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Endodormancy (ED) is a crucial stage in the life cycle of many perennial plants. ED release requires accumulating a certain amount of cold exposure, measured as chilling units. However, the mechanism governing the effect of chilling on ED duration is poorly understood. We used the potato tuber model to investigate the response to chilling as associated with ED release. We measured the accumulation of specific sugars during and after chilling, defined as sugar units. We discovered that ED duration correlated better with sugar units accumulation than chilling units. A logistic function was developed based on sugar units measurements to predict ED duration. Knockout or overexpression of the vacuolar invertase gene (StVInv) unexpectedly modified sugar units levels and extended or shortened ED, respectively. Silencing the energy sensor SNF1‐related protein kinase 1, induced higher sugar units accumulation and shorter ED. Sugar units accumulation induced by chilling or transgenic lines reduced plasmodesmal (PD) closure in the dormant bud meristem. Chilling or knockout of abscisic acid (ABA) 8′‐hydroxylase induced ABA accumulation, in parallel to sweetening, and antagonistically promoted PD closure. Our results suggest that chilling induce sugar units and ABA accumulation, resulting in antagonistic signals for symplastic connection of the dormant bud. Summary statement: Endodormancy (ED) duration of potato tuber correlates better with soluble sugars accumulation (sugar units) than chilling units. Chilling induces sugar units and ABA accumulation, resulting in antagonistic signals for symplastic connection of the dormant bud, determining ED duration. [ABSTRACT FROM AUTHOR]

Water deficit currently acts as one of the largest limiting factors for agricultural productivity worldwide. Additionally, limitation by water scarcity is projected to continue in the future with the further onset of effects of global climate change. As a result, it is critical to develop or breed for crops that have increased water use efficiency and that are more capable of coping with water scarce conditions. However, increased intrinsic water use efficiency (iWUE) typically brings a trade-off with CO2 assimilation as all gas exchange is mediated by stomata, through which CO2 enters the leaf while water vapor exits. Previously, promising results were shown using guard-cell-targeted overexpression of hexokinase to increase iWUE without incurring a penalty in photosynthetic rates or biomass production. Here, two homozygous transgenic tobacco (Nicotiana tabacum) lines expressing Arabidopsis Hexokinase 1 (AtHXK1) constitutively (35SHXK2 and 35SHXK5) and a line that had guard-cell-targeted overexpression of AtHXK1 (GCHXK2) were evaluated relative to wild type for traits related to photosynthesis and yield. In this study, iWUE was significantly higher in GCHXK2 compared with wild type without negatively impacting CO2 assimilation, although results were dependent upon leaf age and proximity of precipitation event to gas exchange measurement. [ABSTRACT FROM AUTHOR]

Sucrose synthase (SuSy) and fructokinase (FRK) work together to control carbohydrate flux in sink tissues. SuSy cleaves sucrose into fructose and UDP-glucose; whereas FRK phosphorylates fructose. Previous results have shown that suppression of the SUS1,3&4 genes by SUS-RNAi alters auxin transport in the shoot apical meristems of tomato plants and affects cotyledons and leaf structure; whereas antisense suppression of FRK2 affects vascular development. To explore the joint developmental roles of SuSy and FRK, we crossed SUS-RNAi plants with FRK2-antisense plants to create double-mutant plants. The double-mutant plants exhibited novel phenotypes that were absent from the parent lines. About a third of the plants showed arrested shoot apical meristem around the transition to flowering and developed ectopic meristems. Use of the auxin reporter DR5::VENUS revealed a significantly reduced auxin response in the shoot apical meristems of the double-mutant, indicating that auxin levels were low. Altered inflorescence phyllotaxis and significant disorientation of vascular tissues were also observed. In addition, the fruits and the seeds of the double-mutant plants were very small and the seeds had very low germination rates. These results show that SUS1,3&4 and FRK2 enzymes are jointly essential for proper meristematic and vascular development, and for fruit and seed development. [ABSTRACT FROM AUTHOR]

The hypocotyls of germinating seedlings elongate in a search for light to enable autotrophic sugar production. Upon exposure to light, photoreceptors that are activated by blue and red light halt elongation by preventing the degradation of the hypocotyl-elongation inhibitor HY5 and by inhibiting the activity of the elongation-promoting transcription factors PIFs. The question of how sugar affects hypocotyl elongation and which cell types stimulate and stop that elongation remains unresolved. We found that overexpression of a sugar sensor, Arabidopsis hexokinase 1 (HXK1), in guard cells promotes hypocotyl elongation under white and blue light through PIF4. Furthermore, expression of PIF4 in guard cells is sufficient to promote hypocotyl elongation in the light, while expression of HY5 in guard cells is sufficient to inhibit the elongation of the hy5 mutant and the elongation stimulated by HXK1. HY5 exits the guard cells and inhibits hypocotyl elongation, but is degraded in the dark. We also show that the inhibition of hypocotyl elongation by guard cells' HY5 involves auto-activation of HY5 expression in other tissues. It appears that guard cells are capable of coordinating hypocotyl elongation and that sugar and HXK1 have the opposite effect of light on hypocotyl elongation, converging at PIF4. Kelly et al. show that Arabidopsis hexokinase1 (HXK1) expressed in guard-cells is sufficient to drive hypocotyl elongation through increasing the activity of PIF4 and auxin level, and competing with the effects of HY5. This study provides insights into how light and sucrose antagonistically coordinate the effort to achieve the height necessary for efficient photosynthetic, autotrophic sugar production. [ABSTRACT FROM AUTHOR]

Crop yield is largely affected by global climate change. Especially periods of heat and drought limit crop productivity worldwide. According to current models of future climate scenarios, heatwaves and periods of drought are likely to increase. Potato, as an important food crop of temperate latitudes, is very sensitive to heat and drought which impact tuber yield and quality. To improve abiotic stress resilience of potato plants, we aimed at co-expressing hexokinase 1 from Arabidopsis thaliana (AtHXK1) in guard cells and SELF-PRUNING 6A (SP6A) using the leaf/stem-specific StLS1 promoter in order to increase water use efficiency as well as tuberization under drought and heat stress. Guard cell-specific expression of AtHXK1 decreased stomatal conductance and improved water use efficiency of transgenic potato plants as has been shown for other crop plants. Additionally, co-expression with the FT-homolog SP6A stimulated tuberization and improved assimilate allocation to developing tubers under control as well as under single and combined drought and heat stress conditions. Thus, co-expression of both proteins provides a novel strategy to improve abiotic stress tolerance of potato plants. [ABSTRACT FROM AUTHOR]

The temporal formation and spatial distribution of stomata on the surface of citrus floral organs and, specifically, on the ovule from which the fruit develops, were analyzed using citrus plants that express green fluorescent protein (GFP) under the guard cell-specific KST1 promoter. Stomata are found on the style, sepal, and anther of the closed flower and on ovules from the stage of anthesis. It has previously been shown that hexokinase (HXK) mediates sugar-sensing in leaf guard cells and stimulates stomatal closure. The activity and response of citrus fruit stomata to sugar-sensing by HXK was examined using plants that express HXK under the KST1 promoter. Those plants are referred to as GCHXK plants. The transpiration of young green GCHXK citrus fruits was significantly reduced, indicating that their stomata respond to sugar similar to leaf stomata. Toward fruit maturation, fruit stomata are plugged and stop functioning, which explains why WT and GCHXK mature yellow fruits exhibited similar water loss. Seeds of the GCHXK plants were smaller and germinated more slowly than the WT seeds. We suggest that the stomata of young green citrus fruits, but not mature yellow fruits, respond to sugar levels via HXK and that fruit stomata are important for proper seed development. [ABSTRACT FROM AUTHOR]

Water is a limiting resource for many land plants. Most of the water taken up by plants is lost to the atmosphere through the stomata, which are adjustable pores on the leaf surface that allow for gas exchange between the plant and the atmosphere. Modulating stomatal activity might be an effective way to reduce plants' water consumption and enhance their productivity under normal, as well as water-limiting conditions. Our recent discovery of stomatal regulation by sugars that is mediated by guard-cell hexokinase (HXK), a sugar-sensing enzyme, has raised the possibility that HXK might be used to increase plant water-use efficiency (WUE; i.e., carbon gain per unit of water). We show here that transgenic tomato and Arabidopsis plants with increased expression of HXK in their guard cells (GCHXK plants) exhibit reduced transpiration and higher WUE without any negative effects on growth under normal conditions, as well as drought avoidance and improved photosynthesis and growth under limited-water conditions. Our results demonstrate that exclusive expression of HXK in guard cells is an effective tool for improving WUE, and plant performance under drought. [ABSTRACT FROM AUTHOR]

Background: Although elevated [CO2] causes an increase of photosynthesis in the short-term, this increase is often attenuated over time due to decreased photosynthetic capacity of the leaf in a process called photosynthetic acclimation to elevated CO2 (PAC). PAC is often accompanied by N deficiency and sink:source imbalance. The aim of this study is to investigate mechanisms that lead to PAC, N deficiency and sink:source imbalance in tomato plants grown in elevated [CO2] and how they are affected by different levels of N treatments.Methods: Two long-term experiment and two short-term experiments were conducted in which tomato plants were grown in chambers with ambient [CO2] and elevated [CO2] combined with different levels of N nutrition. The following parameters were measured: 1) Biomass 2)Leaf N, P and K concentrations, 3) leaf NO3− concentration, 4) Gas exchange 5) Rubisco expression and 6) Leaf starch concentration.Results: Plants grown at e[CO2] had increased biomass and starch, and decreased gas exchange, stomatal conductivity, Rubisco expression, Vcmax, NPK and leaf NO3−. Increasing N fertilization counteracted many of the effects of elevated [CO2].Conclusions: PAC was caused by decreased N uptake or transport coupled with increased growth which leads to N deficiency and a sink:source imbalance. Increased N fertilization counteracted the effect of e[CO2] on photosynthesis, N status, and sink:source imbalance. Furthermore, elevated [CO2] caused stomata to partially close, which accounted for some of the PAC observed. [ABSTRACT FROM AUTHOR]

As plants evolved to function on land, they developed stomata for effective gas exchange, for photosynthesis and for controlling water loss. We have recently shown that sugars, as the end product of photosynthesis, close the stomata of various angiosperm species, to coordinate sugar production with water loss. In the current study, we examined the sugar responses of the stomata of phylogenetically different plant species and species that employ different photosynthetic mechanisms (i.e., C3, C4 and CAM). To examine the effect of sucrose on stomata, we treated leaves with sucrose and then measured their stomatal apertures. Sucrose reduced stomatal aperture, as compared to an osmotic control, suggesting that regulation of stomata by sugars is a trait that evolved early in evolutionary history and has been conserved across different groups of plants. [ABSTRACT FROM AUTHOR]

Metabolic enzymes have been found to play roles in plant development. Sucrose synthase (SUS) is one of the two enzyme families involved in sucrose cleavage in plants. In tomato, six SUS genes have been found. We generated transgenic tomato plants with RNAi suppression of SlSUS1, SlSUS3 and SlSUS4 genes. Independent transgenic lines with RNAi suppression of more than one SUS gene exhibited morphological effects on their cotyledons and leaf structure, but there were no significant effects on their carbohydrate levels, demonstrating that SUS has a developmental function, in addition to its metabolic function. Shoot apices of the transgenic lines showed elevated expression of JAGGED (JAG) and the auxin transporter PIN1. In a PIN1-GFP fusion reporter/SUS-RNAi hybrid, PIN1-GFP patterns were altered in developing leaves (as compared to control plants), indicating that SlSUS suppression alters auxin signaling. These results suggest possible roles for SUS in the regulation of plant growth and leaf morphology, in association with the auxin-signaling pathway. [ABSTRACT FROM AUTHOR]

Sugars affect central aspects of plant physiology, including photosynthesis, stomatal behavior and the loss of water through the stomata. Yet, the potential effects of sugars on plant aquaporins ( AQPs) and water conductance have not been examined. We used database and transcriptional analyses, as well as cellular and whole-plant functional techniques to examine the link between sugar-related genes and AQPs. Database analyses revealed a high level of correlation between the expression of AQPs and that of sugar-related genes, including the Arabidopsis hexokinases 1 ( At HXK1). Increased expression of At HXK1, as well as the addition of its primary substrate, glucose (Glc), repressed the expression of 10 AQPs from the plasma membrane-intrinsic proteins ( PIP) subfamily ( PIP- AQPs) and induced the expression of two stress-related PIP- AQPs. The osmotic water permeability of mesophyll protoplasts of At HXK1-expressing plants and the leaf hydraulic conductance of those plants were significantly reduced, in line with the decreased expression of PIP- AQPs. Conversely, hxk1 mutants demonstrated a higher level of hydraulic conductance, with increased water potential in their leaves. In addition, the presence of Glc reduced leaf water potential, as compared with an osmotic control, indicating that Glc reduces the movement of water from the xylem into the mesophyll. The production of sugars entails a significant loss of water and these results suggest that sugars and At HXK1 affect the expression of AQP genes and reduce leaf water conductance, to coordinate sugar levels with the loss of water through transpiration. [ABSTRACT FROM AUTHOR]

To date, guard cell promoters have been examined in only a few species, primarily annual dicots. A partial segment of the potato (Solanum tuberosum) KST1 promoter (KST1 partial promoter, KST1ppro) has previously been shown to confer guard cell expression in potato, tomato (Solanum lycopersicum), citrus [Troyer citrange (C. sinensisxPoncirus trifoliata)], and Arabidopsis (Arabidopsis thaliana). Here, we describe an extensive analysis of the expression pattern of KST1ppro in eight (previously reported, as well as new) species from five different angiosperm families, including the Solanaceae and the Cucurbitaceae, Arabidopsis, the monocot barley (Hordeum vulgare), and two perennial species: grapevine (Vitis vinifera) and citrus. Using confocal imaging and three-dimensional movies, we demonstrate that KST1ppro drives guard cell expression in all of these species, making it the first dicot-originated guard cell promoter shown to be active in a monocot and the first promoter reported to confer guard cell expression in barley and cucumber (Cucumis sativus). The results presented here indicate that KST1ppro can be used to drive constitutive guard cell expression in monocots and dicots and in both annual and perennial plants. In addition, we show that the KST1ppro is active in guard cells shortly after the symmetric division of the guard mother cell and generates stable expression in mature guard cells. This allows us to follow the spatial and temporal distribution of stomata in cotyledons and true leaves. [ABSTRACT FROM AUTHOR]

Sucrose (a disaccharide made of glucose and fructose) is the primary carbon source transported to sink organs in many plants. Since fructose accounts for half of the hexoses used for metabolism in sink tissues, plant fructokinases (FRKs), the main fructose-phosphorylating enzymes, are likely to play a central role in plant development. However, to date, their specific functions have been the subject of only limited study. The Arabidopsis genome contains seven genes encoding six cytosolic FRKs and a single plastidic FRK. T-DNA knockout mutants for five of the seven FRKs were identified and used in this study. Single knockouts of the FRK mutants did not exhibit any unusual phenotype. Double-mutants of AtFRK6 (plastidic) and AtFRK7 showed normal growth in soil, but yielded dark, distorted seeds. The seed distortion could be complemented by expression of the well-characterized tomato SlFRK1, confirming that a lack of FRK activity was the primary cause of the seed phenotype. Seeds of the double-mutant germinated, but failed to establish on 1/2 MS plates. Seed establishment was made possible by the addition of glucose or sucrose, indicating reduced seed storage reserves. Metabolic profiling of the double-mutant seeds revealed decreased TCA cycle metabolites and reduced fatty acid metabolism. Examination of the mutant embryo cells revealed smaller oil bodies, the primary storage reserve in Arabidopsis seeds. Quadruple and penta FRK mutants showed growth inhibition and leaf wilting. Anatomical analysis revealed smaller trachea elements and smaller xylem area, accompanied by necrosis around the cambium and the phloem. These results demonstrate overlapping and complementary roles of the plastidic AtFRK6 and the cytosolic AtFRK7 in seed storage accumulation, and the importance of AtFRKs for vascular development. [ABSTRACT FROM AUTHOR]

The present study explores the extent to which maternal attachment and teacher–student attachment-like relationships explain the socioemotional adaptation of students with disabilities. Participants consisted of 65 dyads of homeroom teachers and their students (from a middle-to-low-class area in Northern Israel) with learning disabilities (LD), attention deficit hyperactivity disorder (ADHD), and LD/ADHD-co-morbidity (mean age = 10.9). Students were assessed based on the maternal attachment security scale (ASS) and the appraisal of the teacher as a secure base scale. Homeroom teachers completed the student-teacher security scale. Third-party teachers reported on students’ school adaptation. Results indicated that beyond the disability factor, secure teacher-student relationships reduced the students’ externalising problem behaviours and improved student learning proficiencies that are considered to be affected by the adequacy of executive functioning. Additionally, maternal attachment reduced internalising problem behaviours. Implications for the protective role of teachers of students with disabilities and for research on teacher–student relationship quality are discussed. [ABSTRACT FROM PUBLISHER]

Plants have two kinds of fructokinases ( FRKs) that catalyze the key step of fructose phosphorylation, cytosolic and plastidic. The major cytosolic tomato FRK, Sl FRK2, is essential for the development of xylem vessels., In order to study the role of Sl FRK3, which encodes the only plastidic FRK, we generated transgenic tomato ( Solanum lycopersicon) plants with RNAi suppression of Sl FRK3 as well as plants expressing beta-glucoronidase ( GUS) under the Sl FRK3 promoter., GUS staining indicated SlFRK3 expression in vascular tissues of the leaves and stems, including cambium, differentiating xylem, young xylem fibers and phloem companion cells. Suppression of Sl FRK3 reduced the stem xylem area, stem and root water conductance, and whole-plant transpiration, with minor effects on plant development. However, suppression of Sl FRK3 accompanied by partial suppression of Sl FRK2 induced significant growth-inhibition effects, including the wilting of mature leaves. Grafting experiments revealed that these growth effects are imposed primarily by the leaves, whose petioles had unlignified, thin-walled xylem fibers with collapsed parenchyma cells around the vessels. A cross between the Sl FRK2-antisense and Sl FRK3- RNAi lines exhibited similar wilting and anatomical effects, confirming that these effects are the result of the combined suppression of Sl FRK3 and Sl FRK2., These results demonstrate a role of the plastidic Sl FRK3 in xylem development and hydraulic conductance. [ABSTRACT FROM AUTHOR]

Hexokinase (HXK) is a sugar-phosphorylating enzyme involved in sugar-sensing. It has recently been shown that HXK in guard cells mediates stomatal closure and coordinates photosynthesis with transpiration in the annual species tomato and Arabidopsis. To examine the role of HXK in the control of the stomatal movement of perennial plants, we generated citrus plants that express Arabidopsis HXK1 (AtHXK1) under KST1, a guard cell-specific promoter. The expression of KST1 in the guard cells of citrus plants has been verified using GFP as a reporter gene. The expression of AtHXK1 in the guard cells of citrus reduced stomatal conductance and transpiration with no negative effect on the rate of photosynthesis, leading to increased water-use efficiency. The effects of light intensity and humidity on stomatal behavior were examined in rooted leaves of the citrus plants. The optimal intensity of photosynthetically active radiation and lower humidity enhanced stomatal closure of AtHXK1-expressing leaves, supporting the role of sugar in the regulation of citrus stomata. These results suggest that HXK coordinates photosynthesis and transpiration and stimulates stomatal closure not only in annual species, but also in perennial species. [ABSTRACT FROM AUTHOR]

Increasing fructokinase (FRK) activity in cotton (Gossypium hirsutum L.) plants may reduce fructose inhibition of sucrose synthase (Sus) and lead to improved fibre yield and quality. Cotton was transformed with a tomato (Solanum lycopersicum L.) fructokinase gene (LeFRK1) under the control of the CMV 35S promoter. In a greenhouse, the LeFRK1 plants had increased fibre and leaf FRK activity over nonexpressing nulls, but not improved fibre length and strength. Compared with the nulls, LeFRK1 plants yielded 13-100% more seed-cotton mass per boll and more bolls per plant, and therefore more seed cotton and fibre yield per plant. The enhanced yield was related to a greater seed number per boll for LeFRK1 plants. Photosynthetic rates were not appreciably different among genotypes. However, more area per leaf and leaf number (in some instances) for LeFRK1 plants than for nulls enhanced the capacity for C gain. Larger leaf areas for LeFRK1 plants were associated with larger stem diameters. Lower sucrose levels in developing leaves of LeFRK1 plants suggest that LeFRK1 overexpression leads to improved in vivo Sus activity in developing leaves and possibly in developing seeds. The improvement in yield for LeFRK1 plants may also be the result of improvements in photosynthate supply as a consequence of greater leaf area. [ABSTRACT FROM AUTHOR]

Guard cells represent a unique single cell-type system for the study of cellular responses to abiotic and biotic perturbations that affect stomatal movement. Decades of effort through both classical physiological and functional genomics approaches have generated an enormous amount of information on the roles of individual metabolites in stomatal guard cell function and physiology. Recent application of metabolomics methods has produced a substantial amount of new information on metabolome control of stomatal movement. In conjunction with other “omics” approaches, the knowledge-base is growing to reach a systems-level description of this single cell-type. Here we summarize current knowledge of the guard cell metabolome and highlight critical metabolites that bear significant impact on future engineering and breeding efforts to generate plants/crops that are resistant to environmental challenges and produce high yield and quality products for food and energy security. [ABSTRACT FROM AUTHOR]

Our understanding of the cellular role of aquaporins (AQPs) in the regulation of whole-plant hydraulics, in general, and extravascular, radial hydraulic conductance in leaves (Kleaf), in particular, is still fairly limited. We hypothesized that the AQPs of the vascular bundle sheath (BS) cells regulate Kleaf. To examine this hypothesis, AQP genes were silenced using artificial microRNAs that were expressed constitutively or specifically targeted to the BS. MicroRNA sequences were designed to target all five AQP genes from the PLASMA MEMBRANE-INTRINSIC PROTEIN1 (PIP1) subfamily. Our results show that the constitutively silenced PIP1 (35S promoter) plants had decreased PIP1 transcript and protein levels and decreased mesophyll and BS osmotic water permeability (Pf), mesophyll conductance of CO2, photosynthesis, Kleaf, transpiration, and shoot biomass. Plants in which the PIP1 subfamily was silenced only in the BS (SCARECROW:microRNA plants) exhibited decreased mesophyll and BS Pf and decreased Kleaf but no decreases in the rest of the parameters listed above, with the net result of increased shoot biomass. We excluded the possibility of SCARECROW promoter activity in the mesophyll. Hence, the fact that SCARECROW:microRNA mesophyll exhibited reduced Pf, but not reduced mesophyll conductance of CO2, suggests that the BS-mesophyll hydraulic continuum acts as a feed-forward control signal. The role of AQPs in the hierarchy of the hydraulic signal pathway controlling leaf water status under normal and limited-water conditions is discussed. [ABSTRACT FROM AUTHOR]

Stomata control gaseous fluxes between the internal leaf air spaces and the external atmosphere. Guard cells determine stomatal aperture and must operate to ensure an appropriate balance between CO2 uptake for photosynthesis (A) and water loss, and ultimately plant water use efficiency (WUE). A strong correlation between A and stomatal conductance (gs) is well documented and often observed, but the underlying mechanisms, possible signals and metabolites that promote this relationship are currently unknown. In this review we evaluate the current literature on mesophyll-driven signals that may coordinate stomatal behaviour with mesophyll carbon assimilation. We explore a possible role of various metabolites including sucrose and malate (from several potential sources; including guard cell photosynthesis) and new evidence that improvements in WUE have been made by manipulating sucrose metabolism within the guard cells. Finally we discuss the new tools and techniques available for potentially manipulating cell-specific metabolism, including guard and mesophyll cells, in order to elucidate mesophyll-derived signals that coordinate mesophyll CO2 demands with stomatal behaviour, in order to provide a mechanistic understanding of these processes as this may identify potential targets for manipulations in order to improve plant WUE and crop yield. [ABSTRACT FROM AUTHOR]

The basic requirements for plant growth are light, CO2, water, and minerals. However, the absorption and utilization of each of these requires investment on the part of the plant. The primary products of plants are sugars, and the hexose sugars glucose and fructose are the raw material for most of the metabolic pathways and organic matter in plants. To be metabolized, hexose sugars must first be phosphorylated. Only two families of enzymes capable of catalysing the essential irreversible phosphorylation of glucose and fructose have been identified in plants, hexokinases (HXKs) and fructokinases (FRKs). These hexose-phosphorylating enzymes appear to coordinate sugar production with the abilities to absorb light, CO2, water, and minerals. This review describes the long- and short-term effects mediated by HXK and FRK in various tissues, as well as the role of these enzymes in the coordination of sugar production with the absorption of light, CO2, water, and minerals. [ABSTRACT FROM AUTHOR]

Increased expression of the aquaporin NtAQP1, which is known to function as a plasmalemma channel for CO2 and water, increases the rate of both photosynthesis and transpiration. In contrast, increased expression of Arabidopsis hexokinase1 (AtHXK1), a dual-function enzyme that mediates sugar sensing, decreases the expression of photosynthetic genes and the rate of transpiration and inhibits growth. Here, we show that AtHXK1 also decreases root and stem hydraulic conductivity and leaf mesophyll CO2 conductance (gm). Due to their opposite effects on plant development and physiology, we examined the relationship between NtAQP1 and AtHXK1 at the whole-plant level using transgenic tomato plants expressing both genes simultaneously. NtAQP1 significantly improved growth and increased the transpiration rates of AtHXK1-expressing plants. Reciprocal grafting experiments indicated that this complementation occurs when both genes are expressed simultaneously in the shoot. Yet, NtAQP1 had only a marginal effect on the hydraulic conductivity of the double-transgenic plants, suggesting that the complementary effect of NtAQP1 is unrelated to shoot water transport. Rather, NtAQP1 significantly increased leaf mesophyll CO2 conductance and enhanced the rate of photosynthesis, suggesting that NtAQP1 facilitated the growth of the double-transgenic plants by enhancing mesophyll conductance of CO2. [ABSTRACT FROM AUTHOR]

Stomata, composed of two guard cells, are the gates whose controlled movement allows the plant to balance the demand for CO2 for photosynthesis with the loss of water through transpiration. Increased guard-cell osmolarity leads to the opening of the stomata and decreased osmolarity causes the stomata to close. The role of sugars in the regulation of stomata is not yet clear. In this study, we examined the role of hexokinase ( HXK), a sugar-phosphorylating enzyme involved in sugar-sensing, in guard cells and its effect on stomatal aperture. We show here that increased expression of HXK in guard cells accelerates stomatal closure. We further show that this closure is induced by sugar and is mediated by abscisic acid. These findings support the existence of a feedback-inhibition mechanism that is mediated by a product of photosynthesis, namely sucrose. When the rate of sucrose production exceeds the rate at which sucrose is loaded into the phloem, the surplus sucrose is carried toward the stomata by the transpiration stream and stimulates stomatal closure via HXK, thereby preventing the loss of precious water. [ABSTRACT FROM AUTHOR]

Hexose sugars, such as glucose and fructose produced in plants, are ubiquitous in most organisms and are the origin of most of the organic matter found in nature. To be utilized, hexose sugars must first be phosphorylated. The central role of hexose-phosphorylating enzymes has attracted the attention of many researchers, leading to novel discoveries. Only two families of enzymes capable of phosphorylating glucose and fructose have been identified in plants; hexokinases (HXKs), and fructokinases (FRKs). Intensive investigations of these two families in numerous plant species have yielded a wealth of knowledge regarding the genes number, enzymatic characterization, intracellular localization, and developmental and physiological roles of several HXKs and FRKs. The emerging picture indicates that HXK and FRK enzymes found at specific intracellular locations play distinct roles in plant metabolism and development. Individual HXKs were shown for the first time to be dual-function enzymes -- sensing sugar levels independent of their catalytic activity and controlling gene expression and major developmental pathways, as well as hormonal interactions. FRK, on the other hand, seems to play a central metabolic role in vascular tissues, controlling the amounts of sugars allocated for vascular development. While a clearer picture of the roles of these two types of enzymes is emerging, many questions remain unsolved, such as the specific tissues and types of cells in which these enzymes function, the roles of individual HXK and FRK genes, and how these enzymes interact with hormones in the regulation of developmental processes. It is anticipated that ongoing efforts will broaden our knowledge of these important plant enzymes and their potential uses in themodification of plant traits. [ABSTRACT FROM AUTHOR]

Hexose sugars, such as glucose and fructose produced in plants, are ubiquitous in most organisms and are the origin of most of the organic matter found in nature. To be utilized, hexose sugars must first be phosphorylated. The central role of hexose-phosphorylating enzymes has attracted the attention of many researchers, leading to novel discoveries. Only two families of enzymes capable of phosphorylating glucose and fructose have been identified in plants; hexokinases (HXKs) and fructokinases (FRKs). Intensive investigations of these two families in numerous plant species have yielded a wealth of knowledge regarding the genes number, enzymatic characterization, intracellular localization and developmental and physiological roles of several HXKs and FRKs. The emerging picture indicates that HXK and FRK enzymes found at specific intracellular locations play distinct roles in plant metabolism and development. Individual HXKs were shown for the first time to be dual-function enzymes - sensing sugar levels independent of their catalytic activity and also controlling gene expression and major developmental pathways, as well as hormonal interactions. FRK, on the other hand, seems to play a central metabolic role in vascular tissues, controlling the amounts of sugars allocated for vascular development. While a clearer picture of the roles of these two types of enzymes is emerging, many questions remain unsolved, such as the specific tissues and types of cells in which these enzymes function, the roles of individual HXK and FRK genes, and how these enzymes interact with hormones in the regulation of developmental processes. It is anticipated that ongoing efforts will broaden our knowledge of these important plant enzymes and their potential uses in the modification of plant traits. [ABSTRACT FROM AUTHOR]

We examined the concurrent associations between early adolescents’ representations of mother-child attachment relationships and how they process social information in their peer relationships. Attachment representations were examined in a normative sample of 97 males and 88 females (mean age = 10.35 years), using an adaptation of the Attachment Doll Story Completion Task. Structural Equation Modeling was used to assess general latent structures of social-information processing (SIP) orientations across the different SIP steps. As expected, secure mother-child attachment representations were positively associated with prosocial SIP orientation and negatively with antisocial SIP orientation. Avoidant attachment was associated negatively with prosocial and distress expression SIP orientations. Ambivalent attachment was positively associated with distress expression SIP orientation. Disorganized attachment was positively associated with SIP distress expression orientation and with antisocial SIP orientation. Results are discussed as reflecting a generalization of social knowledge and regulation strategies from the attachment system to the affiliative system. [ABSTRACT FROM PUBLISHER]

The article presents a study which examined the danger of transgenic selection and the new roles of Arabidopsis thaliana hexokinase (AtHXK1). It states that tomato (Solanum lycopersicum) plants that overexpress AtHXK1 showed inhibited growth, reduced photosynthesis and accelerated leaf senescence when planted in soil under regular growth conditions. It notes that based on studies, tomato is the only plant species tested which was affected by high level of AtHXK1 expression.

DNA-binding protein phosphatases (DBPs) have been identified as a novel class of plant-specific regulatory factors playing a role in plant-virus interactions. NtDBP1 from tobacco (Nicotiana tabacum) was shown to participate in transcriptional regulation of gene expression in response to virus infection in compatible interactions, and AtDBP1, its closest relative in the model plant Arabidopsis (Arabidopsis thaliana), has recently been found to mediate susceptibility to potyvirus, one of the most speciose taxa of plant viruses. Here, we report on the identification of a novel family of highly conserved small polypeptides that interact with DBP1 proteins both in tobacco and Arabidopsis, which we have designated DBP-interacting protein 2 (DIP2). The interaction of AtDIP2 with AtDBP1 was demonstrated in vivo by bimolecular fluorescence complementation, and AtDIP2 was shown to functionally interfere with AtDBP1 in yeast. Furthermore, reducing AtDIP2 gene expression leads to increased susceptibility to the potyvirus Plum pox virus and to a lesser extent also to Turnip mosaic virus, whereas overexpression results in enhanced resistance. Therefore, we describe a novel family of conserved small polypeptides in plants and identify AtDIP2 as a novel host factor contributing fo resistance to potyvirus in Arabidopsis. [ABSTRACT FROM AUTHOR]

Cytogenetic analysis of DNA from a girl with severe psychomotor retardation revealed a de novo pericentric inversion of chromosome 2: 46,XX,inv(2)(p15q24.2). In order to elucidate the possible role of the inversion in the girl's abnormal phenotype, we analyzed the inversion breakpoints. FISH analysis revealed BAC clones spanning the breakpoints at 2p and 2q of the inversion. Southern blot hybridization with DNA probes from the BAC regions was used to refine the localization of the breakpoints, followed by inverse-PCR which enabled us to sequence the inversion breakpoints. We found a complex chromosomal rearrangement, including five breakpoints, four at 2q and one at 2p joined with minor insertions/deletions of a few bases. The breakpoint at 2p was within the NRXN1 gene that has previously been associated with autism, intellectual disabilities, and psychiatric disorders. In 2q, the breakpoints disrupted two genes, TANC1 and RBMS1; the phenotypic effect of these genes is not currently known. © 2011 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

The sweet melon fruit is characterized by a metabolic transition during its development that leads to extensive accumulation of the disaccharide sucrose in the mature fruit. While the biochemistry of the sugar metabolism pathway of the cucurbits has been well studied, a comprehensive analysis of the pathway at the transcriptional level allows for a global genomic view of sugar metabolism during fruit sink development. We identified 42 genes encoding the enzymatic reactions of the sugar metabolism pathway in melon. The expression pattern of the 42 genes during fruit development of the sweet melon cv Dulce was determined from a deep sequencing analysis performed by 454 pyrosequencing technology, comprising over 350,000 transcripts from four stages of developing melon fruit flesh, allowing for digital expression of the complete metabolic pathway. The results shed light on the transcriptional control of sugar metabolism in the developing sweet melon fruit, particularly the metabolic transition to sucrose accumulation, and point to a concerted metabolic transition that occurs during fruit development. [ABSTRACT FROM AUTHOR]

Sucrose synthase (SUS) plays a role in many contexts of sugar metabolism, including low-oxygen and low-ATP respiration and the synthesis of cellulose. In tomato ( Solanum lycopersicum), as in many plants, SUS is encoded by genes at several independent loci. Here, we report the isolation of a novel tomato SUS ( SlSUS) isoform, SlSUS4, that is homologous to potato SUS isoform 1 ( StSUS1) and also shows greater homology to SUS isoforms of other plants than to the other tomato SUS isoforms. All three tomato isoforms are very similar in genomic structure and sequence, yet each is located on a separate chromosome. Real-time expression analysis of the three distinct isoforms revealed widely varying patterns of expression, in terms of both tissue specificity and overall magnitude of expression. Analysis of SlSUS expression along the tomato stem revealed opposing expression gradients for two of the SlSUS isoforms, in apparent correlation with vascular tissue maturation. Western-blot analysis of SlSUS protein showed an increasing SlSUS concentration gradient along the developmental axis of the tomato stem, with the protein concentrated mainly in the vascular tissue of the stem. These gene expression and protein accumulation patterns indicate that each isoform may play a discrete role in the development of tomato plants, most notably in the development of vascular tissue in the stem. [ABSTRACT FROM AUTHOR]

It has been suggested that LeFRK2, the major fructose-phosphorylating enzyme in tomato plants, may be required for stem xylem development. Yet, we do not know if this enzyme affects the development of individual vessels, whether it affects water conductance, or whether it affects phloem development and sugar transport. Here, we show that suppression of LeFRK2 results in a significant reduction in the size of vascular cells and slows fiber maturation. The vessels in stems of LeFRK2-antisense plants are narrower than in WT plants and have thinner secondary cell walls. Although the cambium produces rounded secondary vessels, these vessels become deformed during the early stages of xylem maturation. Water conductance is then reduced in stems, roots, and leaves, suggesting that LeFRK2 influences xylem development throughout the entire vascular system. Interestingly, the build-up of positive xylem pressure under static (no-flow) conditions was also decreased. Suppression of LeFRK2 reduced the length and width of the sieve elements, as well as callose deposition. To examine the effect of LeFRK2 suppression on phloem transport, we created triple-grafted plants in which a portion of the wild-type stem was replaced with an antisense interstcok, and compared the contents of the transported sugar, sucrose, in the different portions of these stems. Sucrose contents above and within the LeFRK2-antisense interstock were significantly higher than those below the graft. These results show that the antisense interstock restricted the downward movement of sucrose, suggesting that LeFRK2 is required for both phloem and xylem development. [ABSTRACT FROM AUTHOR]

Hexokinase, a hexose-phosphorylating enzyme, has emerged as a central enzyme in sugar-sensing processes. A few HXK isozymes have been identified in various plant species. These isozymes have been classified into two major groups; plastidic (type A) isozymes located in the plastid stroma and those containing a membrane anchor domain (type B) located mainly adjacent to the mitochondria, but also found in the nucleus. Of all the hexokinases that have been characterized to date, the only exception to this rule is a spinach type B HXK (SoHXK1) that, by means of subcellular fractionation, has been localized to the outer membrane of plastids. However, SoHXK1 has a membrane anchor domain that is almost identical to that of the other type B HXKs. To determine the localization of SoHXK1 enzyme by other means, we expressed SoHXK1::GFP fusion protein in tobacco and Arabidopsis protoplasts and compared its localization with that of the Arabidopsis AtHXK1::GFP fusion protein that shares a similar N-terminal membrane anchor domain. SoHXK1::GFP is localized adjacent to the mitochondria, similar to AtHXK1::GFP and all other previously examined type B HXKs. Proteomic analysis had previously identified AtHXK1 on the outside of the mitochondrial membrane. We, therefore, suggest that SoHXK1 enzyme is located adjacent to the mitochondria like the other type B HXKs that share the same N-terminal membrane anchor domain. [ABSTRACT FROM AUTHOR]

Numerous GAST-like genes have been identified in various plant species. All code for small proteins with a conserved C-terminal region in which 12 cysteines are located in exactly the same positions. We have previously identified five gibberellin (GA)-induced GAST1-like genes in petunia, GIP1– 5. GIP2 is expressed in elongating zones, and its suppression in transgenic petunia plants inhibits stem elongation, suggesting a role for the protein in GA-induced cell growth. However, nothing is known about the biochemical activity of GIP2 or any other GAST-like protein. As all contain putative catalytic disulfide bonds (putative redox-active cysteines), we speculated that they might be involved in redox regulation. Expression analysis of GIP2, GIP4 and GIP5 revealed that they are induced by H2O2. To study whether GIP2 modulates H2O2 levels, we generated transgenic petunia plants expressing GIP2 under the regulation of the ubiquitous CaMV 35S promoter. The transgene reduced H2O2 levels in leaves following wounding. It also reduced the levels of H2O2 in guard cells following osmotic stress and ABA treatments, leading to the suppression of stomatal closure. In addition, the transgene promoted stem and corolla elongation. As reactive oxygen species (ROS) are involved in cell elongation, we suggest that GIP2 affects growth by regulating the levels of ROS. As all known GAST-like proteins contain putative redox-active cysteines, they may all act as antioxidants. [ABSTRACT FROM AUTHOR]

Four hexokinase ( LeHXK1–4) and four fructokinase ( LeFRK1–4) genes were identified in tomato plants. Previous GFP fusion studies indicate that the gene product of LeHXK3 is associated with the mitochondria while that of LeHXK4 is located within plastids. In this study we found that the enzyme encoded by the fructokinase gene LeFRK3 is also located within plastids. The presence of LeFrk3 enzyme in plastids raises the question of the origin of fructose in these organelles. The other three FRKs enzymes, LeFrk1&2&4, are located in the cytosol. Unlike LeFrk1&2&4, the two additional HXKs, LeHxk1&2, share a common membrane anchor domain and are associated with the mitochondria similar to LeHxk3. The difference in the locations of the cytoplasmic FRK and HXK isozymes suggests that glucose phosphorylation is confined to defined special intracellular localizations while fructose phosphorylation is less confined. [ABSTRACT FROM AUTHOR]

Sugars induce death of Saccharomyces cerevisiae within a few hours in the absence of additional nutrients to support growth; by contrast, cells incubated in water or in the presence of other nutrients without sugar remain viable for weeks. Here we show that this sugar-induced cell death (SICD) is characterized by rapid production of reactive oxygen species (ROS), RNA and DNA degradation, membrane damage, nucleus fragmentation and cell shrinkage. Addition of ascorbic acid to sugar-incubated cells prevents SICD, indicating that SICD is initiated by ROS. The lack of a protection mechanism against SICD suggests that sugars use to be the limiting nutrients for yeast and are probably depleted before all other nutrients. Being the limiting nutrient, sugars became the growth-stimulating agent, signaling the presence of sufficient nutrients for growth, but in the absence of the complementing nutrients they induce apoptotic death. [Copyright &y& Elsevier]

We have conducted a comprehensive metabolic profiling on tomato (Lycopersicon esculentum) leaf and developing fruit tissue using a recently established gas chromatography-mass spectrometry profiling protocol alongside conventional spectrophotometric and liquid chromatographic methodologies. Applying a combination of these techniques, we were able to identify in excess of 70 small-M[sub r] metabolites and to catalogue the metabolite composition of developing tomato fruit. In addition to comparing differences in metabolite content between source and sink tissues of the tomato plant and after the change in metabolite pool sizes through fruit development, we have assessed the influence of hexose phosphorylation through fruit development by analyzing transgenic plants constitutively overexpressing Arabidopsis hexokinase AtHXK1. Analysis of the total hexokinase activity in developing fruits revealed that both wild-type and transgenic fruits exhibit decreasing hexokinase activity with development but that the relative activity of the transgenic lines with respect to wild type increases with development. Conversely, both point-by-point and principal component analyses suggest that the metabolic phenotype of these lines becomes less distinct from wild type during development. In summary, the data presented in this paper demonstrate that the influence of hexose phosphorylation diminishes during fruit development and highlights the importance of greater temporal resolution of metabolism. [ABSTRACT FROM AUTHOR]

Much interest has been expressed in the last several years in the possibility of developing new oilseed, medicinal and garden crops from wild Cruciferae, for human consumption and industrial uses. One of the promising species is Sinapis alba (white mustard) which is very common throughout Israel. S. alba is very well known in folk traditions in the Mediterranean area as a medicinal and spice plant. As a part of a general survey of Crucifer species in Israel, 280 seed accessions of S. alba were collected. In order to assess biodiversity within the species, several parameters were recorded, and it was found that oil quality-as expressed by fatty-acid profile -and other factors of agronomical importance were affected by the siteof origin. Cultivation of selected accessions under controlled conditions demonstrated the maintenance of biodiversity. The level of genetic variability among eight accessions collected from two geographical locations was analyzed by RAPD (Random Amplified Polymorphic DNA), and a genetic distance between accessions from the two locations was found. Seeds from each accession were subsequently cultivated in three different climatic regions. After one year of cultivation, a diverging effect on the genetic polymorphism was observed. The results indicate the importance of biodiversity conservation in the process of development and evaluation of germplasm of S. alba for the use of man. [ABSTRACT FROM PUBLISHER]

By using immunolocalization and differential extraction methods we show that only apoplastic invertase, but not vacuolar invertase, was present in the mature, sucrose-accumulating L. hirsutum pericarp. In contrast, in the hexose-accumulating L. esculentum fruit, both the apoplastic and vacuolar invertase activities and protein content increase in the mature fruit. Quantitative expression studies of the soluble invertase gene (TIV1) and the apoplastic invertase genes (LINs) showed that only TIV1 gene expression could account for the species and developmental differences of both soluble and insoluble enzyme activity of the pericarp. The expression of the LIN genes encoding for apoplastic tomato invertases was unrelated to the differences in bound enzyme activity and could not account for the rise in bound invertase activity in the mature L. esculentum fruit. Evidence is presented that the bound invertase activity of tomato fruit is also the TIV1 gene product. The presence of apoplastic invertase in the mature sucrose-accumulating L. hirsutum fruit suggests a hydrolysis-resynthesis mechanism of sucrose uptake. In order to test this hypothesis, we studied short- and long-term uptakes of asymmetrically labelled 3H-fructosyl-sucrose accompanied by compartmental analysis of the sugars in attached whole fruits of L. hirsutum and L. esculentum. The results indicate that hydrolysis-resynthesis is slow in the sucrose-accumulating fruit but is not an integral part of an uptake and compartmentation mechanism. [ABSTRACT FROM AUTHOR]

The concurrent association between security of attachment and adaptive functioning at school in middle childhood was examined. A sample of 113 children of 4th and 5th grade filled out a selfreport measure of attachment security (Kerns, Klepac, & Cole, 1996) and were administered the Doll Story Completion task (Bretherton, Ridgeway, & Cassidy, 1990b) modified for use with children in middle childhood to assess the quality and the security of attachment-related representations of the relationship with the mother. According to the latter measure children were classified as secure, avoidant, ambivalent, or disorganised with regard to attachment. Their teachers completed several questionnaires assessing each child's academic achievement, emotional and social adjustment, and frequency of behavioural problems. In addition, each participating class underwent a sociometric procedure. Findings based on correlations and comparisons of attachment groups indicated that secure children showed better adjustment to school as reflected in teachers' reports of scholastic, emotional, social, and behavioural adjustment, as well as in peer-rated social status. Avoidant and disorganised children showed the poorest adjustment. Findings indicated the usefulness of attachment theory in understanding adjustment to the school environment in middle childhood. [ABSTRACT FROM AUTHOR]

Sugars like glucose and fructose induce death of yeast cells within a few hours, in the absence of additional nutrients to support growth, while cells incubated in water remain viable for weeks. This sugar-induced cell death (SlCD) by glucose and fructose required glucose or fructose phosphorylation since yeast cells deficient in hexose phosphorylation did not die. However, when hexose phosphorylation is restored by complementation with Arabidopsis thaliana hexokinase, the cells died. The affinity of A. thaliana hexokinase is about 400 times higher for glucose than for fructose, therefore, A. thaliana hexokinase was further utilized to study the role of hexose phosphorylation in SICD. The rate of SICD of hexokinase-deficient yeast cells expressing A. thaliana hexokinase was significantly slower in fructose than in glucose, indicating that SICD is determined by the rate of hexose phosphorylation. The significance of hexose phosphorylation and its role in SICD is discussed. [ABSTRACT FROM AUTHOR]

Abiotic stresses such as drought and saline water impose major limitations on plant growth. Modulation of stomatal behavior may help plants cope with such stresses by reducing both water loss and salt uptake. Hexokinase (HXK) is a sugar-phosphorylating enzyme involved in guard cells' sugar-sensing, mediating stomatal closure and coordinating photosynthesis with transpiration. We generated transgenic tobacco lines expressing the Arabidopsis hexokinase1 (AtHXK1) under the guard cell-specific promoter KST1 and examined those plants using growth room and greenhouse experiments. The expression of AtHXK1 in tobacco guard cells reduced stomatal conductance and transpiration by about 25% with no negative effects on photosynthesis or growth, leading to increased water-use efficiency. In addition, these plants exhibited tolerance to drought and salt stress due to their lower transpiration rate, indicating that improved stomatal function has the potential to improve plant performance under stress conditions. [ABSTRACT FROM AUTHOR]

A correction to the article "LeFRK2 is required for phloem and xylem differentiation and the transport of both sugar and water" is presented.

A cDNA clone, pAUK1, with an open reading frame (ORF) coding for a hypothetical 164-amino-acid protein was isolated from an Arabidopsis thaliana (L.) Heynh cDNA library. The clone was attached, tail to tail, to the 3′ end of A. thaliana hexokinase cDNA. An almost identical sequence had been previously described as the 5′ untranslated region (5′ UTR) of A. thaliana calmodulin cDNA (ACaM-2). Sequence comparison with three additional A. thaliana truncated cDNA clones which appear in a database (GenBank) supports the conclusion that pAUKl is identical to the 5′ UTR of ACaM-2 and that the 5′ UTR of ACaM-2 is an independent cDNA artificially linked to A. thaliana calmodulin cDNA. [ABSTRACT FROM AUTHOR]

Summary Raffinose and stachyose are ubiquitous galactosyl-sucrose oligosaccharides in the plant kingdom which play major roles, second only to sucrose, in photoassimilate translocation and seed carbohydrate storage. These sugars are initially metabolised by α-galactosidases (α-gal). We report the cloning and functional expression of the first genes, CmAGA1 and CmAGA2 , encoding for plant α-gals with alkaline pH optima from melon fruit (Cucumis melo L.), a raffinose and stachyose translocating species. The alkaline α-gal genes show very high sequence homology with a family of undefined ‘seed imbibition proteins’ (SIPs) which are present in a wide range of plant families. In order to confirm the function of SIP proteins, a representative SIP gene, from tomato, was expressed and shown to have alkaline α-gal activity. Phylogenetic analysis based on amino acid sequences shows that the family of alkaline α-gals shares little homology with the known prokaryotic and eukaryotic α-gals of glycosyl hydrolase families 27 and 36, with the exception of two cross-family conserved sequences containing aspartates which probably function in the catalytic step. This previously uncharacterised, plant-specific α-gal family of glycosyl hydrolases, with optimal activity at neutral-alkaline pH likely functions in key processes of galactosyl-oligosaccharide metabolism, such as during seed germination and translocation of RFO photosynthate. [ABSTRACT FROM AUTHOR]