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Construction and analysis of EST libraries of the trans-polyisoprene producing plant, Eucommia ulmoides Oliver

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Abstract

Eucommia ulmoides Oliver is one of a few woody plants capable of producing abundant quantities of trans-polyisoprene rubber in their leaves, barks, and seed coats. One cDNA library each was constructed from its outer stem tissue and inner stem tissue. They comprised a total of 27,752 expressed sequence tags (ESTs) representing 10,520 unigenes made up of 4,302 contigs and 6,218 singletons. Homologues of genes coding for rubber particle membrane proteins that participate in the synthesis of high-molecular poly-isoprene in latex were isolated, as well as those encoding known major latex proteins (MLPs). MLPs extensively shared ESTs, indicating their abundant expression during trans-polyisoprene rubber biosynthesis. The six mevalonate pathway genes which are implicated in the synthesis of isopentenyl diphosphate (IPP), a starting material of poly-isoprene biosynthesis, were isolated, and their role in IPP biosynthesis was confirmed by functional complementation of suitable yeast mutants. Genes encoding five full-length trans-isoprenyl diphosphate synthases were also isolated, and two among those synthesized farnesyl diphosphate from IPP and dimethylallyl diphosphate, an assumed intermediate of rubber biosynthesis. This study should provide a valuable resource for further studies of rubber synthesis in E. ulmoides.

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Abbreviations

DMAPP:

Dimethylallyl diphosphate

DXP:

Deoxy-xylulose phosphate

FPP:

Farnesyl diphosphate

GGPP:

Geranylgeranyl diphosphate

HMG-CoA:

3-Hydroxy-3-methylglutaryl-CoA

IPP:

Isopentenyl diphosphate

IT:

Inner stem tissue

MEP:

Methylerythritol phosphate

MLP:

Major latex protein

OT:

Outer stem tissue

REF:

Rubber elongation factor

RPMP:

Rubber particle membrane protein

SRPP:

Small rubber particle protein

TIDS:

Trans-isoprenyl diphosphate synthase

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Acknowledgments

This work was partially supported by the New Energy and Industrial Technology Development Organization (NEDO).

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Authors and Affiliations

  1. Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka, 565-0871, Japan

    Nobuaki Suzuki, Takashi Nishikawa, Takeshi Bamba, Ei-ichiro Fukusaki, Akio Kobayashi & Yoshihisa Nakazawa

  2. Department of Bioscience, Nagahama Institute of Bio-Science and Technology, 1266 Tamura, Nagahama, 526-0829, Japan

    Yukio Mukai

  3. Kitasato Institute for Life Sciences, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa, 228-8555, Japan

    Atsushi Yamashita & Masahira Hattori

  4. Department of Information Systems, Graduate School of Information Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara, 630-0101, Japan

    Naotake Ogasawara

  5. Kazusa DNA Research Institute, Kisarazu, Chiba, 292-0818, Japan

    Yoshiyuki Ogata, Nozomu Sakurai, Hideyuki Suzuki & Daisuke Shibata

  6. Technical Research Institute, Hitachi Zosen Corporation, Osaka, 565-0871, Japan

    Hirotaka Uefuji & Yoshihisa Nakazawa

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  1. Nobuaki Suzuki
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  2. Hirotaka Uefuji
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Suzuki, N., Uefuji, H., Nishikawa, T. et al. Construction and analysis of EST libraries of the trans-polyisoprene producing plant, Eucommia ulmoides Oliver. Planta 236, 1405–1417 (2012). https://doi.org/10.1007/s00425-012-1679-x

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