Cloning, Sequence Analysis and Expression Patterns during Seed Germination of a Rapeseed (Brassica napus L.) G-x-S-x-G-motif Lipase Gene

  • Imen GLAIED GHRAM Unité Protéomie Fonctionnelle et Potentiel Nutraceutique de la Biodiversité de Tunisie. Institut des Sciences Biologiques Appliquées de Tunis
  • Hatem BELGUITH Unité Protéomie Fonctionnelle et Potentiel Nutraceutique de la Biodiversité de Tunisie. Institut des Sciences Biologiques Appliquées de Tunis
  • Maha BEN MUSTAPHA Biochemistry and Molecular Biology Laboratory (03/UR/0902), Faculty of Sciences of Bizerte, University of Carthage & Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB), National Institute of Applied Sciences and Technology (INSAT), University of Carthage
  • Issam HIMILA Institut Pasteur de Tunis
  • Balkiss BOUHAOUALA Institut Pasteur de Tunis
  • Oscar VICENTE IBMCP Universidad Politécnica de Valencia, Camino de Vera s/n. 46022 Valencia
  • Jeannette BEN HAMIDA Unité Protéomie Fonctionnelle et Potentiel Nutraceutique de la Biodiversité de Tunisie. Institut des Sciences Biologiques Appliquées de Tunis

Abstract

Lipases catalyze the hydrolysis of ester bonds in triacylglycerides, generating glycerol and free fatty acids. These enzymes are encoded by extremely complex gene families, and appear to fulfil many different biological functions. Although they are present in all types of organisms, available information on plant lipases is still very limited, as compared to their bacterial and animal counterparts. A full-length clone, BnLIP, encoding a putative lipase, has been isolated by PCR amplification of Brassica napus genomic DNA, with oligonucleotide primers derived from the sequence of an Arabidopsis thaliana homologue. The clone included an open reading frame of 1581 bp encoding a polypeptide of 526 amino acids, with a calculated molecular mass of 59.5 kDa. Analysis of the deduced protein sequence, sequence alignment with homologous proteins from related plant species, and a phylogenetic analysis revealed that the BnLIP protein belongs to the ‘classical’ GxSxG-motif lipase family. RT-PCR assays indicated that the BnLIP gene is expressed specifically, but only transiently, during seed germination: the lipase mRNA was not present at detectable levels in ungerminated seeds, was detected only three days after seed imbibition, but its levels decreased rapidly afterwards. No expression was observed in roots, stems or leaves of adult plants. This expression pattern suggests that BnLIP is one of the lipases involved in the hydrolysis of triacylglycerides stored in rapeseed seeds, ultimately providing nutrients and energy to sustain seedling growth until photosynthesis is activated.

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Published
2016-12-14
How to Cite
GLAIED GHRAM, I., BELGUITH, H., BEN MUSTAPHA, M., HIMILA, I., BOUHAOUALA, B., VICENTE, O., & BEN HAMIDA, J. (2016). Cloning, Sequence Analysis and Expression Patterns during Seed Germination of a Rapeseed (Brassica napus L.) G-x-S-x-G-motif Lipase Gene. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 44(2), 435-444. https://doi.org/10.15835/nbha44210472
Section
Research Articles