5-n-Alk(en)ylresorcinol accumulation across developmental stages of triticale through comparative metabolite profiling: implications of fresh and dry basis

Figures & data

Table 1. Developmental stages along life cycle of triticale and test plant parts.

No.	Developmental stage	BBCH Scale^a	Description	DAS^b	Plant parts used
1	First leaf	10	Coleoptile	12	Leaves and roots
2	Two leaves	12	Two leaves unfolded	22	Leaves and roots
3	Flag leaf	39	Flag leaf fully unrolled	40	Leaves and roots
4	Inflorescence	59	Influence fully emerged	71	Leaves, roots spike
5	Late-milky grain	77	Late milk	120	Leaves, roots, spike, and seed
6	Harvest	99	Harvested product	144	Leaves, roots, spike, and seed

^aNomenclature according to the BBCH scale from 00 to 99. ^bDAE: Days after the seed germination.

Table 2. Total AR (TAR) content, total phenolic content (TPC), and DPPH· radical-scavenging capacity of triticale samples at different developmental stages and plant parts from different processed plant materials (i.e. fresh (FW) and dry (DW) processing) and extraction methods (i.e. overnight solvent assisted (OSA) and supercritical fluid (SCF) extraction).

		Fresh weight processing (FW)				Dry weight processing (DW)
Plant parts	PS ^†	Code ^‡	TAR ^§	TPC ^¶	DPPH ^£ IC50	Code ^‡	TAR ^§	TPC ^¶	DPPH ^£ IC50	TAR SCF ^¥
										1	2
Leaf	1	LF1	489.3^A	7844.8^A	237.8	LD1	375.88^A	2264.43^A	544.2	0.35^C	0.38^C
	2	LF2	309.6^C	5044.4^B	229.9	LD2	145.91^E	1713.72^A	697.5	0.01^E	0.10^E
	3	LF3	292.8^CD	2778.6^D	416.8	LD3	83.06^F	716.16^C	767.5	0.56^A	0.37^C
	4	LF4	115.7^F	1586.8^E	334.6	LD4	49.14^FG	451.00^D	858.2	0.41^B	0.42^B
	5	LF5	100.2^G	95.4^J	439.4	LD5	79.19^F	939.94^B	492.4	0.28^D	0.30^D
	6	LF6	80.3^H	559.3^H	806.4	LD6	86.12^F	76.49^E	>1000	0.56^A	0.56^A
Root	1	RF1	57.6^J	720.5^H	637.7	RD1	235.10^D	4600.57^A	>1000	0.20^C	0.71^A
	2	RF2	135.6^F	3634.3^C	494.0	RD2	291.80^C	4088.77^B	>1000	0.17^D	0.24^D
	3	RF3	332.9^C	2794.4^D	860.0	RD3	165.51^E	1069.08^C	>1000	0.17^D	0.18^E
	4	RF4	77.4^H	997.4^F	429.1	RD4	58.73^FG	369.97^E	697.7	0.53^A	0.53^B
	5	RF5	67.3^I	416.1^I	823.2	RD5	68.55^FG	474.48^D	670.7	0.16^D	0.16^E
	6	RF6	62.8^I	117.1^J	485.3	RD6	63.68^FG	136.33^F	>1000	0.46^B	0.48^C
Spike	4	SF4	158.8^E	1490.3^E	463.8	SD4	78.85^F	547.72^A	>1000	0.56^C	0.56^C
	5	SF5	122.0^F	118.9^J	903.3	SD5	156.15^E	514.00^B	856.4	1.09^A	1.13^A
	6	SF6	271.2^CD	62.6^J	>1000	SD6	358.00^B	897.73^B	456.8	0.66^B	0.66^B
Kernel	5	KF5	390.4^B	411.8^I	324.7	KD5	235.65^D	243.76^A	462.0	0.56^A	0.62^B
	6	KF6	76.0^H	844.6^G	255.4	KD6	227.71^D	63.42^B	352.6	(1)^B	0.45^A

^†PS= Developmental stage: 1. First leaf, 2. Two leaves, 3. Flag leaf, 4. Inflorescence, 5. Late-milky grain, and 6. Harvest. ^‡Sample code: the first character means the plant part evaluated: L= Leaf, R= Root, S= Spike, and K= Kernel. The second character means the processing condition: F= Fresh and D= Dry. The third character means the code number for the respective developmental stages (1–6) tested. ^§TAR= Total 5-n-alk(en)ylresorcinol (AR) content expressed as mg Olivetol equivalents per g dry extract (DE) (mg OE/g DM). TPC= Total Phenolic Content in µg gallic acid per equivalents per g dry material (µg GAE/g DM). ^£DPPH· free radical scavenging capacity expressed as half-maximal inhibitory concentration (IC₅₀) in µg/mL. ^¥TARC of DW samples extracted by CO₂ at Supercritical Fluid (SCF) condition using a flow of 10.00 mL/min. SCF pressure/temperature (P/T) conditions: 1. 4000 psi/50°C and 2. 6000 psi/60°C. Different superscript capital letters mean significant differences between samples (p < .05) according to post-hoc Tukey’s test (n = 3).

Figure 1. (a) heatmap of the relative abundance of those AR detected in all samples separating into fresh (FW) and dry (DW) triticale samples. (b) heatmaps of the relative abundance of those AR detected in FW and DW-derived extracts per plant part.

Figure 2. ASCA models relating two factors, i.e. plant material processing prior extraction (fresh weight (FW) and dry weight (DW)) and test developmental stages (i.e. first leaf, two leaves, flag leaf, inflorescence, late-milky grain, and harvest) per plant part: (a) leaves, (b) roots, (c) spikes, and (d) kernels.

Figure 3. MEBA-derived plots of three distinctive AR homologs (C_17:1, C₁₇ and C₂₃) per plant part along developmental stages. DW= dry weight triticale, and FW= fresh weight triticale.

Table 3. Composition and total relative percentage of AR homologs in test kernel samples extracted by SCF and analyzed by GC-MS after silylation-based derivatization.

Sample code^a	AR homolog abundance (%)									TOTAL AR (%)^b
	C15	C17	C19:1	C19	C21:1	C21	C23:1	C23	C25
KF5	0.24	6.76	0	30.8	0.32	31.01	0	8.51	1.97	79.61
KF6	0.24	11.43	1.93	36.56	0.56	38.28	0.21	11.12	1.99	98.40
KD5	0	6.67	0	49.82	0	39.53	0	0	0	96.02
KD6	0	6.33	0	44.95	0	31.73	0	2.1	0	85.11

^aKF5 = fresh kernels at developmental stage 5; KF6 = fresh kernels at developmental stage 6; KD5 = dry kernels at developmental stage 6; KD6 = dry kernels at developmental stage 6; ^bTotal AR as the sum of the AR homolog abundance measured in the GC-MS-derived total ion chromatogram (TIC).

Data availability statement

All data generated or analyzed during this study are included in this published article. The datasets used and analyzed in this study can be found in Supplementary Material.