Affordable Access

Access to the full text

Expression patterns of cp4-epsps gene in diverse transgenic Saccharum officinarum L. genotypes

Authors
  • Imran, Muhammad1, 2
  • Barboza, Andre Luiz3
  • Asad, Shaheen1, 2
  • Khalid, Zafar M.4
  • Mukhtar, Zahid1, 2
  • 1 National Institute for Biotechnology and Genetic Engineering, Agricultural Biotechnology Division, Jhang Road, Faisalabad, Pakistan , Faisalabad (Pakistan)
  • 2 Pakistan Institute of Engineering and Applied Sciences, Nilore, Islamabad, Pakistan , Nilore, Islamabad (Pakistan)
  • 3 Escola Superior Agricultura Luiz De Quiroz, University of Sao Paulo, Laboratory of Genomics and Molecular Biology, Department of Biological Sciences, Piracicaba, SP, Brazil , Piracicaba (Brazil)
  • 4 International Islamic University, Department of Bioinformatics and Biotechnology, Islamabad, Pakistan , Islamabad (Pakistan)
Type
Published Article
Journal
Physiology and Molecular Biology of Plants
Publisher
Springer India
Publication Date
Feb 23, 2019
Volume
25
Issue
3
Pages
779–786
Identifiers
DOI: 10.1007/s12298-019-00650-y
Source
Springer Nature
Keywords
License
Yellow

Abstract

Glyphosate, a functional analogue of phosphoenolpyruvate (PEP), blocks the shikimate pathway by inhibiting the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS; EC 2.5.1.19) through interference with the conversion of (shikimate-3-phosphate) S3P and PEP to 5-enolpyruvylshikimate-3-phosphate (EPSP) and subsequently leads to plant death. This metabolic pathway possesses great potential to be used for development of herbicide resistant transgenic crops and here in this study, we wanted to check the expression potential of CP4-EPSPS gene in various sugarcane genotypes. A synthetic version of CP4-EPSPS gene synthesized commercially, cloned in pGreen0029 vector, was transformed into regenerable embryogenic calli of three different sugarcane cultivars HSF-240, S2003US-778 and S2003US-114 using biolistic gene transfer approach for comparative transcriptional studies. Transgenic lines screened by PCR analysis were subjected to Southern hybridization for checking transgene integration patterns. All the tested lines were found to contain multiple (3–6) insert copies. Putative transgenic plants produced the CP4-EPSPS protein which was detected using immunoblot analysis. The CP4-EPSPS transcript expression detected by qRT-PCR was found to vary from genotype to genotype and is being reported first time. In vitro glyphosate assay showed that transformed plants were conferring herbicide tolerance. It is concluded that different cultivars of sugarcane give variable expression of the same transgene and reasons for this phenomenon needs to be investigated.

Report this publication

Statistics

Seen <100 times