Abstract:Farnesyl diphosphate synthase (FPS) is a key enzyme of isoprenoids biosynthesis. However, knowledge of the FPSs of euphorbiaceous species is limited. In this study, ten FPSs were identified in four euphorbiaceous plants. These FPSs exhibited similar exon/intron structure. The deduced FPS proteins showed close identities and exhibited the typical structure of plant FPS. The members of the FPS family exhibit tissue expression patterns that vary among several euphorbiaceous plant species under normal growth condi… Show more
“…FPPS catalyses the sequential condensation of two molecules of IPP with DMAPP to form C 15 ‐FPP, the immediate precursor for all sesquiterpenes (Cornish, ; Lange et al ., ). But FPPS is also know to accept C 10 ‐GPP as an allylic substrate to generate FPP (Guo et al ., ; Hemmerlin et al ., ). Further sesquiterpenes synthesized in cytoplasm can be derived from plastidial GPP too (Adam et al ., ).…”
SummaryMany aromatic plants, such as spearmint, produce valuable essential oils in specialized structures called peltate glandular trichomes (PGTs). Understanding the regulatory mechanisms behind the production of these important secondary metabolites will help design new approaches to engineer them. Here, we identified a PGT‐specific R2R3‐MYB gene, MsMYB, from comparative RNA‐Seq data of spearmint and functionally characterized it. Analysis of MsMYB‐RNAi transgenic lines showed increased levels of monoterpenes, and MsMYB‐overexpressing lines exhibited decreased levels of monoterpenes. These results suggest that MsMYB is a novel negative regulator of monoterpene biosynthesis. Ectopic expression of MsMYB, in sweet basil and tobacco, perturbed sesquiterpene‐ and diterpene‐derived metabolite production. In addition, we found that MsMYB binds to cis‐elements of MsGPPS.LSU and suppresses its expression. Phylogenetic analysis placed MsMYB in subgroup 7 of R2R3‐MYBs whose members govern phenylpropanoid pathway and are regulated by miR858. Analysis of transgenic lines showed that MsMYB is more specific to terpene biosynthesis as it did not affect metabolites derived from phenylpropanoid pathway. Further, our results indicate that MsMYB is probably not regulated by miR858, like other members of subgroup 7.
“…FPPS catalyses the sequential condensation of two molecules of IPP with DMAPP to form C 15 ‐FPP, the immediate precursor for all sesquiterpenes (Cornish, ; Lange et al ., ). But FPPS is also know to accept C 10 ‐GPP as an allylic substrate to generate FPP (Guo et al ., ; Hemmerlin et al ., ). Further sesquiterpenes synthesized in cytoplasm can be derived from plastidial GPP too (Adam et al ., ).…”
SummaryMany aromatic plants, such as spearmint, produce valuable essential oils in specialized structures called peltate glandular trichomes (PGTs). Understanding the regulatory mechanisms behind the production of these important secondary metabolites will help design new approaches to engineer them. Here, we identified a PGT‐specific R2R3‐MYB gene, MsMYB, from comparative RNA‐Seq data of spearmint and functionally characterized it. Analysis of MsMYB‐RNAi transgenic lines showed increased levels of monoterpenes, and MsMYB‐overexpressing lines exhibited decreased levels of monoterpenes. These results suggest that MsMYB is a novel negative regulator of monoterpene biosynthesis. Ectopic expression of MsMYB, in sweet basil and tobacco, perturbed sesquiterpene‐ and diterpene‐derived metabolite production. In addition, we found that MsMYB binds to cis‐elements of MsGPPS.LSU and suppresses its expression. Phylogenetic analysis placed MsMYB in subgroup 7 of R2R3‐MYBs whose members govern phenylpropanoid pathway and are regulated by miR858. Analysis of transgenic lines showed that MsMYB is more specific to terpene biosynthesis as it did not affect metabolites derived from phenylpropanoid pathway. Further, our results indicate that MsMYB is probably not regulated by miR858, like other members of subgroup 7.
“…HbFPS1 expressed obviously in the laticifers which is possible to involve in natural rubber biosynthesis 43 . The yeast one-hybrid analysis was performed to investigate whether HblMADS24 binds the HbFPS1 promoter.…”
Section: Resultsmentioning
confidence: 99%
“…HbFPS1 expressed obviously in the laticifers which is possible to involve in NR biosynthesis. However, HbFPS2 and HbFPS3 have no cell-type specific expression, and they are likely to act as housekeeping nature to involve in isoprenoid biosynthesis 43 .…”
MADS-box transcription factors possess many functions in plant reproduction and development. However, few MADS-box genes related to secondary metabolites regulation have been identified. In
H
evea
brasiliensis
, natural rubber is a representative
cis
-polyisoprenoids in secondary metabolism which occurs in the rubber laticifer cells, the molecular regulation basis of natural rubber biosynthesis is not clear. Here, a total of 24 MADS-box genes including 4 type I MADS-box genes and 20 type II MADS-box genes were identified in the transcriptome of rubber tree latex. The phylogenetic analysis was performed to clarify the evolutionary relationships of all the 24 rubber tree MADS-box proteins with MADS-box transcription factors from
Arabidopsis thaliana
and
Oryza sativa
. Four type I MADS-box genes were subdivided into Mα (3 genes) and Mβ (1 gene). Twenty type II MADS-box genes were subclassified into MIKC* (8 genes) and MIKC
c
(12 genes). Eight MADS-box genes (
HblMADS3
,
5
,
6
,
7
,
9
,
13
,
23
,
24
) were predominant expression in laticifers. ABA up-regulated the expression of
HblMADS9
, and the expression of
HblMADS3
,
HblMADS5
,
HblMADS24
were up-regulated by MeJA. The function of HblMADS24 was elucidated. HblMADS24 bound
HbFPS1
promoter in yeast and HblMADS24 activated
HbFPS1
promoter in tobacco plants. Moreover, we proposed that HblMADS24 is a transcription activator of
HbFPS1
which taking part in natural rubber biosynthesis.
“…Thus, FPS should be considered as a crucial enzyme in the natural rubber biosynthesis. The rubber tree FPS genes (named HbFPS1 , HbFPS2 , and HbFPS3 ) have been cloned and characterized 11 . The expression of HbFPS1 exhibits a positive correlation with natural rubber biosynthesis 11 , 12 .…”
Section: Introductionmentioning
confidence: 99%
“…The rubber tree FPS genes (named HbFPS1 , HbFPS2 , and HbFPS3 ) have been cloned and characterized 11 . The expression of HbFPS1 exhibits a positive correlation with natural rubber biosynthesis 11 , 12 . Recently two MYB transcription factors (HblMYB19 and HblMYB44) are identified to up-regulate the expression of HbFPS1 13 .…”
Farnesyl pyrophosphate synthase (FPS) is a key enzyme that catalyzes the formation of farnesyl pyrophosphate, the main initiator for rubber chain initiation in Hevea brasiliensis Muell. Arg. The transcriptional regulatory mechanisms of the FPS gene still not well understood. Here, a WRKY transcription factor designated HbWRKY27 was obtained by screening the latex cDNA library applied the HbFPS1 promoter as bait. HbWRKY27 interacted with the HbFPS1 promoter was further identified by individual Y1H and EMSA assays. HbWRKY27 belongs to group IIe WRKY subfamily which contains a typical WRKY domain and C-X5-CX23-HXH motif. HbWRKY27 was localized to the nucleus. HbWRKY27 predominantly accumulated in latex. HbWRKY27 was up-regulated in latex by ethrel, salicylic acid, abscisic acid, and methyl jasmonate treatment. Transient expression of HbWRKY27 led to increasing the activity of the HbFPS1 promoter in tobacco plant, suggesting that HbWRKY27 positively regulates the HbFPS1 expression. Taken together, an upstream transcription factor of the key natural rubber biosynthesis gene HbFPS1 was identified and this study will provide novel transcriptional regulatory mechanisms of the FPS gene in Hevea brasiliensis.
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