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Furthermore, modification of in vitro culture conditions explained DNA sequence variation, demethylation, and de novo methylation in the CHG context, as indicated by analysis of variance. Linear regression indicated that DNA sequence variation was related to de novo DNA methylation in the CHG context. Mediation analysis showed the role of copper ions as a mediator of sequence variation in the CHG context. No other contexts showed a significant sequence variation in mediation analysis. Silver ions did not act as a mediator between any methylation contexts and sequence variation. Thus, incorporating copper ions in the induction medium should be treated with caution.LONELY GUY has been previously characterized in flowering plants to be involved in the direct activation of cytokinins. In this study, the function of the only LONELY GUY gene (CvarLOG1) from unicellular green microalga Chlorella variabilis NC64A has been investigated. CvarLOG1 expressed mainly in the lag and log phases of growth and was confirmed to be a cytokinin-activating enzyme. Overexpression of CvarLOG1 in Chlorella led to extended life in culture by almost 10-20 days, creating a "stay-green" phenotype. In the transformed alga, the cell cycle was lengthened due to delayed entry into the G2/M phase contrary to the known role of cytokinins in stimulating G2/M transition possibly due to excessive levels of this hormone. However, due to the sustained growth and delayed senescence, there was an increase in cell number by 11% and in biomass by 46% at the stationary phase, indicating a potential application for the biofuel industry. The total carbohydrate and lipid yield increased by approximately 30 and 20%, respectively. RNA-Seq-based transcriptomic analysis revealed that the genes associated with light and dark reactions of photosynthesis were upregulated, which may be the reason for the increased biomass. These data show that LOG plays an essential role during the cell cycle and in the functioning of the chloroplast and that the pathway leading to direct activation of cytokinins via LOG is functional in algae.Almost all elevated atmospheric CO2 concentrations (eCO2) studies have not addressed the potential responses of plant growth to different CO2 in daytime and nighttime. The present study was to determine the impact of daytime and/or nighttime eCO2 on growth and quality of mulberry (Morus alba L.), a perennial multipurpose cash plant. Six-month-old mulberry seedlings were hence grown in environmentally auto-controlled growth chambers under four CO2 concentrations (1) ambient CO2 (ACO2, 410 μmol mol-1 daytime/460 μmol mol-1 nighttime), (2) sole daytime elevated CO2 (DeCO2, 710 μmol mol-1/460 μmol mol-1), (3) sole nighttime elevated CO2 (NeCO2, 410 μmol mol-1/760 μmol mol-1), and (4) continuous daytime and nighttime elevated CO2 (D + NeCO2, 710 μmol mol-1/760 μmol mol-1). Plant growth characteristics, nutrient uptake, and leaf quality were then examined after 120 days of CO2 exposure. Compared to control, DeCO2 and (D + N)eCO2 increased plant biomass production and thus the harvest of nutrients and accumulation of leaf carbohydrates (starch, soluble sugar, and fatty acid) and N-containing compounds (free amino acid and protein), though there were some decreases in the concentration of leaf N, P, Mg, Fe, and Zn. NeCO2 had no significant effects on leaf yield but an extent positive effect on leaf nutritional quality due to their concentration increase in leaf B, Cu, starch, and soluble sugar. Meanwhile, (D + N)eCO2 decreased mulberry leaf yield and harvest of nutritious compounds for silkworm when compared with DeCO2. The reason may be associated to N, P, Mg, Fe, and Zn that are closely related to leaf pigment and N metabolism. Therefore, the rational application of mineral nutrient (especially N, P, Fe, Mg, and Zn) fertilizers is important for a sustainable mulberry production under future atmosphere CO2 concentrations.To elucidate the effect of light intensity on the cold response (5°C; 7 days) in Arabidopsis thaliana, we compared the following parameters under standard light (150 μmol m-2 s-1), low light (20 μmol m-2 s-1), and dark conditions membrane damage, photosynthetic parameters, cytokinin oxidase/dehydrogenase (CKX) activity, phytohormone levels, and transcription of selected stress- and hormone-related genes and proteome. The impact of cytokinins (CKs), hormones directly interacting with the light signaling pathway, on cold responses was evaluated using transformants overexpressing CK biosynthetic gene isopentenyl transferase (DEXIPT) or CK degradation gene HvCKX2 (DEXCKX) under a dexamethasone-inducible promoter. In wild-type plants, cold treatment under light conditions caused down-regulation of CKs (in shoots) and auxin, while abscisic acid (ABA), jasmonates, and salicylic acid (SA) were up-regulated, especially under low light. Cold treatment in the dark strongly suppressed all phytohormones, except ABA. DEXIP via ProteomeXchange, identifier PXD020480). Cold stress induced proteins involved in ABA and jasmonate metabolism, antioxidant enzymes, and enzymes of flavonoid and glucosinolate biosynthesis. DEXIPT plants up-regulated phospholipase D and MAP-kinase 4. Cold stress response at the proteome level was similar in all genotypes under optimal light intensity, differing significantly under low light. The data characterized the decisive effect of light-CK cross-talk in the regulation of cold stress responses.Adapting to the omnipresent gravitational field was a fundamental basis driving the flourishing of terrestrial plants on the Earth. Plants have evolved a remarkable capability that not only allows them to live and develop within the Earth's gravity field, but it also enables them to use the gravity vector to guide the growth of roots and shoots, in a process known as gravitropism. Triggered by gravistimulation, plant gravitropism is a highly complex, multistep process that requires many organelles and players to function in an intricate coordinated way. Although this process has been studied for several 100 years, much remains unclear, particularly the early events that trigger the relocation of the auxin efflux carrier PIN-FORMED (PIN) proteins, which presumably leads to the asymmetrical redistribution of auxin. In the past decade, the LAZY gene family has been identified as a crucial player that ensures the proper redistribution of auxin and a normal tropic response for both roots and shoots upon gravistimulation. LAZY proteins appear to be participating in the early steps of gravity signaling, as the mutation of LAZY genes consistently leads to altered auxin redistribution in multiple plant species. The identification and characterization of the LAZY gene family have significantly advanced our understanding of plant gravitropism, and opened new frontiers of investigation into the novel molecular details of the early events of gravitropism. Here we review current knowledge of the LAZY gene family and the mechanism modulated by LAZY proteins for controlling both roots and shoots gravitropism. We also discuss the evolutionary significance and conservation of the LAZY gene family in plants.Molecular characterization of genetically modified plants can provide crucial information for the development of detection and identification methods, to comply with traceability, and labeling requirements prior to commercialization. Detailed description of the genetic modification was previously a challenging step in the safety assessment, since it required the use of laborious and time-consuming techniques. In this study an accurate, simple, and fast method was developed for molecular characterization of genetically modified (GM) plants, following a user-friendly workflow for researchers with limited bioinformatic capabilities. Three GM events from a diverse array of crop species-perennial ryegrass, white clover, and canola-were used to test the approach that exploits long-read sequencing by the MinION device, from Oxford Nanopore Technologies. The method delivered a higher degree of resolution of the transgenic events within the host genome than has previously been possible with the standard Illumina short-range sequencing strategies. The flanking sequences, copy number, and presence of backbone sequences, and overall transgene insertion structure were determined for each of the plant genomes, with the additional identification of moderate-sized secondary insertions that would have previously been missed. The proposed workflow takes only about 1 week from DNA extraction to analyzed result, and the method will complement the existing approaches for molecular characterization of GM plants, since it makes the process faster, simpler, and more cost-effective.Low night temperature (LNT) causes environmental stress and has a severe and negative impact on plant growth and productivity. Synthetic elicitors can regulate plant growth and induce defense mechanisms from this type of stress. Here, we evaluated the effect of the exogenous growth regulator diethyl aminoethyl hexanoate (DA-6) in tomato leaf response to LNT stress. Our results showed that exogenous DA-6 activates the expression of chlorophyll synthesis and photosystem-related genes, and results in higher photosynthetic activity and chlorophyll production. Furthermore, DA-6 can regulate the synthesis of endogenous cytokinin (CTK) and the expression of decomposition genes to stabilize chloroplast structure, reduce oxidative damage, and maintain the photochemical activity of tomato leaves under LNT stress. DA-6 maintains a high level of ABA content and induces the expression of CBF genes, indicating that DA-6 may participate in the cold response signaling pathway and induce the expression of downstream low temperature response genes and accumulation of compatible osmolytes. This study unravels a mode of action by which plant growth regulators can improve low temperature tolerance and provides important considerations for their application to alleviate the harmful effects of cold stress.Powdery mildew (PM), caused by Podosphaera xanthii (Px), is one of the most devastating fungal diseases of melon worldwide. The use of resistant cultivars is considered to be the best and most effective approach to control this disease. In this study, an F2 segregating population derived from a cross between a resistant (wm-6) and a susceptible cultivar (12D-1) of melon was used to map major powdery mildew resistance genes using bulked segregant analysis (BSA), in combination with next-generation sequencing (NGS). A novel quantitative trait locus (QTL) named qCmPMR-12 for resistance to PM on chromosome 12 was identified, which ranged from 22.0 Mb to 22.9 Mb. RNA-Seq analysis indicated that the MELO3C002434 gene encoding an ankyrin repeat-containing protein was considered to be the most likely candidate gene that was associated with resistance to PM. Moreover, 15 polymorphic SNPs around the target area were successfully converted to Kompetitive Allele-Specific PCR (KASP) markers (P less then 0.0001). The novel QTL and candidate gene identified from this study provide insights into the genetic mechanism of PM resistance in melon, and the tightly linked KASP markers developed in this research can be used for marker-assisted selection (MAS) to improve powdery mildew resistance in melon breeding programs.