Comparative RNA-Seq analysis reveals genes associated with masculinization in female Cannabis sativa

Abstract

Main conclusion Using RNA profiling, we identified several silver thiosulfate-induced genes that potentially control the masculinization of female Cannabis sativa plants. Abstract Genetically female Cannabis sativa plants normally bear female flowers, but can develop male flowers in response to environmental and developmental cues. In an attempt to elucidate the molecular elements responsible for sex expression in C. sativa plants, we developed genetically female lines producing both female and chemically-induced male flowers. Furthermore, we carried out RNA-Seq assays aimed at identifying differentially expressed genes responsible for male flower development in female plants. The results revealed over 10,500 differentially expressed genes, of which around 200 potentially control masculinization of female cannabis plants. These genes include transcription factors and other genes involved in male organ (i.e., anther and pollen) development, as well as genes involved in phytohormone signalling and male-biased phenotypes. The expressions of 15 of these genes were further validated by qPCR assay confirming similar expression patterns to that of RNA-Seq data. These genes would be useful for understanding predisposed plants producing flowers of both sex types in the same plant, and help breeders to regulate the masculinization of female plants through targeted breeding and plant biotechnology.

Untargeted Metabolomic Analyses Reveal Chemical Complexity of Dioecious

“…However, it is not possible to discriminate these isomers with the LC stationary phase used in this study. This is the first report of the upregulation of OPDA in Cannabis flowers and given that APA-and auxin-related genes are directly implicated in sex expression, [20,51,55] this compound may represent a molecule of interest for further investigation.…”

Fertilization Following Pollination Predominantly Decreases Phytocannabinoids Accumulation and Alters the Accumulation of Terpenoids in Cannabis Inflorescences

“…It may be the result of altered regulation of synthesis enzymes, for example the upregulation of THCA synthase or CBCA synthase (Laverty et al, 2019). A previous study found over 10,000 genes are differentially expressed upon masculinization of female plants (Adal et al, 2021), but it is not clear how these genes are related to phytocannabinoid expression in the fertilized female plant. A donor-dependent effect was also detected in the specific increase in the Eudesmol family (α-, β-, and γ-Eudesmol) components, which were highly increased in the THC-rich female upon fertilization by the THC-rich male plant (Figure 6A) and a parallel specific increase in Linalool in the CBD-rich female fertilized by the induced male (Figure 6B).…”

Construction of a High-Density Genetic Map and Identification of Leaf Trait-Related QTLs in Chinese Bayberry (Myrica rubra)

“…Low-quality reads (>10% Ns or a PHRED score <10 for at least 50% of bases), and those containing adaptor sequences were removed from the resultant reads using an internally developed script. The expression levels of mapped-paired reads were normalized as reads per kilobase per million mapped reads (RPKM), with the CLC RNA-Seq analysis tool ( Adal et al, 2021 ). The normalized reads were then used for the differential expression analyses in two groups.…”