Sucrose Phosphate Synthase Gene Research Articles

Analysis of sugar components and identification of SPS genes in citrus fruit development.

Sugar is a primary determinant of citrus fruit flavour, but undergoes varied accumulation processes across different citrus varieties owing to high genetic variability. Sucrose phosphate synthase (SPS), a key enzyme in glucose metabolism, plays a crucial role in this context. Despite its significance, there is limited research on sugar component quality and the expression and regulatory prediction of SPS genes during citrus fruit development. Therefore, we analysed the sugar quality formation process in 'Kiyomi' and 'Succosa', two citrus varieties, and performed a comprehensive genome-wide analysis of citrus CsSPSs. We observed that the accumulation of sugar components significantly differs between the two varieties, with the identification of four CsSPSs in citrus. CsSPS sequences were highly conserved, featuring typical SPS protein domains. Expression analysis revealed a positive correlation between CsSPS expression and sugar accumulation in citrus fruits. However, CsSPS expression displays specificity to different citrus tissues and varieties. Transcriptome co-expression network analysis suggests the involvement of multiple transcription factors in shaping citrus fruit sugar quality through the regulation of CsSPSs. Notably, the expression levels of four CsWRKYs (CsWRKY2, CsWRKY20, CsWRKY28, CsWRKY32), were significantly positively correlated with CsSPSs and CsWRKY20 might can activate sugar accumulation in citrus fruit through CsSPS2. Collectively, we further emphasize the potential importance of CsWRKYs in citrus sugar metabolism, our findings serve as a reference for understanding sugar component formation and predicting CsSPS expression and regulation during citrus fruit development.

Comparative Analysis of Sucrose-Regulatory Genes in High- and Low-Sucrose Sister Clones of Sugarcane.

Sugarcane is a significant primitive source of sugar and energy worldwide. The progress in enhancing the sugar content in sugarcane cultivars remains limited due to an insufficient understanding of specific genes related to sucrose production. The present investigation examined the enzyme activities, levels of reducing and non-reducing sugars, and transcript expression using RT-qPCR to assess the gene expression associated with sucrose metabolism in a high-sucrose sugarcane clone (GXB9) in comparison to a low-sucrose sister clone (B9). Sucrose phosphate synthase (SPS), sucrose phosphate phosphatase (SPP), sucrose synthase (SuSy), cell wall invertase (CWI), soluble acid invertase (SAI), and neutral invertase (NI) are essential enzymes involved in sucrose metabolism in sugarcane. The activities of these enzymes were comparatively quantified and analyzed in immature and maturing internodes of the high- and low-sucrose clones. The results showed that the higher-sucrose-accumulating clone had greater sucrose concentrations than the low-sucrose-accumulating clone; however, maturing internodes had higher sucrose levels than immature internodes in both clones. Hexose concentrations were higher in immature internodes than in maturing internodes for both clones. The SPS and SPP enzymes activities were higher in the high-sucrose-storing clone than in the low-sucrose clone. SuSy activity was higher in the low-sucrose clone than in the high-sucrose clone; further, the degree of SuSy activity was higher in immature internodes than in maturing internodes for both clones. The SPS gene expression was considerably higher in mature internodes of the high-sucrose clones than the low-sucrose clone. Conversely, the SuSy gene exhibited up-regulated expression in the low-sucrose clone. The enhanced expression of SPS in the high-sucrose clone compared to the low-sucrose clone suggests that SPS plays a major role in the increased accumulation of sucrose. These findings provide the opportunity to improve sugarcane cultivars by regulating the activity of genes related to sucrose metabolism using transgenic techniques.

Overexpression of ZmSUS1 increased drought resistance of maize (Zea mays L.) by regulating sucrose metabolism and soluble sugar content.

ZmSUS1 improved drought tolerance of maize by regulating sucrose metabolism and increasing soluble sugar content, and endowing transgenic maize with higher relative water content and photosynthesis levels. Sucrose synthase (SUS), a key enzyme of sugar metabolism, plays an important role in the regulation of carbon partitioning in plant, and affects important agronomic traits and abiotic responses to adversity. However, the function of ZmSUS1 in plant drought tolerance is still unknown. In this study, the expression patterns of ZmSUS1 in different tissues and under drought stress were analyzed in maize (Zea mays L.). It was found that ZmSUS1 was highly expressed during kernel development but also in leaves and roots of maize, and ZmSUS1 was induced by drought stress. Homozygous transgenic maize lines overexpressing ZmSUS1 increased the content and activity of SUS under drought stress and exhibited higher relative water content, proline and abscisic acid content in leaves. Specifically, the net photosynthetic rate and the soluble sugar contents including sucrose, glucose, fructose and SUS decomposition products including UDP-glucose (UDP-G) and ADP-glucose (ADP-G) in transgenic plants were significantly improved after drought stress. RNA-seq analysis showed that overexpressing of ZmSUS1 mainly affected the expression level of carbon metabolism-related genes. Especially the expression level of sucrose metabolism-related genes including sucrose phosphatase gene (SPP), sucrose phosphate synthase gene (SPS) and invertase gene (INV) were significantly up-regulated in transgenic maize. Overall, these results suggested that ZmSUS1 improved drought tolerance by regulating sucrose metabolism and increasing the soluble sugar content, and endowing transgenic maize with higher relative water content and photosynthesis levels, which can serve as a new gene candidate for cultivating drought-resistant maize varieties.

Phylogenetic and Expression Analysis of the Sucrose Synthase and Sucrose Phosphate Synthase Gene Family in Potatoes.

Sucrose synthase (SUS) and sucrose phosphate synthase (SPS) are essential in plant sucrose metabolism. The potato is an important crop worldwide, but systematic analyses of the StSUS and StSPS gene families in potatoes are still lacking. Ten sucrose metabolism-related genes were identified in this study. The SUSs and SPSs could each be split into three subgroups through phylogenetic analysis. StSUSIc was the most highly expressed gene in different developmental tissues. Ka/Ks analysis showed that StSUSIb and StSUSIc were subjected to more-significant homozygous selection pressure. Our cis-acting element analysis of the StSUS and StSPS promoter sequences showed four elements: defense- and stress-responsive, hormone-responsive, light-responsive, and transcription factor elements. The expression of StSUS and StSPS genes was found to be regulated by circadian rhythm. In the treatments of 1% to 5% sucrose, glucose, and fructose, the expression of StSUS and StSPS family genes was enhanced by sucrose, but inhibited at high-glucose and fructose concentrations. This study identified six StSUS and four StSPS genes and analyzed their gene structure, conserved motifs, chromosome position, promoter elements, phylogenetic tree, and tissue-specific expression patterns. Our results will motivate more research into the biological process underlying the genes of sucrose metabolism in potatoes.

A Review of the Diverse Genes and Molecules Involved in Sucrose Metabolism and Innovative Approaches to Improve Sucrose Content in Sugarcane

Sugarcane (Saccharum spp. hybrid) is the chief source of sugar and biofuel globally and is prominent among cash crops. Sucrose is the main required product in sugarcane, and many studies have been performed to understand the phenomena of sucrose synthesis, metabolism, and accumulation in sugarcane. However, none of the studies concluded that a single gene is responsible for the sucrose content. Instead, a complex mechanism consisting of several genes, such as sucrose phosphate synthase genes (SPS1, SPS2, SPS4, SPS5), sucrose synthase genes (SuSy1, SuSy2, SuSy4), invertase genes (INV, CWIN, NIN1, CINV2), and phytohormone, trehalose, transcription factor (TF), protein kinase, and sugar transporter genes are working spatiotemporally in sugarcane. Currently, omics approaches like transcriptomics, proteomics, and metabolomics are also being used to explore the sugar metabolism in sugarcane, but integrated transcriptomic, proteomic, and metabolomic studies have been less reported. The results obtained from the integrated analysis of transcriptomics, proteomics, and metabolomics are more reliable because the strong gene expression, received in the form of abundant mRNA, does not guarantee the plentiful existence of associated proteins or their particular activity in the target cells or tissues, which discloses the restraint of single interpretation and stresses the significance of the integrated analysis of transcriptomics, proteomics, and metabolomics. This review highlights different genes and molecules contributing to sugar metabolism at different stages and the significance of omics approaches in explaining sucrose metabolism, especially sucrose accumulation in sugarcane. It is also a vital source of knowledge for sugarcane breeders, particularly associated with sucrose content improvement and bioethanol energy production.

Effects of exogenous melatonin on sugar and organic acid metabolism in early-ripening peach fruits.

To evaluated the effects melatonin (MT) on the sugar and acid metabolism of early-ripening peach fruits, the concentration of 150 μmol/L MT was sprayed on the leaves of peach trees. MT increased the contents of total soluble sugar and sucrose in peach fruits during the whole ripening period, and increased the contents of glucose and sorbitol at the mature stage. During the whole ripening period, MT also increased the activities of sucrose synthase, sucrose phosphate synthase, neutral invertase, and acidic invertase and the relative expression levels of sucrose synthase, sucrose phosphate synthase, neutral invertase, and acidic invertase genes, while decreased the activity of sorbitol oxidase and the relative expression level of sorbitol dehydrogenase to some extent. Moreover, MT decreased the contents of total organic acid, malic acid, and citric acid at mature stage. At mature stage, MT decreased the activities of citrate synthetase and phosphoenolpyruvate carboxylase and the relative expression levels of citrate synthetase and phosphoenolpyruvate carboxylase genes, while increased the relative expression levels of Nicotinamide adenine dinucleotide phosphate (NADP+)-malic enzyme, malate dehydrogenase, and aconitase genes. Therefore, MT promotes the sugar accumulation and organic acid degradation in early-ripening peach fruits.

Sucrose phosphate synthase 8 is required for the remobilization of carbon reserves in rice stems during grain filling.

Carbon reserve remobilization in stems is closely related to rice grain filling. Sucrose phosphate synthase (SPS) is highly associated with carbon reserve remobilization. In this study, we investigated the expression pattern of SPS genes in various rice tissues, and found that SPS8 is the major SPS isoform in rice stems during the grain-filling stage. We then constructed sps8 mutants using the CRISPR/Cas9 system. The SPS activity of the sps8 mutants was markedly reduced in the stems. In addition, the sps8 mutants exhibited significant starch accumulation in stems. 14C-labelling experiments revealed that the remobilization of non-structural carbohydrates from rice stems to grains was impaired in the sps8 mutants. In the sps8 mutants, grain filling was delayed and yield decreased by 15% due to a reduced percentage of ripened grains. RNA sequencing and quantitative PCR analyses indicated that the genes involved in starch synthesis and degradation were up-regulated in the sps8 mutant stems. In addition, the activity of the enzymes involved in starch synthesis and degradation was increased in the sps8 stems. These results demonstrate that SPS8 is required for carbon reserve remobilization from rice stems to grains, and that its absence may enhance 'futile cycles' of starch synthesis and degradation in rice stems.

Comparative expression analysis of sucrose phosphate synthase gene family in a low and high sucrose Pakistani sugarcane cultivars.

Sugarcane is the world's largest cultivated crop by biomass and is the main source of sugar and biofuel. Sucrose phosphate synthase (SPS) enzymes are directly involved in the synthesis of sucrose. Here, we analyzed and compared one of the important gene families involved in sucrose metabolism in a high and low sucrose sugarcane cultivar. A comprehensive in silico analysis of the SoSPS family displayed their phylogenetic relationship, gene and protein structure, miRNA targets, protein interaction network (PPI), gene ontology and collinearity. This was followed by a spatial expression analysis in two different sugarcane varieties. The phylogenetic reconstruction distributed AtSPS, ZmSPS, OsSPS, SoSPS and SbSPS into three main groups (A, B, C). The regulatory region of SoSPS genes carries ABRE, ARE, G-box, and MYC as the most dominant cis-regulatory elements. The PPI analysis predicted a total of 14 unique proteins interacting with SPS. The predominant expression of SPS in chloroplast clearly indicates that they are the most active in the organelle which is the hub of photosynthesis. Similarly, gene ontology attributed SPS to sucrose phosphate synthase and glucosyl transferase molecular functions, as well as sucrose biosynthetic and disaccharide biological processes. Overall, the expression of SPS in CPF252 (high sucrose variety) was higher in leaf and culm as compared to that of CPF 251 (low sucrose variety). In brief, this study adds to the present literature about sugarcane, sucrose metabolism and role of SPS in sucrose metabolism thereby opening up further avenues of research in crop improvement.

Genome-Wide Identification of GmSPS Gene Family in Soybean and Expression Analysis in Response to Cold Stress.

Sucrose metabolism plays a critical role in development, stress response, and yield formation of plants. Sucrose phosphate synthase (SPS) is the key rate-limiting enzyme in the sucrose synthesis pathway. To date, genome-wide survey and comprehensive analysis of the SPS gene family in soybean (Glycine max) have yet to be performed. In this study, seven genes encoding SPS were identified in soybean genome. The structural characteristics, phylogenetics, tissue expression patterns, and cold stress response of these GmSPSs were investigated. A comparative phylogenetic analysis of SPS proteins in soybean, Medicago truncatula, Medicago sativa, Lotus japonicus, Arabidopsis, and rice revealed four families. GmSPSs were clustered into three families from A to C, and have undergone five segmental duplication events under purifying selection. All GmSPS genes had various expression patterns in different tissues, and family A members GmSPS13/17 were highly expressed in nodules. Remarkably, all GmSPS promoters contain multiple low-temperature-responsive elements such as potential binding sites of inducer of CBF expression 1 (ICE1), the central regulator in cold response. qRT-PCR proved that these GmSPS genes, especially GmSPS8/18, were induced by cold treatment in soybean leaves, and the expression pattern of GmICE1 under cold treatment was similar to that of GmSPS8/18. Further transient expression analysis in Nicotiana benthamiana and electrophoretic mobility shift assay (EMSA) indicated that GmSPS8 and GmSPS18 transcriptions were directly activated by GmICE1. Taken together, our findings may aid in future efforts to clarify the potential roles of GmSPS genes in response to cold stress in soybean.

Effects of Extreme Root Restriction on the Nutritional and Flavor Quality, and Sucrose Metabolism of Tomato (Solanum lycopersicum L.)

Root restriction is suitable for horticultural soilless cultivation characterized by high efficiency and quality in the case of high density and low node order pinching. However, little research is available on the mechanism of root restriction improving the flavor and nutritional quality of tomatoes. We investigated the effects of Extreme Root Restriction (ERR, 750 mL/plant) on the content of metabolites, activity of enzymes, and gene expression level involving sucrose metabolism in different clusters of two tomato types. The fruit diameter and single fruit weight of common tomato at CIII were reduced by 5.6% and 14% under ERR, as a result, the fruit uniformity throughout the whole plant was improved. The ERR enhanced the accumulation of metabolites in tomato fruits, such as soluble sugars, amino acids, vitamin C, lycopene, and polyphenol, which was caused by ‘concentration effect’ that occurred with a reduction of fruit size. The activities of enzymes (SS, SPS, NI, AI) at CIII and CIV of cherry tomatoes increased by 3–4 folds under ERR. ERR enhanced accumulation of sucrose, glucose, and fructose in tomato fruits not only by modulating activities of metabolizing enzymes but also by inducing the expression of sucrose metabolism genes, including sucrose synthase genes (SS1, SS3–6) in common tomato, fructokinase genes (FKs), hexokinase genes (HKs), and sucrose phosphate synthase genes (SPSs), in cherry tomato. The above results are expected to provide a theoretical basis for root restriction cultivation techniques and practical guidance for high-quality tomato production in industrialized cultivation.

PvSPS4 is involved in regulation of root sugar balance in common bean under salt stress

Salinity is a stress factor that decreases global agricultural yield. Crops such as rice, tomato, potato, and legumes are susceptible to salt, thus this problem requires urgent attention. Disruption of carbohydrate metabolism can have negative effects on stress tolerance. Sucrose phosphate synthase (SPS) enzymes operate in a key regulatory step in sucrose synthesis. Therefore, they have a significant role in the regulation of sugar metabolism. This study focused on the function of the SPS homolog -pvSPS4- in the roots of legume crop common bean under salinity stress. We previously showed that pvSPS4 expression is root-specific and is upregulated under salt stress in a salt-tolerant common bean genotype. This upregulation was accompanied by an accumulation of sugars in the roots. In the current study, using the same genotype, we generated composite common bean plants with wild-type shoot and pvSPS4 knock-down roots. Composite plants exhibited a more sensitive phenotype under salt stress compared to control and mock plants. pvSPS4 knock-down disturbed the root glucose/sucrose ratio and balance of Ca+2, Mg+2, and K+ in both root and leaves which resulted in a reduction in photosynthetic pigments together with osmoregulation and antioxidant capability. Our results imply that pvSPS4 is an important gene for carbohydrate balance regulation under salt-stress in the common bean root tissues and sets an example for the significance of mainly disregarded roles of SPS genes in sink tissues.

Nontarget Site-Based Resistance to Fenoxaprop-P-ethyl and Candidate Genes Involved in Alopecurus japonicus

Nontarget-site resistance (NTSR) is a complex multigenic trait that is associated with the potential mechanisms of herbicide resistance which pose a serious threat to global crop protection. However, the NTSR mechanisms of Alopecurus japonicus, a malignant weed infesting wheat fields, are less characterized. In this study, we used RNA-sequencing transcriptome and enzyme activity detection to investigate the NTSR mechanisms and candidate genes involved in fenoxaprop-P-ethyl (FE) in a previously identified resistant population compared to the sensitive population of A. japonicus. Transcriptome analysis identified nine upregulated genes, which were constitutively overexpressed and upregulated by FE application in the resistant population, and the results were validated using quantitative real-time PCR. These genes including one cytochrome P450 monooxygenase (P450) gene (CYP75B4), one ATP-binding cassette (ABC) transporter gene (ABCG36), one laccase (LAC) gene (LAC15), one 9-cis-epoxycarotenoid dioxygenase (NCED) gene (NCED5), two purple acid phosphatase (PAP) genes (PAP4, PAP15), one sucrose phosphate synthase (SPS) gene (SPS3), one protein related to disease resistance gene (RGA3) and one immune protein gene (R1B-17). The activity assay of LAC, NCED, PAP and SPS revealed that the activities of these enzymes in the resistant population were significantly higher than those in the sensitive population at 0 h and after FE application at 12 h, 24 h and 72 h. Nevertheless, whether LAC, NCED, PAP and SPS genes were involved in herbicide metabolism needs to be further validated. Our results revealed that CYP, ABC transporter and LAC genes may participate in A. japonicus resistance. These genes identified in the present study provide new insights into the resistance mechanism of weeds in response to herbicide. Our study also implies the complexity of the NTSR mechanisms of weeds.

Sucrose metabolism analysis in a high sucrose sugarcane mutant clone at a mature stage in contrast to low sucrose parental clone through the transcriptomic approach

BackgroundThe significance of sugarcane as a major crop is due to the production of sucrose and a significant source of ethanol. Sucrose phosphate synthase (SPS), Sucrose synthase (SuSy), and cell wall invertase (CWIN) genes regulate sucrose metabolism and accumulation in high sucrose sugarcane; such genes are significant for manipulating sucrose content. The current study explored the genes associated with sucrose metabolism and accumulation in a high sucrose sugarcane mutant clone GXB9 compared to its low sucrose parental clone B9.ResultsA total of 100, 262 differentially expressed genes were obtained, and 69, 637 (69.46%) got annotation in public databases COG, GO, KEGG, KOG, Pfam, eggNOG, NR, and Swiss-Prot. The SPS5 gene was significantly up-regulated in the maturing internodes of GXB9 and downregulated in B9. The SuSy and CWIN genes were up-regulated in the immature internodes; however, no expression was found in the maturing internodes of GXB9. The genes involved in trehalose synthesis were downregulated in GXB9 while up-regulated in B9. The cellulose synthase (CeS) genes were significantly up-regulated in the immature internodes than in maturing internodes of GXB9.ConclusionsThe key finding of this research suggests that the SPS5 gene has a predominant role in enhancing the sucrose accumulation ability of GXB9. No expression of SuSy and CWIN genes in maturing internodes and downregulation of trehalose genes in GXB9 may also have contributed to the higher sucrose accumulation in the stalk of high sucrose mutant. The current finding is a source for sugarcane breeding, particularly improving sucrose content.Graphical

Genome-wide identification and function analysis of the sucrose phosphate synthase MdSPS gene family in apple

Sucrose phosphate synthase (SPS) is a rate-limiting enzyme that works in conjunction with sucrose-6-phosphate phosphatase (SPP) for sucrose synthesis, and it plays an essential role in energy provisioning during growth and development in plants as well as improving fruit quality. However, studies on the systematic analysis and evolutionary pattern of the SPS gene family in apple are still lacking. In the present study, a total of seven MdSPS and four MdSPP genes were identified from the Malus domestica genome GDDH13 v1.1. The gene structures and their promoter cis-elements, protein conserved motifs, subcellular localizations, physiological functions and biochemical properties were analyzed. A chromosomal location and gene-duplication analysis demonstrated that whole-genome duplication (WGD) and segmental duplication played vital roles in MdSPS gene family expansion. The Ka/Ks ratio of pairwise MdSPS genes indicated that the members of this family have undergone strong purifying selection during domestication. Furthermore, three SPS gene subfamilies were classified based on phylogenetic relationships, and old gene duplications and significantly divergent evolutionary rates were observed among the SPS gene subfamilies. In addition, a major gene related to sucrose accumulation (MdSPSA2.3) was identified according to the highly consistent trends in the changes of its expression in four apple varieties (‘Golden Delicious’, ‘Fuji’, ‘Qinguan’ and ‘Honeycrisp’) and the correlation between gene expression and soluble sugar content during fruit development. Furthermore, the virus-induced silencing of MdSPSA2.3 confirmed its function in sucrose accumulation in apple fruit. The present study lays a theoretical foundation for better clarifying the biological functions of the MdSPS genes during apple fruit development.

DETECTION OF GENETICALLY MODIFIED RICE LOCALLY AVAILABLE IN PUNJAB, PAKISTAN

Rice (Oryza sativa L.) is an important cereal crop that provides food to half of the world's population. Pakistan's traditional and premium quality rice is mostly exported to Europe. Transgenic or genetically modified rice (GM RICE) has been developed in the USA, China, and other countries, like herbicide-tolerant Liberty-Link rice (LLRICE06, LLRICE62, LLRICE601) and insect-resistant Bt-63. International and national biosafety regulations allow countries to stop the import of any genetically modified organism (GMO) due to its possible safety concerns. PCR-based methods are recommended for the accurate detection of GM rice. Qualitative PCR is used to detect the presence or absence of GMOs in a rice sample. The objective of this study was to check the presence of GMOs in the rice samples collected from rice-growing areas in Punjab. Eighty-two rice samples were collected randomly from various markets/locations in rice-growing areas of Punjab for this study. Twelve bulks were prepared to represent eighty-two rice samples. The CTAB method was used to extract the DNA from rice samples to analyze it. PCR of rice bulk samples was performed using primers specific for SPS, CP, p35S, tNOS, nptII, Hpt and Bt 63 rice. Positive and negative controls were included in each PCR. Primers specific for chloroplast (CP) and Sucrose phosphate synthase (SPS) genes were used to confirm DNA quality for amplification. Then, primers specific for the cauliflower mosaic virus (CaMV) 35S promoter and the Agrobacterium nopaline synthase (NOS) terminator were used for the detection of GM rice through PCR, neomycin phosphotransferase II (nptII) and Hygromycin phosphotransferase (Hpt) were used as a selectable marker and Bt 63 rice was used in insect resistance. The gel electrophoresis of PCR was conducted using a DNA ladder and then taking of the image was through gel documentation system for visualization and analysis. According to the results, all the rice bulks representing the eighty-two rice samples were negative for p35S, tNOS, nptII, Hygro and Bt 63 rice event. These findings are encouraging and demonstrate that rice grown in kalar track of Punjab, Pakistan, is non-GMO. Such surveillance and monitoring would be largely beneficial to maintain the purity of our traditional non-GM rice and to avoid any possible mixing of GM rice seeds, especially at the time of import. All these efforts will ensure and protect our national share in the international trade of rice.

Transcriptome Analysis on the Underlying Physiological Mechanism of Calcium and Magnesium Resolving “Sugar Receding” in ‘Feizixiao’ Litchi Pulp

The sugar contents of ‘Feizixiao’ litchi (Litchi chinensis Sonn.) decrease at full maturity; calcium (Ca) and magnesium (Mg) foliar fertilizer can resolve this “sugar receding”. To investigate the physiological mechanism of Ca and Mg foliar fertilizer used to resolve the “sugar receding” phenomenon in ‘Feizixiao’ litchi pulp, 16-year-old litchi trees were treated with 0.3% CaCl2 + 0.3% MgCl2 foliar spraying or water as a control. We determined the pulp sugar content over a two-year period in 2020 and 2021. Pulp total RNA was extracted for transcriptome sequencing in 2020, and the expression pattern of 10 differentially expressed genes (DEGs) was verified by real-time PCR in 2020 and 2021. The results showed that the fertilizer treatment significantly increased pulp fructose and total soluble sugar contents at maturity in both years. According to Gene Ontology (GO) functional enrichment analysis, there were 155 DEGs divided into 35 GO categories, among which 49 DEGs were divided into 49 pathways according to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. We isolated sugar-metabolism-related enzyme genes, including sucrose synthase (SS), acid invertase (AI), neutral invertase (NI), sucrose phosphate synthase (SPS), and hexokinase (HK). All sucrose-metabolism-related enzyme (NI, AI, SS, SPS) genes were downregulated, and six of the seven HK genes were downregulated. The expression patterns of the 10 DEGs were verified by real-time PCR, which showed significant linear relationships (r2020 = 0.9127, r2021 = 0.8705). In conclusion, the fertilizer treatment inhibited the synthesis of sucrose and phosphorylation of hexose by downregulating the expression of the SS, SPS, and HK genes, thus increasing the fructose and total soluble sugar contents in ‘Feizixiao’ litchi.

Identification, Characterization and Expression Analysis of the Sucrose Phosphate Synthase Gene Family in Vitis vinifera

Sucrose is the end product of photosynthesis in plants. Sucrose phosphate synthase (SPS) is a plant enzyme involved in sucrose biosynthesis that plays a crucial role in sugar metabolism, plant growth and biomass accumulation. So far, SPS gene family in grape (Vitis vinifera) has not been described. In this study, the genome-wide identification and characterization of SPS genes (VviSPS) in grape were analyzed. The physical and chemical properties, gene structure, evolutionary relationship, conserved domains, protein structure prediction and expression analyses were performed. In the results, total 4 putative VviSPS genes were identified, all of which had similar characteristics in genome and peptide sequences. Meanwhile, the four VviSPS genes also showed similar expression patterns in different tissues. They were mainly active in leaves and pulps, especially in mature leaves and early pulps. In conclusion, these results would provide a reference for the VviSPS gene function analysis in the future.

SlSPS, a Sucrose Phosphate Synthase Gene, Mediates Plant Growth and Thermotolerance in Tomato

Heat stress (HS) has been considered as a severe threat to crop yields in recent years. Sucrose, as a major product of photosynthesis, plays an important role in plant growth and stress response. Sucrose phosphate synthase (SPS) is a key rate-limiting enzyme in the sucrose synthesis pathway in plants. However, its molecular mechanism and signaling pathway remain unclear. In this study, we identified a novel SPS gene (SlSPS) in tomato and generated over-expression and knock-out of SlSPS gene transgenic tomato plants to investigate its biological functions related to the growth and thermotolerance of tomato. Over-expression of SlSPS gene increased the growth and biomass of transgenic tomato plants, such as fresh weight, dry weight, plant height, fruit weight and root length. In contrast, knock-out of SlSPS gene decreased the growth and biomass of transgenic tomato plants. Under heat stress, the survival rates were positively correlated with the expression level of SlSPS gene in different tomato varieties. Furthermore, SlSPS-overexpressing tomato plants showed higher SPS activity and sucrose content and heat stress resistant phenotypes. By comparison, knock-out tomato plants showed lower SPS activity and sucrose content and susceptible to heat stress. The determination of several reference values of oxidative stress parameters were also consistent with their heat resistance of these transgenic plants. In summary, SlSPS gene could positively mediate the growth and thermotolerance in tomato plants.

Effect of Saline–Alkali Stress on Sugar Metabolism of Jujube Fruit

Sugar, an osmoregulatory substance used by plants to adapt to abiotic stresses such as drought and salinity, is one of the most important indexes of fruit quality. In this study, 0–150 mM saline–alkali solutions (NaCl:NaHCO3 = 3:1) were used to irrigate the roots of 10-year-old “Junzao” fruit trees during the growth period to explore the regulation mechanism of different concentrations of saline–alkali stress on sugar and reactive oxygen metabolism in jujube fruit at maturity. The results showed that under low stress (0~90 mM), the contents of sucrose, glucose, and fructose in the jujube fruit and the activities of sucrose phosphate synthase (SPS), sucrose synthase decomposition direction (SS-I), and sucrose synthase synthesis direction (SS-II) increased with increases in stress concentration, results that were consistent with the relative expression trends of the SPS and SS genes; however, the results were reversed under high concentrations (120 and 150 mM). The soluble acid invertase (S-AI) activity decreased with increases in stress concentration under low stress, and the results were reversed with high stress, which was consistent with the relative expression trends of the ZjcINV3, ZjnINV1, and ZjnINV3. Research regarding the response of antioxidant enzymes in fruits under saline–alkali stress showed that only the differences in peroxidase (POD) activity under saline–alkali stress were consistent with sugar accumulation; the proline (PRO), catalase (CAT) decreased and the malondialdehyde (MDA) superoxide dismutase (SOD) increased with increases in saline–alkali stress. These results indicate that the sugar metabolism and antioxidase jointly promote and regulate sugar accumulation in jujube fruits in a low saline–alkali environment.

PpybZIP43 contributes to sucrose synthesis in pear fruits by activating PpySPS3 expression and interacts with PpySTOP1.

Sucrose is an important factor affecting sweetness and flavor in pear fruits, but the molecular mechanism of sucrose synthesis regulation is relatively unknown. Here, we characterized a transcription factor gene from pear (Pyrus pyrifolia Nakai cv. "Hosui") fruits, PpybZIP43, and found that the transient overexpression of PpybZIP43 in pear fruits significantly increased the sucrose content and the relative expression level of sucrose phosphate synthase genes (PpySPS3 and PpySPS8). Subcellular localization analysis in tobacco leaves showed that PpybZIP43 was localized in the nucleus. Yeast one-hybrid, electrophoretic mobility shift assay (EMSA), and dual-luciferase reporter assays indicated that PpybZIP43 was able to activate the expression of PpySPS3 by binding specifically to the G-box (CACGTG) element in the promoter. The protein-protein interaction assays using yeast two-hybrid, bimolecular fluorescence complementation (BiFC), firefly luciferase complementation imaging (LCI), and glutathione S-transferase (GST) pull-down demonstrated that PpybZIP43 could directly interact with PpySTOP1 to form a transcription complex. This study is helpful for understanding the molecular basis of sucrose synthesis and accumulation in pear fruits and provides candidate genes for breeding.