Modulation of cannabis flower characteristics and THC through a biostimulant complex of molasses, Aloe vera extract, and fish-hydrolysate

Wise, Kimber; Simovich, Tomer; Gill, Harsharn; Selby-Pham, Jamie

doi:10.2478/fhort-2025-0022

Figures & Tables

Impact of BC to cannabis flower odour properties. Sensory assessment of cannabis flower odour strength, desirability, and quality by 14 untrained panellists (3 flowers per treatment), measured on a 5-point Likert scale. Data presented as average ± standard deviations, with marginal significance indicated as ‡ for p < 0.1, as determined by paired samples t-tests. BC, biostimulant complex.

Impact of BC to cannabis flower OD strengths. Sensory assessment of cannabis flower OD strengths by 14 untrained panellists (3 flowers per treatment), measured on a 5-point Likert scale. Data presented as average ± standard deviations, with statistical significance indicated as * for p < 0.05, as determined by paired samples t-tests. BC, biostimulant complex; OD, odour descriptor.

Changes to cannabis sugar leaf metabolites from growth with a BC. PCA of sugar leaf metabolite profiles from three biological replicates, with colours indicating treatment 95% confidence regions. BC, biostimulant complex; PCA, principal component analysis.

Assessment of BC impact to cannabis flower N-IR spectra. Average control (blue) and BC (magenta) N-IR spectra of cured cannabis flowers analysed between 4000 cm−1 and 12000 cm−1 (n = 9). Shading indicates wavenumbers with statistical differences, with dark shading representing statistical significance p < 0.05 and light shading representing marginal significance of p < 0.1, as determined by an independent samples t-test. BC, biostimulant complex; N-IR, near-infrared.

Change to cannabis root biochemistry from growth with a BC. PCA of (A) root metabolite profiles and (B) root phytohormone profiles. Average root concentrations ± standard deviation of (C) GA4, (D) Methyl-IAA, and (E) SA, and (F) root chitinase activity, following 1 week of control or BC treatments. All data based on three biological replicates for control and BC, with significant differences indicated as ** for p < 0.01 and marginal significance indicated as ‡ for p < 0.1, as determined by an independent samples t-test. BC, biostimulant complex; GA4, gibberellin A4; IAA, indole-3-acetic acid; Methyl-IAA, methyl indole-3-acetic acid; PCA, principal component analysis; SA, salicylic acid.

Figure S1.

HS profile PCA. HS, head space; PCA, principal component analysis.

Human OR affinity model (model 1). Polynomial regression analysis with 10-fold cross-validation to predict OR affinity from physicochemical measures (n = 83). OR, odorant receptor.

PCA of sensory data (incudes strength, desirability, quality, and all OD strength measures). OD, odour descriptor; PCA, principal component analysis.

Association between observed changed to volatile and OD strength within cannabis flowers. BC, biostimulant complex; OD, odour descriptor.

Elimination half-life (T½) model (model 3). Polynomial regression analysis with 10-fold cross-validation to predict T½ from physicochemical measures (n = 851).

BBB index model (model 4). Polynomial regression analysis with 10-fold cross-validation to predict BBB index from physicochemical measures (n = 83). BBB, blood brain barrier.

sPLS-DA of N-IR spectral bins (bin width = 100 wavelengths). BC, biostimulant complex; N-IR, near infrared; sPLS-DA, sparse partial least squares discriminant analysis.

Reviewer proportions for being helpful for anxiety (model 5). Polynomial regression analysis with 10-fold cross-validation to predict reviewer proportions for being helpful for anxiety use from terpene and THC measures (n = 93). THC, tetrahydrocannabinol.

Reviewer proportions for being helpful for depression (model 6). Polynomial regression analysis with 10-fold cross-validation to predict reviewer proportions for being helpful for depression use from terpene and THC measures (n = 59). THC, tetrahydrocannabinol.

PCA of leaf metabolite profiles. BC, biostimulant complex; PCA, principal component analysis.

PCA of leaf phytohormone profiles. BC, biostimulant complex; PCA, principal component analysis.

PCA of sugar leaf phytohormone profiles. BC, biostimulant complex; PCA, principal component analysis.

Root phytohormone profiles_

Phytohormone	N	Control (ng · g⁻¹ FW)		BC (ng · g⁻¹ FW)		p-value
Phytohormone	N	Mean	SD	Mean	SD	p-value
12-Oxo-phytodienoic acid (OPDA)	3	1591.727	359.115	1606.366	321.241	0.961
Abscisic acid (ABA)	3	23.060	3.139	18.464	3.173	0.149
Brassinolide	3	0.000	−	0.000	−	−
Cinnamic acid (CA)	3	0.000	0.000	14.377	13.219	0.133
Gibberellin A3 (GA3)	3	0.000	−	0.000	−	−
Gibberellin A4 (GA4)	3	11.651	0.436	10.061	0.973	0.061
Indole-3-acetic acid (IAA)	3	33.050	5.401	37.717	4.542	0.316
Indole-3-butyric acid (IBA)	3	0.000	−	0.000	−	−
Indole-3-carboxylic acid (ICA)	3	0.000	0.000	3.094	2.705	0.186 ^a
Jasmonic acid (JA)	3	0.855	0.499	1.438	0.690	0.301
Jasmonic acid-isoleucine (JA-Ile)	3	0.000	−	0.000	−	−
Methyl-indole-3-acetic acid (Methyl-IAA)	3	5.453	1.616	15.399	1.172	< 0.001
Methyl-jasmonic acid (Methyl-JA)	3	0.000	−	0.000	−	−
Salicylic acid (SA)	3	90.327	16.307	317.285	29.904	< 0.001
Zeatin	3	0.000	−	0.000	−	−

Predicted change to reviewer proportions for flowers grown with BC_

Use	Predicted proportion that find it helpful (%)		Fold-change^a	p-value
Use	Control	BC	Fold-change^a	p-value
Anxiety	27	25	0.91	0.069
Depression	41	46	1.10	0.031

Flower odour measures_

Measure	N (participants)	Control		BC		Mean paired difference	p-value
Measure	N (participants)	Mean	SD	Mean	SD	Mean paired difference	p-value
Desirability	14	0.238	0.646	0.143	0.428	−0.095	0.547
Quality	14	0.095	0.546	−0.167	0.387	−0.262	0.103
Strength	14	2.095	0.659	1.595	0.526	−0.500	0.007

Flower volatile comparison from BC treatment_

Peak ID	N	Fold-change ^a	p-value	OR affinity (kcal · mol⁻¹)
Linalool	9	0.74	0.011	−5.1 ^d
2-Norbornanol	9	0.85	0.078 ^b	−5.0 ^c
Borneol	9	0.86	0.092 ^b	−5.5 ^c
p-Cymen-8-ol	9	0.70	0.005	−5.2 ^c
Terpineol	9	0.73	0.012	−5.3 ^c
Ylangene	9	0.69	0.067 ^b	−7.0 ^d
β-Caryophyllene	9	0.70	0.059 ^b	−7.0 ^d
α-Bergamotene	9	0.68	0.064	−5.7 ^c
α-Humulene	9	0.66	0.052 ^b	−6.6 ^c
β-Eudesmene	9	0.64	0.064	−6.3 ^b

Coefficients for prediction of T½ (model 3)_

Term	Coefficient	p-value
Constant	−14.4006229265679497	0.281
Number_of_aliphatic_OH_groups	1.1590455637133770	0.175
Fraction of rotatable bonds	−1.3092721739145663	0.600
Number_of_heterocycles	−1.3096488245410460	0.038
MolarRefractivity	−0.2057843152416829	0.001
Number_of_atoms	0.7292247559072755	0.188
Number_of_bonds	−13.3089067550692555	<0.001
Geometrical radius	−43.7699892535148081	0.074
Zagreb_group_index_1	3.1733985192039618	<0.001
Zagreb_group_index_2	−0.5573206401475080	<0.001
abonds	2.3470943597991503	<0.001
logP_1	−8.4327332210325956	0.004
R2NH	0.4248298509055615	0.189
ROH	−3.0859362430839505	<0.001
RCOR	−51.5963515226930625	<0.001
RCCH	−7.6481737638213767	0.146
Number_of_heterocycles²	−0.2198849622511800	0.252
Geometrical radius²	144.6809633479945774	<0.001
Zagreb_group_index_1²	−0.0002790970137844	<0.001
logP_1²	0.4533710831155871	<0.001
ROH²	0.1898899908268611	0.001
RCOR²	−11.4515975383076043	<0.001
Number of aliphatic OH groups × Fraction of	−9.1291453725480132	0.006
rotatable_bonds
Number_of_aliphatic_OH_groups × Number_of_	1.6012391295871116	<0.001
heterocycles
Number_of_aliphatic_OH_groups × MolarRefractivity	−0.1252444892292043	<0.001
Number_of_aliphatic_OH_groups × Number_of_atoms	0.7474311264807053	< 0.001
Number_of_aliphatic_OH_groups × Zagreb_group_	−0.1230566705429028	<0.001
index_1
Number_of_aliphatic_OH_groups × Zagreb_group_	0.0383547857193748	0.002
index_2
Number_of_aliphatic_OH_groups × abonds	0.0620697739663554	0.081
Number_of_aliphatic_OH_groups × RCOR	1.3592198025075215	0.095
Fraction_of_rotatable_bonds × abonds	−0.3569886783521736	0.188
Fraction_of_rotatable_bonds × ROH	10.8456620807097082	0.001
Fraction_of_rotatable_bonds × RCOR	−11.3502805681771850	0.021
Number_of_heterocycles × Number_of_atoms	−0.2323530073416142	0.029
Number_of_heterocycles × Zagreb_group_index_1	0.1118053774742066	<0.001
Number_of_heterocycles × Zagreb_group_index_2	−0.0598852303340916	<0.001
Number_of_heterocycles × R2NH	−0.1118543997938758	0.368
Number_of_heterocycles × RCOR	4.2346377985582464	<0.001
Number_of_heterocycles × RCCH	15.0866738457708820	0.017
MolarRefractivity × Zagreb_group_index_2	0.0006001834083347	<0.001
MolarRefractivity × logP_l	−0.0476370535331036	0.008
Number of atoms × abonds	−0.0527014722648722	0.058
Number_of_atoms × logP_1	−0.3406604375175311	< 0.001
Number_of_atoms × RCOR	−0.8512257161355755	0.253
Number of bonds × Geometrical radius	12.3554842409376313	< 0.001
Number_of_bonds × abonds	0.0375992769713121	0.234
Number_of_bonds × logP_1	1.0928914576770774	<0.001
Number_of_bonds × RCOR	2.2709827042367157	0.022
Geometrical_radius × Zagreb_group_index_1	−3.0687885400353694	<0.001
Geometrical_radius × Zagreb_group_index_2	0.5826849894258733	<0.001
Geometrical_radius × abonds	−2.0569426785700724	<0.001
Geometrical_radius × logP_1	9.2947190000174178	0.001
Geometrical_radius × RCOR	63.0601826659657618	<0.001
Zagreb_group_index_1 × logP_1	−0.1309816278880524	<0.001
Zagreb_group_index_1 × RCOR	−0.2487231221576796	0.001
Zagreb_group_index_2 × abonds	0.0024055714971864	0.087
Zagreb_group_index_2 × ROH	−0.0081751060510380	<0.001
abonds × logP_1	−0.1538070350082036	<0.001
abonds × R2NH	−0.0306955363982863	0.053
abonds × RCOR	0.4936988943195205	< 0.001
abonds × RCCH	0.8979972845110127	0.003
logP_1 × ROH	0.5768623154995151	<0.001
logP_1 × RCOR	15.0975212136558934	<0.001
logP_1 × RCCH	0.0846813351199347	0.941
R2NH × RCOR	0.0443611523935290	0.949
ROH × RCOR	−0.9626923122968017	0.124
ROH × RCCH	3.5309731402574753	0.029
RCOR × RCCH	−80.7917122680247957	<0.001
Number_of_heterocycles³	0.0575198050118327	0.004
Geometrical radius³	−82.2909815734712282	<0.001
RCOR³	3.3631450065824469	<0.001
Number_of_aliphatic_OH_groups × Fraction_of_ rotatable_bonds × abonds	−0.3063861434585855	0.026
Number_of_aliphatic_OH_groups × Number_of_ heterocycles²	0.2214888423749514	<0.001
Number_of_aliphatic_OH_groups × Number_of_ heterocycles × Zagreb_group_index_2	0.0004197810162712	0.070
Number_of_aliphatic_OH_groups × Number_of_ atoms × abonds	0.0003475317581468	0.056
Number_of_aliphatic_OH_groups × abonds × RCOR	−0.1476876886014743	0.012
Number_of_heterocycles² × RCCH	8.4897847228558074	0.005
Number_of_heterocycles × Number_of_atoms × RCOR	−0.8844730128826180	<0.001
Number_of_heterocycles × Zagreb_group_index_1²	−0.0000016028999970	0.005
Number_of_heterocycles × Zagreb_group_ index_1 × RCOR	0.1669210843082060	< 0.001
Number_of_heterocycles × R2NH × RCOR	−0.3807557871833461	0.049
Number_of_bonds × abonds × logP_1	0.0004754092355291	<0.001
Geometrical_radius × Zagreb_group_index_1 × logP_1	0.0049254113703736	0.007
Geometrical_radius × logP_1 × RCOR	−19.1616364971431494	<0.001
abonds × logP_1²	−0.0130411905685735	<0.001
logP_1² × ROH	−0.0186812881717686	0.007
logP_1² × RCOR	0.2034621762117447	<0.001
logP_1 × RCOR × RCCH	25.0921469271286917	<0.001

Root bioassay measures_

Measure	N	Control (ng · g ¹ FW)		BC (ng · g⁻¹ FW)		p-value
Measure	N	Mean	SD	Mean	SD	p-value
Protein content (mg total protein · g⁻¹ FW)	3	6.689	0.136	6.739	0.705	0.052
Hydrogen peroxide content (μmol H₂O₂ · g⁻¹ FW)	3	9.205	3.727	8.987	1.369	0.184
Peroxidase activity (△Abs₄₇₀ · min⁻¹ · mg⁻¹ total protein)	3	0.000	0.000	0.027	0.047	0.016 ^a
Chitinase activity (μmol GlcNAc released · g⁻¹ total protein)	3	631.444	64.877	710.774	116.034	0.311

Comparison of metabolites in sugar leaves that were changed with significance or marginal significance_

Metabolite	Synonym	N	Fold-change ^a	p-value
(1S,2R)-1-(3,4-Dihydroxyphenyl)-6,7-dihydroxy-N,N’-bis[2-(4-hydroxyphenyl)ethyl]-1,2-dihydro-2,3-naphthalenedicarboxamide (1ξ)-1,5-Anhydro-1-[2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4-oxo-4H-	Cannabisin B	3	0.520	0.006
chromen-8-yl] -D-galactitol	Orientin	3	0.848	0.091
1,5-Anhydro-1-[2-(3,4-dihydroxyphenyl)-5-hydroxy-7-methoxy-4-oxo-4H-chromen-8-yl]hexitol		3	0.753	0.006
1-deoxy-D-altro-heptulo se 7-pho sphate		3	0.801	0.037
1-Hydroxy-3,5,6-trimethoxy-10-methyl-9(10H)-acridinone		3	0.639	0.003
2,3,4,5-Tetrahydroxypentanal		3	0.779	0.040
2-C-methylerythritol 4-phosphate		3	0.632	0.050
3-[3-(beta-D-Glucopyranosyloxy)-2-methoxyphenyl]propanoic acid		3	0.827	0.057
4-(2-Carboxyethyl)-2-methoxyphenyl beta-D-glucopyranosiduronic acid		3	0.728	0.073
4-Hydroxy-5-(3’,4’,5’-trihydroxyphenyl)-valeric acid-O-methyl-O-glucuronide		3	0.835	0.062
4-Oxoproline		3	0.469	0.061
5,6-Dihydroxy-3-(hydroxymethyl)-2-oxo-3-cyclohexen-1-yl 2-hydroxy-6-methylbenzoate		3	0.721	0.076
8-Hydroxy-4-methoxy-7-methyl-7,8-dihydro-5H-furo[2,3-g]isochromen-5-one		3	0.725	0.081
Apigetrin		3	0.755	0.015
Azelaic acid		3	0.518	0.017
Cynaroside		3	0.771	0.016
D-α-Hydroxyglutaric acid		3	0.788	0.082
Gluconic acid		3	0.718	0.002
Hept-2-ulose		3	0.775	0.041
Hinokitiol glucoside		3	0.809	0.018
Luteolin 7-O-malonylglucoside		3	0.828	0.089
N-(2,3,4-Trimethoxybenzoyl)g lycine		3	0.738	0.053
Pantothenic acid		3	0.746	0.076
Salicylic acid		3	0.604	0.094
trans-p-Sinapoyl-Î²-D-glucopyranoside		3	0.814	0.061

Nutrient composition of BC_

Nutrient	BC% w/v
Nitrogen (N) ^a	0.260
Phosphorous (P) ^b	0.045
Potassium (K) ^b	1.023
Calcium (Ca) ^b	0.059
Magnesium (Mg) ^b	0.213
Sulphur (S) ^a	0.440
Silicon dioxide (SiO₂) ^b	0.021
Iron (Fe) ^b	<0.005
Zinc (Zn) ^b	<0.005
Manganese (Mn) ^b	<0.005
Boron (B) ^b	<0.005

Growth and yield measures_

Measure	N	Control		BC		p-value
Measure	N	Mean	SD	Mean	SD	p-value
Height (mm)	8	1164.88	19.86	1148.75	63.26	0.510 ^a
Stem thickness (mm)	8	12.74	1.04	12.88	1.55	0.839
Per-plant flower yield (FW, g)	8	172.78	23.69	166.45	27.73	0.631
FW flower water%	40	72.58	2.60	72.54	2.98	0.947

Comparison of metabolites in leaves that were changed with significance or marginal significance_

Metabolite	N	Fold-change ^a	p-value
(1)-1,5-Anhydro-1-[2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4-oxo-4H-chromen-8-yl]-	3	2.666	0.081 ^a
D-galactitol
(2E)-3-(4-Hydroxy-3,5-dimethoxyphenyl)acrylic (2S,3S,4S,5R)-3,4,5-trihydroxy-	3	2.286	0.063
6-oxotetrahydro-2H-pyran-2-carboxylic anhydride (non-preferred name)
(Z)-Aconitic acid	3	2.626	0.099
13(S)-HOTrE	3	4.080	0.070
1-Caffeoyl-beta-D-glucose	3	3.603	0.012
1-heptadecanoyl-sn-glycero-3-phosphate	3	3.102	0.072
1-O-[(2E)-3-(3,4-Dihydroxyphenyl)-2-propenoyl]-beta-D-glucopyranuronic acid	3	2.785	0.089
2-(2-Carboxyethyl)-4-methyl-5-pentyl-3-furoic acid	3	2.154	0.027
2-(2-Carboxyethyl)-5-hydroxyphenyl hexopyranosiduronic acid	3	3.400	0.014
2-(6,10-Dimethyl-8-oxospiro[4.5]dec-6-en-2-yl)-2-hydroxypropyl hexopyranoside	3	3.752	0.057
3-(Hydroxymethyl)-1-oxo-1H-isochromen-6-yl hexopyranosiduronic acid	3	2.473	0.044
3-Hydroxy-2-{[(2E)-3-(4-hydroxyphenyl)-2-propenoyl]oxy}propanoic acid	3	2.977	0.037
4-(2-Carboxyethyl)-2-methoxyphenyl beta-D-glucopyranosiduronic acid	3	3.842	0.077
4-Oxoproline	3	3.186	0.098
5 – [(5-Hydroxytetrahydro-2-furanyl)methyl]-1,3-benzenediol	3	3.206	0.070
5-[(E)-2-Carboxyvinyl]-2-hydroxyphenyl beta-D-glucopyranosiduronic acid	3	2.836	0.084 ^a
6-Methoxy-2-oxo-3,4-dihydro-2H-chromen-7-yl beta-D-glucopyranoside	3	2.521	0.077
butyrin	3	3.968	0.031
Caffeic acid 3-glucoside	3	2.860	0.068
Cannabigerolic acid	3	2.502	0.065
Corchorifatty acid F	3	5.463	0.084
D-(-)-Fructose	3	2.505	0.026
Dimethyl adipate	3	2.459	0.076
Genistein	3	3.013	0.050
L-Glutamic acid	3	3.646	0.079
Luteolin	3	3.122	0.089
Luteolin 7-O-malonylglucoside	3	2.901	0.063
Methyl 4,5,7,11-tetrahydroxy-2-methyl-2’,10-dioxospiro[9-oxatricyclo[6.3.1.0~1,5~]	3	2.924	0.020
dodecane-6,3’-oxetane]-7-carboxylate
N-feruloylglycine	3	4.763	0.065
p-Coumaric acid glucoside	3	2.305	0.045

OD strengths_

Measure	N (participants)	Control		BC		Paired difference ^a	p-value
Measure	N (participants)	Mean	SD	Mean	SD	Paired difference ^a	p-value
Apple	14	0.024	0.089	0.000	0.000	−0.024	0.323 ^b
Balsamic	14	0.000	0.000	0.000	0.000	−	−
Berry	14	0.000	0.000	0.024	0.089	0.024	0.323 ^b
Butter	14	0.000	0.000	0.000	0.000	−	−
Cheese	14	0.000	0.000	0.000	0.000	−	−
Citrus	14	0.095	0.242	0.167	0.314	0.071	0.503
Clove	14	0.143	0.386	0.071	0.142	−0.071	0.429
Coffee	14	0.095	0.356	0.095	0.356	0.000	1.000
Earthy	14	0.857	1.130	0.786	0.843	−0.071	0.770
Eucalyptus	14	0.500	0.748	0.262	0.437	−0.238	0.152 ^b
Floral	14	0.619	0.856	0.429	0.530	−0.190	0.348
Fruity	14	0.191	0.284	0.024	0.089	−0.167	0.046 ^b
Grape	14	0.000	0.000	0.000	0.000	−	−
Grassy	14	0.738	1.039	0.929	1.095	0.190	0.474
Herbal	14	1.167	1.189	1.119	1.059	−0.048	0.864
Minty	14	0.691	0.947	0.262	0.325	−0.429	0.029 ^b
Nutty	14	0.000	0.000	0.024	0.089	0.024	0.323 ^b
Peach	14	0.000	0.000	0.000	0.000	−	−
Peppery	14	0.191	0.363	0.095	0.275	−0.095	0.327
Petrol	14	0.000	0.000	0.000	0.000	−	−
Pine	14	0.238	0.479	0.095	0.356	−0.143	0.242 ^b
Pineapple	14	0.024	0.089	0.000	0.000	−0.024	0.323 ^b
Rose	14	0.095	0.275	0.048	0.121	−0.048	0.473
Spicy	14	0.310	1.066	0.452	0.966	0.143	0.537
Sweet	14	0.095	0.204	0.095	0.242	0.000	1.000
Tea	14	0.548	0.517	0.643	0.790	0.095	0.671
Tobacco	14	0.262	0.456	0.405	0.669	0.143	0.386
Tropical fruit	14	0.000	0.000	0.000	0.000	−	−
Vanilla	14	0.024	0.089	0.000	0.000	−0.024	0.323 ^b
Woody	14	0.619	0.885	0.714	0.678	0.095	0.682

Comparison of metabolites in roots that were changed with significance or marginal significance_

Metabolite	N	Fold-change ^a	p-value
(2alpha, 3xi, 5alpha, 9alpha, 10beta, 13alpha)-2-Hydroxytaxa-4(20),	3	0.475	0.015
11-diene-5,9,10,13-tetrayl tetraacetate
(2R)-1-{[(2-Aminoethoxy)(hydroxy)phosphoryl]oxy}-3-hydroxy-2-propanyl	3	0.290	0.007
(9Z,12Z,15Z)-9,12,15-octadecatrienoate
(2R)-2-Hydroxy-3-(phosphonooxy)propyl (11Z,14E)-11,14-icosadienoate	3	0.323	0.083
1-(3-Acetyl-2,4,6-trihydroxyphenyl)-1,5-anhydrohexitol	3	12.688	0.068 ^a
1-(3-Cyanophenyl)-3-{[(2R,4S,5S)-5-{[4-(4-fluorophenyl)-1-piperazinyl]methyl}-	3	0.283	0.006
1-azabicyclo[2.2.2]oct-2-yl]methyl}urea
1,5-Anhydro-D-fructo se	3	0.778	0.026
1-Caffeoyl-beta-D-glucose	3	1.422	0.065
1-heptadecanoyl-sn-glycero-3-phosphate	3	0.157	<0.001
1-linoleoyl-sn-glycero-3-phosphoethanolamine	3	0.269	0.028 ^a
1-Oleoyl-2-hydroxy-sn-glycero-3-PE	3	0.328	0.021
2,3,4,5-Tetrahydroxypentanal	3	0.661	0.036
2-[4-(Hydroxymethyl)cyclohexyl]-2-propanyl hexopyranoside	3	0.561	0.029
2-Phenylethyl D-glucopyranoside	3	0.333	0.007
3-(Hydroxymethyl)-1-oxo-1H-isochromen-6-yl hexopyranosiduronic acid	3	0.666	0.067
3,3’-Diisopropyl-6,6’-dimethyl-2,2’,5,5’-biphenyltetrol	3	0.230	0.016
4-(2-Carboxyethyl)-2-methoxyphenyl beta-D-glucopyranosiduronic acid	3	0.428	0.057 ^a
4,7-Dihydroxy-2,2-dimethyl-3,4-dihydro-2H-chromen-5-yl hexopyranosiduronic acid	3	1.927	0.015
4-Hydroxy-5-(3’,4’,5’-trihydroxyphenyl)-valeric acid-O-methyl-O-glucuronide	3	0.617	0.034
4-Undecylbenzenesulfonic acid	3	0.430	0.079 ^a
5,5’-Diisopropyl-2,2’-dimethyl-3,3’,4,4’-biphenyltetrol	3	0.199	0.029
5,6-Dihydroxy-2,6-bis(3-methyl-2-buten-1-yl)-4-(4-methylpentanoyl)-4-cyclohexene-1,	3	0.324	0.045
3-dione
5 – [(5-Hydroxytetrahydro-2-furanyl)methyl]-1,3-benzenediol	3	0.538	0.055
6-Hydroxy-3,4a,5-trimethyl-2-oxo-2,4,4a,5,6,7,8,8a,9,9a-decahydronaphtho[2,3-b]	3	4.263	0.004
furan-4-yl (2Z)-2-methyl-2-butenoate
6-Oxoestriol	3	0.321	0.007
7-Hydroxy-2-oxo-2H-chromen-8-yl beta-D-glucopyranoside	3	0.481	0.011
8-Azabicyclo[3.2.1]octane-1,2,4,7-tetrol	3	0.636	0.081
beta-D-Ethyl glucuronide	3	0.799	0.035
Caffeic acid 3-glucoside	3	0.409	0.050
Cannabidiolic acid	3	0.309	0.040
Cannabigerolic acid	3	0.369	0.045
D-(-)-Ribose	3	0.731	0.049
Hept-2-ulose	3	0.661	0.037
Hinokitiol glucoside	3	0.648	0.020
Hydrocortisone acetate	3	0.455	0.055
Jasmolin II	3	0.203	0.050
L-Glutathione oxidized	3	0.694	0.055
Marinobufagenin	3	0.515	0.083
Methyl (17beta)-3,6-dimethoxyestra-1,3,5(10),6,8-pentaene-17-carboxylate	3	0.260	0.008
Methyl 4,5,7,11-tetrahydroxy-2-methyl-2’,10-dioxospiro[9-oxatricyclo[6.3.1.0~1,5~]	3	2.759	0.008
dodecane-6,3’-oxetane]-7-carboxylate
N-(2,3,4-Trimethoxybenzoyl)g lycine	3	0.796	0.087
N-(Phenylacetyl)aspartic acid	3	4.426	0.001
N-Benzoylaspartic acid	3	2.184	0.016
Nor-9-carboxy-δ9-THC	3	0.290	0.058
Rutin	3	0.386	0.047
Salicylic acid	3	4.900	0.010
Sibrafiban	3	0.571	0.001
Sucrose	3	0.648	0.063

Leaf phytohormone profiles_

Phytohormone	N	Control (ng · g⁻¹ FW)		BC (ng · g⁻¹ FW)		p-value
Phytohormone	N	Mean	SD	Mean	SD	p-value
12-Oxo-phytodienoic acid (OPDA)	3	3419.789	704.776	3701.079	639.307	0.636
Abscisic acid (ABA)	3	411.945	111.064	521.110	78.443	0.237
Brassinolide	3	0.000	−	0.000	−	−
Cinnamic acid (CA)	3	0.000	−	0.000	−	−
Gibberellin A3 (GA3)	3	0.000	−	0.000	−	−
Gibberellin A4 (GA4)	3	0.000	−	0.000	−	−
Indole-3-acetic acid (IAA)	3	10.418	0.995	11.331	0.085	0.252 ^a
Indole-3-butyric acid (IBA)	3	0.000	−	0.000	−	−
Indole-3-carboxylic acid (ICA)	3	1.630	2.823	3.533	3.070	0.473
Jasmonic acid (JA)	3	5.872	2.914	8.948	1.454	0.177
Jasmonic acid-isoleucine (JA-Ile)	3	6.709	6.086	15.753	12.871	0.333
Methyl-indole-3-acetic acid (Methyl-IAA)	3	0.000	−	0.000	−	−
Methyl-jasmonic acid (Methyl-JA)	3	16.757	7.856	9.279	5.344	0.244
Salicylic acid (SA)	3	103.891	17.961	124.315	38.016	0.447
Zeatin	3	9.823	1.625	10.464	1.838	0.674

Pharmacokinetic properties of flower volatiles altered by growth with BC_

Flower volatile	Fold-change^a	p-value	T_½^b (hr)	BBB index
Linalool	0.74	0.011	16.8	8.3^c
2-Norbornanol	0.85	0.078	9.4	0.0^d
Borneol	0.86	0.092	15.1	3.0^d
p-Cymen-8-ol	0.70	0.005	9.2	4.1^d
Terpineol	0.73	0.012	15.5	5.0^d
Ylangene	0.69	0.067	38.4	11.1^c
β-Caryophyllene	0.70	0.059	42.0	13.3^c
α-Bergamotene	0.68	0.064	40.8	8.7^d
α-Humulene	0.66	0.052	43.9	14.2^c
β-Eudesmene	0.64	0.064	42.6	12.9^d

Coefficients for prediction of being helpful for anxiety (model 5)_

Term	Coefficient	p-value
Constant	0.3493779002367680	<0.001
α-Humulene	0.8798417229601690	0.048
β-Limonene	−0.1622691586687510	<0.001
β-Myrcene	−0.1076790958648940	0.030
β-Ocimene	−0.2517684486903870	0.006
Camphene	0.7923380032907140	0.252
γ-Terpinene	−34.1130191417850000	<0.001
Terpinolene	0.1012123365266380	0.088
trans-Nerolidol	−14.4253428834657000	0.035
THC	−0.3715078664606990	0.095
α-Humelene²	−1.4018939392389500	0.043
β-Myrcene²	0.1857526868455400	<0.001
γ-Terpinene²	375.4539239428840000	0.002
trans-Nerolidol²	794.7242808869300000	0.027
α-Humulene × β-Limonene	0.7790401367405530	0.020
α-Humulene × THC	−3.5237601156201400	0.074
β-Myrcene × trans-Nerolidol	63.3048932161677000	0.098
β-Ocimene × THC	0.8707362097038800	0.034
γ-Terpinene × THC	113.5074239826430000	0.014
β-Myrcene × trans-Nerolidol²	−2718.9714110588700000	0.163

Sensory analysis: participant demographics_

Demographic	Frequency	Percentage (%)
Age (years)
19–28	5	36
29–38	5	36
39–48	2	14
>49	2	14
Ethnicity
White or Caucasian	6	43
Asian or Pacific Islander	8	57
Gender
Male	7	50
Female	7	50

ODs associated with altered HS volatiles_

Compound	ODs ^a
2-Norbornanol	Leafy, citrus, herbal, green
Borneol	Balsamic, camphor, earthy, musty, herbal, minty, mouldy, pepper, pine, woody
Linalool	Blueberry, bois de rose, citrus, floral, green, lavender, lemon, orange, rose, sweet, terpenic, waxy, woody
p-Cymen-8-ol	Camphor, cherry, citrus, cool, earthy, musty, floral, fruity, musty, mouldy, sweet, walnut
Terpineol	Anisic, citrus, cool, floral, lemon, lilac, limonene, minty, oily, pine, resin, terpenic, woody
Ylangene	Woody, honey, spices
α-Bergamotene	Woody, tea, warm
α-Humulene	Woody, spices
β-Caryophyllene	Woody, clove, dry, spices, sweet
β-Eudesmene	Herbal

Changes in peak area for HS identified compounds_

Peak ID	N	Fold-change a	p-value
2-Norbornanol	9	0.85	0.078 ^b
2-Pinanol	9	1.03	0.395
Borneol	9	0.86	0.092 ^b
Camphene	9	1.12	0.276
Dehydrosabinene	9	1.16	0.112
delta-3-Carene	9	0.92	0.336
Eucalyptol	9	0.92	0.241
Fenchol	9	0.92	0.164 ^b
Ipsdienol	9	0.80	0.191
Limonene	9	0.96	0.401
Linalool	9	0.74	0.011
Ocimene	9	1.00	0.490
p-Cymen-8-ol	9	0.70	0.005
p-Cymene	9	0.87	0.119
Terpineol	9	0.73	0.012
Terpinolene	9	0.90	0.213
Ylangene	9	0.69	0.067 ^b
α-Bergamotene	9	0.68	0.064
α-Humulene	9	0.66	0.052 ^b
α-Pinene	9	1.11	0.216
α-Terpinene	9	0.91	0.286
β-Caryophyllene	9	0.70	0.059 ^b
β-Eudesmene	9	0.64	0.064
β-Myrcene	9	0.85	0.117
β-Ocimene	9	1.06	0.378
β-Pinene	9	1.08	0.313
γ-Terpinene	9	0.91	0.223

Coefficients for prediction of being helpful for depression (model 6)_

Term	Coefficient	p-value
Constant	0.3532528057435070	<0.001
THC	−0.4747423305432570	0.285
β-Limonene	−0.1008144474280880	0.013
Eucalyptol	−9.4195048772503600	0.023
Linalool	2.3861320207786800	0.127
β-Ocimene	0.0958394623976688	0.125
trans-Nerolidol	32.3303525233572000	0.001
Linalool²	−21.7804178628571000	0.001
THC × Linalool	−15.3357472064389000	0.047
THC × trans-Nerolidol	−134.9349802361570000	0.003
THC × Linalool²	110.8828051662290000	0.001

Coefficients for prediction of BBB index (model 4)_

Term	Coefficient	p-value
Constant	69.9850536035912000	0.077
LogP	16.3556360234803000	0.001
MW	−0.3131349429968060	0.738
Nrot	−25.6219150613349000	0.119
Vol	−1.5903781872639000	<0.001
MW²	0.0084485573872430	0.098
Nrot²	−1.2847296133815200	0.063
Vol²	0.0051074771946313	<0.001
LogP × Nrot	−9.7577696510109300	0.002
LogP × Vol	−0.0726649622079701	0.004
MW × Nrot	−0.0474921788682433	0.900
Nrot × Vol	0.8098155078732790	0.002
MW³	−0.0000290308583602	0.003
Nrot³	0.2621598197638000	0.094
LogP × Nrot × Vol	0.0441943553567649	0.007
MW² × Nrot	−0.0027881504869329	0.146
Nrot × Vol²	−0.0023883590260709	0.003
Nrot⁴	−0.0140373602293692	0.193
MW³ × Nrot	0.0000105403591019	0.006

CBD and THC contents_

Measure	N	Control		BC		p-value
Measure	N	Mean	SD	Mean	SD	p-value
Flower CW (g)	9	1.02	0.18	0.94	0.17	0.355
Flower DW (g)	9	0.88	0.15	0.81	0.15	0.359
Cured flower water (%)	9	13.90	0.53	13.96	0.83	0.865
CBD (%)	9	6.05	0.87	6.44	1.18	0.434
THC (%)	9	0.32	0.42	0.99	0.63	0.016

LR of odour strength from HS – model 2 variables_

Variable	Coefficient	p-value
Constant (1)	−57.1997	0.001
Constant (2)	−51.3960	0.003
Constant (3)	−49.5906	0.003
Constant (4)	−45.8615	0.007
2-Pinanol	−0.00011	0.005
Camphene	−2.2E-06	0.041
Dehydrosabinene	−1.8E-05	0.004
Fenchol	−4.6E-05	0.011
Ipsdienol	7.92E-05	0.022
p-Cymen-8-ol	6.52E-05	0.004
α-Humulene	0.000114	0.003
α-Pinene	5.03E-07	0.002
α-Terpinene	−4.7E-06	0.008
β-Caryophyllene	−4.7E-05	0.001
β-Eudesmene	−6.3E-05	0.047
γ-Terpinene	5.82E-06	0.044

Coefficients for prediction of OR affinity (model 1)_

Term	Coefficient	p-value
Constant	9.3757203453075600	0.022
LogP	0.8116613443435150	0.016
MW	−0.2741951882012750	0.001
Nrot	1.4839802991579400	0.010
LogP²	−0.1400157920570250	0.003
MW²	0.0015895798912707	0.002
Nrot²	−0.4184100716169980	0.059
LogP × Nrot	−0.5997731948635700	0.008
MW³	−0.0000031481780252	0.002
Nrot³	0.0824347258214221	0.021
LogP² × Nrot	0.0765532679846256	0.004
LogP × Nrot²	0.0603271285251953	0.217
Nrot⁴	−0.0028773663882733	0.078
LogP × Nrot³	−0.0098864118642961	0.081

OD detection_

Measure	N (responses)	Control		BC		p-value
Measure	N (responses)	Yes (count)	Yes (%)	Yes (count)	Yes (%)	p-value
Apple	42	1	2	0	0	1.000
Balsamic	42	0	0	0	0	1.000
Berry	42	0	0	1	2	1.000
Butter	42	0	0	0	0	1.000
Cheese	42	0	0	0	0	1.000
Citrus	42	3	7	4	10	1.000
Clove	42	3	7	3	7	1.000
Coffee	42	3	7	3	7	1.000
Earthy	42	16	38	16	38	1.000
Eucalyptus	42	12	29	7	17	0.188
Floral	42	14	33	9	21	0.217
Fruity	42	6	14	1	2	0.043
Grape	42	0	0	0	0	1.000
Grassy	42	14	33	16	38	0.648
Herbal	42	21	50	22	52	0.827
Minty	42	15	36	8	19	0.081
Nutty	42	0	0	1	2	1.000
Peach	42	0	0	0	0	1.000
Peppery	42	6	14	4	10	0.499
Petrol	42	0	0	0	0	1.000
Pine	42	6	14	2	5	0.132
Pineapple	42	1	2	0	0	1.000
Rose	42	3	7	2	5	1.000
Spicy	42	4	10	8	19	0.208
Sweet	42	3	7	2	5	1.000
Tea	42	13	31	13	31	1.000
Tobacco	42	6	14	10	24	0.263
Tropical fruit	42	0	0	0	0	1.000
Vanilla	42	1	2	0	0	1.000
Woody	42	13	31	16	38	0.490

LR of odour strength from HS – model 2 summary_

Measure	p-value
All slopes equal to zero	<0.001
Goodness-of-fit (deviance)	0.966

Sugar leaf phytohormone profiles_

Phytohormone	N	Control (ng · g ¹ FW)		BC (ng · g⁻¹ FW)		p-value
Phytohormone	N	Mean	SD	Mean	SD	p-value
12-Oxo-phytodienoic acid (OPDA)	3	15526.809	9943.563	14466.007	9925.655	0.902
Abscisic acid (ABA)	3	1208.957	362.717	1240.918	580.237	0.939
Brassinolide	3	0.000	−	0.000	−	−
Cinnamic acid (CA)	3	0.000	−	0.000	−	−
Gibberellin A3 (GA3)	3	0.000	−	0.000	−	−
Gibberellin A4 (GA4)	3	0.000	−	0.000	−	−
Indole-3-acetic acid (IAA)	3	0.000	−	0.000	−	−
Indole-3-butyric acid (IBA)	3	0.000	−	0.000	−	−
Indole-3-carboxylic acid (ICA)	3	13.906	1.805	12.147	2.567	0.387
Jasmonic acid (JA)	3	35.496	12.276	33.736	18.240	0.896
Jasmonic acid-isoleucine (JA-Ile)	3	29.395	12.987	20.515	9.265	0.390
Methyl-indole-3-acetic acid (Methyl-IAA)	3	0.000	−	0.000	−	−
Methyl-jasmonic acid (Methyl-JA)	3	5.769	1.783	8.497	2.754	0.223
Salicylic acid (SA)	3	645.416	235.545	489.762	163.708	0.400
Zeatin	3	21.996	9.961	16.841	6.331	0.491

Modulation of cannabis flower characteristics and THC through a biostimulant complex of molasses, Aloe vera extract, and fish-hydrolysate

Figures & Tables

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure S1.

Figure S2.

Figure S3.

Figure S4.

Figure S5.

Figure S6.

Figure S7.

Figure S8.

Figure S9.

Figure S10.

Figure S11.

Figure S12.

Root phytohormone profiles_

Predicted change to reviewer proportions for flowers grown with BC_

Flower odour measures_

Flower volatile comparison from BC treatment_

Coefficients for prediction of T½ (model 3)_

Root bioassay measures_

Comparison of metabolites in sugar leaves that were changed with significance or marginal significance_

Nutrient composition of BC_

Growth and yield measures_

Comparison of metabolites in leaves that were changed with significance or marginal significance_

OD strengths_

Comparison of metabolites in roots that were changed with significance or marginal significance_

Leaf phytohormone profiles_

Pharmacokinetic properties of flower volatiles altered by growth with BC_

Coefficients for prediction of being helpful for anxiety (model 5)_

Sensory analysis: participant demographics_

ODs associated with altered HS volatiles_

Changes in peak area for HS identified compounds_

Coefficients for prediction of being helpful for depression (model 6)_

Coefficients for prediction of BBB index (model 4)_

CBD and THC contents_

LR of odour strength from HS – model 2 variables_

Coefficients for prediction of OR affinity (model 1)_

OD detection_

LR of odour strength from HS – model 2 summary_

Sugar leaf phytohormone profiles_