Reference : Unveiling two new trichome-specific promoters of interest for metabolic engineering i...
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Life sciences : Biotechnology
Unveiling two new trichome-specific promoters of interest for metabolic engineering in Nicotiana tabacum
Pottier, Mathieu mailto [Université de Liège - ULiège > Département des sciences de la vie > Care "PhytoSYSTEMS" >]
Laterre, Raphaëlle []
Van Wessem, Astrid []
Ramirez, Aldana M. []
Herman, Xavier []
Boutry, Marc []
Hachez, Charles []
[en] tobacco ; glandular trichome ; metabolic engineering ; RbcS ; Rubisco small subunit ; CBTS ; CPS ; LTP ; MALD
[en] Nicotiana tabacum has emerged as a suitable host for metabolic engineering of terpenoids and
derivatives in tall glandular trichomes, which actively synthesize and secrete specialized
metabolites. However, implementation of an entire biosynthetic pathway in glandular
trichomes requires the identification of trichome-specific promoters to appropriately drive the
expression of the transgenes needed to set up the desired pathway. In this context, RT-qPCR
analysis was carried out on wild-type N. tabacum plants to compare the expression pattern
and level of NtRbS-T1 and NtMALD1, two newly identified genes expressed in glandular
trichomes, with those of NtCYP71D16, NtCBTS2α, NtCPS2, NtLTP1, which were reported in
the literature to be specifically expressed in glandular trichomes. The latter were previously
investigated separately, preventing any accurate comparison of their expression level. We
show that NtRbcS-T1 and NtMALD1 are specifically expressed in glandular trichomes just
like NtCYP71D16, NtCBTS2α, NtCPS2, while NtLTP1 was also expressed in other leaf tissues
as well as in the stem. Transcriptional fusions of all six promoters to the GUS-VENUS
reporter gene were introduced in N. tabacum by Agrobacterium-mediated transformation.
Most transgenic lines displayed GUS activity in tall glandular trichomes. In some transgenic
lines, except for pNtLTP1:GUS-VENUS, this expression was specific. In other transgenic
lines, GUS expression was extended to other tissues, probably resulting from a position effect
during transgene integration. We discuss alternatives to overcome this lack of tissue
specificity in some transgenic lines, should some of these promoters be used in the context of
metabolic engineering in N. tabacum.
Unveiling two new trichome-specific promoters of interest for metabolic engineering in Nicotiana tabacum

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