African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12488

Full Length Research Paper

Analysis of gene expression by promoter reporter constructs and knockout plants: A case study under phosphorus stress

Kefyalew Negisho1*, Tesfaye Balemi2 and Belayneh Admassu1
1Holetta Agricultural Research Center, National Agricultural Biotechnology Research Programme, Ethiopian Institute of Agricultural Research, Ethiopia. 2Department of Plant Science and Horticulture, College of Agricultural and Veterinary Science, Ambo University, Ethiopia.
Email: [email protected]

  •  Accepted: 21 July 2011
  •  Published: 24 October 2011

Abstract

Promoter reporter constructs were developed using Gateway Cloning Technology for plant non-specific lipid transport proteins (nsLTP1, AT4g12545 and nsLTP2, AT5g46900), MADS-Box (AT4g37940) and leucine rich repeat (LRR, AT2g28970)genes. The purpose of the construct was to investigate gene expression and function under phosphorus (P) stress. Agrobacterium tumefaciens was used to transform Arabidopsis thaliana. Transformed (T1), knockout and wild type ecotype Colombia-0 A. thaliana were investigated. To monitor the gene activity, plants were grown under different conditions: fully inside solid media, on the surface of solid media and hydroponically with 0.5 mM l-1 phosphorus and without phosphorus. Plants were also grown on MS media with selectable marker containing 50 mM l-1Kanamycin. Morphological root assessment was also done by using knockoutplants for the respective promoters. The promoters under study were induced promoter. GUS staining was observed on T1 plants when planted fully inside solid media and on the selectable marker for all the constructs regardless of phosphorus treatments. Mechanical impedance, oxygen deficiency and stress due to selectable marker were the probable reasons for the root specific gene expression. This suggests that these genes were involved in adaptation mechanism for plants under such stress conditions.  

 

Key words: Arabidopsis thaliana, gene expression, kanamycin, knockout plants, LRR (leucine rich repeat), MADS-Box, nsLTPs (non-specific lipid transport proteins), selectable marker.