Biology subjects

Dowling, C. A.

Publications and source records attributed to Dowling, C. A..

5 recordsLinked to original sources

bioRxiv2026

CannSelect: A High-Quality Genotyping Platform for Cannabis sativa

The field of genomics has enabled extraordinary progress in horticultural crop research. However, there is still a need for cost-effective, high-resolution technologies flexible to the diversity found in emerging crops. To this end, we introduce CannSelect, a high-quality genotyping platform for Cannabis sativa. Designed for use in diversity analyses and trait mapping, probe targets were selected from four genotyped diversity panels and a curated gene list. This platform has been used to effectively map day-neutrality in a segregating population to the Autoflower1 locus with average capture efficiencies of 88.5%. With broad genome coverage, demonstrated target specificity, and reproducibility, CannSelect is expected to perform well across the diversity of C. sativa. We describe the methodology used to design CannSelect v1.0 and performance metrics for testing capture efficiency and target alignment in diverse genome assemblies. The CannSelect platform represents a robust and scalable, genome-wide genotyping tool for C. sativa researchers and breeders.

genomics↗

bioRxiv2025

A simple and reliable PCR-based method to differentiate between XX and XY sex genotypes in Cannabis sativa

Cannabis sativa is mainly a dioecious plant, which means that female and male flowers develop on separate individuals, which is controlled by an XY sex determination system: females have two X chromosomes and male plants carry an X and a Y chromosome. C. sativa is a crop that has a wide variety of applications, some of which depend on the sex of the plant. Females are, for example, used for the cannabinoid production, as cannabinoids are produced in the female inflorescence. However, while adult C. sativa individuals present high sexual dimorphism, it is not possible to phenotypically distinguish male and female plants before flowering. Here we present the identification of a highly conserved sex marker, CsPDS5, and the development of a robust, reliable and affordable PCR-based method to determine the sex genotype. In contrast to other sex genotyping methods, our approach relies on a gene polymorphic between the X and Y chromosomes and therefore requires only a single PCR with one pair of primers. The method was tested in 14 cultivars and 6 crossings, with different tissues and developmental stages, on a total of 600 samples, with 100% accuracy. Our assay allows early sex identification and hemp selection, which is useful for both research and industrial purposes. Finally, the pipeline presented here to identify genes polymorphic between the X and Y chromosomes can serve to discover new sex markers, not only in C. sativa, but other dioecious plants and other organisms with 2 sexes.

plant biology↗

bioRxiv2024

FT-like genes in Cannabis and hops: sex specific expression and copy-number variation may explain flowering time variation

Cannabis sativa is a fascinating, yet under-researched, species. To facilitate the global expansion of C. sativa cultivation a greater understanding of flowering time control is crucial. The PEBP gene family consists of universal promoters and repressors of flowering, with homologs of FLOWERING LOCUS T (FT) being highly conserved key regulators of flowering. FT encodes florigen, and balancing the florigen and anti-florigen signals is key for fine-tuning a crops flowering to local climatic conditions. Here, we provide an in-depth characterisation of the PEBP gene family in C. sativa and the closely related species H. lupulus. Phylogenetic analysis reveals expansion of FT and TFL1/CEN clades in the Cannabaceae. The retention of the duplicated PEBP genes likely has functional significance with divergent sequences and expression patterns hinting at signatures of sub-functionalisation. We speculate that duplicated PEBP genes have been crucial to the evolution of photoperiod insensitivity and sexually dimorphic flowering in C. sativa and harnessing the available genetic variation for these traits will be key for establishing this future crop.

plant biology↗

bioRxiv2023

A FLOWERING LOCUS T ortholog is associated with photoperiod-insensitive flowering in hemp (Cannabis sativa L.)

Hemp (Cannabis sativa L.) is an extraordinarily versatile crop, with applications ranging from medicinal compounds to seed oil and fibre products. Cannabis sativa is a short-day plant, and its flowering is tightly controlled by photoperiod. However, substantial genetic variation exists for photoperiod sensitivity in C. sativa, and photoperiod-insensitive ("autoflower") cultivars are available. Using a bi-parental mapping population and bulked segregant analysis, we identified Autoflower2, a 0.5 Mbp locus significantly associated with photoperiod-insensitive flowering in hemp. Autoflower2 contains an ortholog of the central flowering time regulator FLOWERING LOCUS T (FT) from Arabidopsis thaliana which we termed CsFT1. Extensive sequence divergence between alleles of CsFT1 was identified between photoperiod-sensitive and insensitive cultivars of C. sativa, including a duplication of CsFT1 and sequence differences especially in introns. Genotyping of several mapping populations and a diversity panel confirmed a strong correlation between CsFT1 alleles and photoperiod response as well as affirming that at least two independent loci for this agriculturally important trait, Autoflower1 and Autoflower2, exist in the C. sativa gene pool. This study reveals the multiple independent origins of photoperiod insensitivity in C. sativa, supporting the likelihood of a complex domestication history in this species. By integrating the genetic relaxation of photoperiod sensitivity into novel C. sativa cultivars, expansion to higher latitudes will be permitted, thus allowing the full potential of this versatile crop to be reached.

plant biology↗

bioRxiv2022

A protocol for rapid generation cycling (speed breeding) of hemp (Cannabis sativa) for research and agriculture

Hemp (Cannabis sativa) is a highly versatile multi-purpose crop with a multitude of applications, from textiles, biofuel and building material to high-value food products for consumer markets. Furthermore, non-hallucinogenic cannabinoids like cannabidiol (CBD), which can be extracted from female hemp flowers, are potentially valuable pharmacological compounds. In addition, hemp has high carbon sequestration potential due to its rapid growth rate. Therefore, the hemp industry is gaining more traction and breeding hemp cultivars adapted to local climate conditions or bred for specific applications is becoming increasingly important. Here, we present a method for rapid generation cycling (speed breeding) for hemp. The speed breeding protocol makes use of the photoperiod sensitivity of Cannabis. It encompasses vegetative growth of the plants for two weeks under continuous light, followed by flower induction, pollination and seed development for four weeks under short-day conditions and a seed ripening phase under continuous light and water stress. With the protocol introduced here, a generation time of under nine weeks (61 days) from seed to seed can be achieved. Our method furthermore synchronises flowering time of different hemp cultivars, thus facilitating crosses between cultivars. The extremely short generation time will enable hemp researchers and breeders to perform crosses in a time-efficient way and generate new hemp cultivars with defined genetic characteristics in a shorter amount of time.

plant biology↗