Content of cannabinoids in clonally propagated industrial hemp

Figures & data

Table 1. Breeding information on tested hemp accessions and the parent components of hybrids (Carmagnola × K 290 (1565) and Beniko × Carmagnola (1491)).

Cultivar/accession	Origin	Purpose	Genotypic expression	Vegetative cycle (days)	Plant height (cm)	CBD content (%)	THC content (%)
Henola	Poland	Grain	Monoecious	115–120	150–200	0.07–1.00	<0.02
Epsilon 68	France	Fiber/Grain	Monoecious	140–150	250–350	0.51–1.04	0.13–0.03
K290	Ukraine	Fiber/Grain	Dioecious	130–140	300–350	1.25–2.03	0.04
Carmagnola	Italy	Fiber/CBD	Dioecious	140–160	200–250	0.13–3.80	0.01–0.24
Beniko	Poland	Fiber/Grain	Monoecious	140–160	250–300	0.39–1.50	0.01–0.08

Data were obtained from Fibrous Plants Gene Bank INF&MP. K290 is a local landrace from Dnipro province.

(former Dnepropetrovsk province).

Figure 1. a) in vitro regenerated plant (EPS/40), b) acclimatization of plantlets, c) plants growing in the vegetation hall.

Figure 2. a) HPLC profile of EPS/40 genotype at λ = 228 nm, b) HPLC profile of K290/136 genotype at λ = 228 nm. 1—CBD, 2—CBDA (pink colored), 3—Δ⁹-THC, 4—Δ⁹-THCA.

Figure 3. Sum (%) of total cannabinoids (CBDA, CBD, Δ⁹-THCA, Δ⁹-THC, CBC and CBG) in tested genotypes. Mean values in column with the same letter are not significantly different at p = .05 (tukey’s multiple range test). Genotypes: HN—Henola; 1491—Beniko xCarmagnola, 1565—Carmagnola x K 290; EPS—Epsilon 68.

Table 2. Content of CBDA, CBD, ∆⁹-THCA, ∆⁹-THC, CBC, CBG, and CBN in tested genotypes.

No	Genotype	n	Mean content % (±SD) DW
			CBDA	CBD	∆^9-THCA	∆^9-THC	CBC	CBG
1	HN/2	7	0.749 ± 0.082^ab	0.216 ± 0.060^bcd	0.014 ± 0.005^a	0.014 ± 0.005^a	0.034 ± 0.008^cde	<0.01
2	HN/35	8	0.568 ± 0.060^a	0.110 ± 0.028^ab	0.014 ± 0.005^a	<0.01	0.010 ± 0.00^a	<0.01
3	HN/39	10	0.478 ± 0.097^a	0.046 ± 0.021^a	0.018 ± 0.006^a	<0.01	0.010 ± 0.00^a	<0.01
4	HN/64	7	0.619 ± 0.114^a	0.173 ± 0.073^bcd	0.020 ± 0.000^ab	0.010 ± 0.000^a	0.016 ± 0.005^ab	<0.01
5	1491/78	10	2.000 ± 0.283^de	0.208 ± 0.047^bcd	0.052 ± 0.006^abc	0.017 ± 0.005^a	0.038 ± 0.006^e	0.016 ± 0.005^a
6	1491/5	9	3.311 ± 0.554^f	0.249 ± 0.046^cde	0.082 ± 0.014^bc	0.022 ± 0.004^a	0.031 ± 0.003^bcde	0.020 ± 0.006^ab
7	K290/K1	9	2.027 ± 0.571^de	0.266 ± 0.133^de	0.058 ± 0.033^abc	0.018 ± 0.008^a	0.036 ± 0.017^de	0.017 ± 0.005^a
8	K290/136	5	2.176 ± 0.287^de	0.200 ± 0.081^bcd	0.824 ± 0.200^d	0.298 ± 0.080^b	0.036 ± 0.011^cde	0.018 ± 0.005^ab
9	1565/8	7	1.649 ± 0.211^cde	0.106 ± 0.024^ab	0.043 ± 0.005^abc	0.010 ± 0.000^a	0.017 ± 0.005^abc	0.050 ± 0.008^c
10	1565/26	10	1.632 ± 0.360^cd	0.353 ± 0.134^e	0.039 ± 0.017^ab	0.026 ± 0.007^a	0.031 ± 0.01^bcde	0.027 ± 0.005^b
11	EPS/1	9	1.876 ± 0.108^de	0.244 ± 0.054^cd	0.039 ± 0.003^abc	0.017 ± 0.005^a	0.021 ± 0.003^abcd	<0.01
12	EPS/6	8	2.158 ± 0.440^de	0.221 ± 0.064^bcd	0.053 ± 0.014^abc	0.018 ± 0.005^a	0.043 ± 0.017^e	0.020 ± 0.00^ab
13	EPS/10	9	1.860 ± 0.392^de	0.167 ± 0.037^bcd	0.051 ± 0.011^abc	0.013 ± 0.005^a	0.034 ± 0.012^de	<0.01
14	EPS/19	8	1.214 ± 0.121^bc	0.170 ± 0.046^bcd	0.034 ± 0.007^ab	0.012 ± 0.004^a	0.016 ± 0.005^ab	0.010 ± 0.00^a
15	EPS/21	10	2.159 ± 0.175^e	0.151 ± 0.021^bc	0.055 ± 0.010^abc	0.013 ± 0.005^a	0.031 ± 0.011^bcde	<0.01
16	EPS/24	5	3.058 ± 0.324^f	0.252 ± 0.038^cde	0.066 ± 0.009^abc	0.016 ± 0.005^a	0.026 ± 0.005^abcde	<0.01
17	EPS/40	8	4.723 ± 0.542^g	0.269 ± 0.004^de	0.103 ± 0.010^c	0.019 ± 0.010^a	0.069 ± 0.011^f	<0.01

Mean values in column with the same letter are not significantly different at p = .05 (Tukey’s multiple range test). HN – Henola; 1491—Beniko x Carmagnola, 1565—Carmagnola x K 290; EPS – Epsilon 68, n – number of plants.

Table 3. Content of total CBD and total THC, ratio of CBD/THC and ratio of THC/CBD.

Genotype	n	Mean content % (±SD) DW		Ratio of total CBD/THC	Ratio of total THC/CBD
		Total CBD	Total THC
HN/39	10	0.465 ± 0.103^a	0.026 ± 0.006^a	17.88	0.06
HN/35	8	0.608 ± 0.073^a	0.022 ± 0.005^a	27.64	0.04
HN/64	7	0.715 ± 0.153^ab	0.028 ± 0.001^ab	25.54	0.04
HN/2	7	0.872 ± 0.080^ab	0.025 ± 0.005^ab	34.88	0.03
EPS/19	8	1.234 ± 0.140^bc	0.041 ± 0.007^ab	30.10	0.03
1565/8	7	1.552 ± 0.205^cd	0.048 ± 0.004^ab	32.33	0.03
1565/26	10	1.784 ± 0.244^de	0.060 ± 0.012^abc	29.73	0.03
EPS/10	9	1.798 ± 0.368^de	0.058 ± 0.011^abc	31.00	0.03
EPS/1	9	1.889 ± 0.147^de	0.051 ± 0.007^abc	37.04	0.03
1491/78	10	1.962 ± 0.288^de	0.063 ± 0.008^bc	31.14	0.03
K290/K1	9	2.043 ± 0.515^de	0.068 ± 0.025^bcd	30.04	0.03
EPS/21	10	2.044 ± 0.159^de	0.059 ± 0.010^abc	34.64	0.03
K290/136	5	2.108 ± 0.181^de	1.021 ± 0.132^e	2.06	0.48
EPS/6	8	2.113 ± 0.337^e	0.064 ± 0.009^bcd	33.02	0.03
EPS/24	5	3.034 ± 0.321^f	0.074 ± 0.012^bcd	27.49	0.04
1491/5	9	3.153 ± 0.524^f	0.094 ± 0.015^cd	33.54	0.03
EPS/40	8	4.410 ± 0.491^g	0.109 ± 0.010^d	40.46	0.02

HN-Henola. 1491 - Beniko x Carmagnola. 1565 - Carmagnola x K 290; EPS-Epsilon 68; n- number of plants. Values of total CBD and total THC are mean ± SD of three repetitions for chemical analysis. Mean values in column with the same letter are not significantly different at p = .05 (Tukey’s multiple range test).

Table 4. Coefficient of variation calculated for THCA and CBDA.

Genotype	n	CV (%)
		CBDA	THCA
HN/39	10	20.3	33.3
HN/35	8	10.6	35.7
HN/64	7	18.4	0.0
HN/2	7	10.9	35.7
EPS/19	8	10.0	20.6
1565/8	7	12.8	11.6
1565/26	10	22.1	22,1
EPS/10	9	21.1	21.6
EPS/1	9	5.8	7.7
1491/78	10	14.2	11.5
K290/K1	9	28.2	38.7
EPS/21	10	8.1	15.6
K290/136	5	13.2	24.3
EPS/6	8	20.4	26.4
EPS/24	5	10.4	13.6
1491/5	9	16.7	17.1
EPS/40	8	11.5	9.7

HN-Henola. 1491 - Beniko x Carmagnola. 1565 - Carmagnola x K 290; EPS-Epsilon 68; n- number of plants.