Vacuolar Invertase Research Articles

Effects of potassium deficiency on the enzymatic changes in developing cotton fibers

Although effects of potassium (K) on cotton growth have been explored extensively, the effects of K deficiency on the physiological changes closely related to cotton fiber development are lacking. Thus, a 2-year field experiment was conducted with two cotton cultivars (Simian 3 and Siza 3) under 0 kg K2O ha−1 (K deficiency) and 300 kg K2O ha−1 (K sufficiency). The results showed that tonoplast adenosine triphosphatase (V-ATPase), pyrophosphatase (PPase), plasma membrane H+-ATPase (PM H+-ATPase), phosphoenolpyruvate carboxylase (PEPC), sucrose synthesis (SuSy) and vacuolar invertase (V-INV) were highly sensitive to K deficiency. The decreases in those enzymes resulted in low malate and soluble sugar contents, which together with low K concentration declined the driving force for fiber elongation, leading to significantly lower fiber length in the 0 kg K2O ha−1 treatment. The activity of sucrose phosphate synthase (SPS) was obviously increased by K deficiency before 20 days post anthesis (DPA), which could partly explain the acceleration of fiber cellulose synthesis and the increase in fiber strength in the 0 kg K2O ha−1 treatment in the early stage. However, SPS activity was decreased by K deficiency after 20 DPA and SuSy activity was reduced by K deficiency at any sampling date, resulting in low fiber strength in the end. Compared with Simian 3, the enzymes V-ATPase, PPase, PM H+-ATPase, PEPC and SuSy during fiber elongation stage were more sensitive to K deficiency in Siza 3, and the enzymes SuSy and SPS during fiber-thickening stage were more sensitive to K deficiency in Siza 3, which were the important reasons causing greater decreases in final fiber length and final fiber strength for Siza 3 than Simian 3 under K deficiency.

RNAi-mediated silencing of endogenous Vlnv gene confers stable reduction of cold-induced sweetening in potato (Solanum tuberosum L. cv. Désirée)

Potato tubers must be cold-stored to extend their shelf life and maintain an uninterrupted supply chain for food processors. However, a side-effect of low-temperature storage is manifested in terms of cold-induced sweetening (CIS) of potato tubers, which reduces the processing quality and the commercial value of the end-products. RNA interference (RNAi) technology, whereby transgene-derived small interfering RNAs can trigger the homology-based knockdown of cognate host genes and can initiate gene silencing, has been successfully applied in crop improvement through targeted gene knockout in host plants. In the current study, transgenic potato plants (Solanum tuberosum cv. Desiree) were generated, expressing a 300 bp hairpin loop nucleotide sequence targeting the potato vacuolar invertase gene (VInv), under the constitutive Cauliflower mosaic virus 35S promoter. Tubers collected from transgenic lines showed a significant reduction in reducing sugar content after 180 days of cold storage, without showing any measurable off-target effects on plant morphology and tuberization compared to non-transformed control plants. The cold-stored tubers were further assayed for chip color, which showed a fairly light colored quality in the samples originating from RNAi lines. Together with similar effects seen in previously published experiments involving other potato varieties, the Desiree results described here establish the efficacy of using RNAi for the successful reduction of CIS in potato tubers.

Coordinated Regulation of Cold Induced Sweetening in Tetraploid Potato Families by Isozymes of UDP-Glucose Pyrophosphorylase and Vacuolar Acid Invertase

Past investigations have suggested that both UGPase and AcInv activities can be used as markers to screen genetically diverse potato clones for cold induced sweetening resistance (CIS-R). The goal of this study was to define their cooperative interaction in regulating sweetening. Inter- and intra-ploidy hybridizations of good (G) and poor (P) processing 24 or 48 chromosome potato clones were used to create 24 potato families. Potatoes were field grown and 460 progeny (≤20 each family) were stored for five months in the cold (4 C). Tubers from each progeny plant were evaluated for cold induced sweetening resistance (CIS-R) and correlated with the percentage of A-II isozymes of UDP-glucose pyrophosphorylase (UGPase; EC 2.7.7.9); and acid invertase activity (AcInv; EC 3.2.1.26). Each progeny was given a CIS-R score of 1–10 (1-most resistance, 10 least resistance). The families were grouped into four classes based on (1) high or low AcInv activity (low being a SA of 0.30 or less) (2) high or low percentage of A-II isozymes (low being 50% or less), and (3) CIS-R score. In high AcInv families, CIS-R was low regardless of the percentage of A-II isozymes present. In low AcInv activity families, there was a trend for average chip color to improve as the percentage of A-II isozymes increased from 0% to 40%. This increase in CIS-R in low AcInv families is likely due to the kinetic properties unique to the A-II forms of UGPase (principally UGP5) which limit the formation of sucrose via sucrose-6-phosphate synthase (SPS; EC 2.4.1.14). Lower concentrations of sucrose can lead to a decrease in reducing sugar production via vacuolar AcInv and lighter chip and fry colors. In selecting tetraploid parents, for the development of processing potato clones with improved CIS-R, it is recommended they have a basal AcInv SA of 0.30 or less and have A-II isozymes of UGPase.

Effects of nitrogen forms on carbohydrate metabolism and storage‐root formation of sweet potato

AbstractIn this study, we examined the effects of different forms of nitrogen (N) fertilizer on carbohydrate metabolism and storage root formation in sweet potato [Ipomoea batatas L. (Lam.) cv. Shangshu 19 and cv. Jixu 23] in 2015–2016. Two fertilizer treatments, ammonium nitrogen (AN) and amide nitrogen (XN), were applied at 60 kg ha−1 in a two‐factor split‐plot design. The effects of nitrogen form on the morphology of adventitious roots, carbohydrate metabolism in potential storage roots, and number of storage roots per plant in sweet potato were investigated. The results show that during the early growth phase, the AN treatment significantly increased the number of adventitious roots, root tips, root length density, and fresh weight of roots (in pot trials). This treatment also significantly decreased the sucrose concentration of potential storage roots and increased the activities of cell wall, vacuolar, and cytoplasmic invertases. However, XN‐treated potential storage roots showed a relatively high starch concentration, activities of sucrose synthase and ADP‐glucose pyrophosphorylase, and transcription of sporamin genes. At the canopy closure period, the AN treatment significantly increased the number of storage roots of 0.5–5.0 cm in diameter and decreased the number of those > 5 cm in diameter compared to the control. The XN treatment induced the opposite effects. In the harvesting period, the AN treatment produced the highest storage root yield and number of storage roots per plant. Thus, in field trials the AN treatment induced a greater increase in production by increasing the number of storage roots.

Comparison of sucrose metabolism in wheat seedlings during drought stress and subsequent recovery

Sucrose is a dominant sugar transported to the sink organs of a plant where it is metabolized to other compounds or stored. Here, the importance of sucrose metabolism in a drought-tolerant wheat cultivar was compared to a drought-sensitive one. The 4-d-old Triticum aestivum L. seedlings were exposed to drought induced by water cessation for 7 d and recovery after re-watering for further 7 d. Under control conditions, constitutive expression of genes encoding vacuolar invertase (VI) and sucrose synthase (SuS) and activity of sucrose phosphate synthase (SPS) were significantly higher in the tolerant cultivar than in the sensitive one. Drought promoted the expressions of SPS and VI genes in the tolerant cultivar and increased their activities to 175 and 132 %, respectively, of those under control conditions. The activity of SuS and expression of its gene, however, were identical in both cultivars under drought stress. These changes resulted in more remarkable accumulation of sucrose in tolerant than in sensitive cultivar under water stress.

Characterization of Sugar Contents and Sucrose Metabolizing Enzymes in Developing Leaves of Hevea brasiliensis.

Sucrose-metabolizing enzymes in plant leaves have hitherto been investigated mainly in temperate plants, and rarely conducted in tandem with gene expression and sugar analysis. Here, we investigated the sugar content, gene expression, and the activity of sucrose-metabolizing enzymes in the leaves of Hevea brasiliensis, a tropical tree widely cultivated for natural rubber. Sucrose, fructose and glucose were the major sugars detected in Hevea leaves at four developmental stages (I to IV), with starch and quebrachitol as minor saccharides. Fructose and glucose contents increased until stage III, but decreased strongly at stage IV (mature leaves). On the other hand, sucrose increased continuously throughout leaf development. Activities of all sucrose-cleaving enzymes decreased markedly at maturation, consistent with transcript decline for most of their encoding genes. Activity of sucrose phosphate synthase (SPS) was low in spite of its high transcript levels at maturation. Hence, the high sucrose content in mature leaves was not due to increased sucrose-synthesizing activity, but more to the decline in sucrose cleavage. Gene expression and activities of sucrose-metabolizing enzymes in Hevea leaves showed striking differences compared with other plants. Unlike in most other species where vacuolar invertase predominates in sucrose cleavage in developing leaves, cytoplasmic invertase and sucrose synthase (cleavage direction) also featured prominently in Hevea. Whereas SPS is normally responsible for sucrose synthesis in plant leaves, sucrose synthase (synthesis direction) was comparable or higher than that of SPS in Hevea leaves. Mature Hevea leaves had an unusually high sucrose:starch ratio of about 11, the highest reported to date in plants.

Genetic Evidence for the Role of a Rice Vacuolar Invertase as a Molecular Sink Strength Determinant

BackgroundRice is a major crop feeding the majority of the global population, and increasing its sink strength is one of the modes to alleviate the declining availability of food for the rapidly growing world population. We demonstrate a role for an important rice vacuolar invertase isoform, OsINV3, in sink strength determination.ResultsOsINV3 mutants showed shorter panicles with lighter and smaller grains, owing to a smaller cell size on the outer and inner surfaces of the palea and lemma as observed by scanning electron microscopy. Further, strong promoter::GUS expression was observed in the palea, lemma and the rachis branches in the young elongating panicles, which supported the role of OsINV3 in cell expansion and thus, in spikelet size and panicle length determination. Size of the spikelet was found to directly influence the grain weight, which was confirmed by the lack of differences in weights of hulled grain for differently segregated alleles in the heterozygous lines. Assessment of field grown mutants not only revealed a drastic reduction in the percentage of ripened grain, 1000-grain weight and final yield, but also significantly reduced partitioning of assimilates to the panicles, whereby the total dry weight remained unaffected. Determination of the non-structural carbohydrate contents revealed a lower hexose-to-sucrose ratio in the panicles of the mutants from panicle initiation to 10 days after heading, a stage that identifies as the critical pre-storage phase of grain filling, whereas the starch contents were not affected. In addition, strong promoter::GUS expression was observed in the dorsal end of ovary during the pre-storage phase until 6 days after flowering, highlighting a function for OsINV3 in monitoring the initial grain filling stage.ConclusionsOsINV3 was found to regulate spikelet size by playing a key role in cell expansion, driving the movement of assimilates for grain filling by modulating the hexose-to-sucrose ratio, contributing in grain weight determination and thus, the grain yield.

Sugar metabolism reprogramming in a non-climacteric bud mutant of a climacteric plum fruit during development on the tree.

We investigated sugar metabolism in leaves and fruits of two Japanese plum (Prunus salicina Lindl.) cultivars, the climacteric Santa Rosa and its bud sport mutant the non-climacteric Sweet Miriam, during development on the tree. We previously characterized differences between the two cultivars. Here, we identified key sugar metabolic pathways. Pearson coefficient correlations of metabolomics and transcriptomic data and weighted gene co-expression network analysis (WGCNA) of RNA sequencing (RNA-Seq) data allowed the identification of 11 key sugar metabolism-associated genes: sucrose synthase, sucrose phosphate synthase, cytosolic invertase, vacuolar invertase, invertase inhibitor, α-galactosidase, β-galactosidase, galactokinase, trehalase, galactinol synthase, and raffinose synthase. These pathways were further assessed and validated through the biochemical characterization of the gene products and with metabolite analysis. Our results demonstrated the reprogramming of sugar metabolism in both leaves and fruits in the non-climacteric plum, which displayed a shift towards increased sorbitol synthesis. Climacteric and non-climacteric fruits showed differences in their UDP-galactose metabolism towards the production of galactose and raffinose, respectively. The higher content of galactinol, myo-inositol, raffinose, and trehalose in the non-climacteric fruits could improve the ability of the fruits to cope with the oxidative processes associated with fruit ripening. Overall, our results support a relationship between sugar metabolism, ethylene, and ripening behavior.

Phylloxeration effects on the sink activity and assimilation rate in phylloxera (Daktulosphaira vitifoliaeFitch) infested grapevines (Vitisspp.)

Grape phylloxera (Daktulosphaira vitifoliae Fitch) induces root galls on tips of susceptible and partially resistant rootstocks. The monophagous parasite affects the host by inhibition of water- and mineral uptake, sink-source translocation, changes in secondary metabolisms and manipulation of root growth. Grafted phylloxerated vines rarely show severe plant damage. Compensation effects on leaf galled vines have been proposed, however no information is available for root galled vines. In two experiments we aimed to determine effects of root phylloxeration on the primary metabolism of Vitis spp. We hypothesise that affected vines respond with a compensation strategy consisting of an increased root growth, leaf respiration and photosynthetic activity. Experiment 1 was conducted to analyse phylloxeration effects on carbohydrate mobility within grafted vines (V. vinifera L. 'Pinot Noir' on SO4). Experiment 2 analyzed potential phylloxeration effects on vines under nitrogen deficiency on Teleki 5C (V. berlandieri × V. riparia). Assessed parameters included plant phenology, quantitative plant growth, levels of macro and micro nutrients, photosynthetic activity, gas exchange and activity of enzymes related to carbohydrate mobility (sucrose synthase, vacuolar invertase, cell wall invertase). Our results showed an increased root mass in combination with elevated carbon and nitrogen levels in the root system of phylloxerated vines. Gas exchange and enzymatic activity of enzymes related to carbohydrate mobility were increased upon phylloxeration. However shoot and berry growths were not reduced. In conclusion these results show that more carbohydrates are imported in the root system upon phylloxeration without reductions of shoot and berry growth indicating a compensation strategy of the host. However it is unclear whether grape phylloxera actively triggers the import of carbohydrates to the root system by potential effectors or the host counteracts the lack of nutrients withdrawn by the parasite.

The ripening disorder berry shrivel affects anthocyanin biosynthesis and sugar metabolism in Zweigelt grape berries

Main conclusionTimescale analyses suggest the berry shrivel (BS) disorder is induced before veraison with strong effects on anthocyanin biosynthesis, and minor effects on sugar transport and metabolism.Berry shrivel (BS)-affected grapes have low sugar contents, high acidity, less anthocyanins and flaccid berries. To date no pathogenic causes are known, and studies to elucidate the molecular basis leading to symptom induction and development are limited. Here we present a study on pre-symptomatic as well as symptomatic BS berries to characterize early metabolic changes, with focus on anthocyanin biosynthesis and sugars metabolism. Healthy and BS berries from six sampling time points were used (BBCH79–BBCH89). Our objectives are (1) to search for the beginning of BS-related physiological processes; (2) to search for key enzymes and sugar transporters involved in BS induction and development and (3) to understand the consequences on polyphenol biosynthesis. We employed high performance anion exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) and liquid chromatography–tandem mass spectrometry (LC–MS/MS) technology for sugar and polyphenol analyses, respectively. Additionally we conducted expression analyses (qPCR) of key genes and enzymatic activity assays. Our results show that BS-related processes start before veraison, as determined by slightly reduced hexose contents and reduced expression levels of a vacuolar invertase (VviGIN1), two monosaccharide transporters (VviTMT2, VviTMT3) and the anthocyanin biosynthesis (VviUFGT, VviMYBA1/2) genes. Lower amounts of delphinidin and cyanidin glycosidic forms were determined, while caftaric acid, quercetin-3-O-glucuronide and (+)-catechin were increased in BS berries. Although not all results were conclusive, especially for the sugar metabolism, our data provide important knowledge to improve the understanding of the highly complex berry shrivel ripening disorder.

Exogenous cytokinin application to Actinidia chinensis var. deliciosa \u2018Hayward\u2019 fruit promotes fruit expansion through water uptake

Exogenous application of a cytokinin-like compound forchlorfenuron (CPPU) can promote fruit growth, although often at the expense of dry matter (DM), an important indicator of fruit quality. Actinidia chinensis var. deliciosa ‘Hayward’ fruit are very responsive to CPPU treatments, but the mechanism underlying the significant fruit weight increase and associated decrease in DM is unclear. In this study, we hypothesised that CPPU-enhanced growth increases fruit carbohydrate demand, but limited carbohydrate supply resulted in decreased fruit DM. During fruit development, CPPU effects on physical parameters, metabolites, osmotic pressure and transcriptional changes were assessed under conditions of both standard and a high carbohydrate supply. We showed that CPPU increased fruit fresh weight but the dramatic DM decrease was not carbohydrate limited. Enhanced glucose and fructose concentrations contributed to an increase in soluble carbohydrate osmotic pressure, which was correlated with increased water accumulation in CPPU-treated fruit and up-regulation of water channel aquaporin gene PIP2.4 at 49 days after anthesis. Transcipt analysis suggested that the molecular mechanism contributing to increased glucose and fructose concentrations was altered by carbohydrate supply. At standard carbohydrate supply, the early glucose increase in CPPU fruit was associated with reduced starch synthesis and increased starch degradation. When carbohydrate supply was high, the early glucose increase in CPPU fruit was associated with a general decrease in starch synthesis but up-regulation of vacuolar invertase and fructokinase genes. We conclude that CPPU affected fruit expansion by increasing the osmotically-driven water uptake and its effect was not carbohydrate supply-limited.

Etiolated Stem Branching Is a Result of Systemic Signaling Associated with Sucrose Level.

The potato (Solanum tuberosum) tuber is a swollen stem. Sprouts growing from the tuber nodes represent loss of apical dominance and branching. Long cold storage induces loss of tuber apical dominance and results in secondary branching. Here, we show that a similar branching pattern can be induced by short heat treatment of the tubers. Detached sprouts were induced to branch by the heat treatment only when attached to a parenchyma cylinder. Grafting experiments showed that the scion branches only when grafted onto heat- or cold-treated tuber parenchyma, suggesting that the branching signal is transmitted systemically from the bud-base parenchyma to the grafted stem. Exogenous supply of sucrose (Suc), glucose, or fructose solution to detached sprouts induced branching in a dose-responsive manner, and an increase in Suc level was observed in tuber parenchyma upon branching induction, suggesting a role for elevated parenchyma sugars in the regulation of branching. However, sugar analysis of the apex and node after grafting showed no distinct differences in sugar levels between branching and nonbranching stems. Vacuolar invertase is a key enzyme in determining the level of Suc and its cleavage products, glucose and fructose, in potato parenchyma. Silencing of the vacuolar invertase-encoding gene led to increased tuber branching in combination with branching-inducing treatments. These results suggest that Suc in the parenchyma induces branching through signaling and not by excess mobilization from the parenchyma to the stem.

Differential expression of vacuolar and defective cell wall invertase genes in roots and seeds of metalliferous and non-metalliferous populations of Rumex dentatus under copper stress

Acid invertase activities in roots and young seeds of a metalliferous population (MP) of Rumex dentatus were previously observed to be significantly higher than those of a non-metalliferous population (NMP) under Cu stress. To date, no acid invertase gene has been cloned from R. dentatus. Here, we isolated four full-length cDNAs from the two populations of R. dentatus, presumably encoding cell wall (RdnCIN1 and RdmCIN1 from the NMP and MP, respectively) and vacuolar invertases (RdnVIN1 and RdmVIN1 from the NMP and MP, respectively). Unexpectedly, RdnCIN1 and RdmCIN1 most likely encode special defective invertases with highly attenuated sucrose-hydrolyzing capacity. The transcript levels of RdmCIN1 were significantly higher than those of RdnCIN1 in roots and young seeds under Cu stress, whereas under control conditions, the former was initially lower than the latter. Unexpected high correlations were observed between the transcript levels of RdnCIN1 and RdmCIN1 and the activity of cell wall invertase, even though RdnCIN1 and RdmCIN1 do not encode catalytically active invertases. Similarly, the transcript levels of RdmVIN1 in roots and young seeds were increased under Cu stress, whereas those of RdnVIN1 were decreased. The high correlations between the transcript levels of RdnVIN1 and RdmVIN1 and the activity of vacuolar invertase indicate that RdnVIN1 and RdmVIN1 might control distinct vacuolar invertase activities in the two populations. Moreover, a possible indirect role for acid invertases in Cu tolerance, mediated by generating a range of sugars used as nutrients and signaling molecules, is discussed.

Suppression of the vacuolar invertase gene delays senescent sweetening in chipping potatoes.

Potato chip processors require potato tubers that meet quality specifications for fried chip color, and color depends largely upon tuber sugar contents. At later times in storage, potatoes accumulate sucrose, glucose, and fructose. This developmental process, senescent sweetening, manifests as a blush of color near the center of the fried chip, becomes more severe with time, and limits the storage period. Vacuolar invertase (VInv) converts sucrose to glucose and fructose and is hypothesized to play a role in senescent sweetening. To test this hypothesis, senescent sweetening was quantified in multiple lines of potato with reduced VInv expression. Chip darkening from senescent sweetening was delayed by about 4 weeks for tubers with reduced VInv expression. A strong positive correlation between frequency of dark chips and tuber hexose content was observed. Tubers with reduced VInv expression had lower hexose to sucrose ratios than controls. VInv activity contributes to reducing sugar accumulation during senescent sweetening. Sucrose breakdown during frying may contribute to chip darkening. Suppressing VInv expression increases the storage period of the chipping potato crop, which is an important consideration, as potatoes with reduced VInv expression are entering commercial production in the USA. © 2017 Society of Chemical Industry.

Comparative Transcriptome Analysis Reveals Critical Function of Sucrose Metabolism Related-Enzymes in Starch Accumulation in the Storage Root of Sweet Potato.

The starch properties of the storage root (SR) affect the quality of sweet potato (Ipomoea batatas (L.) Lam.). Although numerous studies have analyzed the accumulation and properties of starch in sweet potato SRs, the transcriptomic variation associated with starch properties in SR has not been quantified. In this study, we measured the starch and sugar contents and analyzed the transcriptome profiles of SRs harvested from sweet potatoes with high, medium, and extremely low starch contents, at five developmental stages [65, 80, 95, 110, and 125 days after transplanting (DAP)]. We found that differences in both water content and starch accumulation in the dry matter affect the starch content of SRs in different sweet potato genotypes. Based on transcriptome sequencing data, we assembled 112336 unigenes, and identified several differentially expressed genes (DEGs) involved in starch and sucrose metabolism, and revealed the transcriptional regulatory network controlling starch and sucrose metabolism in sweet potato SRs. Correlation analysis between expression patterns and starch and sugar contents suggested that the sugar–starch conversion steps catalyzed by sucrose synthase (SuSy) and UDP-glucose pyrophosphorylase (UGPase) may be essential for starch accumulation in the dry matter of SRs, and IbβFRUCT2, a vacuolar acid invertase, might also be a key regulator of starch content in the SRs. Our results provide valuable resources for future investigations aimed at deciphering the molecular mechanisms determining the starch properties of sweet potato SRs.

TAI vacuolar invertase orthologs: the interspecific variability in tomato plants (Solanum section Lycopersicon).

Understanding the genetic mechanisms underlying carbohydrate metabolism can promote the development of biotechnological advances in fruit plants. The flesh tomato fruit represents an ideal system for examining the role of sucrose cleavage enzymes in fruit development, and wild tomato species differing in storage sugars serve as an excellent research material for this purpose. Plant vacuolar invertase is a key enzyme of sucrose metabolism in the sink organs. In the present study, we identified complete gene sequences encoding the TAI vacuolar invertase in 11 wild and one cultivated tomato accessions of the Solanum section Lycopersicon. The average level of interspecific polymorphism in TAI genes was 8.58%; however, in the green-fruited tomatoes, the TAI genes contained 100 times more SNPs than those in the red-fruited accessions. The TAI proteins demonstrated 8% variability, whereas the red-fruited species had none. A TAI-based phylogenetic tree revealed two main clusters containing self-compatible and self-incompatible species, which concurs with the previous crossability-based division and demonstrates that the TAI genes reflect the evolutionary relationships between the red- and green-fruited tomatoes. Furthermore, we detected differential expression patterns of the TAI genes in the fruits of wild and cultivated tomatoes, which corresponded to sugar composition. The polymorphism analysis of the TAI acid invertases of Solanum section Lycopersicon species will contribute to the understanding of the genetic potential of TAI genes to impact tomato breeding through genetic engineering of the carbohydrate composition in the fruit.

Water deficit effects on carbon metabolism in cotton fibers during fiber elongation phase

The accumulation level of soluble carbohydrates in fibers during flowering may affect the final fiber length in upland cotton (Gossypium hirsutum L.). The objective of this study was to determine the relationship of carbohydrate levels and carbon metabolism enzyme activities in fibers to fiber elongation under water deficit stress. A pot study was conducted in 2013 with treatments consisting of control (well-watered) and water stress using two cotton genotypes differing in drought tolerance, namely S011 (drought sensitive) and T886 (drought tolerant). Water stress treatment was designed as withholding water from the pots until stomatal closure followed by limited water supply for 25 days where water-stressed plants received 40% of optimum quantity of water. The results showed that the glucose, fructose, sucrose and starch concentrations decreased in water-stressed fibers compared to the control. The degree of decline in hexose and total soluble sugar concentrations under water deficit stress was higher in the sensitive genotype (S011) than the tolerant genotype (T886). Similar changes were also observed in vacuolar invertase (VIN) and sucrose synthase (SuSy) activities. The reduction in hexose concentration might be due to decreased sucrose-cleaving enzyme activities especially VIN activity. The study showed that drought stress decreased VIN and SuSy activities, and hence hexose concentration in fiber cells, which resulted in a slower fiber elongation compared to the control.

Predictive Markers for Cold-Induced Sweetening Resistance in Cold Stored Potatoes (Solanum tuberosum L.)

An approach has been developed to screen a large number of potato clones for cold induced sweetening (CIS) resistance in breeding programs. Two key enzymes responsible for reducing sugar accumulation during cold storage were identified. Clones with the A-II isozymes of UDP-glucose pyrophosphorylase coupled with low activity of vacuolar acid invertase enzyme had increased resistance to CIS by forming less suc, which is subsequently hydrolyzed to the undesirable reducing sugars, glc and fru. Six named cultivars and 192 genetically diverse clones from various breeding programs in USA were analyzed over two years for the two key enzymes and sugar concentration in cold stored tubers. The predictability for CIS resistance during cold storage was 94% both years. Clones classified as class A accumulated low concentration of reducing sugar glc during cold storage. It is suggested that these two predictor enzymes can be used for screening parents and selections in potato breeding program.

QTL mapping of potato chip color and tuber traits within an autotetraploid family

Cultivated potato (Solanum tuberosum L.) is a highly heterozygous autotetraploid crop species, and this creates challenges for traditional line development and molecular breeding. Recent availability of a single-nucleotide polymorphism (SNP) array with 8303 features and software packages for linkage and association mapping in autotetraploid species present new opportunities for the identification of genomic regions that contribute to high-value traits in cultivated potato. A biparental tetraploid potato population was evaluated across three field seasons and storage trials in order to identify quantitative trait loci (QTL) for multiple tuber traits including fried chip color after 5.5–7.2 °C storage. Tetra-allelic dosage information was used to construct a genetic linkage map that covered 1041 cM and contained 2095 SNP markers with a median marker interval of 0.4 cM. A total of 41 QTL were identified for flower color, tuber yield, tuber number per plant, tuber weight, tuber size, and chip color after various storage regimes. Moderate effect QTL for chip color at 3 months were identified that co-localized with candidate genes vacuolar invertase (VInv), invertase inhibitor (INH2), and apoplastic invertase (Inv ap -b). A separate QTL for chip color after 6 months of storage was identified in the short arm of chromosome 2, and this locus may contribute to variation in senescent sweetening resistance. QTL for tuber weight, length, and width co-localized with a known QTL for plant maturity on chromosome 5. Genome-wide association mapping using a polyploid model detected the tuber size QTL and identified a number of candidate SNPs, but was unable to detect markers significantly associated with chip color.

Enhanced vacuolar invertase activity and capability for carbohydrate import in GA-treated inflorescence correlate with increased fruit set in grapevine

Application of exogenous gibberellin (GA) has been widely used to improve fruit set in grapevine, although the underlying molecular mechanism remains unknown. In this work, GA application at 10 days before anthesis promoted fruit set of grapevine “Fenghou” (Vitis vinifera × V. labrusca) and increased sugar content and acid invertase activity in the ovary, especially vacuolar invertase (VIN) activity during fruit set, compared with the control. However, the GA biosynthesis inhibitor paclobutrazol (PAC) had the opposite effect. Tissue-specific expression analysis and in situ hybridization showed that transcripts of three acid invertase genes were primarily detected in the phloem of the flower and fruitlet. In contrast to the expression of the cell wall invertase gene, which was only induced before blooming, transcript accumulation of the VIN genes VvGIN1 and VvGIN2 was enhanced during fruit set by GA application. Nevertheless, PAC treatment reduced VvGIN1 transcript accumulation after anthesis, but had no significant effect on the VvGIN2 transcript level at the same time point. Interestingly, the effect of GA and PAC application on expression of VvPIN1-like and VvPIN3-like (auxin transporters) was similar to that on VvGIN1 after blooming. In addition, the inducibility of the VvGIN1 and VvGIN2 promoters by GA was demonstrated using transient expression assays in tobacco leaves. These results indicated that enhanced VIN activity and carbohydrate import into grapevine fruitlets by GA application contribute to the increased fruit set through increasing sink strength and sugar signaling activities, possibly by regulating polar auxin transport.