RESUMO
Potato (Solanum tuberosum L.) is cultivated worldwide for its underground tubers, which provide an important part of human nutrition and serve as a model system for belowground storage organ formation. Similar to flowering, stolon-expressed FLOWERING LOCUS T-like (FT-like) protein SELF-PRUNING 6A (StSP6A) plays an instrumental role in tuberization by binding to the bZIP transcription factors StABI5-like 1 (StABL1) and StFD-like 1 (StFDL1), causing transcriptional reprogramming at the stolon subapical apices. However, the molecular mechanism regulating the widely conserved FT-bZIP interactions remains largely unexplored. Here, we identified a TCP transcription factor StAST1 (StABL1 and StSP6A-associated TCP protein 1) binding to both StSP6A and StABL1. StAST1 is specifically expressed in the vascular tissue of leaves and developing stolons. Silencing of StAST1 leads to accelerated tuberization and a shortened life cycle. Molecular dissection reveals that the interaction of StAST1 with StSP6A and StABL1 attenuates the formation of the alternative tuberigen activation complex (aTAC). We also observed StAST1 directly activates the expression of potato GA 20-oxidase gene (StGA20ox1) to regulate GA responses. These results demonstrate StAST1 functions as a tuberization repressor by regulating plant hormone levels; our findings also suggest a mechanism by which the widely conserved FT-FD genetic module is fine-tuned.
Assuntos
Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Tubérculos , Solanum tuberosum , Fatores de Transcrição , Solanum tuberosum/genética , Solanum tuberosum/metabolismo , Solanum tuberosum/fisiologia , Solanum tuberosum/crescimento & desenvolvimento , Tubérculos/genética , Tubérculos/crescimento & desenvolvimento , Tubérculos/metabolismo , Tubérculos/fisiologia , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genéticaRESUMO
Photoperiod plays a critical role in controlling the formation of sexual or vegetative reproductive organs in potato. Although StPHYF-silenced plants overcome day-length limitations to tuberize through a systemic effect on tuberigen StSP6A expression in the stolon, the comprehensive regulatory network of StPHYF remains obscure. Therefore, the present study investigated the transcriptomes of StPHYF-silenced plants and observed that, in addition to known components of the photoperiodic tuberization pathway, florigen StSP3D and other flowering-related genes were activated in StPHYF-silenced plants, exhibiting an early flowering response. Additionally, grafting experiments uncovered the long-distance effect of StPHYF silencing on gene expression in the stolon, including the circadian clock components, flowering-associated MADSs, and tuberization-related regulatory genes. Similar to the AtFT-AtAP1 regulatory module in Arabidopsis, the present study established that the AP1-like StMADS1 functions downstream of the tuberigen activation complex (TAC) and that suppressing StMADS1 inhibits tuberization in vitro and delays tuberization in vivo. Moreover, the expression of StSP6A was downregulated in StMADS1-silenced plants, implying the expression of StSP6A may be feedback-regulated by StMADS1. Overall, these results reveal that the regulatory network of StPHYF controls flowering and tuberization and targets the crucial tuberization factor StMADS1 through TAC, thereby providing a better understanding of StPHYF-mediated day-length perception during potato reproduction.
Assuntos
Arabidopsis , Fitocromo , Solanum tuberosum , Fitocromo/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Solanum tuberosum/metabolismo , Transcriptoma , Tubérculos/metabolismo , Folhas de Planta/metabolismo , Fotoperíodo , Arabidopsis/genética , Reprodução , Regulação da Expressão Gênica de Plantas/genéticaRESUMO
Transitory starch and vacuolar sugars function as highly dynamic pools of instantly accessible metabolites in plant leaf cells. Their metabolic regulation is critical for plant survival. The tonoplast sugar transporters (TSTs), responsible for sugar uptake into vacuoles, regulate cellular sugar partitioning and vacuolar sugar accumulation. However, whether TSTs are involved in leaf transient starch turnover and plant growth is unclear. Here, we found that suppressing StTST3.1 resulted in growth retardation and pale green leaves in potato plants. StTST3.1-silenced plants displayed abnormal chloroplasts and impaired photosynthetic performance. The subcellular localization assay and the oscillation expression patterns revealed that StTST3.1 encoded a tonoplast-localized protein and responded to photoperiod. Moreover, RNA-seq analyses identified that starch synthase (SS2 and SS6) and glucan water, dikinase (GWD), were downregulated in StTST3.1-silenced lines. Correspondingly, the capacity for starch synthesis and degradation was decreased in StTST3.1-silenced lines. Surprisingly, StTST3.1-silenced leaves accumulated exceptionally high levels of maltose but low levels of sucrose and hexose. Additionally, chlorophyll content was reduced in StTST3.1-silenced leaves. Analysis of chlorophyll metabolic pathways found that Non-Yellow Coloring 1 (NYC1)-like (NOL), encoding a chloroplast-localized key enzyme that catalyzes the initial step of chlorophyll b degradation, was upregulated in StTST3.1-silenced leaves. Transient overexpression of StNOL accelerated chlorophyll b degradation in tobacco leaves. Our results indicated that StTST3.1 is involved in transitory starch turnover and chlorophyll metabolism, thereby playing a critical role in normal potato plant growth.
Assuntos
Solanum tuberosum , Amido , Amido/metabolismo , Vacúolos/metabolismo , Plantas/metabolismo , Folhas de Planta/metabolismo , Clorofila/metabolismo , Maltose/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulação da Expressão Gênica de PlantasRESUMO
Phytohormones and their interactions play critical roles in Solanum tuberosum (potato) tuberization. The stimulatory role of jasmonic acid (JA) in tuber development is well established because of its significant promotion of tuber initiation and tuber bulking. However, the dynamics and potential function of JA signalling in potato tuberization remain largely unknown. The present study investigated the role of the JAZ1 subtype, a suppressor of JA signalling, in potato tuberization. Using 35S:StJAZ1-like-GUS as a reporter, we showed that JA signalling was attenuated from the bud end to the stem end shortly after tuber initiation. Overexpression of StJAZ1-like suppressed tuber initiation by restricting the competence for tuber formation in stolon tips, as demonstrated by grafting an untransformed potato cultivar to the stock of StJAZ1-like-overexpressing transgenic potato plants (StJAZ1-like ox). In addition, transcriptional profiling analysis revealed that StJAZ1-like modulates the expression of genes associated with transcriptional regulators, cell cycle, cytoskeleton and phytohormones. Furthermore, we showed that StJAZ1-like is destabilised upon treatment with abcisic acid (ABA), and the attenuated tuberization phenotype in StJAZ1-like ox plants can be partially rescued by ABA treatment. Altogether, these results revealed that StJAZ1-like-mediated JA signalling plays an essential role in potato tuberization.
Assuntos
Ciclopentanos/metabolismo , Oxilipinas/metabolismo , Tubérculos/metabolismo , Proteínas Repressoras/metabolismo , Transdução de Sinais , Solanum tuberosum/metabolismo , Regulação da Expressão Gênica de Plantas , Reguladores de Crescimento de Plantas/genética , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas , Plantas Geneticamente Modificadas/genética , Proteínas Repressoras/genética , Solanum tuberosum/genética , Fatores de Transcrição/metabolismo , TranscriptomaRESUMO
Potato (Solanum tuberosum L.) maturity involves several important traits, including the onset of tuberization, flowering, leaf senescence, and the length of the plant life cycle. The timing of flowering and tuberization in potato is mediated by seasonal fluctuations in photoperiod and is thought to be separately controlled by the FLOWERING LOCUS T-like (FT-like) genes SELF-PRUNING 3D (StSP3D) and SELF-PRUNING 6A (StSP6A). However, the biological relationship between these morphological transitions that occur almost synchronously remains unknown. Here, we show that StABI5-like 1 (StABL1), a transcription factor central to abscisic acid (ABA) signaling, is a binding partner of StSP3D and StSP6A, forming an alternative florigen activation complex and alternative tuberigen activation complex in a 14-3-3-dependent manner. Overexpression of StABL1 results in the early initiation of flowering and tuberization as well as a short life cycle. Using genome-wide chromatin immunoprecipitation sequencing and RNA-sequencing, we demonstrate that AGAMOUS-like and GA 2-oxidase 1 genes are regulated by StABL1. Phytohormone profiling indicates an altered gibberellic acid (GA) metabolism and that StABL1-overexpressing plants are insensitive to the inhibitory effect of GA with respect to tuberization. Collectively, our results suggest that StABL1 functions with FT-like genes to promote flowering and tuberization and consequently life cycle length in potato, providing insight into the pleiotropic functioning of the FT gene.
Assuntos
Solanum tuberosum , Flores/fisiologia , Regulação da Expressão Gênica de Plantas , Fotoperíodo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ligação Proteica , Solanum tuberosum/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
Abscisic acid (ABA) is critical in drought tolerance and plant growth. Group A protein type 2C phosphatases (PP2Cs) are negative regulators of ABA signaling and plant adaptation to stress. Knowledge about the functions of potato group A PP2Cs is limited. Here, we report that the potato group A PP2C StHAB1 is broadly expressed in potato plants and strongly induced by ABA and drought. Suppression of StHAB1 enhanced potato ABA sensitivity and drought tolerance, whereas overexpression of the dominant mutant StHAB1G276D compromised ABA sensitivity and drought tolerance. StHAB1 interacts with almost all ABA receptors and the Snf1-Related Kinase OST1. Suppressing StHAB1 and overexpressing StHAB1G276D alter potato growth morphology; notably, overexpression of StHAB1G276D causes excessive shoot branching. RNA-sequencing analyses identified that the auxin efflux carrier genes StPIN3, StPIN5, and StPIN8 were up-regulated in StHAB1G276D-overexpressing axillary buds. Correspondingly, the auxin concentration was reduced in StHAB1G276D-overexpressing axillary buds, consistent with the role of auxin in repressing lateral branch outgrowth. The expression of BRANCHED1s (StBRC1a and StBRC1b) was unchanged in StHAB1G276D-overexpressing axillary buds, suggesting that StHAB1G276D overexpression does not cause axillary bud outgrowth via regulation of BRC1 expression. Our findings demonstrate that StHAB1 is vital in potato drought tolerance and shoot branching.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Solanum tuberosum , Ácido Abscísico/metabolismo , Proteínas de Arabidopsis/metabolismo , Fosfoproteínas Fosfatases/metabolismo , Arabidopsis/genética , Solanum tuberosum/genética , Solanum tuberosum/metabolismo , Resistência à Seca , Ácidos Indolacéticos/metabolismo , Secas , Regulação da Expressão Gênica de Plantas , Fatores de Transcrição/metabolismoRESUMO
SELF-PRUNING 6A (SP6A), a homolog of FLOWERING LOCUS T (FT), has been identified as tuberigen in potato. StSP6A is a mobile signal synthesized in leaves and transmitted to the stolon through phloem, and plays multiple roles in the growth and development of potato. However, the global StSP6A protein interaction network in potato remains poorly understood. In this study, BK-StSP6A was firstly used as the bait to investigate the StSP6A interaction network by screening the yeast two-hybrid (Y2H) library of potato, resulting in the selection of 200 independent positive clones and identification of 77 interacting proteins. Then, the interaction between StSP6A and its interactors was further confirmed by the Y2H and BiFC assays, and three interactors were selected for further expression analysis. Finally, the expression pattern of Flowering Promoting Factor 1.1 (StFPF1.1), No Flowering in Short Days 1 and 2 (StNFL1 and StNFL2) was studied. The three genes were highly expressed in flowers or flower buds. StFPF1.1 exhibited an expression pattern similar to that of StSP6A at the stolon swelling stages. StPHYF-silenced plants showed up-regulated expression of StFPF1.1 and StSP6A, while expression of StNFL1 and StNFL2 was down-regulated in the stolon. The identification of these interacting proteins lays a solid foundation for further functional studies of StSP6A.
Assuntos
Solanum tuberosum , Flores/metabolismo , Regulação da Expressão Gênica de Plantas , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Tubérculos/genética , Solanum tuberosum/metabolismoRESUMO
Plants program their meristem-associated developmental switches for timely adaptation to a changing environment. Potato (Solanum tuberosum L.) tubers differentiate from specialized belowground branches or stolons through radial expansion of their terminal ends. During this process, the stolon apex and closest axillary buds enter a dormancy state that leads to tuber eyes, which are reactivated the following spring and generate a clonally identical plant. The potato FLOWERING LOCUS T homolog SELF-PRUNING 6A (StSP6A) was previously identified as the major tuber-inducing signal that integrates day-length cues to control the storage switch. However, whether some other long-range signals also act as tuber organogenesis stimuli remains unknown. Here, we show that the florigen SELF PRUNING 3D (StSP3D) and FLOWERING LOCUS T-like 1 (StFTL1) genes are activated by short days, analogously to StSP6A. Overexpression of StSP3D or StFTL1 promotes tuber formation under non-inductive long days, and the tuber-inducing activity of these proteins is graft transmissible. Using the non-tuber-bearing wild species Solanum etuberosum, a natural SP6A null mutant, we show that leaf-expressed SP6A is dispensable for StSP3D long-range activity. StSP3D and StFTL1 mediate secondary activation of StSP6A in stolon tips, leading to amplification of this tuberigen signal. StSP3D and StFTL1 were observed to bind the same protein partners as StSP6A, suggesting that they can also form transcriptionally active complexes. Together, our findings show that additional mobile tuber-inducing signals are regulated by the photoperiodic pathway.
Assuntos
Solanum tuberosum , Solanum tuberosum/genética , Solanum tuberosum/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Folhas de Planta/metabolismo , Fatores de Transcrição/metabolismo , Tubérculos/genética , Tubérculos/metabolismoRESUMO
Cold-induced sweetening (CIS), the undesirable sugar accumulation in cold-stored potato (Solanum tuberosum L.) tubers, is a severe postharvest issue in the potato processing industry. Although the process of sucrose hydrolysis by vacuolar invertase during potato CIS is well understood, there is limited knowledge about the transportation of sucrose from the cytosol to the vacuole during postharvest cold storage. Here, we report that among the three potato tonoplast sugar transporters (TSTs), StTST1 exhibits the highest expression in tubers during postharvest cold storage. Subcellular localization analysis demonstrates that StTST1 is a tonoplast-localized protein. StTST1 knockdown decreases reducing sugar accumulation in tubers during low-temperature storage. Compared to wild-type, potato chips produced from StTST1-silenced tubers displayed significantly lower acrylamide levels and lighter color after cold storage. Transcriptome analysis manifests that suppression of StTST1 promotes starch synthesis and inhibits starch degradation in cold-stored tubers. We further establish that the increased sucrose content in the StTST1-silenced tubers might cause a decrease in the ABA content, thereby inhibiting the ABA-signaling pathway. We demonstrate that the down-regulation of ß-amylase StBAM1 in StTST1-silenced tubers might be directly controlled by ABA-responsive element-binding proteins (AREBs). Altogether, we have shown that StTST1 plays a critical role in sugar accumulation and starch metabolism regulation during postharvest cold storage. Thus, our findings provide a new strategy to improve the frying quality of cold-stored tubers and reduce the acrylamide content in potato chips.
RESUMO
The chloroplast, the energy organelle unique to plants and green algae, performs many functions, including photosynthesis and biosynthesis of metabolites. However, as the most critical tuber crop worldwide, the chloroplast proteome of potato (Solanum tuberosum) has not been explored. Here, we use Percoll density gradient centrifugation to isolate intact chloroplasts from leaves of potato cultivar E3 and establish a reference proteome map of potato chloroplast by bottom-up proteomics. A total of 1834 non-redundant proteins were identified in the chloroplast proteome, including 51 proteins encoded by the chloroplast genome. Extensive sequence-based localization prediction revealed over 62% of proteins to be chloroplast resident by at least one algorithm. Sixteen proteins were selected to evaluate the prediction result by transient fluorescence assay, which confirmed that 14 were distributed in distinct internal compartments of the chloroplast. In addition, we identified 136 phosphorylation sites in 61 proteins encoded by chloroplast proteome. Furthermore, we reconstruct the snapshots along starch metabolic pathways in the two different types of plastids by a comparative analysis between chloroplast and previously reported amyloplast proteomes. Altogether, our results establish a comprehensive proteome map with post-translationally modified sites of potato chloroplast, which would provide the theoretical principle for the research of the photosynthesis pathway and starch metabolism.
RESUMO
Which sugar transporter regulates sugar accumulation in tubers is largely unknown. Accumulation of reducing sugar (RS) in potato (Solanum tuberosum L.) tubers negatively affects the quality of tubers undergoing the frying process. However, little is known about the genes involved in regulating RS content in tubers at harvest. Here, we have identified two tonoplast sugar transporter (TST) 3-type isoforms (StTST3.1 and StTST3.2) in potato. Quantitative real-time PCR results indicate that StTST3.1 and StTST3.2 possess distinct expression patterns in various potato tissues. StTST3.2 was found to be the expressed TST3-type isoform in tubers. Further subcellular localization analysis revealed that StTST3.2 was targeted to the tonoplast. Silencing of StTST3.2 in potato by stable transformation resulted in significantly lower RS content in tubers at harvest or after room temperature storage, suggesting StTST3.2 plays an important role in RS accumulation in tubers. Accordingly, compared with the unsilenced control, potato chips processed from StTST3.2-silenced tubers exhibited lighter color and dramatically decreased acrylamide production at harvest or after room temperature storage. In addition, we demonstrated that silencing of StTST3.2 has no significant effect on potato growth and development. Thus, suppression of StTST3.2 could be another effective approach for improving processing quality and decreasing acrylamide content in potato tubers.