Two of the abundant transcripts encoding type 2 metallothionein (MT) proteins designated as MET2a and MET2b were selected in our previous study due to their high abundance (16.05 %) in the suppression subtractive hybridization library and their involvement in fruit development and maturation. The present study involves the isolation of the full-length cDNA encoding MET2a and MET2b from the ripening oil palm fruit mesocarp, examining their expression pattern compared to the other two previously reported type-3 MT members (MT3-A and MT3-B) in various oil palm organs including different vegetative and reproductive tissues. The full-length cDNA sequences of MET2a and MET2b were 571 and 553 bp and they were designated as EgMT2a and EgMT2b, respectively. The sequences of the EgMT2a and EgMT2b were then compared for sequence similarities in the database using both BLASTN and BLASTX programs. Their sequences were homologous (67 -77 %) with several type-2 MTs in plants. All four MT encoding genes were differentially expressed in the ripening oil palm mesocarp tissues, but undetectable in the vegetative tissues examined. All MT genes examined were significantly up-regulated in the mature developmental stages of oil palm fruit mesocarp, except for EgMT2b which was expressed only at 17 weeks after anthesis. The type 2 MT proteins are related to a greater degree to the late fruit-ripening stage than the type 3 MT proteins consistent with their reported functions in homeostasis or detoxification. The findings in the present study contribute to better understanding the molecular mechanisms involved in fruit ripening in oil palm.

Abscisic acid (ABA) is an important phytohormone involved in the abiotic stress resistance in plants. The ABA-responsive element (ABRE) binding factors play significant roles in the plant development and response to abiotic stresses, but none so far have been isolated and characterized from the oil palm. Two ABA-responsive cDNA clones, named EABF and EABF1, were isolated from the oil palm fruits using yeast one-hybrid system. The EABF had a conserved AP2/EREBP DNA-binding domain (DNA-BD) and a potential nuclear localization sequence (NLS). No previously known DNA-BD was identified from the EABF1 sequence. The EABF and EABF1 proteins were classified as DREB/CBF and bZIP family members based on the multiple sequence alignment and phylogenetic analysis. Both proteins showed ABRE-binding and transcriptional activation properties in yeast. Furthermore, both proteins were able to trans-activate the down-stream expression of the LacZ reporter gene in yeast. An electrophoretic mobility shift assay revealed that in addition to the ABRE sequence, both proteins could bind to the DRE sequence as well. Transcriptional analysis revealed that the expression of EABF was induced in response to the ABA in the oil palm fruits and leaves, but not in roots, while the EABF1 was constitutively induced in all tissues. The expressions of both genes were strongly induced in fruits in response to the ABA, ethylene, methyl jasmonate, drought, cold and high-salinity treatments, indicating that the EABF and EABF1 might act as connectors among different stress signal transduction pathways. Our results indicate that the EABF and EABF1 are novel stress-responsive transcription factors, which are involved in the abiotic stress response and ABA signaling in the oil palm and could be used for production of stress-tolerant transgenic crops.

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