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The molecular and biochemical basis for varietal variation in sesquiterpene content in melon (Cucumis melo L.) rinds

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Abstract

A combined chemical, biochemical and molecular study was conducted to understand the differential accumulation of volatile sesquiterpenes in melon fruits. Sesquiterpenes were present mainly in the rinds of climacteric varieties, and a great diversity in their composition was found among varieties. Sesquiterpenes were generally absent in non-climacteric varieties. Two climacteric melon varieties, the green-fleshed ‘Noy Yizre'el’, and the orange-fleshed ‘Dulce’ were further examined. In ‘Noy Yizre'el’ the main sesquiterpenes accumulated are δ-cadinene, γ-cadinene and α-copaene, while α-farnesene is the main sesquiterpene in ‘Dulce’. Sesquiterpene synthase activities, mainly restricted to rinds of mature fruits, were shown to generate different sesquiterpenes in each variety according to the compositions found in rinds. EST melon database mining yielded two novel cDNAs coding for members of the Tps gene family termed CmTpsNY and CmTpsDul respectively, that are 43.2% similar. Heterologous expression in E. coli of CmTpsNY produced mainly δ-copaene, α-copaene, β-caryophyllene, germacrene D, α-muurolene, γ-cadinene, δ-cadinene, and α-cadinene, while CmTpsDul produced α-farnesene only. CmTpsNY was mostly expressed in ‘Noy Yizre'el’ rind while CmTpsDul expression was specific to ’Dulce’ rind. None of these genes was expressed in rinds of the non-climacteric ‘Tam Dew’ cultivar. Our results indicate that different sesquiterpene synthases encoded by different members of the Tps gene family are active in melon varieties and this specificity modulates the accumulation of sesquiterpenes. The genes are differentially transcriptionally regulated during fruit development and according to variety and are likely to be associated with chemical differences responsible for the unique aromas of melon varieties.

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Acknowledgements

This work was partially supported by the ‘ARO Center for the Improvement of Cucurbit Fruit Quality’, by the Israel Ministry of Science Grant and by grant Nos. IS-3333-02 and IS-3877-06 of BARD, the United States-Israel Binational Agricultural Research and Development Fund and by the EU project MetaPhor Food-CT-2006-03622. Publication No. 122/2007 of the Agricultural Research Organization, Bet Dagan Israel. We thank Uzi Ravid, Olga Larkov, Uzi Sa’ar and Fabian Baumkoler and Eti Braindes for assistance with sample preparation and GC-MS analysis. We also thank Eyal Koren, Maya Lotan-Pompan, Tamar Zakai and Rachel Ofir for assistance with figure preparation and scientific writing.

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Authors and Affiliations

  1. Institute of Plant Sciences, Newe Ya’ar Research Center, Agricultural Research Organization, P.O. Box 1021, Ramat Yishay, 30095, Israel

    Vitaly Portnoy, Yael Benyamini, Einat Bar, Rotem Harel-Beja, Joseph Burger, Yaakov Tadmor, Efraim Lewinsohn & Nurit Katzir

  2. Department of Biology, Technion-Israel Institute of Technology, Haifa, 32000, Israel

    Shimon Gepstein

  3. United States Department of Agriculture and Boyce Thompson Institute for Plant Research, Cornell University, Tower Road, Ithaca, NY, 14853, USA

    James J. Giovannoni

  4. Institute of Plant Sciences, The Volcani Center, Agricultural Research Organization, P.O. Box 6, Bet Dagan, 76100, Israel

    Arthur A. Schaffer

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  1. Vitaly Portnoy
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  2. Yael Benyamini
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  3. Einat Bar
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  4. Rotem Harel-Beja
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Correspondence to Efraim Lewinsohn.

Additional information

Vitaly Portnoy, Yael Benyamini and Einat Bar have equally contributed to the work.

The nucleotide sequence data reported here appears in the GenBank nucleotide database under the accession numbers EU158098 and EU158099.

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11103_2008_9296_MOESM1_ESM.tif

Figure S1: Mass spectra of products formed by the expressed sesquiterpene synthase CmTpsNY and CmTpsDul with FDP as substrate. Cell-free extracts prepared from the flesh and rind of 'Noy Yizre’el' and 'Dulce' respectively were incubated with FDP under the assay conditions. Product identification was done by GC-MS according to the retention time and compared to their the mass spectrum (upper chromatogram, A–D, E–H) to literature (lower chromatogram, A–D, E–H). . The products are: A—α-Copaene, B—β-Caryophyllene, C—Germacrene D, D—α-Muurolene, E—g-Cadinene, F—d-Cadinene, G—a Cadinene. The product formed by 'Dulce' extracts (left panels) was α-farnesene (H). (TIF 59 kb)

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Portnoy, V., Benyamini, Y., Bar, E. et al. The molecular and biochemical basis for varietal variation in sesquiterpene content in melon (Cucumis melo L.) rinds. Plant Mol Biol 66, 647–661 (2008). https://doi.org/10.1007/s11103-008-9296-6

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