## 4 Materials and Methods
### Subcellular Localization Analysis Using the Rice Greening Protoplasts  
The preparation of rice protoplasts and their transfection using polyethylene glycol (PEG) were performed as previously described [38] using the leaves of 7 day-old-seedlings. The leaves of rice (_Oryza sativa_ L. _Japonic_ cv. \"Imi\") seedlings were segmented to be digested in Cellulase R-10 and Macerozyme R-10 enzyme solution (Yakult Honsha, Japan) for the removal of cell walls. 10\({}^{6}\) isolated protoplast cells were transfected with 5 \(\upmu\)g of each plasmid with an equal volume of 40% PEG-3350 solution (Sigma, St. Louis, MO, USA) containing 0.5 M mannitol and 100 mM CaCl\({}_{2}\). After the overnight incubation in the dark at 28 \({}^{\circ}\)C, fluorescence signals from the transfected protoplasts were observed and imaged using a Carl Zeiss LSM700 inverted confocal microscope (Carl Zeiss, Oberkochen, Germany). The sub-organelle marker constructs OsPSY2-mCherry [38], G-rb and px-rb [39] were used to identify plasticglobules, golgi complexes and peroxisomes, respectively. Mitochondria were probed by MitoTracker(tm) Red CMXRos (Invitrogen, Carlsbad, CA, USA) and the auto-fluorescent signals of chlorophylls were detected at excitation and emission wavelengths of 555 and \(>\)650 nm, respectively. The following are available online at [[http://www.mdpi.com/1422-0067/21/23/8927/s1](http://www.mdpi.com/1422-0067/21/23/8927/s1)]([http://www.mdpi.com/1422-0067/21/23/8927/s1](http://www.mdpi.com/1422-0067/21/23/8927/s1)). Figure S1. The biosynthetic schematics of plant _trans_-preurltransferases (PTs) using various allylic donors. Figure S2. The gene expression profiling data of an _OsFPSS_ (LOC_Os04g56210) extracted from RiceXPro ([[http://ricexpro.dna.affrc.go.jp/](http://ricexpro.dna.affrc.go.jp/)]([http://ricexpro.dna.affrc.go.jp/](http://ricexpro.dna.affrc.go.jp/))). Figure S3. Subcellular localization of three rice Group I _trans_-PTs. Table S1. Primer list for expression profiling of PT genes Table S2. Primers for cloning into pDONR221-Kz-Ndel-sGFP. Table S3. The list of plant _trans_-preurltransferases analyzed in phylogenetic tree analysis. Table S4. The list of plant _trans_-preurltransferases analyzed in the multiple sequence alignment. Table S5. The differential amino acid residues of \"three floors\" motifs in the families of Groups I, II and III. Table S6. Spatial and temporal expression of rice _trans_-PT genes. Table S7. Expression profiles of rice _trans_-PT genes in MeJA, ABA or GA-treated rice leaves. [PERSON]; [PERSON]; [PERSON] Terpenoid synthase structures: A so far incomplete view of complex catalysis. _Nat. Prod. Rep._**2012**, _29_, 1153-1175. [CrossRef]
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