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Key Message: Two major QTLs for cold tolerance in pumpkin were localised, and CmoERF017 was identified as a key candidate gene within these QTLs via RNA-seq. Functional analysis revealed that CmoERF017 was a positive regulator of pumpkin in response to low-temperature stress. Low temperature is a key environmental factor that affects the protected cultivation of cucumber (Cucumis sativus L.) in winter, and the cold tolerance of cucumber/pumpkin-grafted seedlings depends on the rootstock. Pumpkin (Cucurbita spp.) has a well-developed root system, high resistance and wide adaptation, commonly used as rootstock for cucumber to improve the cold tolerance of grafted seedlings. This study used two high-generation inbred lines of Cucurbita moschata with significant differences in cold tolerance. We identified key candidate genes within the major cold tolerance QTL of rootstocks using QTL-seq and RNA-seq and investigated the function and molecular mechanisms of these genes in response to low-temperature stress. Results showed that QTL-seq located two cold tolerance QTLs, qCII-1 and qCII-2, while RNA-seq located 28 differentially expressed genes within these QTLs. CmoERF017 was finally identified as a key candidate gene. Functional validation results indicated that CmoERF017 is a positive regulator of pumpkin in response to low-temperature stress and affected root ABA synthesis and signalling by directly regulating the expression of SDR7 and ABI5. This study identified a key gene for low-temperature stress tolerance in rootstock pumpkin and clarified its role in the molecular mechanism of hormone-mediated plant cold tolerance. The study findings enrich the theoretical understanding of low-temperature stress tolerance in pumpkin and are valuable for the selection and breeding of cold-tolerant varieties of pumpkin used for rootstocks., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The globally changing climatic condition is increasing the incidences of drought in several parts of the world. This is predicted and already shown to not only impact plant growth and flower development, but also plant-pollinator interactions and the pollination success of entomophilous plants. However, there is a large gap in our understanding of how drought affects the different flowers and pollen transfer among flowers in sexually polymorphic species. Here, we evaluated in monoecious Styrian oil pumpkin, and separately for female and male flowers, the responses of drought stress on flower production, petal size, nectar, floral scent and visitation by bumblebee pollinators. Drought stress adversely affected all floral traits studied, except floral scent. Although both flower sexes were adversely affected by drought stress, the effects were more severe on female flowers, with most of the female flowers even aborted before opening. The drought had negative effects on floral visitation by the pollinators, which generally preferred female flowers. Overall, our study highlights that the two flower sexes of a monoecious plant species are differently affected by drought stress and calls for further investigations to better understand the cues used by the pollinators to discriminate against male flowers and against flowers of drought-stressed plants., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Drought is an increasingly important consequence of climate change. Drought often causes plants to alter patterns of resource allocation, which in turn can affect how plants interact with other species. How these altered interactions subsequently influence plant reproductive success remains incompletely understood and may depend on the degree of specialization exhibited by antagonists and mutualists. Specialist pollinators, for example, are dependent on floral resources from their obligate hosts and under drought conditions may thus indiscriminately visit these hosts (at least in certain circumstances). Generalist pollinators, in contrast, may only forage on host plants in good condition, given that they can forage on other plant species. We tested this hypothesis and its consequences for plant reproduction in squash (Cucurbita pepo) grown along an experimental moisture gradient ranging from dry (growth and flowering compromised) to wet conditions. Floral visitation increased with plant soil moisture for generalist honey bees but was independent of plant soil moisture for specialist squash bees. Pollen production increased with plant soil moisture, and fluorescent pigments placed on flowers revealed that pollinators primarily moved pollen from male flowers on well-watered plants to the stigmas of female flowers on well-watered plants. Seed set increased with increasing plant soil moisture but, notably, was higher in bee-pollinated plants compared to plants pollinated by hand with an even mix of pollen from plants grown at either end of the experimental moisture gradient. These results suggest that superior pollen rewards, perhaps combined with selective foraging by generalists, enhanced reproductive success in C. pepo when plant soil moisture was high and more generally illustrate that pollinator behavior may contribute to how drought conditions affect plant reproduction., (© 2023 The Authors. Ecology published by Wiley Periodicals LLC on behalf of The Ecological Society of America.)

Heat shock protein 70s (HSP70s) are highly conserved proteins that are involved in stress responses. These chaperones play pivotal roles in protein folding, removing the extra amounts of oxidized proteins, preventing protein denaturation, and improving the antioxidant system activities. This conserved family has been characterized in several crops under drought stress conditions. However, there is no study on HSP70s in pumpkin ( Cucurbita moschata ). Therefore, we performed a comprehensive analysis of this gene family, including phylogenetic relationship, motif and gene structure analysis, gene duplication, collinearity, and promoter analysis. In this research, we found 21 HSP70s that were classified into five groups (from A to E). These genes were mostly localized in the cytoplasm, chloroplast, mitochondria, nucleus, and endoplasmic reticulum (ER). We could observe more similarity in closely linked subfamilies in terms of motifs, the number of introns/exons, and the corresponding cellular compartments. According to the collinearity analysis, gene duplication had occurred as a result of purifying selection. The results showed that the occurrence of gene duplication for all nine gene pairs was due to segmental duplication (SD). Synteny analysis revealed a closer relationship between pumpkin and cucumber than pumpkin and Arabidopsis. Promoter analysis showed the presence of various cis-regulatory elements in the up-stream region of the HSP70 genes, such as hormones and stress-responsive elements, indicating a potential role of this gene family in stress tolerance. We furtherly performed the gene expression analysis of the HSP70s in pumpkin under progressive drought stress. Pumpkin is widely used as a rootstock to improve stress tolerance, as well as fruit quality of cucumber scion. Since stress-responsive mobile molecules translocate through vascular tissue from roots to the whole plant body, we used the xylem of grafted materials to study the expression patterns of the HSP70 (potentially mobile) gene family. The results indicated that all CmoHSP70s had very low expression levels at 4 days after stress (DAS). However, the genes showed different expression patterns by progressing he drought period. For example, the expression of CmoHSP70 -4 (in subgroup E) and CmoHSP70 -14 (in subgroup C) sharply increased at 6 and 11 DAS, respectively. However, the expression of all genes belonging to subgroup A did not change significantly in response to drought stress. These findings indicated the diverse roles of this gene family under drought stress and provided valuable information for further investigation on the function of this gene family, especially under stressful conditions.

Global pollinator decline is a major concern. Several factors-climate change, land-use change, the reduction of flowers, pesticide use, and invasive species-have been suggested as the reasons. Despite being a potential reason, the effect of ants on flowers received less attention. The consequences of ants being attracted to nectar sources in plants vary depending upon factors like the nectar source's position, ants' identity, and other mutualists interacting with the plants. We studied the interaction between flower-visiting ants and pollinators in Cucurbita maxima and compared the competition exerted by native and invasive ants on its pollinators to examine the hypothesis that the invasive ants exacerbate more interference competition to pollinators than the native ants. We assessed the pollinator's choice, visitation rate, and time spent/visit on the flowers. Regardless of species and nativity, ants negatively influenced all the pollinator visitation traits, such as visitation rate and duration spent on flowers. The invasive ants exerted a higher interference competition on the pollinators than the native ants did. Despite performing pollination in flowers with generalist pollination syndrome, ants can threaten plant-pollinator mutualism in specialist plants like monoecious plants. A better understanding of factors influencing pollination will help in implementing better management practices.

Floral nectar is a sugary solution produced by nectaries to attract and reward pollinators. Nectar metabolites, such as sugars, are synthesized within the nectary during secretion from both pre-stored and direct phloem-derived precursors. In addition to sugars, nectars contain nitrogenous compounds such as amino acids; however, little is known about the role(s) of nitrogen (N) compounds in nectary function. In this study, we investigated N metabolism in Cucurbita pepo (squash) floral nectaries in order to understand how various N-containing compounds are produced and determine the role of N metabolism in nectar secretion. The expression and activity of key enzymes involved in primary N assimilation, including nitrate reductase (NR) and alanine aminotransferase (AlaAT), were induced during secretion in C. pepo nectaries. Alanine (Ala) accumulated to about 35% of total amino acids in nectaries and nectar during peak secretion; however, alteration of vascular nitrate supply had no impact on Ala accumulation during secretion, suggesting that nectar(y) amino acids are produced by precursors other than nitrate. In addition, nitric oxide (NO) is produced from nitrate and nitrite, at least partially by NR, in nectaries and nectar. Hypoxia-related processes are induced in nectaries during secretion, including lactic acid and ethanolic fermentation. Finally, treatments that alter nitrate supply affect levels of hypoxic metabolites, nectar volume and nectar sugar composition. The induction of N metabolism in C. pepo nectaries thus plays an important role in the synthesis and secretion of nectar sugar., (© 2020 Society for Experimental Biology and John Wiley & Sons Ltd.)

Cyclocephalini beetles of the genus Cyclocephala (Coleoptera: Melolonthidae: Dynastinae) use flowers of some plants as food, shelter, and mating sites. However, little is known about floral scent chemistry involved in this interaction. Here we show that a sesquiterpene alcohol mediates attraction of Cyclocephala paraguayensis Arrow, on bottle gourd flowers, Lagenaria siceraria (Cucurbitaceae). Both males and females started to aggregate on the flowers at twilight; after that, mating began and remained for the entire night. GC-FID/EAD analysis of the L. siceraria floral scent collected in the field revealed that only the major constituent of the airborne volatiles elicited electroantennographic responses on male and female antennae of C. paraguayensis. This compound was identified as (3S,6E)-nerolidol, which was tested in two field trapping trials in Brazil. In the first bioassay, traps baited with nerolidol (mix of isomers) captured significantly more adult C. paraguayensis than control traps. In the second field trial, catches in traps baited with a mixture of isomers or enantiopure nerolidol were significantly higher than captures in control traps, but the treatments did not differ significantly. Analysis from the gut content of adult C. paraguayensis showed the presence of pollen, suggesting that they also use bottle gourd flowers for their nourishment. Taken together, these results suggest that (3S,6E)-nerolidol plays an essential role in the reproductive behavior of C. paraguayensis by eliciting aggregation, mating, and feeding., Competing Interests: The authors declare no conflict of interest.

The remote sensing of a plant's physiological state is a key problem of precision agriculture. The photochemical reflectance index (PRI), which is based on the intensities of the reflected light at 531 and 570 nm, is an important tool for the remote sensing of photosynthetic processes in plants. In particular, the PRI can be strongly connected with the non-photochemical quenching of chlorophyll fluorescence (NPQ) and the quantum yield of photosystem II (Ф PSII ); however, this connection is dependent on illumination, the intensity of stressor actions, the time scale of measurements, etc. The aim of the present work was to analyze the connection of PRI with the energy-dependent component of NPQ (NPQ F ) and Ф PSII under heating and soil drought conditions. Pea, wheat, and pumpkin seedlings, which were grown under controlled conditions, were investigated. A PAM fluorometer Dual-PAM-100 and spectrometer S-100 were used for measurements of photosynthetic parameters and PRI, respectively. It was shown that heat stress increased the NPQ F and the magnitude of light-induced changes in PRI (ΔPRI) and decreased Ф PSII in pea seedlings. The decreased Ф PSII and increased ΔPRI were observed in wheat after heating, but significant changes in NPQ F were absent; the significant decrease in Ф PSII was observed in pumpkin seedlings, while there were no significant changes in the other parameters. ΔPRI and NPQ F after heating were significantly correlated. However, a significant correlation of the absolute values of PRI with photosynthetic parameters was absent. The soil drought increased NPQ F and the magnitude of ΔPRI and decreased Ф PSII in peas. ΔPRI was strongly correlated with photosynthetic parameters, but this correlation was absent for the absolute value of PRI. Thus, ΔPRI is strongly connected with the magnitude of NPQ F and can be used as an estimator of this parameter.

Background: Crop quality and yield are affected by abiotic and biotic stresses, and heat shock transcription factors (Hsfs) are considered to play important roles in regulating plant tolerance under various stresses. To investigate the response of Cucurbita moschata to abiotic stress, we analyzed the genome of C. moschata., Results: In this research, a total of 36 C. moschata Hsf (CmHsf) members were identified and classified into three subfamilies (I, II, and III) according to their amino acid sequence identity. The Hsfs of the same subfamily usually exhibit a similar gene structure (intron-exon distribution) and conserved domains (DNA-binding and other functional domains). Chromosome localization analysis showed that the 36 CmHsfs were unevenly distributed on 18 of the 21 chromosomes (except for Cm_Chr00, Cm_Chr08 and Cm_Chr20), among which 18 genes formed 9 duplicated gene pairs that have undergone segmental duplication events. The Ka/Ks ratio showed that the duplicated CmHsfs have mainly experienced strong purifying selection. High-level synteny was observed between C. moschata and other Cucurbitaceae species., Conclusions: The expression profile of CmHsfs in the roots, stems, cotyledons and true leaves revealed that the CmHsfs exhibit tissue specificity. The analysis of cis-acting elements and quantitative real-time polymerase chain reaction (qRT-PCR) revealed that some key CmHsfs were activated by cold stress, heat stress, hormones and salicylic acid. This study lays the foundation for revealing the role of CmHsfs in resistance to various stresses, which is of great significance for the selection of stress-tolerant C. moschata.

Native bees provide essential pollination services in both natural and managed ecosystems. However, declines in native bee species highlight the need for increased understanding of land management methods that can promote healthy, persistent populations and diverse communities. This can be challenging and costly using traditional scientific methods, but citizen science can overcome many limitations. In this study, we examined the distribution and abundance of an agriculturally important wild bee species, the squash bee (Eucera (Peponapis) pruinosa, Hymenoptera: Apidae). They are ground nesting, specialist bees that depend on cultivated varieties of Cucurbita (squash, pumpkins, gourds). The intimate relationship between squash bees and their host plants suggests that they are likely sensitive to farm management practices, particularly those that disturb the soil. In this study, citizen scientists across Michigan used a survey to submit field management and bee observation data. Survey results indicated that squash bees occupy a wide geographic range and are more abundant in farms with reduced soil disturbance. Citizen science provided an inexpensive and effective method for examining impacts of farm management practices on squash bees and could be a valuable tool for monitoring and conserving other native pollinators., Competing Interests: The authors have declared that no competing interests exist.

Genomic prediction has become an increasingly popular tool for hybrid performance evaluation in plant breeding mainly because that it can reduce cost and accelerate a breeding program. In this study, we propose a systematic procedure to predict hybrid performance using a genomic selection (GS) model that takes both additive and dominance marker effects into account. We first demonstrate the advantage of the additive-dominance effects model over the only additive effects model through a simulation study. Based on the additive-dominance model, we predict genomic estimated breeding values (GEBVs) for individual hybrid combinations and their parental lines. The GEBV-based specific combining ability (SCA) for each hybrid and general combining ability (GCA) for its parental lines are then derived to quantify the degree of midparent heterosis (MPH) or better-parent heterosis (BPH) of the hybrid. Finally, we estimate the variance components resulting from additive and dominance gene action effects and heritability using a genomic best linear unbiased predictor (g-BLUP) model. These estimates are used to justify the results of the genomic prediction study. A pumpkin ( spp.) data set is given to illustrate the provided procedure. The data set consists of 320 parental lines with 61,179 collected single nucleotide polymorphism (SNP) markers; 119, 120, and 120 phenotypic values of hybrids on three quantitative traits within maxima Duchesne; and 89, 111, and 90 phenotypic values of hybrids on the same three quantitative traits within Dechesne., (© 2019 The Author(s).)

This study investigates the effect of SPIONs (superparamagnetic iron oxide nanoparticles, ∼12.5 nm in size) on summer squash plant (Cucurbita pepo) in the presence and absence of supplementary iron (Fe(II)-EDTA). The plants were grown in nutrient solution with different iron sources: (i) Fe(II)-EDTA, (ii) without Fe(II)-EDTA (iii) SPIONs only, and (iv) Fe(II)-EDTA with SPIONs. Plant growth and development were assessed after 20 days of soaking by measuring phenological parameters such as plant biomass, chlorophyll content, amount of carotenoids, and the catalase enzyme activity. Transmission electron microscopy, inductively coupled plasma atomic emission spectroscopy, X-ray diffraction, and vibrating sample magnetometer methods were used to detect uptake and translocation of SPIONs in plant tissues. Our results showed that SPIONs treatment (without Fe(II)-EDTA) caused growth retardation and decreased the plant biomass and chlorophyll content. Hence, they are not efficient sources to compensate for iron demand of squash plant. Electron microscopy observations, magnetization and elemental analyses revealed that SPIONs are taken-up by plant roots but not translocate to upper organs. In roots, SPIONs use a symplastic route for intercellular transfer. These findings suggest that as an iron source, SPIONs alone are not efficient for plant growth, but can contribute it together with Fe(II)-EDTA., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Numerous studies on short term effects of copper-based nanomaterials on plants have been published, however investigations with plants grown in a complex soil medium are lacking. In this study Grey Zucchini (Cucurbita pepo) was grown in an environmental growth chamber using a 1:1 (v/v) potting mix native soil mixture amended with Kocide 3000, nCuO, bCuO, or Cu NPs. After 3 weeks Cu concentrations in the root, stem, and leaves of treated plants were significantly higher than control plants. This increase in Cu concentration did not adversely affect plant growth or chlorophyll production. The activity ascorbate peroxidase (APX) in the roots tissues of plants treated with Kocide 3000, nCuO, and bCuO decreased by at least 45%. Catalase (CAT) activity in root tissues of plants treated with 50 mg/kg of Cu NP decreased by 77%, while those treated at 200 mg/kg were reduced by 80%, compared to controls. The activity of APX and CAT in the leaves of all treated plants remained similar to control plants. Based on the endpoints used in this study, with the exception of a decrease in the accumulation of Zn and B in the roots, the exposure of zucchini to the tested copper compounds resulted in no negative effects., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Approaches that enable high-throughput, non-destructive measurement of plant traits are essential for programs seeking to improve crop yields through physiological breeding. However, many key traits still require measurement using slow, labor-intensive, and destructive approaches. We investigated the potential to retrieve key traits associated with leaf source-sink balance and carbon-nitrogen status from leaf optical properties. Structural and biochemical traits and leaf reflectance (500-2400 nm) of eight crop species were measured and used to develop predictive 'spectra-trait' models using partial least squares regression. Independent validation data demonstrated that the models achieved very high predictive power for C, N, C:N ratio, leaf mass per area, water content, and protein content (R2>0.85), good predictive capability for starch, sucrose, glucose, and free amino acids (R2=0.58-0.80), and some predictive capability for nitrate (R2=0.51) and fructose (R2=0.44). Our spectra-trait models were developed to cover the trait space associated with food or biofuel crop plants and can therefore be applied in a broad range of phenotyping studies., (Published by Oxford University Press on behalf of the Society for Experimental Biology 2019.)

Background: Zucchini fruit set can be limited due to unfavourable environmental conditions in off-seasons crops that caused ineffective pollination/fertilization. Parthenocarpy, the natural or artificial fruit development without fertilization, has been recognized as an important trait to avoid this problem, and is related to auxin signalling. Nevertheless, differences found in transcriptome analysis during early fruit development of zucchini suggest that other complementary pathways could regulate fruit formation in parthenocarpic cultivars of this species. The development of next-generation sequencing technologies (NGS) as RNA-sequencing (RNA-seq) opens a new horizon for mapping and quantifying transcriptome to understand the molecular basis of pathways that could regulate parthenocarpy in this species. The aim of the current study was to analyze fruit transcriptome of two cultivars of zucchini, a non-parthenocarpic cultivar and a parthenocarpic cultivar, in an attempt to identify key genes involved in parthenocarpy., Results: RNA-seq analysis of six libraries (unpollinated, pollinated and auxin treated fruit in a non-parthenocarpic and parthenocarpic cultivar) was performed mapping to a new version of C. pepo transcriptome, with a mean of 92% success rate of mapping. In the non-parthenocarpic cultivar, 6479 and 2186 genes were differentially expressed (DEGs) in pollinated fruit and auxin treated fruit, respectively. In the parthenocarpic cultivar, 10,497 in pollinated fruit and 5718 in auxin treated fruit. A comparison between transcriptome of the unpollinated fruit for each cultivar has been performed determining that 6120 genes were differentially expressed. Annotation analysis of these DEGs revealed that cell cycle, regulation of transcription, carbohydrate metabolism and coordination between auxin, ethylene and gibberellin were enriched biological processes during pollinated and parthenocarpic fruit set., Conclusion: This analysis revealed the important role of hormones during fruit set, establishing the activating role of auxins and gibberellins against the inhibitory role of ethylene and different candidate genes that could be useful as markers for parthenocarpic selection in the current breeding programs of zucchini.

Tissue tolerance to salinity stress is a complex physiological trait composed of multiple 'sub-traits' such as Na+ compartmentalization, K+ retention, and osmotic tolerance. Previous studies have shown that some Cucurbita species employ tissue tolerance to combat salinity and we aimed to identify the physiological and molecular mechanisms involved. Five C. maxima (salt-tolerant) and five C. moschata (salt-sensitive) genotypes were comprehensively assessed for their salt tolerance mechanisms and the results showed that tissue-specific transport characteristics enabled the more tolerant lines to deal with the salt load. This mechanism was associated with the ability of the tolerant species to accumulate more Na+ in the leaf vein and to retain more K+ in the leaf mesophyll. In addition, C. maxima more efficiently retained K+ in the roots when exposed to transient NaCl stress and it was also able to store more Na+ in the xylem parenchyma and cortex in the leaf vein. Compared with C. moschata, C. maxima was also able to rapidly close stomata at early stages of salt stress, thus avoiding water loss; this difference was attributed to higher accumulation of ABA in the leaf. Transcriptome and qRT-PCR analyses revealed critical roles of high-affinity potassium (HKT1) and intracellular Na+/H+ (NHX4/6) transporters as components of the mechanism enabling Na+ exclusion from the leaf mesophyll and Na+ sequestration in the leaf vein. Also essential was a higher expression of NCED3s (encoding 9-cis-epoxycarotenoid dioxygenase, a key rate-limiting enzyme in ABA biosynthesis), which resulted in greater ABA accumulation in the mesophyll and earlier stomata closure in C. maxima.

Plant salt tolerance can be improved by grafting onto salt-tolerant rootstocks. However, the underlying signaling mechanisms behind this phenomenon remain largely unknown. To address this issue, we used a range of physiological and molecular techniques to study responses of self-grafted and pumpkin-grafted cucumber plants exposed to 75 mM NaCl stress. Pumpkin grafting significantly increased the salt tolerance of cucumber plants, as revealed by higher plant dry weight, chlorophyll content and photochemical efficiency (Fv/Fm), and lower leaf Na+ content. Salinity stress resulted in a sharp increase in H2O2 production, reaching a peak 3 h after salt treatment in the pumpkin-grafted cucumber. This enhancement was accompanied by elevated relative expression of respiratory burst oxidase homologue (RBOH) genes RbohD and RbohF and a higher NADPH oxidase activity. However, this increase was much delayed in the self-grafted plants, and the difference between the two grafting combinations disappeared after 24 h. The decreased leaf Na+ content of pumpkin-grafted plants was achieved by higher Na+ exclusion in roots, which was driven by the Na+/H+ antiporter energized by the plasma membrane H+-ATPase, as evidenced by the higher plasma membrane H+-ATPase activity and higher transcript levels for PMA and SOS1. In addition, early stomatal closure was also observed in the pumpkin-grafted cucumber plants, reducing water loss and maintaining the plant's hydration status. When pumpkin-grafted plants were pretreated with an NADPH oxidase inhibitor, diphenylene iodonium (DPI), the H2O2 level decreased significantly, to the level found in self-grafted plants, resulting in the loss of the salt tolerance. Inhibition of the NADPH oxidase-mediated H2O2 signaling in the root also abolished a rapid stomatal closure in the pumpkin-grafted plants. We concluded that the pumpkin-grafted cucumber plants increase their salt tolerance via a mechanism involving the root-sourced respiratory burst oxidase homologue-dependent H2O2 production, which enhances Na+ exclusion from the root and promotes an early stomatal closure.

The grape mealybug, Pseudococcus maritimus (Ehrhorn; Hemiptera: Pseudococcidae), is the primary vector of Grapevine Leafroll associated Viruses (GLRaVs) in Washington State vineyards. Rearing laboratory colonies of grape mealybug has proven difficult. Several host plants were tested to determine their suitability for use as an alternate host plants for laboratory colonies of grape mealybug. Of the plants tested, colonies of grape mealybug were successful on pumpkin (Cucurbita pepo cv 'Connecticut field') leaves and vines. Mealybugs were able to develop from the crawler stage, through developmental instars, and adults were able to reproduce viable offspring. To date this is the only successful study to raise grape mealybugs on a cucurbit. Pumpkin appears to be a viable alternate host plant for laboratory colonies of the grape mealybug.

Plants response to symbiosis with arbuscular mycorrhizal fungi (AMF) under water stress is important to agriculture. Under abiotic stress conditions native fungi are more effective than exotics in improving plant growth and water status. Mycorrhization efficiency is related to soil fungi development and energy cost-benefit ratio. In this study, we assessed the effect on growth, water status and energy metabolism of Cucurbita pepo var. pepo when inoculated with native AMF from the Sonoran desert Mexico (mixed isolate and field consortium), and compared with an exotic species from a temperate region, under drought, low and high salinity conditions. Dry weights, leaf water content, water and osmotic potentials, construction costs, photochemistry and mycorrhization features were quantified. Under drought and low salinity conditions, the mixed isolate increased plant growth and leaf water content. Leaf water potential was increased only by the field consortium under drought conditions (0.5-0.9MPa). Under high salinity, the field consortium increased aerial dry weight (more than 1g) and osmotic potential (0.54MPa), as compared to non-mycorrhized controls. Plants inoculated with native AMF, which supposedly diminish the effects of stress, exhibited low construction costs, increased photochemical capacity, and grew larger external mycelia in comparison to the exotic inoculum., (Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)

Metals and metalloids in soil could be transferred into reproductive organs and floral rewards of hyperaccumulator plants and influence their reproductive success, yet little is known whether non-hyperaccumulator plants can translocate heavy metals from soil into their floral organs and rewards (i.e., nectar and pollen) and, if so, whether plant reproduction will be affected. In our studies, summer squash (Cucurbita pepo L. cv. Golden Apple) was exposed to heavy-metal treatments during bud stage to investigate the translocation of soil-supplemented zinc, copper, nickel and lead into its floral organs (pistil, anther and nectary) and rewards (nectar and pollen) as well as floral metal accumulation effects on its reproduction. The results showed that metals taken up by squash did translocate into its floral organs and rewards, although metal accumulation varied depending on different metal types and concentrations as well as floral organ/reward types. Mean foraging time of honey bees to each male and female flower of squash grown in metal-supplemented soils was shorter relative to that of plants grown in control soils, although the visitation rate of honeybees to both male and female flowers was not affected by metal treatments. Pollen viability, pollen removal and deposition as well as mean mass per seed produced by metal-treated squash that received pollen from plants grown in control soils decreased with elevated soil-supplemented metal concentrations. The fact that squash could translocate soil-supplemented heavy metals into floral organs and rewards indicated possible reproductive consequences caused either directly (i.e., decreasing pollen viability or seed mass) or indirectly (i.e., affecting pollinators' visitation behavior to flowers) to plant fitness., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Leon Chua, the discoverer of a memristor, theoretically predicted that voltage gated ion channels can be memristors. We recently found memristors in different plants such as the Venus flytrap, Mimosa pudica, Aloe vera, apple fruits, and in potato tubers. There are no publications in literature about the existence of memristors in seeds. The goal of this work was to discover if pumpkin seeds might have memristors. We selected Cucurbita pepo L., cv. Cinderella, Cucurbita maxima L. cv Warty Goblin, and Cucurbita maxima L., cv. Jarrahdale seeds for this analysis. In these seeds, we found the presence of resistors with memory. The analysis was based on cyclic voltammetry where a memristor should manifest itself as a nonlinear two-terminal electrical element, which exhibits a pinched hysteresis loop on a current-voltage plane for any bipolar cyclic voltage input signal. Dry dormant pumpkin seeds have very high electrical resistance without memristive properties. The electrostimulation by bipolar sinusoidal or triangular periodic waves induces electrical responses in imbibed pumpkin seeds with fingerprints of memristors. Tetraethylammonium chloride, an inhibitor of voltage gated K(+) channels, transforms a memristor to a resistor in pumpkin seeds. NPPB (5-Nitro-2-(3-phenylpropylamino)benzoic acid) inhibits the memristive properties of imbibed pumpkin seeds. The discovery of memristors in pumpkin seeds creates a new direction in the understanding of electrophysiological phenomena in seeds.

The genus Cucurbita (squashes, pumpkins, gourds) contains numerous domesticated lineages with ancient New World origins. It was broadly distributed in the past but has declined to the point that several of the crops' progenitor species are scarce or unknown in the wild. We hypothesize that Holocene ecological shifts and megafaunal extinctions severely impacted wild Cucurbita, whereas their domestic counterparts adapted to changing conditions via symbiosis with human cultivators. First, we used high-throughput sequencing to analyze complete plastid genomes of 91 total Cucurbita samples, comprising ancient (n = 19), modern wild (n = 30), and modern domestic (n = 42) taxa. This analysis demonstrates independent domestication in eastern North America, evidence of a previously unknown pathway to domestication in northeastern Mexico, and broad archaeological distributions of taxa currently unknown in the wild. Further, sequence similarity between distant wild populations suggests recent fragmentation. Collectively, these results point to wild-type declines coinciding with widespread domestication. Second, we hypothesize that the disappearance of large herbivores struck a critical ecological blow against wild Cucurbita, and we take initial steps to consider this hypothesis through cross-mammal analyses of bitter taste receptor gene repertoires. Directly, megafauna consumed Cucurbita fruits and dispersed their seeds; wild Cucurbita were likely left without mutualistic dispersal partners in the Holocene because they are unpalatable to smaller surviving mammals with more bitter taste receptor genes. Indirectly, megafauna maintained mosaic-like landscapes ideal for Cucurbita, and vegetative changes following the megafaunal extinctions likely crowded out their disturbed-ground niche. Thus, anthropogenic landscapes provided favorable growth habitats and willing dispersal partners in the wake of ecological upheaval.

Previously, we found that the amelioration of photosynthetic capacity by bottle gourd (Lagenaria siceraria Standl.) rootstock in watermelon seedlings (Citrullus lanatus [Thunb.] Mansf.) with salt treatment might be closely related to the enzymes in Calvin cycle such as ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) (Yang et al., 2012). We confirmed this and showed more details in this study that improved photosynthesis of watermelon plants by bottle gourd rootstock was associated with the decreased stomata resistance and the increased photochemical activity and photosynthetic metabolism with or without 100mM NaCl stress for 3 days. The analysis of gas exchange parameters showed that self-grafted plants suffered serious non-stomatal limitation to photosynthesis under salt stress while rootstock-grafted plants were mainly affected by stomata limitation in stress conditions. Further, results showed that NaCl stress markedly reduced the chlorophyll content, damaged the structure of photosynthetic apparatus, and inhibited photochemical activity and CO2 assimilation in self-grafted plants. In contrast, rootstock-grafting increased the chlorophyll content, especially chlorophyll b, and minimized the harmful effects on photosystem II (PSII) reaction center and the thylakoids structure induced by NaCl stress. Furthermore, rootstock-grafting enhanced the content and activity of Rubisco and thus elevated carbon fixation in the leaves of watermelon scions under salt stress. The gene expressions of enzymes related to ribulose-1,5-bisphosphate (RuBP) regeneration were also up-regulated by rootstock and this probably guaranteed the sufficient supply of RuBP for the operation of Calvin cycle in watermelon scions under salt stress. Thus, bottle gourd rootstock promoted photosynthesis by the activation of stomatal and non-stomatal abilities, especially the regulation of a variety of photosynthetic enzymes, including Rubisco in grafted watermelon plants under NaCl stress., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Despite the success of breeding programmes focused on increasing fruit size, relatively little is known about the anatomical and physiological changes required to increase reproductive allocation. To address this gap in knowledge, we compared fruit/ovary anatomy, vascular structure and phloem transport of two varieties of giant pumpkins, and their smaller fruited progenitor under controlled environmental conditions. We also modelled carbon transport into the fruit of competitively grown plants using data collected in the field. There was no evidence that changes in leaf area or photosynthetic capacity impacted fruit size. Instead, giant varieties differed in their ovary morphology and contained more phloem on a cross-sectional area basis in their petioles and pedicels than the ancestral variety. These results suggest that sink activity is important in determining fruit size and that giant pumpkins have an enhanced capacity to transport carbon. The strong connection observed between carbon fixation, phloem structure and fruit growth in field-grown plants indicates that breeding for large fruit has led to changes throughout the carbon transport system that could have important implications for how we think about phloem transport velocity and carbon allocation., (© 2014 John Wiley & Sons Ltd.)

A pressure chamber method was used to examine differences in the root to shoot transfer and xylem sap solubility of caffeine (log Kow=-0.07), triclocarban (log Kow=3.5-4.2) and endosulfan (log Kow=3.8-4.8) for zucchini (cucurbita pepo ssp pepo), squash (cucurbita pepo ssp ovifera), and soybean (glycine max L.). Transpiration stream concentration factors (TSCF) for caffeine (TSCF=0.8) were statistically equivalent for all plant species. However, for the more hydrophobic endosulfan and triclocarban, the TSCF values for zucchini (TSCF=0.6 and 0.4, respectively) were 3 and 10 times greater than the soybean and squash (TSCF=0.2 and 0.05, respectively). The difference in TSCF values was examined by comparing the measured solubilities of caffeine, endosulfan and triclocarban in deionized water to those in soybean and zucchini xylem saps using a modified shake flask method. The measured solubility of organic contaminants in xylem sap has not previously been reported. Caffeine solubilities in the xylem saps of soybean and zucchini were statistically equal to deionized water (21500mgL(-1)) while endosulfan and triclocarban solubilities in the zucchini xylem sap were significantly greater (0.43 and 0.21mgL(-1), respectively) than that of the soybean xylem sap (0.31 and 0.11mgL(-1), respectively) and deionized water (0.34 and 0.11mgL(-1), respectively). This suggests that the enhanced root to shoot transfer of hydrophobic organics reported for zucchini is partly due to increased solubility in the xylem sap. Further xylem sap characterization is needed to determine the mechanism of solubility enhancement., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Pumpkin (Cucurbita pepo L.) pollen is starchy, sucrose-poor and recalcitrant, features opposite to those of several model species; therefore, some differences in carbohydrate metabolism could be expected in this species. By studying pumpkin recalcitrant pollen, the objective was to provide new biochemical evidence to improve understanding of how carbohydrate metabolism might be involved in pollen functioning in advanced stages. Four stages were analysed: immature pollen from 1 day before anthesis, mature pollen, mature pollen exposed to the environment for 7 h, and pollen rehydrated in a culture medium. Pollen viability, water and carbohydrate content and activity of enzymes involved in carbohydrate metabolism were quantified in each stage. Pollen viability and water content dropped quickly after dehiscence, as expected. The slight changes in carbohydrate concentration and enzyme activity during pollen maturation contrast with major changes recorded with ageing and rehydration. Pumpkin pollen seems highly active and closely related to its surrounding environment in all the stages analysed; the latter is particularly evident among insoluble sucrolytic enzymes, mainly wall-bound acid invertase, which would be the most relevant for sucrose cleavage. Each stage was characterised by a particular metabolic/enzymatic profile; some particular features, such as the minor changes during maturation, fast sucrolysis upon rehydration or sharp decrease in insoluble sucrolytic activity with ageing seem to be related to the lack of dormancy and recalcitrant nature of pumpkin pollen., (© 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.)

The striped cucumber beetle [Acalymma vittatum (F.)] is a specialist pest of cucurbits throughout its range in the United States and Canada. Improved integrated pest management options are needed across the pest's range, especially on organic farms where there are few effective controls. Trap cropping in cucurbits is an option, but there are significant challenges to the technique. Because cucurbit flowers are highly attractive to the beetles, four field experiments tested whether cultivar and phenology interact to preferentially aggregate beetles. The first experiment tested the hypothesis that cucurbit flowers were more attractive to striped cucumber beetles than was foliage. The second experiment tested whether there were differences in beetle aggregation between two relatively attractive cultivars. The third and fourth experiments were factorial designs with two plant cultivars and two levels of flowering to specifically test for an interaction of cultivar and flowering. Results indicated that flowers were more attractive than foliage, beetle aggregation was affected by plant cultivar, and that there was an interaction of cultivar with flowering. We conclude that a single cultivar may be sufficient to serve as a generic trap crop to protect a wide variety of cucurbits., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Long-distance intercellular electrical signals, including variation potential (VP) in higher plants, are a potential mechanism of coordinate functional responses in different plant cells under action of stressors. VP, which is caused by damaging factors (e.g., heating, crushing), is transient depolarization with an irregular shape. It can include a long-term depolarization and fast impulse depolarization ('AP-like' spikes). Mechanisms of VP generation and propagation are still under investigation. It is probable that VP is a local electrical response induced by propagation of hydraulic wave and (or) chemical agent. Both hypotheses are based on numerous experimental results but they predict VP velocities which are not in a good accordance with speed of variation potential propagation. Thus combination of hydraulic and chemical signals is the probable mechanism of VP propagation. VP generation is traditionally connected with transient H(+)-ATPase inactivation, but AP-like spikes are also connected with passive ions fluxes. Ca(2+) influx is a probable mechanism which triggers H(+)-ATPase inactivation and ions channels activation at VP.

Grafting onto salt-tolerant pumpkin rootstock can increase cucumber salt tolerance. Previous studies have suggested that this can be attributed to pumpkin roots with higher capacity to limit the transport of Na(+) to the shoot than cucumber roots. However, the mechanism remains unclear. This study investigated the transport of Na(+) in salt-tolerant pumpkin and salt-sensitive cucumber plants under high (200 mM) or moderate (90 mM) NaCl stress. Scanning ion-selective electrode technique showed that pumpkin roots exhibited a higher capacity to extrude Na(+), and a correspondingly increased H(+) influx under 200 or 90 mM NaCl stress. The 200 mM NaCl induced Na(+)/H(+) exchange in the root was inhibited by amiloride (a Na(+)/H(+) antiporter inhibitor) or vanadate [a plasma membrane (PM) H(+) -ATPase inhibitor], indicating that Na(+) exclusion in salt stressed pumpkin and cucumber roots was the result of an active Na(+)/H(+) antiporter across the PM, and the Na(+)/H(+) antiporter system in salt stressed pumpkin roots was sufficient to exclude Na(+) X-ray microanalysis showed higher Na(+) in the cortex, but lower Na(+) in the stele of pumpkin roots than that in cucumber roots under 90 mM NaCl stress, suggesting that the highly vacuolated root cortical cells of pumpkin roots could sequester more Na(+), limit the radial transport of Na(+) to the stele and thus restrict the transport of Na(+) to the shoot. These results provide direct evidence for pumpkin roots with higher capacity to limit the transport of Na(+) to the shoot than cucumber roots., (© 2014 Scandinavian Plant Physiology Society.)

The accumulation and trophic transfer of nanoparticle (NP) or bulk CeO2 through a terrestrial food chain was evaluated. Zucchini (Cucurbita pepo L.) was planted in soil with 0 or 1228 μg/g bulk or NP CeO2. After 28 d, zucchini tissue Ce content was determined by ICP-MS. Leaf tissue from each treatment was used to feed crickets (Acheta domesticus). After 14 d, crickets were analyzed for Ce content or were fed to wolf spiders (family Lycosidae). NP CeO2 significantly suppressed flower mass relative to control and bulk treatments. The Ce content of zucchini was significantly greater when exposure was in the NP form. The flowers, leaves, stems, and roots of zucchini exposed to bulk CeO2 contained 93.3, 707, 331, and 119,000 ng/g, respectively; NP-exposed plants contained 153, 1510, 479, and 567 000 ng/g, respectively. Crickets fed NP CeO2-exposed zucchini leaves contained significantly more Ce (33.6 ng/g) than did control or bulk-exposed insects (15.0-15.2 ng/g). Feces from control, bulk, and NP-exposed crickets contained Ce at 248, 393, and 1010 ng/g, respectively. Spiders that consumed crickets from control or bulk treatments contained nonquantifiable Ce; NP-exposed spiders contained Ce at 5.49 ng/g. These findings show that NP CeO2 accumulates in zucchini at greater levels than equivalent bulk materials and that this greater NP intake results in trophic transfer and possible food chain contamination.

Shoot-root communication is involved in plant stress responses, but its mechanism is largely unknown. To determine the role of roots in stress tolerance, cucumber (Cucumis sativus) shoots from plants with roots of their own or with figleaf gourd (Cucurbita ficifolia, a chilling-tolerant species) or luffa (Luffa cylindrica (L.) M. Roem., a heat-tolerant species) rootstocks were exposed to low (18/13°C), optimal (27/22°C) and high (36/31°C) temperatures, respectively. Grafting onto figleaf gourd and luffa rootstocks significantly alleviated chilling and heat-induced reductions, respectively, in biomass production and CO(2) assimilation capacity in the shoots, while levels of lipid peroxidation and protein oxidation were decreased. Figleaf gourd and luffa rootstocks upregulated a subset of stress-responsive genes involved in signal transduction (MAPK1 and RBOH), transcriptional regulation (MYB and MYC), protein protection (HSP45.9 and HSP70), the antioxidant response (Cu/Zn-SOD, cAPX and GR), and photosynthesis (RBCL, RBCS, RCA and FBPase) at low and high growth temperatures, respectively, and this was accompanied by increased activity of the encoded enzymes and reduced glutathione redox homeostasis in the leaves. Moreover, Heat Shock Protein 70 (HSP70) expression in cucumber leaves was strongly induced by the luffa rootstock at the high growth temperature but slightly induced by the figleaf gourd rootstock at low or high growth temperatures. These results indicate that rootstocks could induce significant changes in the transcripts of stress-responsive and defense-related genes, and the ROS scavenging activity via unknown signals, especially at stressful growth temperatures, and this is one of mechanisms involved in the grafting-induced stress tolerance., (© 2014 Scandinavian Plant Physiology Society.)

Production of cucurbit crops presents growers with numerous challenges. Several severe pests and diseases can be managed through the use of rotation, trap cropping, mechanical barriers, such as row covers, and chemical applications. However, considerations must also be made for pollinating insects, as adequate pollination affects the quantity and quality of fruit. Insecticides may negatively affect pollinators; a concern enhanced in recent years due to losses in managed Apis melifera L. colonies. Row covers can be used in place of chemical control before pollination, but when removed, pests have access to fields along with the pollinators. If pollination services of native bees could be harnessed for use under continuous row covers, both concerns could be balanced for growers. The potential of two bee species which specialize on cucurbit flowers, Peponapis pruinosa Say and Xenoglossa strenua Cresson, were assessed under continuous row covers, employed over acorn squash. Experimental treatments included plots with either naturally or artificially introduced bees under row covers and control plots with row covers either permanently removed at crop flowering, or employed continuously with no added pollinating insects. Pests in plots with permanently removed row covers were managed using standard practices used in certified organic production. Marketable yields from plots inoculated with bees were indistinguishable from those produced under standard practices, indicating this system would provide adequate yields to growers without time and monetary inputs of insecticide applications. Additionally, application of this technique was investigated for muskmelon production and discussed along with considerations for farm management.

Potential decreases in crop yield from reductions in bee-mediated pollination services threaten food production demands of a growing population. Many fruit and vegetable growers supplement their fields with bee colonies during crop bloom. The extent to which crop production requires supplementary pollination services beyond those provided by wild bees is not well documented. Pumpkin, Cucurbita pepo L., requires bee-mediated pollination for fruit development. Previous research identified the common eastern bumble bee, Bombus impatiens (Cresson), as the most efficient pumpkin pollinator. Two concomitant studies were conducted to examine pollination deficits in New York pumpkin fields from 2011 to 2013. In the first study, fruit weight, seed set, and B. impatiens visits to pumpkin flowers were compared across fields supplemented with B. impatiens colonies at a recommended stocking density of five colonies per hectare, a high density of 15 colonies per hectare, or not supplemented with bees. In the second study, fruit weight and seed set of pumpkins that received supplemental pollen through hand-pollination were compared with those that were open-pollinated by wild bees. Results indicated that supplementing pumpkin fields with B. impatiens colonies, regardless of stocking density, did not increase fruit weight, seed set, or B. impatiens visits to pumpkin flowers. Fruit weight and seed set did not differ between hand- and open-pollinated treatments. In general, we conclude that pumpkin production in central New York is not limited by inadequate pollination services provided by wild bees and that on average, supplementation with B. impatiens colonies did not improve pumpkin yield.

Singlet oxygen, a harmful reactive oxygen species, can be quantified with the substance 2,2,6,6-tetramethylpiperidine (TEMP) that reacts with singlet oxygen, forming a stable nitroxyl radical (TEMPO). TEMPO has earlier been quantified with electron paramagnetic resonance (EPR) spectroscopy. In this study, we designed an ultra-high-performance liquid chromatographic-tandem mass spectrometric (UHPLC-ESI-MS/MS) quantification method for TEMPO and showed that the method based on multiple reaction monitoring (MRM) can be used for the measurements of singlet oxygen from both nonbiological and biological samples. Results obtained with both UHPLC-ESI-MS/MS and EPR methods suggest that plant thylakoid membranes produce 3.7 × 10(-7) molecules of singlet oxygen per chlorophyll molecule in a second when illuminated with the photosynthetic photon flux density of 2000 μmol m(-2 ) s(-1)., (© 2014 The American Society of Photobiology.)

Unlabelled: •, Premise of the Study: Floral traits play important roles in pollinator attraction and defense against floral herbivory. However, plants may experience trade-offs between conspicuousness to pollinators and herbivore attraction. Comparative studies provide an excellent framework to examine the role of multiple traits shaping mutualist and antagonist interactions.•, Methods: To assess whether putative defensive and attractive traits predict species interactions, we grew 20 different Cucurbitaceae species and varieties in the field to measure interactions with pollinators and herbivores and in the greenhouse to assess trait variation. Cucurbits are characterized by the production of cucurbitacins, bitter nonvolatile terpenoids that are effective against generalist herbivores but can attract specialist beetles. We determined whether plant traits such as cucurbitacins predict herbivore resistance and pollinator attraction using an information-theoretic approach.•, Key Results: Mutualists and floral antagonists were attracted to the same cucurbit varieties once they flowered. However, rather than cucurbitacin concentration, we found that the size of the flower and volatile emissions of floral sesquiterpenoids explained both pollinator and floral herbivore visitation preference across cucurbit taxa. This pattern held across cucurbit taxa and within the Cucurbita genus.•, Conclusions: Surprisingly, floral sesquiterpenoid volatiles, which are associated with direct defense, indirect defense, and attraction, rather than defense traits such as cucurbitacins, appeared to drive interactions with both pollinators and floral herbivores across cucurbit taxa. Identifying the relevant plant traits for attraction and deterrence is important in this economically valuable crop, particularly if pollinators and floral herbivores use the same plant traits as cues., (© 2014 Botanical Society of America, Inc.)

Microorganisms are ubiquitous and thought to regulate host populations. Although microorganisms can be pathogenic and affect components of fitness, few studies have examined their effects on wild plant populations. As individual traits might not contribute equally to changes in population growth rate, it is essential to examine the entire life cycle to determine how microorganisms affect host population dynamics. In this study, we used data from common garden experiments with plants from three Cucurbita pepo populations exposed to three virus treatments. These data were used to parameterize a deterministic matrix model, which allowed us to estimate the effect of virus on components of fitness and population growth rate. Virus did not reduce fruit number, but population growth rates varied among virus treatments and wild C. pepo populations. The effect of virus on population growth rate depended on virus species and wild C. pepo population. Contributions of life-history transitions and life-history traits to population growth rates varied among populations and virus treatments. However, this population-virus interaction was not evident when examining individual components of fitness. Thus, caution must be used when interpreting the effects of changes in individual traits, as single traits do not always predict population-level change accurately., (© 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.)

• The stress-related phytohormones, salicylic acid (SA) and abscisic acid (ABA), and the three jasmonates, jasmonic acid (JA), cis-12-oxo-phytodienoic acid (cis-OPDA), and (+)-7-iso-jasmonoyl-L-isoleucine (JA-Ile), were investigated in phloem and xylem exudates of Cucurbita maxima. • Phloem and xylem exudates were separately collected and analysed via liquid chromatography-mass spectrometry. • We show direct evidence for all three jasmonates, ABA, and SA in both phloem and xylem exudates of C. maxima. JA and JA-Ile concentrations are higher in xylem (JA: c(xylem) ≈ 199.5 nM, c(phloem) ≈ 43.9 nM; JA-Ile: c(xylem) ≈ 7.9 nM, c(phloem) ≈ 1.6 nM), whereas ABA and SA concentrations are higher in phloem exudates (ABA: c(xylem) ≈ 37.1 nM, c(phloem) ≈ 142.6 nM; SA: c(xylem) ≈ 61.6 nM, c(phloem) ≈ 1319 nM). During bacteria-derived flagellin 22 (flg22)-triggered remote root-to-shoot signalling, phytohormone concentration changed rapidly both in phloem and xylem. • The unequal distribution of phytohormones suggests that phloem and xylem have distinct roles in defence responses. Our data shed light on systemic phytohormone signalling and help explain how plants cope with environmental challenges by lateral exchange between phloem and xylem. Our analysis is a starting point for further investigations of how phytohormones contribute to phloem- and xylem-based defence signalling., (© 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.)

This study aimed to evaluate the behavior of zucchini (Cucurbita pepo L.) and cucumber (Cucumis sativus L.) under boron (B) excess. Plants were grown under greenhouse conditions in a sandy soil-peat mixture using a nutrient solution containing 0.2 (control), 10 and 20 mg L(-1) B. Visible symptoms were quantified and leaf B accumulation, gas exchanges, chlorophyll (Chl) a fluorescence, malondialdehyde by-products and antioxidants were investigated 20 days after the beginning of the treatments. Boron toxicity induced oxidative load and leaf necrotic burns coupled with the reduction of leaf growth and biomass accumulation in both species. Boron excess resulted in a decrease of Chl a/b ratio, potential (Fv/Fm) and actual (ΦPSII) PSII quantum efficiency, photosynthetic rate (Pn), stomatal conductance (gs), and transpiration (E) as well. A general stimulation of the antioxidant enzymes ascorbate peroxidase, catalase and superoxide dismutase was observed, and a significant increase in the oxidized form of ascorbate and glutathione was evidenced for treated plants of both species. A difference between the two species was observed: C. pepo appeared to be more sensitive to B stress being damaged at all B concentration. C. sativus grown at 10 mg L(-1) B in nutrient solution showed some down-regulated mechanisms, i.e. increase in Chl b content and a good photochemical PSII efficiency as well as a higher amount of constitutive antioxidant molecules, that, however, are not sufficient to contrast the negative effects of B.

Background: We have identified a kind of parthenocarpy in zucchini squash which is associated with an incomplete andromonoecy, i.e. a partial conversion of female into bisexual flowers. Given that andromonoecy in this and other cucurbit species is caused by a reduction of ethylene production in the female flower, the associated parthenocarpic development of the fruit suggested the involvement of ethylene in fruit set and early fruit development., Results: We have compared the production of ethylene as well as the expression of 13 ethylene biosynthesis and signalling genes in pollinated and unpollinated ovaries/fruits of two cultivars, one of which is parthenocarpic (Cavili), while the other is non-parthenocarpic (Tosca). In the latter, unpollinated ovaries show an induction of ethylene biosynthesis and ethylene signal transduction pathway genes three days after anthesis, which is concomitant with the initiation of fruit abortion and senescence. Fruit set and early fruit development in pollinated flowers of both cultivars and unpollinated flowers of Cavili is coupled with low ethylene biosynthesis and signalling, which would also explain the partial andromonoecy in the parthenocarpic genotype. The reduction of ethylene production in the ovary cosegregates with parthenocarpy and partial andromonoecy in the selfing progeny of Cavili. Moreover, the induction of ethylene in anthesis (by ethephon treatments) reduced the percentage of bisexual parthenocarpic flowers in Cavili, while the inhibition of ethylene biosynthesis or response (by AVG and STS treatments) induces not only andromonoecy but also the parthenocarpic development of the fruit in both cultivars., Conclusions: Results demonstrate that a reduction of ethylene production or signalling in the zucchini flower is able to induce fruit set and early fruit development, and therefore that ethylene is actively involved in fruit set and early fruit development. Auxin and TIBA treatments, inducing fruit set and early fruit development in this species, also inhibit ethylene production and the expression of ethylene biosynthesis and response genes. A model is presented that discusses the crosstalk between ethylene and auxin in the control of fruit set and early fruit development in zucchini squash.

An important aspect in risk assessment of nanoparticles (NPs) is to understand their environmental interactions. We used hydroponic plant cultures to study nanoparticle-plant-root interaction and translocation and exposed wheat and pumpkin to suspensions of uncoated CeO2-NP for 8d (primary particle size 17-100 nm, 100 mg L(-1)) in the absence and presence of fulvic acid (FA) and gum arabic (GA) as representatives of different types of natural organic matter. The behavior of CeO2-NPs in the hydroponic solution was monitored regarding agglomeration, sedimentation, particle size distribution, surface charge, amounts of root association, and translocation into shoots. NP-dispersions were stable over 8d in the presence of FA or GA, but with growing plants, changes in pH, particle agglomeration rate, and hydrodynamic diameter were observed. None of the plants exhibited reduced growth or any toxic response during the experiment. We found that CeO2-NPs translocated into pumpkin shoots, whereas this did not occur in wheat plants. The presence of FA and GA affected the amount of CeO2 associated with roots (pure>FA>GA) but did not affect the translocation factor. Additionally, we could confirm via TEM and SEM that CeO2-NPs adhered strongly to root surfaces of both plant species., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Collection of cucurbit exudates from cut petioles has been a powerful tool for gaining knowledge on phloem sap composition without full notion of the complex exudation mechanism. Only few publications explicitly mentioned that exudates were collected from the basal side of the cut, which exudes more copiously than the apical side. This is surprising since only exudation from the apical side is supposedly driven by phloem pressure gradients. Composition of carbohydrates and pH values at both wounding sides are equal, whereas protein concentration is higher at the basal side. Apparently, exudation is far more complex than just the delivery of phloem sap. Xylem involvement is indicated by lower protein concentrations after elimination of root pressure. Moreover, dye was sucked into xylem vessels owing to relaxation of negative pressure after cutting. The lateral water efflux from the vessels increases turgor of surrounding cells including sieve elements. Simultaneously, detached parietal proteins (PP1/PP2) induce occlusion of sieve plates and cover wound surface. If root pressure is strong enough, pure xylem sap can be collected after removal of the occlusion plug at the wound surface. The present findings provide a mechanism of sap exudation in Cucurbita maxima, in which the contribution of xylem water is integrated., (© 2012 Blackwell Publishing Ltd.)

The present work has developed a methodology based on thermography and sonic anemometry for studying the microclimate in Mediterranean greenhouses equipped with air heaters and polyethylene distribution ducts to distribute the warm air. Sonic anemometry allows us to identify the airflow pattern generated by the heaters and to analyze the temperature distribution inside the greenhouse, while thermography provides accurate crop temperature data. Air distribution by means of perforated polyethylene ducts at ground level, widely used in Mediterranean-type greenhouses, can generate heterogeneous temperature distributions inside the greenhouse when the system is not correctly designed. The system analyzed in this work used a polyethylene duct with a row of hot air outlet holes (all of equal diameter) that expel warm air toward the ground to avoid plant damage. We have observed that this design (the most widely used in Almería's greenhouses) produces stagnation of hot air in the highest part of the structure, reducing the heating of the crop zone. Using 88 kW heating power (146.7 W ∙ m(-2)) the temperature inside the greenhouse is maintained 7.2 to 11.2 °C above the outside temperature. The crop temperature (17.6 to 19.9 °C) was maintained above the minimum recommended value of 10 °C.

Taking 4 different Cucurbita maxima x C. moschata rootstocks for cucumber (Cucumis sativus) as test materials, a solution culture experiment was conducted to study their growth and antioxidative enzyme activities under the stresses of Ca(NO3)2 and NaCl, with the salt stress tolerance of the rootstocks evaluated by subordinate function. At 30 mmol x L(-1) of Ca (NO3)2 or 45 mmol x L(-1) of NaCl, the growth of the rootstock seedlings was improved; but at 60 and 120 mmol x L(-1) of Ca(NO3)2 or 90 and 180 mmol x L(-1) of NaCl, the growth and the antioxidative systems of the seedlings were inhibited, and the salt injury index of 'Qingzhen No. 1' was the smallest, with the decrement of biomass and SOD, POD and CAT activities and the increment of relative conductance being significantly lower than those of the others. Under the stress of high concentration Ca(NO3)2, the SOD, POD and CAT activities of test rootstocks were higher, and the salt injury index and relative conductance were lower, as compared with those under high concentration NaCl, suggesting that the damage of Ca(NO3)2 stress to cucumber-grafted rootstock were smaller than that of NaCl stress. Among the 4 rootstocks, 'Qingzhen No. 1' had the strongest salt stress tolerance, followed by 'Zuomu Nangua', 'Fengyuan Tiejia', and 'Chaoba Nangua'.

Environmental changes threaten plant-pollinator mutualisms and their critical ecosystem service. Drivers such as land use, invasions and climate change can affect pollinator diversity or species encounter rates. However, nitrogen deposition, climate warming and CO(2) enrichment could interact to disrupt this crucial mutualism by altering plant chemistry in ways that alter floral attractiveness or even nutritional rewards for pollinators. Using a pumpkin model system, we show that these drivers non-additively affect flower morphology, phenology, flower sex ratios and nectar chemistry (sugar and amino acids), thereby altering the attractiveness of nectar to bumble bee pollinators and reducing worker longevity. Alarmingly, bees were attracted to, and consumed more, nectar from a treatment that reduced their survival by 22%. Thus, three of the five major drivers of global environmental change have previously unknown interactive effects on plant-pollinator mutualisms that could not be predicted from studies of individual drivers in isolation., (© 2012 Blackwell Publishing Ltd/CNRS.)

Many organisms face challenges in avoiding predation while searching for mates. For plants, emitting floral fragrances to advertise reproductive structures could increase the attraction of detrimental insects along with pollinators. Very few studies have experimentally evaluated the costs and benefits of fragrance emission with explicit consideration of how plant fitness is affected by both pollinators and florivores. To determine the reproductive consequences of increasing the apparency of reproductive parts, we manipulated fragrance, pollination, and florivores in the wild Texas gourd, Cucurbita pepo var. texana. With enhanced fragrance we found an increase in the attraction of florivores, rather than pollinators, and a decrease in seed production. This study is the first to demonstrate that enhanced floral fragrance can increase the attraction of detrimental florivores and decrease plant reproduction, suggesting that florivory as well as pollination has shaped the evolution of floral scent.

Plant responses to physiological function disorders are called symptoms and they are caused principally by pathogens and nutritional deficiencies. Plant symptoms are commonly used as indicators of the health and nutrition status of plants. Nowadays, the most popular method to quantify plant symptoms is based on visual estimations, consisting on evaluations that raters give based on their observation of plant symptoms; however, this method is inaccurate and imprecise because of its obvious subjectivity. Computational Vision has been employed in plant symptom quantification because of its accuracy and precision. Nevertheless, the systems developed so far lack in-situ, real-time and multi-symptom analysis. There exist methods to obtain information about the health and nutritional status of plants based on reflectance and chlorophyll fluorescence, but they use expensive equipment and are frequently destructive. Therefore, systems able of quantifying plant symptoms overcoming the aforementioned disadvantages that can serve as indicators of health and nutrition in plants are desirable. This paper reports an FPGA-based smart sensor able to perform non-destructive, real-time and in-situ analysis of leaf images to quantify multiple symptoms presented by diseased and malnourished plants; this system can serve as indicator of the health and nutrition in plants. The effectiveness of the proposed smart-sensor was successfully tested by analyzing diseased and malnourished plants.

Male bees can be abundant at flowers, particularly floral hosts of those bee species whose females are taxonomic pollen specialists (oligolecty). Contributions of male bees to host pollination are rarely studied directly despite their prevalence in a number of pollination guilds, including those of some crop plants. In this study, males of the oligolectic bee, Peponapis pruinosa Say, were shown to be effective pollinators of summer squash, Cucurbita pepo L. Seven sequential visits from male P. pruinosa maximized squash fruit set and growth. This number of male visits accumulated during the first hour of their foraging and mate searching at flowers soon after sunrise. Pollination efficacy of male P. pruinosa and their abundances at squash flowers were sufficient to account for most summer squash production at our study sites, and by extrapolation, to two-thirds of all 87 North American farms and market gardens growing squashes that were surveyed for pollinators by collaborators in the Squash Pollinators of the Americas Survey. We posit that the substantial pollination value of male Peponapis bees is a consequence of their species' oligolecty, their mate seeking strategy, and some extreme traits of Cucurbita flowers (massive rewards, flower size, phenology).

A pot experiment with artificial shading was conducted to study the effects of soil moisture content and light intensity on the plant growth and leaf physiological characteristics of squash variety "Jingyingyihao". Under all test soil moisture conditions, 30% shading promoted the growth of "Jingyingyihao", with the highest yield at 70% - 80% soil relative moisture contents. 70% shading inhibited plant growth severely, only flowering and not bearing fruits, no economic yield produced. In all treatments, there was a similar water consumption trend, i. e., both the daily and the total water consumption decreased with increasing shading and decreasing soil moisture content. Among all treatments, 30% shading and 70% - 80% soil relative moisture contents had the highest water use efficiency (2.36 kg mm(-1) hm(-2)) and water output rate (1.57 kg mm(-1) hm(-2)). The net photosynthetic rate, transpiration rate, stomatal conductance, and chlorophyll content of squash leaves decreased with increasing shading, whereas the intercellular CO2 concentration was in adverse. The leaf protective enzyme activity and proline content decreased with increasing shading, and the leaf MAD content decreased in the order of 70% shading, natural radiation, and 30% shading. Under the three light intensities, the change characteristics of squash leaf photosynthesis, protective enzyme activity, and proline and MAD contents differed with the increase of soil relative moisture content.

The objectives of this work were to determine the squash entomofauna in the region of Viçosa, Minas Gerais state, to study their behavior on flowers and their importance for pollination, verifying the role of each pollinator. The most common species were Trigona spinipes (Fabricius), Trigona hyalinata (Lepeletier), Apis mellifera (L.) and Melipona quadrifasciata (Lepeletier). The visitation behavior of A. mellifera, M. quadrifasciata, and Bombus morio (Swederus) were similar. They visited flowers for nectar collection, positioning themselves vertically between the corolla and the sexual structures of the flowers, with the back directed toward the floral axis, which permitted the removal of pollen from the anthers of flowers with stamens and its deposition on the stigma of flowers with pistils, being considered therefore effective pollinators. Trigona spinipes and T. hyalinata foraged in groups, preventing other species from landing on the flowers which they occupied. Due to their small body size and only infrequent contact with the sexual structures of the flowers, these species are considered occasional pollinators. The number of fruits produced differed between freely visited flowers, those prevented from receiving visits and those visited only a single time by M. quadrifasciata, B. morio, A. mellifera, T. hyalinata or T. spinipes. Flowers prevented from receiving visits or visited only once by T. spinipes did not produce fruits. The remaining pollination systems led to fruitification, with open pollination or a single visit from either M. quadrifasciata or B. morio leading to most fruit production.

Sieve elements are one of the least understood cell types in plants. Translocation velocities and volume flow to supply sinks with photoassimilates greatly depend on the geometry of the microfluidic sieve tube system and especially on the anatomy of sieve plates and sieve plate pores. Several models for phloem translocation have been developed, but appropriate data on the geometry of pores, plates, sieve elements, and flow parameters are lacking. We developed a method to clear cells from cytoplasmic constituents to image cell walls by scanning electron microscopy. This method allows high-resolution measurements of sieve element and sieve plate geometries. Sieve tube-specific conductivity and its reduction by callose deposition after injury was calculated for green bean (Phaseolus vulgaris), bamboo (Phyllostachys nuda), squash (Cucurbita maxima), castor bean (Ricinus communis), and tomato (Solanum lycopersicum). Phloem sap velocity measurements by magnetic resonance imaging velocimetry indicate that higher conductivity is not accompanied by a higher velocity. Studies on the temporal development of callose show that small sieve plate pores might be occluded by callose within minutes, but plants containing sieve tubes with large pores need additional mechanisms.