Fruit ripening depends on the accurate control of ripening-related genes expression, with histone deacetylases (HDACs) playing crucial roles in transcriptional regulation. However, the functions of HDACs in fruit maturation remain largely unexplored. Here, we show that SlHDA7 acts as a suppressor of fruit ripening and functions as an H4ac HDAC in tomato. Deletion of SlHDA7 accelerated fruit ripening, while overexpression of SlHDA7 delayed the maturation process. Additionally, ethylene production and carotenoid biosynthesis significantly increased in slhda7 mutant fruits but decreased in SlHDA7-overexpressing fruits. Furthermore, SlHDA7 repress the expression of ethylene production and signaling, carotenoid metabolism, cell wall modification, and transcriptional regulation-related genes. RT-qPCR and ChIP-qPCR analyses indicated that SlHDA7 may deacetylate H4ac, leading to reduced transcript levels of ACO1, GGPPS2, Z-ISO, EXP1, and XYL1 mRNA, consequently suppressing fruit ripening. Moreover, SlHDA7 suppresses fruit ripening by targeting specific ripening-associated transcription factors (TFs) like RIN, FUL1, and ERF.E1, ultimately leading to delayed ripening and prolonged fruit shelf life. In summary, our findings indicate that SlHDA7 negatively modulates tomato fruit maturation by adjusting H4ac levels of these ripening-associated genes and key TFs.
Both the phyllosphere and rhizosphere are inhabited by different kinds of microorganisms that are closely related to plant growth and health. However, it is not clear whether disease-resistant cultivars shape the microbiome to facilitate disease resistance. In this study, significant differences were found in the aboveground and belowground bacterial communities of disease-resistant and disease-susceptible cultivars grown in the same kiwifruit orchard. The phyllosphere of the resistant cultivar ‘Wanjin’ showed greater enrichment of Pseudomonas spp. and Sphingomonas spp. than the susceptible cultivar ‘Donghong’. The rhizosphere microbes of ‘Wanjin’ were less affected by field location, with significantly greater bacterial abundance than those of ‘Donghong’ and more bacteria with potential biocontrol properties. Pseudomonas syringae pv. actinidiae (Psa) infection significantly affected the microbiome of the phyllosphere of kiwifruit plants, especially that of ‘Donghong’. Resistant and susceptible kiwifruit cultivars exhibit distinct beneficial microbial recruitment strategies under Psa challenge. The phyllosphere of ‘Donghong’ in Jinzhai was enriched with Sphingomonas spp. and Pantoea spp. under Psa infection, while the rhizosphere of ‘Wanjin’ was enriched with Sphingomonas spp. and Novosphingobium spp. We further identified five key biomarkers within the microbial community associated with Psa infection. Inoculation experiments showed that Lysobacter sp. R34, Stenotrophomonas sp. R31, Pseudomonas sp. R10 and RS54, which were isolated from belowground compartments of ‘Wanjin’, could positively affect plant performance under Psa challenge. The combination use of Pseudomonas sp. R10 and Stenotrophomonas sp. R31 significantly improve the management of kiwifruit canker. Our findings provided novel insights into soil-microbe-plant interactions and the role of microbes in plant disease resistance and susceptibility.
Propagation through cuttings is a well-established and effective technique for plant multiplication. This study explores the regeneration of poplar roots using spatial transcriptomics to map a detailed developmental trajectory. Mapping of the time-series transcriptome data revealed notable alterations in gene expression during root development, particularly in the activation of cytokinin-responsive genes. Our analysis identified six distinct clusters during the second and third stages, each corresponding to specific anatomical regions with unique gene expression profiles. Auxin response cis-elements (AuxREs) were prevalent in the promoters of these cytokinin-responsive genes, indicating a regulatory interplay between auxin and cytokinin. Pseudo-temporal trajectory analysis mapped the differentiation from cambium cells to root primordium cells, revealing a complex pattern of cell differentiation. SAC56 and LOS1 emerged as potential novel biomarkers for enhancing root regeneration, with distinct spatial expression patterns confirmed by in situ hybridization. This comprehensive spatial analysis enhances our understanding of the molecular interactions driving root regeneration and provides insights for improving plant propagation techniques.
Asteraceae is the largest family of dicotyledons and includes Chrysanthemum and Helianthus, two important genera of ornamental plants. The genus Chrysanthemum consists of more than 30 species and contains many economically important ornamental, medicinal, and industrial plants. To more effectively promote Chrysanthemum research, we constructed the CGD, a Chrysanthemum genome database containing a large amount of data and useful tools. The CGD hosts well-assembled reference genome data for six Chrysanthemum species. These genomic data were fully annotated by comparison with various protein and domain data. Transcriptome data for nine different tissues, five flower developmental stages, and five treatments were subsequently added to the CGD. A fully functional ‘RNA data’ module was designed to provide complete and visual expression profile data. In addition, the CGD also provides many of the latest bioinformatics analysis tools, such as the efficient sgRNA search tool for Chrysanthemum. In conclusion, the CGD provides the latest, richest, and most complete multi-omics resources and powerful tools for Chrysanthemum. Collectively, the CGD will become the central gateway for Chrysanthemum genomics and genetic breeding research and will aid in the study of polyploid evolution.
Boosting plant immunity is an effective alternative to pesticides. However, environmental variations, accentuated by climate change, can compromise immunity. The robustness of a trait corresponds to the absence (or low level) of variation in that trait in the face of an environmental change. Here, we examined two types of robustness, robustness of immunity mean and robustness of immunity variation, and proposed nine quantitative robustness estimators. We characterized the immunity of a set of accessions representative of the natural diversity of pepper (Capsicum annuum L.), to two major pathogens: the oomycete Phytophthora capsici Leon. and potato virus Y. For each pathogen, we measured the immunity of accessions in two contrasting environments in terms of temperature. For each type of robustness and each pathogen, the impact of temperature change on immunity varied between accessions. The robustness estimators proved to be complementary and differed in terms of heritability and ability to discriminate accessions. A positive and significant correlation was observed between immunity and robustness. There was no significant relationship between the robustness of immunity to the two pathogens, but some accessions showed high immunity and robustness against both pathogens. These results justify the need to consider both immunity and robustness to environmental variations in order to select varieties adapted to current and future climate conditions. Phenotypic robustness should also be considered when assessing the “value of sustainable cultivation and use” of future plant varieties, particularly during the application process for protection rights granted from the European Community Plant Variety Office.
Soluble sugars contribute to the taste and flavor of citrus fruit. Potassium (K), known as a quality element, plays key roles in improving sugar accumulation and fruit quality, but the mechanism is largely unknown. This study aims to elucidate how K improves sugar accumulation by regulating carbon flow from source leaves to fruit in Newhall navel orange. We found that optimal fruit K concentrations around 1.5% improved sugar accumulation and fruit quality in citrus. K application increased the strength of both sink and source, as indicated by the increased fruit growth rate, enzyme activities and expression levels of key genes involved in sucrose (Suc) metabolism in fruit and leaf. K application also facilitated Suc transport from source leaves to fruit, as confirmed by the enhanced 13C-Suc level in fruit. Furthermore, we found that navel orange used the symplastic pathway for transporting Suc from source leaves to fruit, and K application enhanced symplastic loading, as demonstrated by the intensified carboxyfluorescein signal and increased plasmodesmata density in leaves. The findings reveal that K stimulates fruit sugar accumulation by increasing carbon flow from source leaves to fruit in Newhall navel orange.
Chloroplasts play a crucial role in essential processes, such as photosynthesis and the synthesis of primary and diverse secondary metabolites. Recent studies have also highlighted their significance linked to phytohormone production in plant immunity, especially SA and JA. Ubiquitination, a key posttranslational modification, usually leads to target protein degradation, which acts as a signal for remodeling the proteome via the induction of protein endocytosis or targeting to other membrane associated systems. Previously, the potato E3 ligase StRFP1 was shown to enhance resistance against Phytophthora infestans, but its mechanism remained unclear. Here, we demonstrate that StRFP1 interacted with the dually localized plastid glucose 6-phosphate transporter StGPT1 on the endoplasmic reticulum (ER). Transiently expressed StGPT1-GFP located on the chloroplast and ER in plant cells. Overexpression of StGPT1 enhances late blight resistance in potato and Nicotiana benthamiana, activates immune responses, including ROS bursts and up-regulation of PTI marker genes. The resistance function of StGPT1 seems to be related to its dual localization. Remarkably, StRFP1 ubiquitinates StGPT1 at the ER, possibly due to its merely transient function in peroxisomes, leading to apparent accumulation in chloroplasts. Our findings point to a novel mechanism by which a plant E3 ligase contributes to immunity via interacting with dually targeted GPT1 at the ER of plant cells.
Phosphorus (P) is the macronutrients essential for the development and growth of plants, but how external inorganic phosphate (Pi) level and signaling affect tea plant growth and characteristic secondary metabolite biosynthesis are not understood. Theanine is major secondary metabolites, and its contents largely determine tea favor and nutrition qualities. Here, we found theanine contents in tea leaves and roots declined as Pi concentration increased in tea plants after Pi feeding. The transcriptome analysis of global gene expression in tea leaves under Pi feeding suggested a wide range of genes involved in Pi/N transport and responses were altered. Among them, CsSPX3 and CsPHL7 transcript levels in response to Pi feeding to tea plants, their expression patterns were generally opposite to these of major theanine biosynthesis genes, indicating possible regulatory correlations. Biochemical analyses showed that CsSPX3 interacted with CsPHL7, and CsPHL7 negatively regulated theanine biosynthesis genes CsGS1 and CsTS1. Meanwhile, VIGS and transient overexpression systems in tea plants verified the functions of CsSPX3 and CsPHL7 in mediating Pi-feeding-repressed theanine biosynthesis. This study offers fresh insights into the regulatory mechanism underlying Pi repression of theanine biosynthesis, and the CsSPX3-CsPHL7-CsGS1/CsTS1 module plays a role in high Pi inhibition of theanine production in tea leaves. It has an instructional significance for guiding the high-quality tea production in tea garden fertilization.
Rose (Rosa rugosa ) petals are rich in diverse secondary metabolites, which have important physiological functions as well as great economic values. Currently, it remains unclear how saline and/or alkaline stress(es) influence the accumulation of secondary metabolites in rose. In this study, we analyzed the transcriptome and metabolite profiles of rose petals under aline-alkali stress and uncovered the induction mechanism underlying major metabolites. Dramatic changes were observed in the expression of 1363 genes and the abundances of 196 metabolites in petals in response to saline-alkali stress. These differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) are mainly associated with flavonoid and terpenoid metabolism and the reconstruction of cell walls. Of them, TERPENE SYNTHASE 31 (TPS31 ) overexpression in tobacco leaves driven by its own promoter resulted in significant alterations in the levels of diverse terpenoids, which were differentially influenced by saline-alkali stress. An integrated analysis of metabolomic and transcriptomic data revealed a high correlation between the abundances of flavonoids/terpenoids and the expression of the transcription factor MYB5. MYB5 may orchestrate the biosynthesis of sesquiterpenoids and proanthocyanidins through direct regulation of TPS31 and ANR expression under aline-alkali stress. Our finding facilitates improving the bioactive substance accumulation of rose petals by metabolic engineering.
In cannabis seedlings, the initiation of solitary flowers is photoperiod-independent. However, when cannabis reaches the adult stage, short-day photoperiod (SD) triggers branching of the shoot apex and a reduction in internode length, leading to development of a condensed inflorescence. We demonstrate that SD affects cannabis plants in two distinct phases: the first includes rapid elongation of the internodes and main stem,and occurring from Day 5 to Day 10 of plant cultivation under SD; in the second phase,elongation of newly developed internodes ceases, and a condensed inflorescence is formed. Exposure of plants to alternating photoperiods revealed that inflorescence onset requires at least three consecutive days of SD, and SD is consistently required throughout inflorescence maturation to support its typical condensed architecture. This photoperiod-dependent morphogenesis was associated with a decrease in gibberellin (GA4) and auxin levels in the shoot apex. Reverting the plants to a long-day photoperiod (LD) increased GA4 and auxin levels, leading to inflorescence disassembly, internode elongation, and subsequent resumption of LD growth patterns. Similar developmental patterns were observed under SD following the application of exogenous GA (and not auxin), which also impeded inflorescence development. Nevertheless, additional studies will help to further evaluate auxin’s role in these developmental changes. We propose a crucial role for GA in sexual reproduction and inflorescence development in female cannabis by mediating photoperiod signaling in the inflorescence tissues.
Camelina (Camelina sativa), an allohexaploid species, is an emerging aviation biofuel crop that has been the focus of resurgent interest in recent decades. To guide future breeding and crop improvement efforts, the community requires a deeper comprehension of subgenome dominance, often noted in allopolyploid species, “alongside an understanding of the genetic diversity” and population structure of material present within breeding programs. We conducted population genetic analyses of a C. sativa diversity panel, leveraging a new genome, to estimate nucleotide diversity and population structure, and analyzed for patterns of subgenome expression dominance among different organs. Our analyses confirm that C. sativa has relatively low genetic diversity and show that the SG3 subgenome has substantially lower genetic diversity compared to the other two subgenomes. Despite the low genetic diversity, our analyses identified 13 distinct subpopulations including two distinct wild populations and others putatively representing founders in existing breeding populations. When analyzing for subgenome composition of long non-coding RNAs, which are known to play important roles in (a)biotic stress tolerance, we found that the SG3 subgenome contained significantly more lincRNAs compared to other subgenomes. Similarly, transcriptome analyses revealed that expression dominance of SG3 is not as strong as previously reported and may not be universal across all organ types. From a global analysis, SG3 “was only significant higher expressed” in flower, flower bud, and fruit organs, which is an important discovery given that the crop yield is associated with these organs. Collectively, these results will be valuable for guiding future breeding efforts in camelina.
Next-generation sequencing (NGS) library construction often requires high-quality DNA extraction, precise adjustment of DNA concentration, and restriction enzyme digestion to reduce genome complexity, which results in increased time and cost in sample preparation and processing. To address these challenges, a PCR-based method for rapid NGS library preparation, named dpMIG-seq, has been developed and proven effective for high-throughput genotyping. However, the application of dpMIG-seq has been limited to diploid and polyploid species with disomic inheritance. In this study, we obtained genome-wide single nucleotide polymorphism (SNP) markers for tetraploid blueberry to evaluate genotyping and downstream analysis outcomes. Comparison of genotyping qualities inferred across samples with different DNA concentrations and multiple bioinformatics approaches revealed high accuracy and reproducibility of dpMIG-seq-based genotyping, with Pearson’s correlation coefficients between replicates in the range of 0.91 to 0.98. Furthermore, we demonstrated that dpMIG-seq enables accurate genotyping of samples with low DNA concentrations. Subsequently, we applied dpMIG-seq to a tetraploid F1 population to examine the inheritance probability of parental alleles. Pairing configuration analysis supported the random meiotic pairing of homologous chromosomes on a genome-wide level. On the other hand, preferential pairing was observed on chr-11, suggesting that there may be an exception to the random pairing. Genotypic data suggested quadrivalent formation within the population, although the frequency of quadrivalent formation varied by chromosome and cultivar. Collectively, the results confirmed applicability of dpMIG-seq for allele dosage genotyping and are expected to catalyze the adoption of this cost-effective and rapid genotyping technology in polyploid studies.
SENSITIVE TO PROTON RHIZOTOXICITY 1 (STOP1) is a core transcription factor that regulates the expression of aluminum (Al) resistance genes to manage Al toxicity in plants. However, the genome-wide roles of SlSTOP1 in the Al stress response of tomato (Solanum lycopersicum) remain largely unknown. Here, we report that SlSTOP1 is crucial for Al tolerance in tomato, as loss-of-function mutants of SlSTOP1 displayed hypersensitivity to Al stress. Aluminum stress had no effect on SlSTOP1 mRNA expression, but promoted accumulation of SlSTOP1 protein in the nucleus. Through integrated DNA affinity purification sequencing and RNA sequencing analysis, we identified 39 SlSTOP1-targeted Al-responsive genes, some of which are homologous to known Al resistance genes in other plant species, suggesting that these SlSTOP1-targeted genes play essential roles in Al resistance in tomato. Furthermore, using peak enrichment analysis of SlSTOP1-targeted sequences, we identified a cis-acting element bound by SlSTOP1 and validated this finding via dual-luciferase reporter and electrophoretic mobility shift assay (EMSA). Additionally, we demonstrated SlHAK5 is one of direct targets of SlSTOP1 and functionally characterized it in terms of Al stress tolerance. Compared with wild-type plants, Slhak5 mutants developed by CRISPR/Cas9 technology presented increased sensitivity to Al stress, which was associated with reduced citrate secretion from the roots. Together, our findings demonstrate that SlSTOP1 directly interacts with cis-acting elements located in the promoters of target genes involved in diverse pathways contributing to Al resistance in tomato.