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Trichomes are specialized hair-like structures in the epidermal cells of the above-ground parts of plants and help to protect them from pests and pathogens, and produce valuable metabolites. Chrysanthemum morifolium , which is used in tea products, has both ornamental and medicinal value; however, it is susceptible to infection by the fungus Alternaria alternata , which can result in substantial economic losses. Increasing the density of glandular trichomes enhances disease resistance and improves the production of medicinal metabolites in chrysanthemums, and jasmonate (JA) is known to promote the formation of trichomes in various plants. However, it remains unclear whether glandular trichomes in chrysanthemums are regulated by JA. In addition, grafting, a technique that can improve plant resistance to biotic stresses, has been poorly examined for its impact on glandular trichomes, terpenoids, and disease resistance. In this study, we demonstrate that grafting with Artemisia vulgaris rootstocks improves the resistance of chrysanthemum scions to A. alternata. Heterografted chrysanthemums exhibited higher trichome density and terpenoid content compared to self-grafted counterparts. Transcriptome analysis highlighted the significant role of CmJAZ1-like in disease resistance in heterografted chrysanthemums. Lines overexpressing CmJAZ1-like exhibited sensitivity to A. alternata , and this was characterized by reduced glandular trichome density and limited terpenoid content. Conversely, CmJAZ1-like silenced lines exhibited resistance to A. alternata and showed increased glandular trichome density and terpenoid content. Higher JA content was found in the heterografted chrysanthemum scions compared to self-grafted ones. Furthermore, we established that JA promoted the development of glandular trichomes and the synthesis of terpenoids while also inducing the degradation of CmJAZ1-like proteins in chrysanthemums. Our findings suggest that higher JA increases trichome density and terpenoid content, thereby enhancing resistance to A. alternata by regulating CmJAZ1-like in heterografted chrysanthemums. [ABSTRACT FROM AUTHOR]

Ethylene-responsive factors (ERF) play an important role in plant responses to waterlogging stress. However, the function and mechanism of action of ERFVIII in response to waterlogging stress remain poorly understood. In this study, we found that expression of the ERF VIIIa gene CmERF4 in chrysanthemum was induced by waterlogging stress. CmERF4 localized to the nucleus when expressed in tobacco leaves. Yeast two-hybrid and luciferase assays showed that CmERF4 is a transcriptional inhibitor. CmERF4 overexpression in chrysanthemum reduced plant waterlogging tolerance, whereas overexpression of the chimeric activator CmERF4 -VP64 reversed its transcriptional activity, promoting higher waterlogging tolerance than that observed in wild-type plants, indicating that CmERF4 negatively regulates waterlogging tolerance. Transcriptome profiling showed that energy metabolism and reactive oxygen species (ROS) pathway-associated genes were differentially expressed between CmERF4 -VP64 and wild-type plants. RT–qPCR analysis of selected energy metabolism and reactive oxygen species-related genes showed that the gene expression patterns were consistent with the expression levels obtained from RNA-seq analysis. Overall, we identified new functions of CmERF4 in negatively regulating chrysanthemum waterlogging tolerance by modulating energy metabolism and ROS pathway genes. [ABSTRACT FROM AUTHOR]

The floral transition from vegetative to reproductive growth is pivotal in the plant life cycle. NUTRITION RESPONSE AND ROOT GROWTH (OsNRRa), as a CONSTANS, CONSTANS-LIKE, TOC1 (CCT) domain protein, delays flowering in rice, and an orthologous protein, CmNRRa, inhibits flowering in chrysanthemum; however, the underlying mechanism remains unknown. In this study, using yeast two-hybrid screening, we identified the 14-3-3 protein family member Cm14-3-3µ as a CmNRRa-interacting protein. A combination of bimolecular fluorescence complementation, pull-down, and co-immunoprecipitation assays was performed to confirm the physical interaction between CmNRRa and Cm14-3-3µ. In addition, expression analysis showed that CmNRRa but not Cm14-3-3µ responded to the diurnal rhythm, whereas both genes were highly expressed in leaves. Moreover, the function of Cm14-3-3µ in flowering time regulation was similar to that of CmNRRa. Furthermore, CmNRRa repressed chrysanthemum FLOWERING LOCUS T - like 3 (CmFTL3) and an APETALA 1 (AP1)/ FRUITFULL (FUL)-like gene (CmAFL1) but induced TERMINAL FLOWER1 (CmTFL1) directly by binding to their promoters. Cm14-3-3µ enhanced the ability of CmNRRa to regulate the expression of these genes. These findings suggest that there is a synergistic relationship between CmNRRa and Cm14-3-3µ in flowering repression in chrysanthemum. [ABSTRACT FROM AUTHOR]

Melatonin, the most widely distributed hormone in nature, plays important roles in plants. Many physiological processes in plants are linked to melatonin, including seed germination, anisotropic cell growth, and senescence. Compared with animals, different plants possess diverse melatonin biosynthetic pathways and regulatory networks. Whereas melatonin biosynthesis in animals is known to be regulated by ambient signals, little is known about how melatonin biosynthesis in plants responds to environmental signals. Plants are affected by numerous environmental factors, such as light, temperature, moisture, carbon dioxide, soil conditions, and nutrient availability at all stages of development and in different tissues. Melatonin content exhibits dynamic changes that affect plant growth and development. Melatonin plays various species-specific roles in plant responses to different environmental conditions. However, much remains to be learned, as not all environmental factors have been studied, and little is known about the mechanisms by which these factors influence melatonin biosynthesis. In this review, we provide a detailed, systematic description of melatonin biosynthesis and signaling and of the roles of melatonin in plant responses to different environmental factors, providing a reference for in-depth research on this important issue. [ABSTRACT FROM AUTHOR]

Stem mechanical strength is one of the most important agronomic traits that affects the resistance of plants against insects and lodging, and plays an essential role in the quality and yield of plants. Several transcription factors regulate mechanical strength in crops. However, mechanisms of stem strength formation and regulation remain largely unexplored, especially in ornamental plants. In this study, we identified an atypical bHLH transcription factor CmHLB (HLH PROTEIN INVOLVED IN LIGNIN BIOSYNTHESIS) in chrysanthemum, belonging to a small bHLH sub-family — the PACLOBUTRAZOL RESISTANCE (PRE) family. Overexpression of CmHLB in chrysanthemum significantly increased mechanical strength of the stem, cell wall thickness, and lignin content, compared with the wild type. In contrast, CmHLB RNA interference lines exhibited the opposite phenotypes. RNA-seq analysis indicated that CmHLB promoted the expression of genes involved in lignin biosynthesis. Furthermore, we demonstrated that CmHLB interacted with Chrysanthemum KNOTTED ARABIDOPSIS THALIANA7 (CmKNAT7) through the KNOX2 domain, which has a conserved function, i.e. it negatively regulates secondary cell wall formation of fibres and lignin biosynthesis. Collectively, our results reveal a novel role for CmHLB in regulating lignin biosynthesis by interacting with CmKNAT7 and affecting stem mechanical strength in Chrysanthemum. [ABSTRACT FROM AUTHOR]

Flavones are among the major colorless pigments synthesized through branches of the flavonoid pathway in plants. However, due to the absence of a gene encoding flavone synthase (FNS) in the model plant Arabidopsis thaliana species, the regulatory mechanism of FNS-catalyzed flavone biosynthesis has rarely been studied in plants. Here, it was found that flavones play a predominant role in the elimination of excess reactive oxygen species (ROS) at high temperatures in colorless plant organs. A novel atypical subgroup 7 (SG7) R2R3-MYB transcription factor, CmMYB012, was found to be induced in response to prolonged high temperatures and to inhibit flavone biosynthesis by directly regulating CmFNS. Moreover, CmMYB012 was also found to inhibit anthocyanin biosynthesis by suppressing the expression of CmCHS, CmDFR, CmANS, and CmUFGT. CmMYB012 overexpression exerted a negative influence on plant fitness and pink flower color formation, while CmMYB012 suppression had the opposite effect in response to high temperatures. Our findings provide new insights into the mechanisms by which high temperatures regulate the metabolism of flavones and anthocyanins to affect plant fitness and flower color formation. [ABSTRACT FROM AUTHOR]

The Mildew Resistance Locus O (MLO) gene family has been investigated in many species. However, there are few studies on chrysanthemum MLO genes. We report in this study that CmMLO17 in Chrysanthemum morifolium was upregulated after Alternaria alternata infection. Silencing of CmMLO17 by artificial microRNA resulted in reduced susceptibility of chrysanthemum to A. alternata infection. Genes in the abscisic acid (ABA) and Ca2+ signaling pathways were upregulated in the CmMLO17-silenced line R20 compared to the wild-type plants. We speculated that CmMLO17-silenced plants had a faster and stronger defense response that was mediated by the ABA and Ca2+ signaling pathways, resulting in reduced susceptibility of chrysanthemum to A. alternata infection. In addition, a candidate gene, CmKIC, that may interact with CmMLO17 was discovered by the yeast two-hybrid assay. The interaction between CmMLO17 and CmKIC was confirmed using the yeast two-hybrid assay and bimolecular fluorescence complementation (BiFC) analysis. CmMLO17 and CmKIC were both located on the plasma membrane, and CmKIC was also located on the nucleus. CmKIC overexpression increased the susceptibility of chrysanthemum to A. alternata, whereas CmKIC silencing resulted in reduced susceptibility. Therefore, CmMLO17 and CmKIC may work together in C. morifolium to support the growth of A. alternata. The results of this study will provide insight into the potential function of MLO and improve the understanding of plant defense responses to necrotrophic pathogens. [ABSTRACT FROM AUTHOR]

The CmBBX8-CmFTL1 regulatory module is a key determinant in the transition from vegetative growth to reproductive development in summer-flowering chrysanthemum. However, the detailed regulatory mechanism of CmBBX8-mediated flowering remains elusive. In this study, we revealed that RADICAL-INDUCED CELL DEATH 1 (CmRCD1) physically associated with CmBBX8 through bimolecular fluorescence complementation (BiFC), pulldown and Coimmunoprecipitation (CoIP) assays. Furthermore, the RCD1-SRO1-TAF4 (RST) domain of CmRCD1 and the B-box of CmBBX8 mediated their interaction. In addition, Luciferase (LUC) assays and electrophoretic mobility shift assay (EMSAs) showed that CmRCD1 repressed the transcriptional activity of CmBBX8 and interfered with its binding to the CmFTL1 promoter, thereby leading to delayed flowering in the summer chrysanthemum 'Yuuka'. These results provide insight into the molecular framework of CmRCD1-CmBBX8-mediated flowering in chrysanthemum. [ABSTRACT FROM AUTHOR]

Chrysanthemum (Chrysanthemum morifolium) is an ideal model species for studying petal morphogenesis because of the diversity in the flower form across varieties; however, the molecular mechanisms underlying petal development are poorly understood. Here, we show that the brassinosteroid transcription factor BRI1-EMS-SUPPRESSOR 1 (CmBES1) in chrysanthemum (C. morifolium cv. Jinba) is important for organ boundary formation because it represses organ boundary identity genes. Chrysanthemum plants overexpressing CmBES1 displayed increased fusion of the outermost ray florets due to the loss of differentiation of the two dorsal petals, which developed simultaneously with the ventral petals. RNA-seq analysis of the overexpression lines revealed potential genes and pathways involved in petal development, such as CUP-SHAPED COTYLEDON (CUC2), CYCLOIDEA 4 (CYC4), genes encoding MADS-box transcription factors and homeodomain-leucine zippers (HD-Zips) and auxin pathway-related genes. This study characterizes the role of CmBES1 in ray floret development by its modulation of flower development and boundary identity genes in chrysanthemum. [ABSTRACT FROM AUTHOR]

Chrysanthemum is frequently attacked by aphids, which greatly hinders the growth and ornamental value of this plant species. WRKY transcription factors play an important role in the response to biotic stresses such as pathogen and insect stresses. Here, chrysanthemum CmWRKY53 was cloned, and its expression was induced by aphid infestation. To verify the role of CmWRKY53 in resistance to aphids, CmWRKY53 transgenic chrysanthemum was generated. CmWRKY53 was found to mediate the susceptibility of chrysanthemum to aphids. The expression levels of secondary metabolite biosynthesis genes, such as peroxidase- and polyphenol oxidase-encoding genes, decreased in CmWRKY53-overexpressing (CmWRKY53-Oe) plants but dramatically increased in chimeric dominant repressor (CmWRKY53-SRDX) plants, suggesting that CmWRKY53 contributes to the susceptibility of chrysanthemum to aphids, possibly due to its role in the regulation of secondary metabolites. [ABSTRACT FROM AUTHOR]

The switch from vegetative growth to reproductive growth is a key event in the development of a plant. Here, the product of the chrysanthemum gene CmMYB2, an R2R3 MYB transcription factor that is localized in the nucleus, was shown to be a component of the switching mechanism. Plants engineered to overexpress CmMYB2 flowered earlier than did wild-type plants, while those in which CmMYB2 was suppressed flowered later. In both the overexpression and RNAi knockdown plants, a number of genes encoding proteins involved in gibberellin synthesis or signaling, as well as in the response to photoperiod, were transcribed at a level that differed from that in the wild type. Both yeast two-hybrid and bimolecular fluorescence complementation assays revealed that CmMYB2 interacts with CmBBX24, a zinc-finger transcription factor known to regulate flowering by its influence on gibberellin synthesis. [ABSTRACT FROM AUTHOR]

Fog computing has become an emerging environment that provides data storage, computing and some other services on the edge of network. It not only can acquire data from terminal devices, but also can provide computing services to users by opening computing resources. Compared with cloud computing, fog devices can collaborate to provide users with powerful computing services through resource allocation. However, as many of fog devices are not monitored, there are some security problems. For example, since fog server processes and maintains user information, device information, task parameters and so on, fog server is easy to perform illegal resource allocation for extra benefits. In this paper, we propose a secure computing resource allocation framework for open fog computing. In our scheme, the fog server is responsible for processing computing requests and resource allocations, and the cloud audit center is responsible for auditing the behaviors of the fog servers and fog nodes. Based on the proposed security framework, our proposed scheme can resist the attack of single malicious node and the collusion attack of fog server and computing devices. Furthermore, the experiments show our proposed scheme is efficient. For example, when the number of initial idle service devices is 40, the rejection rate of allocated tasks is 10% and the total number of sub-tasks is changed from 150 to 200, the total allocation time of our scheme is only changed from 15 ms to 25 ms; additionally, when the task of 5000 order matrix multiplication is tested on 10 service devices, the total computing time of our scheme is |$\sim$| 250 s, which is better than that of single computer (where single computer needs more than 1500 s). Therefore, our proposed scheme has obvious advantages when it faces some tasks that require more computational cost, such as complex scientific computing, distributed massive data query, distributed image processing and so on. [ABSTRACT FROM AUTHOR]

The Asteraceae family is at the forefront of the evolution due to frequent hybridization. Hybridization is associated with the induction of widespread genetic and epigenetic changes and has played an important role in the evolution of many plant taxa. We attempted the intergeneric cross Chrysanthemum morifolium × Leucanthemum paludosum. To obtain the success in cross, we have to turn to ovule rescue. DNA profiling of the amphihaploid and amphidiploid was investigated using amplified fragment length polymorphism, sequence-related amplified polymorphism, start codon targeted polymorphism, and methylation-sensitive amplification polymorphism (MSAP). Hybridization induced rapid changes at the genetic and the epigenetic levels. The genetic changes mainly involved loss of parental fragments and gaining of novel fragments, and some eliminated sequences possibly from the noncoding region of L. paludosum. The MSAP analysis indicated that the level of DNA methylation was lower in the amphiploid (∼45%) than in the parental lines (51.5–50.6%), whereas it increased after amphidiploid formation. Events associated with intergeneric genomic shock were a feature of C. morifolium × L. paludosum hybrid, given that the genetic relationship between the parental species is relatively distant. Our results provide genetic and epigenetic evidence for understanding genomic shock in wide crosses between species in Asteraceae and suggest a need to expand our current evolutionary framework to encompass a genetic/epigenetic dimension when seeking to understand wide crosses. [ABSTRACT FROM AUTHOR]

The kinesins are a family of microtubule-based motor proteins that move directionally along microtubules and are involved in many crucial cellular processes, including cell elongation in plants. Less is known about kinesins directly regulating gene transcription to affect cellular physiological processes. Here, we describe a rice (Oryza sativa) mutant, gibberellin   -deficient dwarf1 (gdd1), that has a phenotype of greatly reduced length of root, stems, spikes, and seeds. This reduced length is due to decreased cell elongation and can be rescued by exogenous gibberellic acid (GA3) treatment. GDD1 was cloned by a map-based approach, was expressed constitutively, and was found to encode the kinesin-like protein BRITTLE CULM12 (BC12). Microtubule cosedimentation assays revealed that BC12/GDD1 bound to microtubules in an ATP-dependent manner. Whole-genome microarray analysis revealed the expression of ent -kaurene oxidase (KO2), which encodes an enzyme involved in GA biosynthesis, was downregulated in gdd1. Electrophoretic mobility shift and chromatin immunoprecipitation assays revealed that GDD1 bound to the element ACCAACTTGAA in the KO2 promoter. In addition, GDD1 was shown to have transactivation activity. The level of endogenous GAs was reduced in gdd1 , and the reorganization of cortical microtubules was altered. Therefore, BC12/GDD1, a kinesin-like protein with transcription regulation activity, mediates cell elongation by regulating the GA biosynthesis pathway in rice. [ABSTRACT FROM AUTHOR]

Through sos3 (salt overly sensitive 3) suppressor screening, two allelic suppressor mutants that are weak alleles of the strong sos3 suppressor sos3hkt1-1 were recovered. Molecular characterization identified T-DNA insertions in the distal promoter region of the Arabidopsis thaliana HKT1 (AtHKT1, At4g10310) in these two weak sos3 suppressors, which results in physical separation of a tandem repeat from the proximal region of the AtHKT1 promoter. The tandem repeat is approximately 3.9 kb upstream of the ATG start codon and functions as an enhancer element to promote reporter gene expression. A putative small RNA target region about 2.6 kb upstream of the ATG start codon is heavily methylated. CHG and CHH methylation but not CG methylation is significantly reduced in the small RNA biogenesis mutant rdr2, indicating that non-CG methylation in this region is mediated by small RNAs. Analysis of AtHKT1 expression in rdr2 suggests that non-CG methylation in the putative small RNA target region represses AtHKT1 expression in shoots. The DNA methylation-deficient mutant met1-3 has nearly complete loss of total cytosine methylation in the putative small RNA target region and is hypersensitive to salt stress. The putative small RNA target region and the tandem repeat are essential for maintaining AtHKT1 expression patterns crucial for salt tolerance. [ABSTRACT FROM AUTHOR]

Cultivated chrysanthemum (Chrysanthemum morifolium Ramat.) is an economically important ornamental plant species grown worldwide. However, the origin of the genus Chrysanthemum remains unclear. This study was conducted in the Hengduan Mountains, Yunnan Province. We took advantage of a special geographic region where the southernmost species of Ajania and the highest altitude population of Chrysanthemum indicum coexist to investigate their evolutionary origins. Diversity analysis of 9 populations of 5 species that came from 3 genera was carried out based on morphological traits and SRAP markers. Furthermore, topographical and ecological analyses and surveys of the vegetation communities in the plots were carried out for correlation analysis, and past data were used to reconstruct the ancient topography and vegetation to estimate the migration path and divergence time. We found that Chrysanthemum and Ajania were closely related based on the smooth transition states among marginal female florets and their common pollination system. The genetic relationship between Phaeostigma and Chrysanthemum was relatively distant, and Ajania was between them. Low light intensity and relatively humid habitats may be driving the elongation and evolution of marginal female florets. We found that Chrysanthemum and related genera were largely restricted to stony topographies at an altitude of ~3000 m.a.s.l. and in specialized alpine coniferous (Pinus) and broad-leaved (Quercus) mixed forest marginal communities. These stony topographies have become ecological islands of refuge for these species in the current interglacial period. The Hengduan Mountains play a key role in the evolution, divergence, and survival of Chrysanthemum and its allies. [ABSTRACT FROM AUTHOR]

Background Human babesiosis is a common zoonosis caused by Babesia and is attracting an increasing concern worldwide. The natural course of babesiosis infection and how the human immune system changes during the course of babesiosis infection are not clear. Methods We followed up 1 case infected with Babesia venatorum for 5 years. The patient was immune-intact and received no standard treatment. Clinical data were obtained from medical records. Microbiological tests, ribonucleic acid (RNA) sequence, and serum cytokines and chemokines were detected at different time points. Results The patient was confirmed as B venatorum infection based on his tick-bite history, clinical manifestations, and positive results of microbiological tests. The parasitemia of the patient persisted for approximately 2 months. With flu-like symptoms aggravating, most cytokines and chemokines in RNA and protein levels increased progressively and reached the peak when fever occurred; and their concentrations decreased to baseline during the same time as clearance of babesia parasites. Conclusions Babesia venatorum infection could take a mild self-limited course in immune-intact individuals. The natural changes of most cytokines and chemokines demonstrated very similar trends, which correlated with blood parasitemia and clinical manifestations. Cytokine profiles involving multiple inflammatory cytokines might be a good indicator of babesia infection. [ABSTRACT FROM AUTHOR]

Chrysanthemum (Chrysanthemum morifolium Ramat.) is a leading flower with applied value worldwide. Developing new chrysanthemum cultivars with novel characteristics such as new flower colors and shapes, plant architectures, flowering times, postharvest quality, and biotic and abiotic stress tolerance in a time- and cost-efficient manner is the ultimate goal for breeders. Various breeding strategies have been employed to improve the aforementioned traits, ranging from conventional techniques, including crossbreeding and mutation breeding, to a series of molecular breeding methods, including transgenic technology, genome editing, and marker-assisted selection (MAS). In addition, the recent extensive advances in high-throughput technologies, especially genomics, transcriptomics, proteomics, metabolomics, and microbiomics, which are collectively referred to as omics platforms, have led to the collection of substantial amounts of data. Integration of these omics data with phenotypic information will enable the identification of genes/pathways responsible for important traits. Several attempts have been made to use emerging molecular and omics methods with the aim of accelerating the breeding of chrysanthemum. However, applying the findings of such studies to practical chrysanthemum breeding remains a considerable challenge, primarily due to the high heterozygosity and polyploidy of the species. This review summarizes the recent achievements in conventional and modern molecular breeding methods and emerging omics technologies and discusses their future applications for improving the agronomic and horticultural characteristics of chrysanthemum. [ABSTRACT FROM AUTHOR]

MYB transcription factors are widely involved in the development of and physiological processes in plants. Here, we isolated the chrysanthemum R2R3-MYB family transcription factor CmMYB15, a homologous gene of AtMYB15. It was demonstrated that CmMYB15 expression was induced by aphids and that CmMYB15 could bind to AC elements, which usually exist in the promoter of lignin biosynthesis genes. Overexpression of CmMYB15 in chrysanthemum enhanced the resistance of aphids. Additionally, the content of lignin and the expression of several lignin biosynthesis genes increased. In summary, the results indicate that CmMYB15 regulates lignin biosynthesis genes that enhance the resistance of chrysanthemum to aphids. [ABSTRACT FROM AUTHOR]