Vacuolar Invertase Research Articles

Sustained carbon import supports sugar accumulation and anthocyanin biosynthesis during fruit development and ripening in blueberry (Vaccinium ashei)

Fruit ripening is a highly coordinated process involving molecular and biochemical changes that collectively determine fruit quality. The underlying metabolic programs and their transitions leading to fruit ripening remain largely under-characterized in blueberry (Vaccinium sp.), which exhibits atypical climacteric behavior. In this study, we focused on sugar, acid and anthocyanin metabolism in two rabbiteye blueberry cultivars, Premier and Powderblue, during fruit development and ripening. Concentrations of the three major sugars, sucrose (Suc), glucose (Glc), and fructose (Fru) increased steadily during fruit development leading up to ripening, and increased dramatically by around 2-fold in ‘Premier’ and 2- to 3-fold in ‘Powderblue’ during the final stage of fruit ripening. Starch concentration was very low throughout fruit development in both cultivars indicating that it does not serve the role of a major transitory carbon (C) storage form in blueberry fruit. Together, these patterns indicate continued import of C, likely in the form of Suc, throughout blueberry fruit development. Concentrations of the predominant acids, malate and quinate, decreased during ripening, and may contribute to increased shikimate biosynthesis which, in-turn, allows for downstream phenylpropanoid metabolism leading to anthocyanin synthesis. Consistently, anthocyanin concentrations were highest in fully ripened blue fruit. Weighted gene co-expression network analysis (WGCNA) was performed using a ‘Powderblue’ fruit ripening transcriptome and targeted fruit metabolite concentration data. A ‘dark turquoise’ module positively correlated with sugars and anthocyanins, and negatively correlated with acids (malate, quinate), was identified. Gene Ontology (GO) enrichment analysis of this module identified transcripts related to sugar, acid, and phenylpropanoid metabolism pathways. Among these, increased transcript abundance of a VACUOLAR INVERTASE during ripening was consistent with sugar storage in the vacuole. In general, transcript abundance of the glycolysis pathway genes was upregulated during ripening. The transcript abundance of PHOSPHOENOLPYRUVATE (PEP) CARBOXYKINASE increased during fruit ripening and was negatively correlated with malate concentration, suggesting increased malate conversion to PEP, which supports anthocyanin production during fruit ripening. This was further supported by the co-upregulation of several anthocyanin biosynthesis-related genes. Together, this study provides insights into important metabolic programs, and their underlying gene expression patterns during fruit development and ripening in blueberry.

Sucrose induces flowering by degradation of the floral repressor Ghd7 via K48-linked polyubiquitination in rice.

Sucrose functions as a signaling molecule in several metabolic pathways as well as in various developmental processes. However, the molecular mechanisms by which sucrose regulates these processes remain largely unknown. In the present study, we demonstrate that sucrose promotes flowering by mediating the stability of a regulatory protein that represses flowering in rice. Exogenous application of sucrose promoted flowering by inducing florigen gene expression. Reduction of sucrose levels in the phloem through genetic modifications, such as the overexpression of the vacuolar invertase OsVIN2 or the mutation of OsSUT2, a sucrose transporter, delayed flowering. Analysis of relative transcript levels of floral regulatory genes showed that sucrose activated Ehd1 upstream of the florigen, with no significant effect on the expression of other upstream genes. Examination of protein stability after sucrose treatment of major floral repressors revealed that the Ghd7 protein was specifically degraded. The Ghd7 protein interacted with the E3 ligase IPA INTERACTING PROTEIN1 (IPI1), and sucrose-induced K48-linked polyubiquitination of Ghd7 via IPI1, leading to protein degradation. Mutants defective in IPI1 delayed flowering, confirming its role in modulating proteins involved in flowering. We conclude that sucrose acts as a signaling molecule to induce flowering by promoting Ghd7 degradation via IPI1.

A vacuolar invertase gene SlVI modulates sugar metabolism and postharvest fruit quality and stress resistance in tomato

Abstract Sugars act as signaling molecules to modulate various growth processes and enhance plant tolerance to various abiotic and biotic stresses. Moreover, sugars contribute to the post-harvest flavor in fleshy fruit crops. To date, the regulation of sugar metabolism and its effect in plant growth, fruit ripening, postharvest quality and stress resistance remains not fully understood. In this study, we investigated the role of tomato gene encoding a vacuolar invertase, hydrolyzing sucrose to glucose and fructose. SlVI is specifically expressed during the tomato fruit ripening process. We found that overexpression of SlVI resulted in increased leaf size and early flowering, while knockout of SlVI led to increased fruit sucrose content, enhanced fruit firmness, and elevated resistance of postharvest fruit to Botrytis cinerea. Moreover, the content of naringenin and total soluble solids was significantly increased in SlVI knockout fruit at postharvest stage. Transcriptome analysis showed a negative feedback regulation triggered by sucrose accumulation in SlVI knockout fruit resulting in a downregulation of BAM3 and AMY2, which are critical for starch degradation. Moreover, genes associated with cell wall, cutin, wax, and flavonoid biosynthesis and pathogen resistance were upregulated in SlVI knockout fruit. Conversely, the expression levels of genes involved in cell wall degradation were decreased in knockout fruit. These results are consistent with the enhanced postharvest quality and resistance. Our findings not only provide new insights into the relationship between tomato fruit sucrose content and postharvest fruit quality, but also suggest new strategies to enhance fruit quality and extend postharvest shelf life.

Reduction in Cold-Induced Sweetening by Cas9 Endonuclease-Mediated Knockout of the POTATO VACUOLAR INVERTASE 1 Gene in the Cultivar ‘Symfonia’

Reduction in Cold-Induced Sweetening by Cas9 Endonuclease-Mediated Knockout of the POTATO VACUOLAR INVERTASE 1 Gene in the Cultivar ‘Symfonia’

Genome-wide analysis of the passion fruit invertase gene family reveals involvement of PeCWINV5 in hexose accumulation

BackgroundInvertases (INVs) are key enzymes in sugar metabolism, cleaving sucrose into glucose and fructose and playing an important role in plant development and the stress response, however, the INV gene family in passion fruit has not been systematically reported.ResultsIn this study, a total of 16 PeINV genes were identified from the passion fruit genome and named according to their subcellular location and chromosome position. These include six cell wall invertase (CWINV) genes, two vacuolar invertase (VINV) genes, and eight neutral/alkaline invertase (N/AINV) genes. The gene structures, phylogenetic tree, and cis-acting elements of PeINV gene family were predicted using bioinformatics methods. Results showed that the upstream promoter region of the PeINV genes contained various response elements; particularly, PeVINV2, PeN/AINV3, PeN/AINV5, PeN/AINV6, PeN/AINV7, and PeN/AINV8 had more response elements. Additionally, the expression profiles of PeINV genes under different abiotic stresses (drought, salt, cold temperature, and high temperature) indicated that PeCWINV5, PeCWINV6, PeVINV1, PeVINV2, PeN/AINV2, PeN/AINV3, PeN/AINV6, and PeN/AINV7 responded significantly to these abiotic stresses, which was consistent with cis-acting element prediction results. Sucrose, glucose, and fructose are main soluble components in passion fruit pulp. The contents of total soluble sugar, hexoses, and sweetness index increased significantly at early stages during fruit ripening. Transcriptome data showed that with an increase in fruit development and maturity, the expression levels of PeCWINV2, PeCWINV5, and PeN/AINV3 exhibited an up-regulated trend, especially for PeCWINV5 which showed highest abundance, this correlated with the accumulation of soluble sugar and sweetness index. Transient overexpression results demonstrated that the contents of fructose, glucose and sucrose increased in the pulp of PeCWINV5 overexpressing fruit. It is speculated that this cell wall invertase gene, PeCWINV5, may play an important role in sucrose unloading and hexose accumulation.ConclusionIn this study, we systematically identified INV genes in passion fruit for the first time and further investigated their physicochemical properties, evolution, and expression patterns. Furthermore, we screened out a key candidate gene involved in hexose accumulation. This study lays a foundation for further study on INV genes and will be beneficial on the genetic improvement of passion fruit breeding.

Genome-Wide Isolation of VIN Gene Family and Functional Identification of HpVIN4 in Red Pitaya (Hylocereus polyrhizus)

Soluble sugars, including glucose, fructose and sucrose, are the most important determinants that affect the flavor and quality of red pitaya (Hylocereus polyrhizus) fruit. Vacuolar invertase (VIN), which catalyzes sucrose hydrolysis into glucose and fructose, is a key type of enzyme responsible for soluble sugar metabolism in plant growth and development. Herein, we conducted genome-wide identification, gene expression analysis, subcellular localization and an enzymatic properties assay for the VIN-encoding genes from red pitaya. During red pitaya fruit development towards ripening, the enzymatic activities of VIN showed an up-regulated trend towards ripening. In total, four isoforms (HpVIN1–4) of the VIN-encoding gene were identified from the pitaya genome. Sequence alignment results revealed that the HpVIN1, HpVIN3 and HpVIN4 proteins contained essential motifs for targeting the vacuole and conserved motifs or residues responsible for sucrose binding and hydrolysis. Gene expression pattern analyses revealed that the level of HpVIN4 was obviously increasing during fruit development and acted as the most abundant VIN isoform towards ripening. Subcellular localization detection via transient expression in Arabidopsis thaliana mesophyll protoplasts revealed that the HpVIN4 protein was localized in the vacuole. Growth complementation tests of heterologous expression in the invertase-deficient baker’s yeast strain suggested that the HpVIN4 protein had a sucrose hydrolysis activity and could restore the yeast growth in vivo. The identification of enzymatic properties in vitro demonstrated that the HpVIN4 protein could degrade sucrose into glucose and fructose with an optimum pH of 4.0. Specifically, the HpVIN4 protein had an estimated Km value of 5.15 ± 1.03 mmol·L−1 for sucrose hydrolysis. Ultimately, this study provides a comprehensive understanding of the potential roles of VINs during fruit development and towards ripening and provides functional gene resources for regulating soluble sugar accumulation in red pitaya fruit.

A mechanohydraulic model supports a role for plasmodesmata in cotton fiber elongation.

Plant cell growth depends on turgor pressure, the cell hydrodynamic pressure, which drives expansion of the extracellular matrix (the cell wall). Turgor pressure regulation depends on several physical, chemical, and biological factors, including vacuolar invertases, which modulate osmotic pressure of the cell, aquaporins, which determine the permeability of the plasma membrane to water, cell wall remodeling factors, which determine cell wall extensibility (inverse of effective viscosity), and plasmodesmata, which are membrane-lined channels that allow free movement of water and solutes between cytoplasms of neighboring cells, like gap junctions in animals. Plasmodesmata permeability varies during plant development and experimental studies have correlated changes in the permeability of plasmodesmal channels to turgor pressure variations. Here, we study the role of plasmodesmal permeability in cotton fiber growth, a type of cell that increases in length by at least three orders of magnitude in a few weeks. We incorporated plasmodesma-dependent movement of water and solutes into a classical model of plant cell expansion. We performed a sensitivity analysis to changes in values of model parameters and found that plasmodesmal permeability is among the most important factors for building up turgor pressure and expanding cotton fibers. Moreover, we found that nonmonotonic behaviors of turgor pressure that have been reported previously in cotton fibers cannot be recovered without accounting for dynamic changes of the parameters used in the model. Altogether, our results suggest an important role for plasmodesmal permeability in the regulation of turgor pressure.

SMS2, a Novel Allele of OsINV3, Regulates Grain Size in Rice.

Grain size has an important effect on rice yield. Although several key genes that regulate seed size have been reported in rice, their molecular mechanisms remain unclear. In this study, a rice small grain size 2 (sms2) mutant was identified, and MutMap resequencing analysis results showed that a 2 bp insertion in the second exon of the LOC_Os02g01590 gene resulted in a grain length and width lower than those of the wild-type Teqing (TQ). We found that SMS2 encoded vacuolar acid invertase, a novel allele of OsINV3, which regulates grain size. GO and KEGG enrichment analyses showed that SMS2 was involved in endoplasmic reticulum protein synthesis, cysteine and methionine metabolism, and propionic acid metabolism, thereby regulating grain size. An analysis of sugar content in young panicles showed that SMS2 reduced sucrose, fructose, and starch contents, thus regulating grain size. A haplotype analysis showed that Hap2 of SMS2 had a longer grain and was widely present in indica rice varieties. Our results provide a new theoretical basis for the molecular and physiological mechanisms by which SMS2 regulates grain size.

Transgenerational Seed Exposure to Elevated CO2 Involves Stress Memory Regulation at Metabolic Levels to Confer Drought Resistance in Wheat.

Drought is the worst environmental stress constraint that inflicts heavy losses to global food production, such as wheat. The metabolic responses of seeds produced overtransgenerational exposure to e[CO2] to recover drought's effects on wheat are still unexplored. Seeds were produced constantly for four generations (F1 to F4) under ambient CO2 (a[CO2], 400 μmol L-1) and elevated CO2 (e[CO2], 800 μmol L-1) concentrations, and then further regrown under natural CO2 conditions to investigate their effects on the stress memory metabolic processes liable for increasing drought resistance in the next generation (F5). At the anthesis stage, plants were subjected to normal (100% FC, field capacity) and drought stress (60% FC) conditions. Under drought stress, plants of transgenerational e[CO2] exposed seeds showed markedly increased superoxide dismutase (16%), catalase (24%), peroxidase (9%), total antioxidants (14%), and proline (35%) levels that helped the plants to sustain normal growth through scavenging of hydrogen peroxide (11%) and malondialdehyde (26%). The carbohydrate metabolic enzymes such as aldolase (36%), phosphoglucomutase (12%), UDP-glucose pyrophosphorylase (25%), vacuolar invertase (33%), glucose-6-phosphate-dehydrogenase (68%), and cell wall invertase (17%) were decreased significantly; however, transgenerational seeds produced under e[CO2] showed a considerable increase in their activities in drought-stressed wheat plants. Moreover, transgenerational e[CO2] exposed seeds under drought stress caused a marked increase in leaf Ψw (15%), chlorophyll a (19%), chlorophyll b (8%), carotenoids (12%), grain spike (16%), hundred grain weight (19%), and grain yield (10%). Hence, transgenerational seeds exposed to e[CO2] upregulate the drought recovery metabolic processes to improve the grain yield of wheat under drought stress conditions.

Battling arid adversity: unveiling the resilience of cotton in the face of drought and innovative mitigation approaches

Climate change has had significant impacts on agriculture, particularly on cotton production, where drought has emerged as a major threat worldwide. Long and intense dry periods in cotton-growing regions have become more frequent and severe. Drought stress severely affects various aspects of cotton plants, including chlorophyll pigments, carbohydrate metabolism, and enzyme activities related to fiber development, such as vacuolar invertase and sucrose synthase. Furthermore, drought stress disrupts the movement of nutrients toward the reproductive tissues in cotton, resulting in compromised pollen function, propagative failure, and fiber characteristics. To tackle these issues, scientists have made advancements in creating drought-resistant cotton varieties through transgenic methods or molecular breeding techniques, genome editing, CRISPR/Cas9, utilizing quantitative trait loci (QTL). Moreover, the application of plant growth regulators and mineral elements has displayed the potential to improve cotton’s ability to endure drought stress while also enhancing fiber yield and quality. These approaches activate stress-responsive signaling pathways, which could contribute to mitigating reproductive failure and improving fiber characteristics. While the impact of drought stress on cotton plants has been extensively studied, the variations in fiber quality resulting from drought stress are not yet completely understood. Current research has been focused on unraveling the mechanisms underlying these changes, including the physiological, biochemical, and molecular alterations during the multiplicative growth phase that contribute to poor fiber development. Understanding these mechanisms will facilitate the development of novel strategies to alleviate the adverse impact of worldwide weather changes on cotton growth and fiber quality. This research focuses on the drought stress in cotton cultivation and explores its different effects on cotton morphology, physiology, crop yield, and fiber characteristics as well as mechanisms by which cotton exhibits drought tolerance and highlights innovative strategies to mitigate drought stress.

Identification of potato varieties suitable for cold storage and reconditioning: A safer alternative to anti-sprouting chemicals for potato sprouting control

Low temperature storage as an alternative to anti-sprouting chemicals in potato storage may induce reducing sugars (RS) accumulation (i.e. glucose and fructose) in potato tubers. This phenomenon is called “cold induced sweetening” (CIS) and occurs in certain varieties. CIS leads to a decrease in the organoleptic qualities and darkening of processed potato and the accumulation of toxic molecules such as acrylamide. To identify potato varieties suitable for storage at low temperatures, we screened six commercial processing varieties: Lady Claire (LC), Verdi, Kiebitz (KB), Pirol, Agria and Markies for their CIS characteristics and sprout-forming potential after storage at 4 °C and 8 °C. Our findings reveal that 4 °C storage allows for efficient sprout reduction in all six tested varieties for up to 4.5 months of storage. Three CIS-resistant varieties, namely Verdi, Lady Claire and Kiebitz, were identified as able to be stored for up to four months at 4 °C with limited increase in glucose content. Conversely, Pirol, Agria and Markies showed an increase in glucose content with a decrease in storage temperature and can be considered as CIS-susceptible varieties. After processing into crisps, the CIS-susceptible varieties displayed poor crisp color quality (brown to black color crisps) after storage for two months at 4 °C compared to the storage at 8 °C, whereas the CIS-resistant varieties had good crisp color quality (pale yellow color crisps) after storage at both 4 and 8 °C. Interestingly, the trends of total RS and/or glucose content in the CIS-resistant and in the CIS-susceptible varieties were correlated with the trends in Vacuolar Invertase (VInv) gene expression for most varieties, as well as with the trends in acrylamide content after processing. In addition, reconditioning of Markies variety after storage at 4 °C by gradually increasing the temperature to 15 °C resulted in a significant decrease of VInv transcript levels (reduction of 80 %), acrylamide content (reduction of 75 %) and glucose content when compared to a storage at 4 °C without reconditioning. Those results demonstrate that the reconditioning technique is a key factor for a sustainable potato storage and for improving the quality of processed potatoes.

Molecular dissection of an intronic enhancer governing cold-induced expression of the vacuolar invertase gene in potato.

Potato (Solanum tuberosum) is the third most important food crop in the world. Potato tubers must be stored at cold temperatures to minimize sprouting and losses due to disease. However, cold temperatures strongly induce the expression of the potato vacuolar invertase gene (VInv) and cause reducing sugar accumulation. This process, referred to as "cold-induced sweetening," is a major postharvest problem for the potato industry. We discovered that the cold-induced expression of VInv is controlled by a 200 bp enhancer, VInvIn2En, located in its second intron. We identified several DNA motifs in VInvIn2En that bind transcription factors involved in the plant cold stress response. Mutation of these DNA motifs abolished VInvIn2En function as a transcriptional enhancer. We developed VInvIn2En deletion lines in both diploid and tetraploid potato using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9)-mediated gene editing. VInv transcription in cold-stored tubers was significantly reduced in the deletion lines. Interestingly, the VInvIn2En sequence is highly conserved among distantly related Solanum species, including tomato (Solanum lycopersicum) and other non-tuber-bearing species. We conclude that the VInv gene and the VInvIn2En enhancer have adopted distinct roles in the cold stress response in tubers of tuber-bearing Solanum species.

Genome-wide identification and characterization of the sucrose invertase gene family in Hemerocallis citrina.

Sucrose invertase is an important catalytic enzyme that is widely distributed in plants and can irreversibly hydrolyze sucrose into fructose and glucose. Daylily is an important perennial flower worldwide and a traditional vegetable in East Asia. Previous studies have suggested that sucrose invertase is involved in the aging of daylily flowers. However, knowledge about the number, physicochemical properties, and expression patterns of daylily sucrose invertases is still lacking. Identifying the daylily sucrose invertase family genes in the genome is highly important for understanding phylogenetic evolution and determining the genetic function of sucrose invertase. To obtain basic knowledge about the number, classification, sequence composition, and physicochemical properties of sucrose invertases in daylily, bioinformatics software was used to analyze the genome of Hemerocallis citrina (H. citrina), and the basic properties of sucrose invertase genes and proteins were obtained. Then, combined with transcriptome data from flower organs at different developmental stages, the expression patterns of each gene were clarified. Finally, the reliability of the transcriptome data was verified by quantitative real-time polymerase chain reaction (PCR). Through software analysis, 35 sucrose invertases were identified from the H. citrina genome and named HcINV1-HcINV35; these enzymes belong to three subfamilies: cell wall invertases, vacuolar invertases, and chloroplast invertases. The amino acid composition, motif types, promoter composition, gene structure, protein physicochemical properties, gene chromosomal localization, and evolutionary adaptability of daylily invertases were determined; these results provided a comprehensive understanding of daylily invertases. The transcriptome expression profile combined with fluorescence quantitative reverse transcription-polymerase chain reaction (RT‒PCR) analysis suggested that almost all daylily invertase genes were expressed in flower organs, but even genes belonging to the same subfamily did not exhibit the same expression pattern at different developmental stages, suggesting that there may be redundancy or dissimilation in the function of daylily sucrose invertases.

Synthetic bacterial community derived from Astragalus mongholicus and plant-plant interactions inhibit cadmium uptake by modulating gene expression, antioxidant system and carbohydrate metabolism under cadmium contaminated soil

In this study we evaluated an alternative strategy to inhibit cadmium (Cd) uptake with synthetic bacterial community (SynCom) in cereal-legume intercropping under Cd contaminated soil. The results exhibited that intercropping coupled with SynCom inoculation promoted wheat biomass and inhibits Cd uptake in shoot (−65.05%), root (−34.66%) and grain (−74.45%) over monoculture without inoculation under Cd stress. However, antioxidant enzyme activities were significantly increased with SynCom inoculation in wheat grown on Cd contaminated soil irrespective of intercropping. SynCom inoculation considerably increased cytoplasmic invertase, vacuolar invertase, hexokinase, phosphoglucoisomerase, glucose-6-phosphate dehydrogenase, phosphofructokinase and SuSy while decreased the activities of ADP-Glucose pyrophosphorylase, aldolase, fructokinase, phosphoglucomutase and UDP-glucose pyrophorylase irrespective of intercropping under Cd stress. Furthermore, the expressions of gene encoding heavy metal stress were significantly decreased with the SynCom inoculation in roots. The expression of TaHMA3-b in root was downregulated by SynCom inoculation in wheat grown under Cd stress. The expressions of gene encoding heavy metal stress in shoot were downregulated by SynCom inoculation over other treatments. These findings not only offered useful information for future study of interaction mechanisms between intercropping and SynCom inoculation, but also demonstrated a suitable technological approach for remediation coupled with agro-production in mildly to substantially Cd polluted soil.

Foliar spraying magnesium promotes soluble sugar accumulation by inducing the activities of sucrose biosynthesis and transport in citrus fruits

Accumulation of soluble sugars promotes the formation of fruit nutritional quality and flavor while proper application of Magnesium (Mg) can increase the sugar content in citrus fruits. However, the underlying mechanism is still unclear. In the present study by using ‘Huagan No.2′ (Citrus reticulate cv. Huagan No.2) as research material, foliar spraying of MgSO4 with two or three times was performed during the fruit expansion period. Results indicated that foliar spraying of Mg significantly increased the total soluble solids (TSS), as well as the contents of sucrose, fructose and glucose in fruits; moreover, the activities of sucrose metabolizing enzymes including sucrose synthase (SS), sucrose phosphate synthesis (SPS) and acid invertase (AI), as well as the transcript levels of SS2, SPS1–3 and vacuolar acid invertase gene (VINV) were significantly increased in the Mg-treated fruits. In addition, the transcript levels of the sucrose transport-related gene (SUT3) and the vacuolar pyrophosphate proton pump type I genes (CsVPP-1 and CsVPP-2) in the fruits were also significantly increased by Mg treatment. These results were further verified by soaking the fruit-bearing branches in 0.5% MgSO4 solution. In conclusion, foliar spraying Mg with two or three times promotes soluble sugar accumulation in fruits through enhancing the sink strength by inducing the activity of sucrose synthase via inducing SS2 transcript level, and improving sucrose transport ability by inducing transcript levels of SUT3, CsVPP-1, CsVPP-2 and VINV.

The Responses of Sucrose Metabolism and Carbon Translocation in Tomato Seedlings under Different Light Spectra.

Light plays a dominant role in the biosynthesis and accumulation of photosynthetic products. However, the metabolism and translocation of photosynthetic products in plants under different light spectra remain elusive. In this study, tomato (Solanum lycopersicum L.) seedlings were treated with different light spectra delivered by light-emitting diodes (LEDs) with the same photosynthetic photon flux density at 300 μmol m-2 s-1, including monochromatic red (660 nm, R), blue (450 nm, B), sun-like white (W, 380-780 nm), or a combination of R and B lights (R:B = 1:1, RB). Compared with W, the biomass distribution ratio for leaves under R, B, and RB decreased by 5.01-9.53%, while the ratio for stems and roots increased by 3.71-6.92% and 0.14-2.81%, respectively. The photosynthetic carbon distribution expressed as 13C enrichment was higher in stems and roots under RB and R, while B led to more 13C transported from leaves and enriched in stems when compared with W. Meanwhile, RB led to significant increases in the activities of phosphate synthase (SPS), sucrose synthase (SS), vacuolar acid invertase (VI), and neutral invertase (NI). The R was more efficient in increasing the activity of SPS and SS, while B was more effective in promoting the activity of VI and NI. The transcript levels of SPS, SS3, NI6, and VI were upregulated under R, B, and RB. However, the transcript patterns of SPS, SS3, NI6, and VI were not consistent with the changes in their encoded enzymes, especially the transcript patterns of SPS and SS3. Our study suggests that the red- and blue-light-induced long-distance and short-distance transport of photosynthetic products in plants, respectively, might result from different regulation of sucrose-metabolizing enzymes from transcriptional and post-transcriptional levels.

Molecular physiology for the increase of soluble sugar accumulation in citrus fruits under drought stress

To investigate the mechanism for drought promoting soluble sugar accumulation will be conducive to the enhancement of citrus fruit quality as well as stress tolerance. Fruit sucrose mainly derives from source leaves. Its accumulation in citrus fruit cell vacuole involves in two processes of unloading in the fruit segment membrane (SM) and translocating to the vacuole of fruit juice sacs (JS). Here, transcript levels of 47 sugar metabolism- and transport-related genes were compared in fruit SM or JS between drought and control treatments. Results indicated that transcript levels of cell wall invertase genes (CwINV2/6) and sucrose synthase genes (SUS2/6) in the SM were significantly increased by the drought. Moreover, transcript levels of SWEET genes (CsSWEET1/2/4/5/9) and monosaccharide transporter gene (CsPMT3) were significantly increased in SM under drought treatment. On the other hand, SUS1/3 and vacuolar invertase (VINV) transcript levels were significantly increased in JS by drought; CsPMT4, sucrose transporter gene 2 (CsSUT2), tonoplast monosaccharide transporter gene 2 (CsTMT2), sugar transport protein gene 1 (CsSTP1), two citrus type I V-PPase genes (CsVPP1, and CsVPP2) were also significantly increased in drought treated JS. Collectively, the imposition of drought stress resulted in more soluble sugar accumulation through enhancing sucrose download by enhancing sink strength- and transport ability-related genes, such as CwINV2/6, SUS2/6, CsSWEET1/2/4/5/9, and CsPMT3, in fruit SM, and soluble sugar storage ability by increasing transcript levels of genes, such as CsPMT4, VINV, CsSUT2, CsTMT2, CsSTP1, CsVPP1, and CsVPP2, in fruit JS.

Genome-Wide Identification and Expression Patterns of Cucumber Invertases and Their Inhibitor Genes.

Invertases and their inhibitors play important roles in sucrose metabolism, growth and development, signal transduction, and biotic and abiotic stress tolerance in many plant species. However, in cucumber, both the gene members and functions of invertase and its inhibitor families remain largely unclear. In this study, in comparison with the orthologues of Citrullus lanatus (watermelon), Cucumis melo (melon), and Arabidopsis thaliana (Arabidopsis), 12 invertase genes and 12 invertase inhibitor genes were identified from the genome of Cucumis sativus (cucumber). Among them, the 12 invertase genes were classified as 4 cell wall invertases, 6 cytoplasmic invertases, and 2 vacuolar invertases. Most invertase genes were conserved in cucumber, melon, and watermelon, with several duplicate genes in melon and watermelon. Transcriptome analysis distinguished these genes into various expression patterns, which included genes CsaV3_2G025540 and CsaV3_2G007220, which were significantly expressed in different tissues, organs, and development stages, and genes CsaV3_7G034730 and CsaV3_5G005910, which might be involved in biotic and abiotic stress. Six genes were further validated in cucumber based on quantitative real-time PCR (qRT-PCR), and three of them showed consistent expression patterns as revealed in the transcriptome. These results provide important information for further studies on the physiological functions of cucumber invertases (CSINVs) and their inhibitors (CSINHs).

Disruptions of sugar utilization and carbohydrate metabolism in rice developing anthers aggravated heat stress-induced pollen abortion

High temperature (HT) stress at reproductive stage is one of most important environment negatively affecting spikelet fertility and rice yield. In this study, the effect of HT exposure on the sugar composition and carbohydrate metabolism in developing anthers and its relation to floret fertility and pollen viability were investigated by different temperature regimes under well-controlled climatic condition. Result showed that HT exposure during microspore development significantly reduced the starch deposition in developing anther and evidently disrupted the spatial distribution of sugar and starch concentrations in different compartments of rice anther, with the higher ratio of sucrose to hexose concentrations in HT-stressed anthers relative to the control ones. Under HT exposure, the amount of starch deposition in the fraction of sporophytic tissues dropped evidently, while the concentrations of sucrose and starch in anther wall tissue enhanced significantly, suggesting that HT exposure impaired the translocation of sucrose from the anther wall tissue to the sporophytic tissues inside rice anther. Furthermore, we presented possible contribution of various genes and key enzymes involving in sugar conversion and carbohydrate metabolism in developing anther to the formation of HT-induced pollen abortion by disrupting the sugar utilization in HT-stressed anther. HT exposure suppressed the activities of cell wall and vacuolar invertase, hexokinase, and ADP-glucose pyrophosphorylase in developing anther, while it was opposite for the effect of HT exposure on sucrose synthase and fructokinase. HT-induced suppression of OsCWIN3 in the anther walls might be strongly responsible for the HT-induced impairments of sugar utilization in HT-stressed anthers.

Coordinated regulation of sucrose and lignin metabolism for arrested silk elongation under drought stress in maize

The delay in silking due to drought stress generates a greater anthesis-silking interval in maize, causing compromised kernel set and thus catastrophic loss of grain yield, which is primarily attributed to the lower silk elongation. However, few studies are done in the understanding of the physiochemical and molecular mechanism that regulate silk elongation in response to drought stress. Thus, a field experiment was conducted to examine silk transcriptomic and metabolomic responses to drought stress using the maize cultivar of AN591. A total of 757 differently expressed genes (DEGs) and 80 differently expressed metabolites (DEMs) were identified in maize silk grown in the drought stress treatment compared to the control treatment. These DEGs and DEMs were enriched in 191 and 36 Kyoto Encyclopedia of genes and genomes (KEGG) pathways, respectively, most being associated with primary and secondary metabolism. Starch and sucrose metabolism (SSM) pathway in primary metabolism and phenylpropanoid biosynthesis (PB) pathway in secondary metabolism were identified in common in both transcriptome and metabolome. Most of DEGs in the SSM pathway were increased in expression, while the DEM were decreased in expression. Conversely, most of DEGs in the PB pathway were decreased in expression, while DEMs were increased in expression. This could be explained by the negative correlation between DEGs and DEMs. In the SSM pathway, there was a significant increase in sucrose content, but a significant decrease in fructose and glucose in response to drought stress, which was attributed to the decreased activities in cell wall invertase (CWIN), vacuolar invertase (VIN), and sucrose synthase (SuSy). Silk lignin content in PB pathway was in a significant decrease in drought treatment compared to the control, due to the decreased activity of peroxidase (POD) that is the key enzyme in the final stage of lignin biosynthesis. In conclusion, this study identified key genes or metabolites involved in sucrose and lignin metabolism which appears to be regulating silk elongation response to drought stress.