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Genomic structure and characterization of a lipase class 3 gene and promoter from oil palm

  • Original Papers
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Biologia Plantarum

Abstract

Lipase class 3 is part of the triacylglycerol lipase family involved in lipid degradation, esterification, and transesterification processes in plants. In this study, a lipase class 3 gene and promoter from oil palm (Elaeis guineensis Jacq.) were isolated and characterized by Northern blot, Southern blot, oil palm genome sequence, and transient expression GUS assay. The full-length lipase class 3 (FLL1) deduced polypeptide encoded 483 amino acids and was identical to that deduced from lipase (EgLip1) cDNA (GI: 409994625). It contained the lipase consensus sequence, GxSxG motif, and a putative catalytic triad and had a 3-dimensional protein model similar to that of a lipase from Giberella zeae with a 50 % identity. The Northern blot and reverse transcription polymerase chain reaction (RT-PCR) show that FLL1 was predominantly expressed in the mesocarp and the expression increased as fruits reached maturity. A lower expression was detected in germinated seedlings and especially in roots. The expression of FLL1 was also enhanced in the mesocarp of cold treated fruits. A high oil accumulation in the mesocarp during fruit development makes this tissue a suitable target for a genetic modification, hence the isolation of the FLL1 promoter. The transient expression of the β-glucuronidase (GUS) gene driven by the FLL1 promoter detected the GUS expression in mesocarp slices, especially in vascular bundles. This suggests the potential role of using the promoter as tool to direct the expression of a transgene to the mesocarp of transgenic oil palm.

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Abbreviations

CARE:

cis-acting regulatory elements

FFA:

free fatty acids

FLL1:

full-length lipase class 3

GUS:

β-glucuronidase

LD-PCR:

long distance polymerase chain reaction

LRE:

light responsive elements

LTRE:

low temperature responsive elements

ORF:

open reading frame

RACE:

rapid amplification of cDNA ends

REG:

regulatory element groups

RT-PCR:

reverse transcription polymerase chain reaction

SDS:

sodium dodecyl sulphate

SSC:

sodium chloride-sodium citrate

TAG:

triacylglycerides

TSS:

transcription start site

UTR:

untranslated region

WAA:

weeks after anthesis

Y-patch:

pyrimidine patch

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Authors and Affiliations

  1. Persiaran Institute, Bandar Baru Bangi, Malaysian Palm Oil Board, No 6, 43000, Kajang Selangor, Malaysia

    A. W. Nurniwalis, R. Zubaidah, H. Zulkifli, M. A. Mohamad Arif, K. L. Chan & G. K. A. Parveez

  2. Faculty of Agriculture and Institute of Tropical Agriculture, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    A. Siti Nor Akmar

  3. School of Biosciences, University of Nottingham Malaysia Campus, Jalan Broga, 43500, Semenyih, Selangor, Malaysia

    A. W. Nurniwalis & F. J. Massawe

Authors
  1. A. W. Nurniwalis
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  2. R. Zubaidah
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  3. A. Siti Nor Akmar
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  4. H. Zulkifli
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  5. M. A. Mohamad Arif
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  6. F. J. Massawe
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  8. G. K. A. Parveez
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Correspondence to A. W. Nurniwalis.

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Acknowledgments: We thank the Director General of MPOB for permission to publish this paper. We would also like to thank Pn Aminah Shuib for a technical help, Chan Pek Lan for a constructive assistance, Dr. Meilina Ong-Abdullah for the supply of oil palm male and female inflorescences total RNA, all personnel in the Gene Function Laboratory, MPOB, for their support and continuous help, and Dr Ariffin Darus, Dr. Ooi Siew Eng, and Dr. Abrizah Othman for technical editing the paper.

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Nurniwalis, A.W., Zubaidah, R., Siti Nor Akmar, A. et al. Genomic structure and characterization of a lipase class 3 gene and promoter from oil palm. Biol Plant 59, 227–236 (2015). https://doi.org/10.1007/s10535-015-0500-7

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  • DOI: https://doi.org/10.1007/s10535-015-0500-7

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