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JXB Advance Access originally published online on June 1, 2007
Journal of Experimental Botany 2007 58(10):2429-2440; doi:10.1093/jxb/erm093
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© The Author [2007]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

Insights into rubber biosynthesis from transcriptome analysis of Hevea brasiliensis latex

Keng-See Chow1,*, Kiew-Lian Wan2,3, Mohd. Noor Mat Isa2, Azlina Bahari1, Siang-Hee Tan4 {dagger}, K Harikrishna4 {dagger} and Hoong-Yeet Yeang1

1Biotechnology Unit, Rubber Research Institute of Malaysia, Malaysian Rubber Board, RRIM Experiment Station, 47000 Sungai Buloh, Selangor, Malaysia
2Malaysia Genome Institute, UKM-MTDC Smart Technology Centre, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia
3School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia
4Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia

* To whom correspondence should be addressed. E-mail: kschow{at}lgm.gov.my

Hevea brasiliensis is the most widely cultivated species for commercial production of natural rubber (cis-polyisoprene). In this study, 10 040 expressed sequence tags (ESTs) were generated from the latex of the rubber tree, which represents the cytoplasmic content of a single cell type, in order to analyse the latex transcription profile with emphasis on rubber biosynthesis-related genes. A total of 3441 unique transcripts (UTs) were obtained after quality editing and assembly of EST sequences. Functional classification of UTs according to the Gene Ontology convention showed that 73.8% were related to genes of unknown function. Among highly expressed ESTs, a significant proportion encoded proteins related to rubber biosynthesis and stress or defence responses. Sequences encoding rubber particle membrane proteins (RPMPs) belonging to three protein families accounted for 12% of the ESTs. Characterization of these ESTs revealed nine RPMP variants (7.9–27 kDa) including the 14 kDa REF (rubber elongation factor) and 22 kDa SRPP (small rubber particle protein). The expression of multiple RPMP isoforms in latex was shown using antibodies against REF and SRPP. Both EST and quantitative reverse transcription-PCR (QRT-PCR) analyses demonstrated REF and SRPP to be the most abundant transcripts in latex. Besides rubber biosynthesis, comparative sequence analysis showed that the RPMPs are highly similar to sequences in the plant kingdom having stress-related functions. Implications of the RPMP function in cis-polyisoprene biosynthesis in the context of transcript abundance and differential gene expression are discussed.

Key words: cis-polyisoprene, EST, Hevea brasiliensis, latex transcriptome, REF, rubber biosynthesis, rubber particle proteins, SRPP

{dagger} Present address: Sime Darby Technology Centre, Jalan Tandang, 46050 Petaling Jaya, Selangor, Malaysia

Received 13 November 2006; Revised 16 March 2007 Accepted 4 April 2007


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