Hemp (Cannabis sativa L.) has recently become an important crop due to the growing market demands for products containing cannabinoids. Unintended cross-pollination of C. sativa crops is one of the most important threats to cannabinoid production and has been shown to reduce cannabinoid yield. Ploidy manipulation has been used in other crops to improve agronomic traits and reduce fertility; however, little is known about the performance of C. sativa polyploids. In this study, colchicine was applied to two proprietary, inbred diploid C. sativa inbred lines, ‘TS1-3’ and ‘P163’, to produce the tetraploids ‘TS1-3 (4x)’ and ‘P163 (4x)’. The diploid, triploid, and tetraploid F1 hybrids from ‘TS1-3’ × ‘P163’, ‘TS1-3 (4x)’ × ‘P163’, and ‘TS1-3 (4x)’ × ‘P163 (4x)’ were produced to test their fertilities, crossing compatibilities, and yields. The results indicated a reduction in fertility in the triploids and the tetraploids, relative to their diploid counterparts. When triploids were used as females, seed yields were less than 2% compared to when diploids were used as females; thus, triploids were determined to be female infertile. The triploids resulting from the crosses made herein displayed increases in biomass and inflorescence weight compared to the diploids created from the same parents in a field setting. Statistical increases in cannabinoid concentrations were not observed. Lastly, asymmetric crossing compatibility was observed between the diploids and the tetraploids of the genotypes tested. The results demonstrate the potential benefits of triploid C. sativa cultivars in commercial agriculture.
Female hemp plants are desired in floral hemp operations due to their higher cannabinoid contents. To produce feminized seeds, a critical step of inducing fertile male flowers on female plants is performed. In feminized seed production, freshly mixed STS (silver thiosulfate + sodium thiosulfate) is applied to female plants as an ethylene inhibitor to induce male flowers. However, the short-shelf stability of the STS buffer can cause difficulty in the application and inconsistent results. Alternative methods with improved accessibility and stable buffers will be beneficial for the hemp industry and hemp breeders. A commercially available floriculture product, Chrysal ALESCO®, contains silver nitrate, the same active ingredient as STS but with increased shelf stability. This study compares Chrysal ALESCO® to the traditional STS standard methods for male flower induction on female plants and their pollen quality. The two treatments were applied to six female hemp accessions with three replicates investigated, and the male flower counts and pollen quality were compared. No statistically significant difference was discovered in their male flower counts; the STS-treated plant produced an average of 478.18 male flowers, and the Chrysal ALESCO®-treated plant produced an average of 498.24 male flowers per plant. Fluorescein diacetate (FDA) and acetocarmine stains were used to investigate the pollen quality (non-aborted rate) of two chosen genotypes. FDA-stained pollen of Chrysal ALESCO® showed a significantly higher non-aborted rate than the pollen of traditional STS-treated plants (p < 0.001); however, only a marginally higher non-aborted rate was discovered by acetocarmine staining (p = 0.0892). In summary, Chrysal ALESCO® performed equally to traditional STS treatment at male flower counts and better or equally in pollen quality. With better shelf stability and easy application, ALESCO® can be a viable alternative option for stimulating male flowers on female hemp plants.
Hibiscus syriacus, azalea, is an important woody ornamental shrub planted throughout many temperate and subtropical regions of the world. However, flower size is smaller in this species than some of its relatives. To increase flower size, interspecific hybridization has been used, and such hybrid cultivars are usually characterized by larger flowers, increased vigor, diverse leaf shapes, and reduced fertility. Our earlier studies have shown that these hybrid cultivars could backcross with H. syriacus when used as male parents. To understand the breeding potential of these hybrid cultivars, two popular tetraploid hybrid cultivars, ‘Lohengrin’ and ‘Resi’, were used as pollen parents to backcross several tetraploid H. syriacus cultivars. As a result, 28.76% and 64.4% of ‘Lohengrin’ and ‘Resi’ progenies exhibited larger flowers than both of their parents. Interestingly, 14 of 18 progenies of ‘Resi’ were putative hexaploids, whereas 19 tested ‘Lohengrin’ progenies were tetraploid. Because putative hexaploid progenies were only observed among progenies of ‘Resi’, this hybrid cultivar appears to produce unreduced gametes. In addition, among the 14 putative hexaploids derived from ‘Resi’, 11 had larger flowers than both of their parents and their tetraploid siblings (p < 0.05). The 45S rDNA and 5S rDNA locus segregation among those BC1F1 progenies was tested by fluorescent in situ hybridization (FISH), and the wide range of 45S rDNA signal numbers among siblings indicated that these aneuploids resulted from unequal segregation or chromosome rearrangement. Chromosome counting confirmed aneuploidy among BC1F1 progenies. Ploidy diversity and aneuploidy have been known to contribute to various elements of morphological diversity, such as larger flower size and reduced fertility, which are important in ornamental plant breeding. The present study demonstrated the breeding potential of interspecific Hibiscus cultivars for increasing ploidy level and flower size.
Strawberries (Fragaria × ananassa) are highly valued for their colorful fruit and flavorful taste. Anthocyanins provide much of the red fruit color, and the flavor is highly affected by soluble sugars and non-volatile organic acids. These fruit composition parameters impact consumer decisions. In this study, strawberry fruits from 17 commercial cultivars and advanced selections were collected weekly from replicated trials at three locations in North Carolina. The relative effects of the location and germplasm on fruit composition were determined, including the soluble solid concentration (SSC), titratable acidity (Tacid), and total anthocyanin content (TAC). The breeding criteria of at least 8.0% SSC and 0.80% Tacid were met by eight and six genotypes, respectively and five of these met both criteria. The fruit TAC ranged from 24.0 to 45.7 mg pelargonidin-3-O-glucoside (P3G) equivalents/100 g FWT. P3G was the dominant pigment in all genotypes, followed by pelargonidin-3-O-rutinoside (P3R). As harvest dates advanced, pH, TAC, P3G, P3R, and pelargonidin-3-O-(6″-malonylglucoside) (P3MG) generally decreased, while Tacid, SSC, and cyanidin-3-O-glucoside (C3G) increased. Composition of field-grown strawberries in this mid-Atlantic location were more influenced by the genotype and harvest date than by the growing location.
Hemp ( Cannabis sativa L.) is a primarily short-day crop grown for grain, fiber, and secondary metabolites. Although terminal flowering of most non-domesticated hemp is regarded as photoperiod sensitive, limited germplasm is available for the development of day-neutral hemp. Day-neutral plants experience flower maturation independently of the short-day photoperiod cue which is typically triggered by photoperiods of less than 14 hours. The day-neutral trait is the subject of increased commercial interest for the purpose of breeding varieties with accelerated flowering time or cultivars that do not require labor- and cost-intensive light deprivation production systems. Genetic markers for the photoperiodic response would help breeders make early selection in the design of suitable cultivars for specific environments and cultivation calendars. Limited refereed information has been produced regarding the genetic regulation of photoperiodism in C. sativa . A population of 318 F2 individuals segregating for day-neutral flowering was developed, phenotyped, and genotyped. Genome-wide association analysis identified markers associated with the day-neutral trait indicating that a single recessive gene controlling photoperiod sensitivity is located within a large region of Chromosome 1. Flanking region sequence data of linked SNP markers was used in the development and validation of a TaqMan-based qPCR assay for the day-neutral trait. A genetic linkage map was produced, and QTL mapping identified two additional markers on Chromosome 1. Candidate genes, TARGET OF EARLY ACTIVATION TAGGED (TOE)/APETALA2 (AP2) , and PSEUDO-RESPONSE REGULATOR 3 (PRR3) , may work together to impact phase transition and photoperiodic flowering respectively and have key domains that are disrupted in day-neutral plants.
A-genome Arachis species (AA; 2n = 2x = 20) are commonly used as secondary germplasm sources in cultivated peanut breeding, Arachis hypogaea L. (AABB; 2n = 4x = 40), for the introgression of various biotic and abiotic stress resistance genes. Genome doubling is critical to overcoming the hybridization barrier of infertility that arises from ploidy-level differences between wild germplasm and cultivated peanuts. To develop improved genome doubling methods, four trials of various concentrations of the mitotic inhibitor treatments colchicine, oryzalin, and trifluralin were tested on the seedlings and seeds of three A-genome species, A. cardenasii, A. correntina, and A. diogoi. A total of 494 seeds/seedlings were treated in the present four trials, with trials 1 to 3 including different concentrations of the three chemical treatments on seedlings, and trial 4 focusing on the treatment period of 5 mM colchicine solution treatment of seeds. A small number of tetraploids were produced from the colchicine and oryzalin gel treatments of seedlings, but all these tetraploid seedlings reverted to diploid or mixoploid states within six months of treatment. In contrast, the 6-h colchicine solution treatment of seeds showed the highest tetraploid conversion rate (6–13% of total treated seeds or 25–40% of surviving seedlings), and the tetraploid plants were repeatedly tested as stable tetraploids. In addition, visibly and statistically larger leaves and flowers were produced by the tetraploid versions of these three species compared to their diploid versions. As a result, stable tetraploid plants of each A-genome species were produced, and a 5 mM colchicine seed treatment is recommended for A-genome and related wild Arachis species genome doubling.