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葡萄萜类环化酶:两个倍半萜合酶cDNA的功能鉴定,分别编码(+)-瓦伦烯合酶和(-)-吉马烯D合酶,以及单萜和倍半萜合酶在葡萄花和浆果中的表达

Vitis vinifera terpenoid cyclases: functional identification of two sesquiterpene synthase cDNAs encoding (+)-valencene synthase and (-)-germacrene D synthase and expression of mono- and sesquiterpene synthases in grapevine flowers and berries.

作者信息

Lücker Joost, Bowen Pat, Bohlmann Jörg

机构信息

Biotechnology Laboratory, Department of Botany, University of British Columbia, 6174 University Boulevard, Vancouver, BC, V6T 1Z3, Canada.

出版信息

Phytochemistry. 2004 Oct;65(19):2649-59. doi: 10.1016/j.phytochem.2004.08.017.

Abstract

Valencene is a volatile sesquiterpene emitted from flowers of grapevine, Vitis vinifera L. A full-length cDNA from the cultivar Gewürztraminer was functionally expressed in Escherichia coli and found to encode valencene synthase (VvVal). The two major products formed by recombinant VvVal enzyme activity with farnesyl diphosphate (FPP) as substrate are (+)-valencene and (-)-7-epi-alpha-selinene. Grapevine valencene synthase is closely related to a second sesquiterpene synthase from this species, (-)-germacrene D synthase (VvGerD). VvVal and VvGerD cDNA probes revealed strong signals in Northern hybridizations with RNA isolated from grapevine flower buds. Transcript levels were lower in open pre-anthesis flowers, flowers after anthesis, or at early onset of fruit development. Similar results were obtained using a third probe, (-)-alpha-terpineol synthase, a monoterpenol synthase. Sesquiterpene synthase and monoterpene synthase transcripts were not detected in the mesocarp and exocarp during early stages of fruit development, but transcripts hybridizing with VvVal appeared during late ripening of the berries. Sesquiterpene synthase transcripts were also detected in young seeds.

摘要

甜橙烯是葡萄(Vitis vinifera L.)花朵释放出的一种挥发性倍半萜。从品种琼瑶浆中获得的一个全长cDNA在大肠杆菌中实现了功能表达,发现其编码甜橙烯合酶(VvVal)。以法尼基二磷酸(FPP)为底物,重组VvVal酶活性形成的两种主要产物是(+)-甜橙烯和(-)-7-表-α-芹子烯。葡萄甜橙烯合酶与该物种的另一种倍半萜合酶,即(-)-吉马烯D合酶(VvGerD)密切相关。VvVal和VvGerD cDNA探针在与从葡萄花芽中分离的RNA进行的Northern杂交中显示出强信号。在开花前开放的花朵、开花后的花朵或果实发育早期,转录水平较低。使用第三种探针,即单萜醇合酶(-)-α-松油醇合酶,也获得了类似的结果。在果实发育早期,中果皮和外果皮中未检测到倍半萜合酶和单萜合酶转录本,但在浆果成熟后期出现了与VvVal杂交的转录本。在幼嫩种子中也检测到了倍半萜合酶转录本。

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