Triploid Atractylodes chinensis (DC.) Koidz. (A. chinensis) is a valuable genetic resource for medicinal plant breeding, and there is a demand to develop an efficient in vitro regeneration protocol to resolve triploid sterility in production. In this study, the effects of sterilization, bud type, 6-benzyladenine (6-BA) combined with 1-naphthaleneacetic acid (NAA) or indole-3-butyric acid (IBA) and transplanting materials on shoot regeneration, rooting and growth were explored. The genetic stability of regenerated plants was verified using flow cytometry (FCM) and simple sequence repetition (SSR) molecular markers. When axillary buds were sterilized with sodium hypochlorite for 30 min and cultured on Murashige and Skoog (MS) + 1.5 mg·L-1 6-BA + 0.4 mg·L-1 NAA medium, the regeneration rate was 61.33%. Strong roots occurred on 1/2 MS combined with 3% sucrose and 0.6 mg·L-1 IBA medium. Rooting plantlets transplanted into a mixture of peat soil: perlite (1:1, v/v), the survival rate up to 90.00%. Histological analyses revealed that shoot meristematic cells derived from cambium cells developed into the apical meristem and formed shoots. FCM analysis determined that the ploidy levels of 20 random regenerated plants were coincident with explants, the number and size of bands obtained by 6 pairs of SSR polymorphic primers were consistent. The results of this study show that our in vitro adventitious regeneration and plantlet transplantation protocol of triploid A. chinensis was efficient and produced strong genetic stability.

Triploid Atractylodes chinensis (DC.) Koidz. (A. chinensis) is a valuable genetic resource for medicinal plant breeding, and there is a demand to develop an e cient in vitro regeneration protocol to resolve triploid sterility in production. In this study, the effects of sterilization, bud type, 6-benzyladenine (6-BA) combined with 1-naphthaleneacetic acid (NAA) or indole-3-butyric acid (IBA) and transplanting materials on shoot regeneration, rooting and growth were explored. The genetic stability of regenerated plants was veri ed using ow cytometry (FCM) and simple sequence repetition (SSR) molecular markers. When axillary buds were sterilized with sodium hypochlorite for 30 min and cultured on Murashige and Skoog (MS) + 1.5 mg•L -1 6-BA + 0.4 mg•L -1 NAA medium, the regeneration rate was 61.33%. Strong roots occurred on 1/2 MS combined with 3% sucrose and 0.6 mg•L -1 IBA medium. Rooting plantlets transplanted into a mixture of peat soil: perlite (1:1, v/v), the survival rate up to 90.00%. Histological analyses revealed that shoot meristematic cells derived from cambium cells developed into the apical meristem and formed shoots. FCM analysis determined that the ploidy levels of 20 random regenerated plants were coincident with explants, the number and size of bands obtained by 6 pairs of SSR polymorphic primers were consistent. The results of this study show that our in vitro adventitious regeneration and plantlet transplantation protocol of triploid A. chinensis was e cient and produced strong genetic stability. Key MessageIn this study, an e cient and genetically stable adventitious regeneration of triploid A. chinensis was established for the rst time.

Background Panax Ginseng is a perennial and semi-shady herb with tremendous medicinal value. Due to its unique botanical characteristics, ginseng is vulnerable to various abiotic factors during its growth and development, especially in high temperatures. Proteins encoded by 14-3-3 genes form a highly conserved protein family that widely exists in eukaryotes. The 14-3-3 family regulates the vital movement of cells and plays an essential role in the response of plants to abiotic stresses, including high temperatures. Currently, there is no relevant research on the 14-3-3 genes of ginseng. Methods The identification of the ginseng 14-3-3 gene family was mainly based on ginseng genomic data and Hidden Markov Models (HMM). We used bioinformatics-related databases and tools to analyze the gene structure, physicochemical properties, cis-acting elements, gene ontology (GO), phylogenetic tree, interacting proteins, and transcription factor regulatory networks. We analyzed the transcriptome data of different ginseng tissues to clarify the expression pattern of the 14-3-3 gene family in ginseng. The expression level and modes of 14-3-3 genes under heat stress were analyzed by quantitative real-time PCR (qRT-PCR) technology to determine the genes in the 14-3-3 gene family responding to high-temperature stress. Results In this study, 42 14-3-3 genes were identified from the ginseng genome and renamed PgGF14-1 to PgGF14-42. Gene structure and evolutionary relationship research divided PgGF14s into epsilon (ε) and non-epsilon (non-ε) groups, mainly located in four evolutionary branches. The gene structure and motif remained highly consistent within a subgroup. The physicochemical properties and structure of the predicted PgGF14 proteins conformed to the essential characteristics of 14-3-3 proteins. RNA-seq results indicated that the detected PgGF14s existed in different organs and tissues but differed in abundance; their expression was higher in roots, stems, leaves, and fruits but lower in seeds. The analysis of GO, cis-acting elements, interacting proteins, and regulatory networks of transcription factors indicated that PgGF14s might participate in physiological processes, such as response to stress, signal transduction, material synthesis-metabolism, and cell development. The qRT-PCR results indicated PgGF14s had multiple expression patterns under high-temperature stress with different change trends in several treatment times, and 38 of them had an apparent response to high-temperature stress. Furthermore, PgGF14-5 was significantly upregulated, and PgGF14-4 was significantly downregulated in all treatment times. This research lays a foundation for further study on the function of 14-3-3 genes and provides theoretical guidance for investigating abiotic stresses in ginseng.