Table I. Major coumaroyl-, caffeoyl- and feruloyl-containing metabolites measured by UPLC-TOF-MS in methanol-water leaf extracts of N. attenuata wild-type and irMYB8 plants.

Rosette leaves of wild-type and irMYB8 plants were wounded (W) with a fabric pattern wheel, immediately treated with M. sexta oral secretions (OS), and harvested 24 h later. Elemental formulas and relative mass errors (in ppm) were calculated using Smart Formula from the UPLC-TOF-MS operating software (see “Materials and Methods”). Candidate formulas were ranked according to both mass deviation and isotope pattern accuracy reflected in the σ value. MS/MS+ spectra for some of the reported parent ions have been published by our group (Gaquerel et al., 2010), and the strategy used for compound annotation is summarized into Supplemental Text S1. Asterisks indicate significant differences (unpaired t test, performed on log₂-transformed data) in the relative intensity of hydroxycinnamic acid-containing metabolites between W+OS-elicited wild-type leaves and untreated wild-type controls (24 h; WT W+OS versus WT Ctrl) and between irMYB8 leaves induced by W+OS compared with identically treated wild-type leaves (24 h; irMYB8 W+OS versus WT W+OS): * P < 0.05, ** P < 0.001, *** P < 0.0001. Different fonts indicate either significant increases (boldface) or decreases (italics) due to W+OS elicitation or MYB8 silencing. Numbers in the compound name column refer to the different annotation levels using the four levels of the metabolite annotation nomenclature proposed by the Metabolome Standard Initiative. Indexes after nitrogen atoms indicate that structural rearrangements during in-source or collision-induced dissociation-MS/MS fragmentation did not allow the unequivocal assignment of the phenylpropanoid residues to the N1, N5, or N10 position of spermidine. N.V., Not visible in Figure 1 but presented in Supplemental Figure S4; NS, not significant.

	P (Unpaired t Test)		Fold Change
Label in Figure 1	WT W+OS Versus WT Ctrl	irMYB8 W+OS Versus WT W+OS	WT W+OS > WT Ctrl	irMYB8 W+OS > WT W+OS	Retention Time	Precursor m/z	Ion Type	Elemental Formula	Error	Abbreviated Name	Annotation
					s				ppm
Co1	*	**	3.98	0.00	337	m/z 235.143	[M+H]+	C13H19N2O2+	4.1	CoP	2N -Coumaroylputrescine isomer 1
Co2	***	***	8.48	0.00	410	m/z 235.143	[M+H]+	C13H19N2O2+	4.1	CoP	2N -Coumaroylputrescine  isomer 2
Co3	NS	NS	–	–		–	–	–	–	Unk	4Unknown
Co4	*	**	9.58	2.13	575	m/z 361.097	[M+H]+	C11H21O13+	0.9	Unk	4Unknown
Co5	*	*	1.48	1.71	612	m/z 339.106	[M+H]+	C16H19O8+	1.3	CoQ	3O -Coumaroylquinic acid  isomer 1
Co6	NS	**	1.11	1.87	700	m/z 339.110	[M+H]+	C16H19O8+	2.8	CoQ	3O -Coumaroylquinic acid  isomer 2
Co7	***	***	0.05	0.25	780	m/z 498.261	[M+H]+	C27H36N3O6+	0.8	CoSiS	N′,N″-Coumaroyl, sinapoyl spermidine isomer 1
C1	**	**	2.72	0.01	254	m/z 251.138	[M+H]+	C13H19N2O3+	3.6	CP	1N -Caffeoylputrescine  isomer 1
C2	**	***	5.74	0.00	333	m/z 251.139	[M+H]+	C13H19N2O3+	3.6	CP	1N -Caffeoylputrescine  isomer 2
C3	NS	**	0.90	1.12	373	m/z 401.122	[M+H]+	C13H19N3O7+	0.3	Unk	4Unknown
C4	NS	NS	1.41	0.93	460	m/z 355.103	[M+H]+	C16H19O9+	2.2	CQ	3O -Caffeoylquinic acid  isomer 1
C5	NS	NS	0.98	0.93	490	m/z 355.109	[M+H]+	C16H19O9+	1.2	CGA	13-O -Caffeoylquinic acid  isomer 2 (CGA)
C6	NS	NS	0.96	0.92	517	m/z 355.104	[M+H]+	C16H19O9+	1.4	CQ	3O -Caffeoylquinic acid  isomer 3
C7	*	NS	0.83	0.94	586	m/z 355.104	[M+H]+	C16H19O9+	1.4	CQ	3O -Caffeoylquinic acid  isomer 4
C8 & C9	NS	**	0.80	0.06	612	m/z 470.229	[M+H]+	C25H32N3O6+	2	DCS	2N′,N″-Dicaffeoylspermidine isomers 2 and 3
C10	*	***	1.77	0.08	646	m/z 470.230	[M+H]+	C25H32N3O6+	2	DCS	2N′,N″-Dicaffeoylspermidine  isomer 4
C11	**	**	5.68	0.01	687	m/z 347.196	[M+H]+	C19H27N2O4+	0.3	Unk	3Unknown putrescine metabolite isomer 1
C11	**	**	1.80	0.03	687	m/z 484.244	[M+H]+	C26H34N3O6+	0.3	CFS	2N′,N″-Caffeoyl, feruloyl spermidine isomer 1
C12	**	**	4.22	0.00	731	m/z 347.198	[M+H]+	C19H27N2O4+	3.3	Unk	3Unknown putrescine  metabolite isomer 2
C12	***	**	2.38	0.03	730	m/z 484.245	[M+H]+	C26H34N3O6+	1.2	CFS	2N′,N″-Caffeoyl, feruloyl spermidine isomer 2
C13	***	**	4.88	0.05	751	m/z 484.245	[M+H]+	C26H34N3O6+	1.2	CFS	2N′,N″-Caffeoyl, feruloyl spermidine isomer 3
C14	NS	**	1.07	0.19	899	m/z 566.286	[M+H]+	C31H40N3O7+	0.1	Unk	3Unknown spermidine  metabolite
C15	*	*	2.91	0.13	1,019	m/z 580.300	[M+H]+	C32H42N3O7+	0.3	Unk	3Unknown spermidine  metabolite
F1	***	***	4.38	0.01	263	m/z 322.212	[M+H]+	C17H28N3O3+	0.7	MFS	2N -Feruloylspermidine  isomer 1
F2	***	***	13.92	0.01	338	m/z 322.212	[M+H]+	C17H28N3O3+	0.7	MFS	2N -Feruloylspermidine  isomer 2
F3	**	***	17.51	0.17	388	m/z 265.153	[M+H]+	C14H21N2O3+	4.5	FP	2N -Feruloylputrescine  isomer 1
F4	***	***	25.71	0.19	464	m/z 265.153	[M+H]+	C14H21N2O3+	3.8	FP	2N -Feruloylputrescine  isomer 2
F5	*	NS	1.71	1.22	661	m/z 369.119	[M+H]+	C17H21O9+	0.6	FQ	3O -Feruloylquinic acid  isomer 1
F6	*	NS	1.74	1.31	731	m/z 369.121	[M+H]+	C17H21O9+	3.6	FQ	3O -Feruloylquinic acid  isomer 2
F7	NS	NS	1.93	0.04	823	m/z 498.260	[M+H]+	C27H36N3O6+	0.7	DFS	2N′,N″-Di-feruloyl-spermidine  isomers 1 and 2
F8	**	*	2.43	0.03	847	m/z 498.259	[M+H]+	C27H36N3O6+	0.7	DFS	2N′,N″-Di-feruloyl-spermidine  isomer 3
N.V.	***	**	0.16	0.00	218	m/z 292.202	[M+H]+	C16H26N3O2+	0.4	MCoS	2N -Coumarylspermidine  isomer 1
N.V.	NS	**	0.06	0.00	289	m/z 292.201	[M+H]+	C16H26N3O2+	0.4	MCoS	2N -Coumarylspermidine  isomer 2
N.V.	NS	**	1.20	0.00	175	m/z 308.195	[M+H]+	C16H26N3O3+	0.9	MCS	2N Caffeoylspermidine  isomer 1
N.V.	***	**	11.08	0.00	229	m/z 308.197	[M+H]+	C16H26N3O3+	0.9	MCS	2N Caffeoylspermidine  isomer 2
N.V.	*	**	0.31	0.03	572	m/z 470.229	[M+H]+	C25H32N3O6+	0.9	DCS	2N′,N″-Dicaffeoylspermidine  isomer 1
N.V.	NS	***	0.80	0.00	658	m/z 454.233	[M+H]+	C25H32N3O5+	1.7	CoCS	2N -Coumaryl, caffeoyl spermidine isomer 1
N.V.	NS	**	0.30	0.00	702	m/z 454.233	[M+H]+	C25H32N3O5+	1.7	CoCS	2N -Coumaryl, caffeoyl spermidine isomers 2 and 3
N.V.	*	***	0.10	0.01	725	m/z 454.235	[M+H]+	C25H32N3O5+	2.5	CoCS	2N -Coumaryl, caffeoyl spermidine isomer 4