Agricultural Sciences in China
2008, 7(5): 521-534
© 2008, CAAS. All rights reserved. Published by Elsevier Ltd.
May 2008
Expression in Arabidopsis of a Strawberry Linalool Synthase Gene Under
the Control of the Inducible Potato PI2 Promoter
YANG Li-mei
1, 2, 3
, Per Mercke
3
, Joop J A van Loon
2
, FANG Zhi-yuan
1
, Marcel Dicke
2
and Maarten A Jongsma
3
1
Institute of Vegetables & Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
2
Laboratory of Entomology, Wageningen University, P.O. Box 8031, 6700 EH Wageningen, The Netherlands
3
Plant Research International, P.O. Box 16, 6700 AA Wageningen, The Netherlands
Abstract
To investigate the role of inducible linalool in Arabidopsis-insect interactions, the FaNES1 linalool synthase (LIS) cDNA
from strawberry with plastid targeting and a synthetic intron (LIS’) was placed under the control of the wound inducible
proteinase inhibitor 2 (PI2) promoter from potato. The construct pBin-P
PI2
-LIS’ was transformed to Arabidopsis thaliana
ecotype Columbia 0. Kanamycin resistant T
0
seedlings were confirmed for the presence and transcription of the LIS’ gene by
PCR analysis on genomic DNA and by RT-PCR analysis on RNA. Genomic and RT-PCR products were sequenced to confirm
correct splicing of the synthetic intron. The expression of active linalool synthase by the P
PI2
-LIS’ gene construct in the
transgenic lines was assessed by measuring linalool emission using solid phase micro-extraction (SPME) GC-MS measurements
after induction with methyl jasmonate. Among 30 tested independent T
2
transgenic lines, 10 exhibited linalool production.
Linalool expression could be induced by methyl jasmonate treatment, but not by diamondback moth larvae.
Key words: Aradidopsis, linalool synthase, potato proteins inhibitor, inducible promoter, methyl jasmonate, Plutella
xylostella, induction, expression
INTRODUCTION
Terpenoids comprise one of the largest families of
compounds produced by plants. They have many
functions, and for example act as hormones, mediators
of polysaccharide synthesis, photosynthetic pigments
(chlorophyll and carotenoids), and membrane
components (Chappell 1995). They also mediate
interactions between plants and their environment: They
are involved in interactions between plants and
microorganisms, between plants and herbivorous
arthropods or between plants and carnivorous
arthropods (Cseke et al. 1998; Aharoni et al . 2003;
2006; Kappers et al. 2005). Moreover, plants may emit
terpenoids that attract insect pollinators (Dobson 1993;
Knudsen et al. 1993).
Linalool (3,7-dimethyl-1,6-octadien-3-ol), a C10
monoterpene volatile, is present in the floral fragrance
of diverse plant families. Due to its sweet, pleasant
fragrance it is used extensively by the flavour and
fragrance industry. Linalool mediates interactions
among plants and other organisms, e.g., it has been
shown to act either as an attractant or a repellent/
deterrent to a broad spectrum of pollinators, herbivores
and parasitoids (Dicke et al . 1990; Raguso and
Pichersky 1999; Wei and Kang 2006).
For the purpose of our research, the latter aspect is
of special interest. Attraction was documented for
insects such as the tarnished plant bug (Chinta et al.
Received 20 January, 2007 Accepted 21 April, 2008
Correspondence Maarten A Jongsma, E-mail: maarten.jongsma@wur.nl, Fax: +31-317-477109, Tel: +31-317-480932, Mobile: +31-630451274