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  • Author or Editor: Lauren E. Kurtz x
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To maximize yield, cannabidiol (CBD) hemp producers prefer female plants, and this is accomplished by using expensive feminized seed, vegetatively propagated female clones, or by removing male plants from dioecious seed lots. Hemp pollen drifts long distances on wind, and pollination of females reduces CBD content. Induction of triploidy is a common strategy used by plant breeders to produce sterile cultivars of agricultural crops. Triploid (3n) hemp, with three sets of chromosomes, was developed by crossing naturally diploid (2n) hemp with tetraploid (4n) hemp. Tetraploid plants used to create triploids were produced using pregerminated seeds and the mitotic spindle inhibitor colchicine. Seedlings from seeds of ‘Abacas’ × [(‘Otto2’ × ‘BaOx’) × (‘BaOx’ × ‘Colorado Cherry’)] treated with 0.05% colchicine or 0.02% colchicine for 12 hours and longer were significantly shorter than controls and ≤1 cm tall at 10 days after sowing. Surviving seedlings exhibited thickened cotyledons and hypocotyls, which indicated a potential change in ploidy. Tetraploid induction ranged from 26% to 64% for pregerminated seeds of five different hemp cultivars (Abacus × Wife, Cherry Wine, Mountain Mango, Wife, and Youngsim10) treated with 0.05% colchicine for 12 hours. Tetraploids had nearly twice the DNA content as diploids according to flow cytometric analysis. Tetraploid ‘Wife’ had larger stomates and reduced stomatal density compared with diploid ‘Wife’. Four triploid ‘Wife’ genotypes produced from crossing tetraploid ‘Wife’ with diploid ‘Wife’ were acclimated to greenhouse conditions after embryo rescue. DNA content and stomate size of triploid ‘Wife’ was intermediate between the parents. This is the first report of triploid plants of hemp. Future research will evaluate the sterility of triploid hemp.

Open Access

Feminized hemp seed producers often use selfing to maintain a strain name; however, selfing may lead to inferior plants for cannabidiol (CBD) production. Using three different hemp strains as parents [Candida (CD-1), Dinamed CBD, and Abacus], two outcrosses [Candida (CD-1) × Abacus and Dinamed CBD × Candida (CD-1)] and one self-cross [Candida (CD-1) × Candida (CD-1)] were conducted to produce feminized seed. Progeny from the self-cross were significantly smaller and had less yield than outcrossed progeny. Selfed progeny were variegated and highly variable for total dry weight and floral dry weight. Discriminant analysis of principal components (DAPC) using amplified fragment length polymorphism (AFLP) separated the three progeny populations and showed that outcrossed populations clustered closer to the maternal parent, possibly the result of a maternal effect. Analysis of molecular variance (AMOVA) indicated that most variation (74.5%) was within populations, because the progeny from all three populations are half-siblings of each other. The selfed progeny population had lower expected heterozygosity (He = 0.085) than each of the outcrossed progeny populations (He ≈ 0.10). These results suggest that selfed progeny may demonstrate inbreeding depression resulting from enhanced expression of homozygous recessive traits. It may be beneficial for feminized seed producers to use outcrossing instead of selfing to generate feminized seed for CBD production.

Open Access

There is demand for micropropagated Cannabis sativa liner plants, because they are uniform, vigorous, and pathogen free; however, availability is limited because of challenges with in vitro culture decline and ex vitro rooting. Ex vitro rooting success of microcuttings was evaluated for ‘Abacus’ and ‘Wife’ when cultures were 6, 9, 12, 15, and 18 weeks old from initiation. Microcuttings of ‘Wife’ harvested from 6, 9, and 12-week-old cultures rooted at or above 80%, but rooting declined to 50% and 30% for 15- and 18-week-old cultures, respectively. Rooting for ‘Abacus’ remained relatively constant between 47% and 70% for microcuttings harvested from 6- to 18-week-old cultures. ‘Wife’ plants grown from microcuttings, stem cuttings, and retip cuttings (cuttings taken from new shoots on recently micropropagated plants) had equivalent total shoot length, number of shoots, and flower dry weight, whereas micropropagated ‘Abacus’ plants had less shoot length and flower dry weight than plants from stem cuttings. However, when micropropagated ‘Abacus’ plants were provided an extra week of vegetative growth to reach an initial size equivalent to stem and retip plants, all plants performed the same. Propagation method did not change cannabinoid content for both ‘Abacus’ and ‘Wife’. Retip cuttings of ‘Abacus’ and ‘Wife’ rooted at 76% to 81% without rooting hormone, which is comparable to rates reported for stem cuttings of C. sativa treated with rooting hormone. Propagators should consider retipping to expand their liner production, because retips root well and possess the same desirable attributes as micropropagated plants.

Open Access