Project description:Panax ginseng C.A. Meyer is a precious Chinese herbal medicine belonging to the Araliaceae family. Because ginseng is extremely taboo, it seriously restricts the sustainable development of the ginseng industry. The allelopathic effect of triterpenoid saponins is one of the important incentives for continuous cropping disorders. However, there are few studies on the mechanism of allelopathic autotoxicity of triterpenoid ginsenosides. Therefore, this study took ginseng hairy roots as the research object, exogenously added ginsenoside Ro to regulate the growth of hairy roots, and measured the changes in its physiological and saponin content. It was found that ginseng hairy roots grew significantly under Ro-0.5 mg/L stress. Inhibition, the specificity of root tip cell injury was obvious, promoted the accumulation of endogenous hormones IAA and SA, and inhibited the accumulation of ABA and JA. The accumulation of ginsenosides was significantly inhibited (except Rg3) under Ro-0.5 mg/L stress. The mRNA analysis of the Ro-0.5 mg/L group and CK group showed that the differentially expressed genes were mostly concentrated in the hormone signal transduction pathway. The genes ARF7 and EFM were up-regulated, and XTH23 and ZOX1 were down-regulated, which can be used as important candidates for hormone-responsive continuous cropping disorders. Gene. 74 differentially expressed miRNAs were identified based on miRNA sequencing, of which 22 were up-regulated and 52 were down-regulated. The target genes of ptc-miR156k_L+1, mtr-miR156b-5p, gma-miR156a_R+1, and mtr-miR156e were all TRINITY_DN14567_c0_g4, TRINITY_DN14567_c0_g4 is a gene in the plant hormone signal transduction pathway, and the four miRNAs are all negatively correlated with mRNA, indicating that ptc-miR156k_L+1, mtr-miR156b-5p, gma-miR156a_R+1, and mtr-miR156e are very likely involved in ginseng Response to continuous cropping disorders and regulation of ginsenoside synthesis. Our findings will provide a useful platform for unraveling the continuous cropping barrier of ginseng and new insights into the genetic engineering of plant stress responses.
Project description:The objectives of this study encompassed a thorough exploration of the potential implications of protein profiling in hairy roots, specifically focusing on optimizing and enhancing C. asiatica organ cell biofactories. In this pursuit, we categorized established C. asiatica hairy root lines according to their capacity for centelloside production, classifying them into HIGH, MID, or LOW categories. For comparative analysis, wild adventitious (Adv) roots were extracted from in vitro C. asiatica seedlings and cultivated in solid MS medium at 25°C in complete darkness, serving as control specimens. This meticulous, label-free proteomic analysis enabled the successful identification of several proteins. Our research substantially builds upon and extends the findings presented by Alcalde et al. (2022) (DOI: 10.3389/fpls.2022.1001023). In their study, distinctive morphological and metabolic variations were noted among different C. asiatica hairy root lines. Such differences are presumably attributable to the random incorporation of a selective set of genes from the T-DNA, with particular emphasis on the rol and aux genes.
Project description:Arctic alpine species experience extended periods of cold and unpredictable conditions during flowering. Thus, often, alpine plants use both sexual and asexual means of reproduction to maximise fitness and ensure reproductive success. We used the arctic alpine perennial Arabis alpina to explore the role of prolonged cold exposure on adventitious rooting. We exposed plants to 4 °C for different durations and scored the presence of adventitious roots on the main stem and axillary branches. Our physiological studies demonstrated the presence of adventitious roots after 21 weeks at 4 °C saturating the effect of cold on this process. Notably, adventitious roots on the main stem developingin specific internodes allowed us to identify the gene regulatory network involved in the formation of adventitious roots in cold using transcriptomics. These data and histological studies indicated that adventitious roots in A. alpina stems initiate during cold exposure and emerge after plants experience growth promoting conditions. While the initiation of adventitious root was not associated with changes of DR5 auxin response and free endogenous auxin level in the stems, the emergence of the adventitious root primordia was. Using the transcriptomic data, we discerned the sequential hormone responses occurring in various stages of adventitious root formation and identified supplementary pathways putatively involved in adventitious root emergence, such as glucosinolate metabolism. Together, our results highlight the role of low temperature during clonal growth in alpine plants and provide insights on the molecular mechanisms involved at distinct stages of adventitious rooting.
Project description:We developed a method to synchronize the induction of lateral roots in primary and adventitious roots of Zea mays, and used it to perform a genome-wide transcriptome analysis of the pericycle cells in front of the phloem poles during lateral root initiation. Lateral roots were induced in primary and adventitious roots of Maize. For the primary root, plants were germinated and grown 64 hours in NPA 50 µM, and then transfered to NAA 50 µM. For the adventitious roots, plants were germinated and grown in water for 6 days, then tranfered 4 days in NPA 25 µM, and finally transfered to NAA 25 µM. For all these roots, pericycle cells located in front of the phloem poles in segments of roots located between 5 and 10 mm distance from the root tip were isolated using laser capture microdissection after cryosection. Material was sampled at 0 hours (NPA) and after 2, 3 and 4 hours of NAA treatment, for both the primary and adventitious roots and also after 6 hours and 9 hours of NAA treatment for the adventitious roots.
Project description:In hairy roots of the model legume Medicago truncatula the saponin production can be elicited by methyl jasmonate treatment. To identify genes potentially involved in saponin biosynthesis or its regulation we carried out transcript profiling by RNA-Seq of M. truncatula hairy roots treated with 100 μM of methyl jasmonate (dissolved in ethanol) for two or 24 hours. As control, M. truncatula hairy roots treated with an equivalent amount of ethanol were profiled.
Project description:This dataset belongs to a set of three RNA-Seq experiments that were carried out to study the regulation of monoterpenoid indole alkaloid production in the medicinal plant Catharanthus roseus. For this dataset, C. roseus hairy roots overexpressing the well-known MIA biosynthesis regulator ORCA3 were analyzed by RNA-Seq. As control, C. roseus hairy roots expressing GUS were used. Each analyzed sample consisted of an independent hairy root line; three hairy root lines per construct were analyzed.
Project description:In this research we have made tomato hairy roots expressing R. irregularis nuclear-lokalized effector proteins, RiSP749, GLOIN707, GLOIN781, and GFP as a control, under the XVE promoter. Hairy roots were cultured for 3 weeks after which the expression of the effectors was induced with estradiol. 24 h after estradiol treatment, roots were harvested and RNA was used for RNA sequening
Project description:We investigated root associated fungi in young Norway spruce (Picea abies) cuttings rooted from slow- and fast-growing trees showing variable growth rate in long-term field experiments and compared their roots’ gene expression patterns five and 18 months after adventitious root initiation. Gene expression patterns of adventitious roots could not be systematically linked with the growth phenotype at the initiation of root formation, and thus fundamental differences in the receptiveness of fungal symbionts could not be assumed.