yun and Mahmood, Maziah (2016) Heat and hydrolytic enzymes treatment improved the Agrobacterium-mediated transformation of recalcitrant indica rice (Oryza sativa L.). Plant Cell, Tissue and Organ Culture, 125 (1). pp. 183-190. ISSN 0167-6857; ESSN: 1573-5044

Abstract / Synopsis

Reported Agrobacterium transformation efficiency of indica rice shoot apices varied from 0.4 to 13.8 %. In order to improve the transformation efficiency, modification of transformation protocols through heat and hydrolytic enzyme treatments on rice shoot apices were carried out. Transient expression study using reporter genes revealed that shoot apices heat treated for 3 min at 42 °C during bacterial immersion showed improved GFP (63.0 %) and GUS (42.5 %) expressions per plant as compared to standard protocol (34.0 % GFP and 36.25 % GUS). Shoot apices pre-treated with hydrolytic enzymes containing macerase, pectinase and cellulase at concentration ratio of 1:1:1 (w/v) also demonstrated high percentage of transient GFP (40.0 %) and GUS (35.0 %) expressions per plant. PCR analyses further confirmed the presence of GFP and GUS genes in the transformants. Stable expressions of GFP and GUS were also obtained in multiple shoots of regenerated shoot apices after 4 weeks of culturing in shoot proliferation media without hygromycin. In conclusion, the transformation efficiencies were improved significantly when heat (15.83 %) and hydrolytic enzymes (16.67 %) were applied as individual treatments as compared to the standard transformation method which only accounted for 5.83 %.


Download File

[img]
Preview
PDF (Abstract)
Heat and hydrolytic enzymes treatment improved the Agrobacterium-mediated transformation of recalcitrant indica rice (Oryza sativa L.).pdf

Download (37kB) | Preview

Additional Metadata

Item Type: Article
Divisions: Faculty of Agriculture
Faculty of Biotechnology and Biomolecular Sciences
Institute of Tropical Agriculture
DOI Number: https://doi.org/10.1007/s11240-015-0926-9
Publisher: Springer
Keywords: Agrobacterium; Heat; Hydrolytic enzymes; Rice; Shoot apices
Depositing User: Nabilah Mustapa
Date Deposited: 18 May 2016 12:55
Last Modified: 18 May 2016 12:55
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1007/s11240-015-0926-9
URI: http://psasir.upm.edu.my/id/eprint/43352
Statistic Details: View Download Statistic

Actions (login required)

View Item View Item