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Variability of vitamin E content in pumpkin seeds (Cucurbita pepo L.).

Pumpkin (Cucurbita pepo L.) seed oil is a common salad oil which is produced in the southern parts of Austria, Slovenia and Hungary. It is dark green and has a high content of free fatty acids. Due to its colour, the oil cannot be used for cooking. The content of vitamin E, especially gamma-tocopherol, is very high. The oil content of the pumpkin seed is about 50%. The seed itself can be eaten. Therefore a pumpkin variety with high vitamin E content is desirable. The aim of this work was to find a variety of Cucurbita pepo which has a high oil yield and a high vitamin E content. A total of 100 breeding lines were tested for their tocopherol content. The tocopherols and tocotrienols are extracted with hexane and analysed by NP-HPLC/FLD with hexane/dioxan (96/4) as eluent, with fluorescence detection at 292/335 nm. The gamma-tocopherol content, which is about 5-10 times as much as that of alpha-tocopherol varies over a broad range (41-620 mg/kg dry pumpkin seeds). Beta- and delta-tocopherol are found at low levels.

Chromatography, High Pressure Liquid↗

Development of recombinant inbred lines and QTL analysis of plant height and fruit shape-related traits in Cucurbita pepo L.

UNLABELLED: Zucchini (Cucurbita pepo subsp. pepo) stands as an economically vital crop in China. In zucchini breeding, plant architectural patterns and fruit morphological characteristics serve as pivotal traits. In this study, we employed quantitative trait locus (QTL) analysis using recombinant inbred lines (RILs) derived from two distinct inbred lines, JinGL (subsp. ovifera) and HM-S2 (subsp. pepo), in conjunction with a high-density genetic map. Our investigation focused on ten QTLs associated with six horticulturally significant traits, including hypocotyl length (HL), plant height (PH), and four fruit-related traits: fruit length (FL), fruit diameter (FD), fruit shape index (FSI), and fruit weight (FW). The QTLs governing HL and PH were mapped to Chr03/LG10 and named qhl3.1 and qph3.1, respectively. The candidate gene Cp4.1LG10g05910/CpDw for qph3.1 was successfully identified. Additionally, three novel QTLs related to fruit size and shape were discovered. Among them, qfsi8.1/qfl8.1, demarcated by Marker238258 and Marker240069 on Chromosome 08/Linkage group 17 (Chr08/LG17), is a new major QTL regulating the fruit shape of zucchini. Through genomic insertion-deletion (InDel) and qRT-PCR analyses, we predicted genes within the qfsi8.1/qfl8.1 candidate interval, uncovering Cp4.1LG17g02030/CpIAA12 and Cp4.1LG17g02010/CpCalB as potential candidate genes. We developed molecular markers tightly linked to qph3.1 and qfl8.1 and validated them in 171 and 224 Cucurbita pepo germplasms, achieving accuracy rates of 96% and 100%, respectively. This study deepens our understanding of the genetic basis of key traits and provides valuable references for molecular breeding in Cucurbita pepo. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11032-025-01592-y.

Cucurbita pepo↗

Variability of fatty acid content in pumpkin seeds (Cucurbita pepo L.).

Pumpkin (Cucurbita pepo L.) seed oil is a common salad oil which is produced in Slovenia, Hungary and the southern parts of Austria. It is dark green and has a high content of free fatty acids. The seed itself can be eaten. Due to its colour and the foam formation, the oil cannot be used for cooking. The content of vitamin E, especially gamma-tocopherol, is very high. The oil content of the pumpkin seed is about 50%. The variability in the oil content is very high resulting from a broad genetic diversity. Thus a breeding programme for increasing the oil productivity is very promising. The four dominant fatty acids are palmitic, stearic, oleic and linoleic acids. These four fatty acids make up 98 +/- 0.13% of the total amount of fatty acids, others being found at levels well below 0.5%.

Austria↗

Assessment of genetic relationships in Cucurbita pepo (Cucurbitaceae) using DNA markers.

Cucurbita pepo (pumpkin, squash, gourd), an economically important species of the Cucurbitaceae, is extremely variable in fruit characteristics. The objective of the present study was to clarify genetic relationships across a broad spectrum of the C. pepo gene pool, with emphasis on domesticates, using AFLP, ISSR and SSR markers. Forty-five accessions were compared for presence or absence of 448 AFLP, 147 ISSR, and 20 SSR bands, their genetic distances (GDs) were estimated and UPGMA cluster analysis was conducted. The results obtained from these three marker systems were highly correlated (P << 0.001). Clustering was in accordance with the division of C. pepo into three subspecies, fraterna, texana and pepo, with the first two less distant to one another than to the last one. Within the clusters, sub-clustering occurred in accordance with fruit shape and size. The subsp. texana cluster consisted of six sub-clusters, one each for the representatives of its five cultivar-groups (Acorn, Crookneck, Scallop, Straightneck and Ovifera Gourd) and wild gourds. Within the subsp. pepo cluster, the representatives of two cultivar-groups (Zucchini and Orange Gourd) formed distinct sub-clusters and the representatives of two other groups (Cocozelle and Vegetable Marrow) tended to sub-cluster separately from one another but formed an assemblage with the representatives of the remaining group (Pumpkin). Within-group GDs were less than corresponding between-group GDs in nearly all comparisons. The smallest-fruited accession, 'Miniature Ball', appears to occupy a genetically central position within C. pepo.

Cucurbita↗

Subspecies-level variation in the phytoextraction of weathered p,p'-DDE by Cucurbita pepo.

Previous studies indicate that Cucurbita pepo can phytoextract highly weathered persistent organic pollutants (POPs) from soil and translocate large quantities to aerial tissues. To investigate intraspecific variability in uptake potential, a field study was conducted to quantify the phytoextraction of weathered p,p'-DDE by 21 cultivar varieties of summer squash from two distinct subspecies, C. pepo ssp texana and C. pepo ssp pepo. Significant differences exist between the two subspecies, with average root and stem to soil bioconcentration factors (BCF, dry weight ratio of contaminant concentration in the vegetation to that in the soil) of 7.22 and 5.40 for ssp pepo and of 2.37 and 0.454 for spp texana, respectively. The amounts of weathered p,p-DDE extracted from the soil by ssp pepo and ssp texana were 0.301 and 0.065%, respectively, with maximum values within each subspecies of 0.780 and 0.182%, respectively. The quantities of 14 inorganic elements were determined in both the soil and tissues (roots, stems, leaves, and fruit) of all 21 cultivar varieties. Phosphorus concentrations in the tissues of ssp pepo were 14 (fruit)-73% (stems) greater than those of ssp texana. These data support our hypothesis that the unique ability of certain cultivars of C. pepo to phytoextract highly weathered POPs from soil is the result of low molecular weight organic acid exudation as a unique phosphorus acquisition mechanism.

Biodegradation, Environmental↗

Crystallization and preliminary X-ray crystallographic studies of mavicyanin from Cucurbita pepo medullosa.

Mavicyanin isolated from Cucurbita pepo medullosa is a glycosylated protein containing a single polypeptide chain of 109 amino-acid residues and is a member of the phytocyanin subclass of cupredoxins. Non-glycosylated recombinant mavicyanin, which was expressed in Escherichia coli, was crystallized by the hanging-drop vapour-diffusion method with ammonium sulfate as the precipitant at pH 5.5. The crystals belonged to the hexagonal space group P6(1) (or P6(5)), with unit-cell parameters a = 64.0, c = 245.0 A, four molecules per asymmetric unit and a solvent content of 59%. X-ray diffraction data were collected to 1.6 A resolution. To solve the structure of mavicyanin, the MAD method as well as a Patterson search method using the structure of stellacyanin as a starting model are presently being utilized.

Ammonium Sulfate↗

Genetic diversity of a germplasm collection of Cucurbita pepo using SRAP and AFLP markers.

Cucurbita pepo is a highly polymorphic species. The cultivars can be grouped into eight morphotypes in two subspecies, ssp. pepo and ssp. ovifera. A collection of 69 accessions representative of the morphotypes and some unclassified types was used for analysing the morphological and molecular diversity of this species. This collection includes commercial cultivars and Spanish landraces, which represent the great diversification of types that have arisen in Europe after this species arrived from America. For the molecular variability studies, two PCR-based systems were employed, AFLP and SRAP, which preferentially amplify ORFs. Principal coordinates analysis and cluster analysis using the UPGMA method clearly separate the accessions into the two subspecies through the use of both markers. However, the gene diversity and the genetic identity values among morphotypes and subspecies varied between the two marker systems. The information given by SRAP markers was more concordant to the morphological variability and to the evolutionary history of the morphotypes than that of AFLP markers. In ssp. ovifera, the accessions of the different morphotypes were basically grouped according to the fruit colour. This may indicate different times of development and also the extent of breeding in the accessions used. This study has allowed identification of new types that can be employed for the development of new cultivars. The landraces of the spp. ovifera, used as ornamental in Europe, have proved to be of great interest for preserving the diversity of C. pepo.

Cluster Analysis↗

A lectin from the exudate of the fruit of the vegetable marrow (Cucurbita pepo) that has a specificity for beta-1,4-linked N-acetylglucosamine oligosaccharides.

Lectins are present in the exudate (presumably from the phloem) of the fruits of three species of the Cucurbitaceae, namely vegetable marrow (Cucurbita pepo), melon (Cucumis melo) and cucumber (Cucumis sativus). They are all strongly inhibited in their activities by chitin oligosaccharides, but only weakly by N-acetylglucosamine. Glycopeptides from soya-bean agglutinin and fetuin are also strong inhibitors of Cucurbita pepo lectin, indicating that it interacts with internal N-acetylglucosamine residues. The lectin from Cucurbita pepo fruit was purified by affinity chromatography by using chitin oligosaccharides covalently attached to Sepharose. The lectin is not a glycoprotein, and it consists of a single polypeptide chain of about 20,000 mol.wt. It is a major protein (18% of the total) of the phloem exudate and it is postulated that it may have an anti-parasitic function.

Acetylglucosamine↗

Artificial elevation of glutathione affects symptom development in ZYMV-infected Cucurbita pepo L. plants.

Styrian oil pumpkin seedlings (Cucurbita pepo L. subsp. pepo var. styriaca GREB: .) were treated for 48 h with 1 mM OTC (L-2-oxothiazolidine-4-carboxylic acid) in order to artificially increase cellular glutathione content. They were inoculated with zucchini yellow mosaic virus (ZYMV) 10 days later. The effects of OTC treatment and ZYMV infection on glutathione levels were examined at the subcellular level by immunogold labeling of glutathione using a transmission electron microscope (TEM). These effects were further tested at the whole-tissue level by high performance liquid chromatography (HPLC). Such tests were carried out a) on roots, cotyledons and the first true leaves immediately after OTC treatment in order to analyze to which extent OTC increases glutathione levels in different cell compartments as well as in the whole organ; and b) in older and younger leaves and in roots three weeks after ZYMV inoculation in order to study how possible effects of OTC on symptom development would correlate with glutathione levels at the subcellular level and in the whole organ. Immunocytological and biochemical investigations revealed that, 48 h after OTC treatment, glutathione content had increased in all investigated organs, up to 144% in peroxisomes of cotyledons. Three weeks after ZYMV inoculation, glutathione labeling density had significantly increased within intact cells of infected leaves, up to 124% in the cytosol of younger leaves. Roots showed decreased amounts of glutathione in the TEM. Biochemical studies revealed that OTC treatment resulted in 41 and 51% higher glutathione content in older and younger ZYMV-infected leaves, respectively, in comparison to untreated and ZYMV-infected plants. Evaluation of symptom development at this point revealed that all untreated ZYMV-infected plants had symptoms, whereas only 42% of OTC-treated ZYMV-infected plants showed signs of symptoms. Quantification of ZYMV particles revealed that all organs of OTC-treated and ZYMV-infected plants contained significantly decreased amounts of ZYMV particles over a period of five weeks when compared to the same organs of untreated ZYMV-infected plants. We can conclude that OTC treatment and subsequently elevated glutathione contents within Styrian oil pumpkin plants led to a strong decrease in virus content, which was accompanied by a suppression of ZYMV-induced symptoms as well as reduced and delayed symptom development within plants exhibiting symptoms.

Antiviral Agents↗

Purification, characterization and subcellular localization of a type-1 ribosome-inactivating protein from the sarcocarp of Cucurbita pepo.

The flesh of the fruit of Cucurbita pepo contains a type-1 ribosome-inactivating protein (RIP), which we named pepocin. Pepocin was purified to apparent homogeneity by acid fractionation, ion-exchange chromatography and adsorption chromatography. The protein was found to have a molecular mass of 26 kDa and a pI of about 9.9. It does not contain glycosidic linkages. The protein inhibits protein synthesis in a rabbit-reticulocyte lysate with an IC50 (concentration causing 50% inhibition) of 15.4 pM, and depurinates 28S rRNA in the ribosomes of the lysate in a manner identical to that of ricin A-chain and other RIP. The enzyme is also active on wheat-germ ribosomes and on Escherichia coli ribosomes. The sequence of the N-terminal 20 amino acids of the protein reveals a close relationship to other RIP. Immunoelectron-microscopic localization of pepocin in the sarcocarp shows that the protein is predominantly localized in intercellular spaces. In addition, the immunolocalized signals are observed in leaf intercellular spaces.

Amino Acid Sequence↗

Identification of the proteins responsible for SAR DNA binding in nuclear matrix of Cucurbita pepo.

The nuclear matrices from White bush (Cucurbita pepo var. patisonina) cell nuclei have been isolated using three methods: I, standard procedure involving extraction of cell nuclei with 2 M NaCl and 1% Triton X-100; II, the same with pre-treatment of cell nuclei with 0.5 mM CuSO4 (stabilisation step); and III, method with extraction by lithium diiodosalicylate (LIS), and compared the polypeptide pattern. The isolated matrices specifically bind SAR DNA derived from human beta-interferon gene in the exogenous SAR binding assay and in the gel mobility shift assay. Using IgG against the 32 kDa endonuclease we have found in the DNA-protein blot assay that this protein is one of the proteins binding SAR DNA. We have identified three proteins with molecular mass of 65 kDa, 60 kDa and 32 kDa which are responsible for SAR DNA binding in the gel mobility shift assay experiments.

Antigens, Nuclear↗

Soil amendments, plant age, and intercropping impact p,p'-DDE bioavailability to Cucurbita pepo.

Field experiments were conducted to optimize the phytoextraction of weathered p,p'-DDE (p,p'-dichlorodiphenyldichloroethylene) by Cucurbita subspecies. The effects of two soil amendments, mycorrhizae or a biosurfactant, on p,p'-DDE accumulation was determined. Also, p,p'-DDE uptake was assessed during plant growth (12, 26, 38, and 62 d), and cultivars that accumulate weathered p,p'-DDE were intercropped with cultivars known not to have that ability. Cucurbita pepo L. ssp. pepo accumulated large amounts of the contaminant, having stem bioconcentration factors, amounts of p,p'-DDE translocated, and contaminant phytoextraction that were 14, 9.9, and 5.0 times greater than C. pepo L. ssp. ovifera (L.) D.S. Decker, respectively. During 62 d, the stem BCF (bioconcentration factor) for p,p'-DDE in subspecies pepo remained constant and the total amount of contaminant accumulated was correlated with plant biomass (r(2) = 0.86). For subspecies ovifera, the stem BCF was highest at 12 d (1.5) but decreased to 0.39 by 62 d, and p,p'-DDE removal was not correlated with plant biomass. Mycorrhizal inoculation increased p,p'-DDE accumulation by both subspecies by an average 4.4 times. For subspecies pepo, mycorrhizae increased the percentage of contaminant extracted from 0.72 to 2.1%. Biosurfactant amendment also enhanced contaminant accumulation by both subspecies, although treatment reduced subspecies ovifera biomass by 60%. The biosurfactant had no effect on the biomass of subspecies pepo, increased the average contaminant concentration by 3.6-fold, and doubled the overall amount of p,p'-DDE removed from the soil. Soil amendments that enhance the mobility of weathered persistent organic pollutants will significantly increase the amount of contaminant phytoextraction by Cucurbita pepo.

Biodegradation, Environmental↗

Pleiotropic mutation in a tendril TCP gene underlies the yield-enhancing multiple-flowering trait in summer squash (Cucurbita pepo).

Crop yield is a focal point in plant breeding. Regulation of lateral budding through apical dominance was a central target of crop domestication, directly affecting crop production. The young fruits of Cucurbita pepo, summer squash, are produced on plants characterized by apical dominance and differentiation of a single flower bud per leaf axil. A single recessive mutation, mf, results in differentiation of more than one flower per leaf axil, thereby directly increasing production because of the continual day-to-day harvest of the summer squash crop. Positional cloning of the Cucurbita pepo mf (Cpmf) gene denoted a frameshift mutation in a TCP transcription factor, Cp4.1LG13g07780, as causative for the increase in axillary flowering. Cpmf is an ortholog of a tendril-development TCP gene in other cucurbits, and likewise, the recessive allele of Cpmf is associated with distorted tendril development. Gene function is context dependent, and we propose that multiple flowering is a unique pleiotropic attribute of mutation in a tendril-development gene of C. pepo. Characterization of a C. pepo collection confirmed a significant association of the Cpmf mutation with multiple flowering and showed that the mutant allele is absent in ancestral C. pepo and one of its two cultivated subspecies. The beneficial mutation occurred and was selected after the domestication of the other subspecies, during its cultivation for young fruit production. We demonstrate the discovery of a causative yield-increasing sequence variant and its practical utilization in breeding. Our findings provide a molecular target for creation of high-yielding, multiple-flowering summer squash cultivars through marker-assisted breeding or precise genome editing.

Cucurbita↗

UV-C irradiation reduces microbial populations and deterioration in Cucurbita pepo fruit tissue.

Tissue slices of zucchini squash (Cucurbita pepo L., cv. Tigress) fruit were exposed to ultraviolet-C (UV-C) radiation from germicidal lamps for 1, 10 or 20 min; however, only 10 and 20 min UV-C exposure significantly reduced microbial activity and deterioration during subsequent storage at 5 or 10 degrees C. UV-C treated slices had higher respiration rates than controls; however, the ethylene production of the slices was not affected by UV-C treatments. Slight UV-C irradiation damage (reddish brown discoloration) was detected on the surface of 10 and 20 min-treated slices after 12 days of storage at 10 degrees C. Slices stored at 5 degrees C did not show UV-C damage. Chilling injury was not observed until after 20 days of storage at 5 degrees C. The symptoms of chilling injury appeared as dried sunken brown spots on the surface of cortex tissue. UV-C treatments did not affect the degree of chilling injury during storage at 5 degrees C. UV-C treatment also had no consistent effect on sugar or malic acid concentrations. The most pronounced effect of UV-C irradiation was to retard microbial growth thereby providing a basis for the frequently observed delay in senescence and subsequent deterioration in fruit tissues.

Journal Article↗

Environmental variation influences the magnitude of inbreeding depression in Cucurbita pepo ssp. texana (Cucurbitaceae).

We grew inbred and outcrossed Cucurbita pepo ssp. texana plants and measured inbreeding depression for several male and female fitness traits 4 years in a row in adjacent fields at the same field station under the same cultivation conditions. We found that the magnitude of inbreeding depression varied from 0.16 to 0.53 from year to year and that those traits which were most affected tended to vary with year. We also grew inbred and outcrossed C. pepo ssp. texana plants in two adjacent fields differing only in the presence of nitrogen fertilizer to examine the effect of nutrient limitation as a form of environmental stress on the magnitude of inbreeding depression. We found that inbreeding depression was more severe in the unfertilized field. Overall, this study illustrates the notion that any estimate of inbreeding depression represents a single point in a cluster of possible estimates that can vary (often dramatically) with growing conditions.

Adaptation, Physiological↗

Azide-binding studies reveal type 3 copper heterogeneity in ascorbate oxidase from the green zucchini squash (Cucurbita pepo).

Titration of native ascorbate oxidase from green zucchini squash (Cucurbita pepo) with azide in 0.1 M-phosphate buffer, pH 6.8, exhibits a biphasic spectral behaviour. Binding of the anion with 'high affinity' (K greater than 5000 M-1) produces a broad increase of absorption in the 400-500 nm region (delta epsilon approximately 1000 M-1.cm-1) and c.d. activity in the 300-450 nm region, whereas azide binding with 'low affinity' (K approximately 100 M-1) is characterized by an intense absorption band at 420 nm (delta epsilon = 6000 M-1.cm-1), corresponding to negative c.d. activity and a decrease of absorption at 330 nm (delta epsilon = -2000 M-1.cm-1). The high-affinity binding involves a minor fraction of the protein containing Type 3 copper in the reduced state, and the spectral features of this azide adduct can be eliminated by treatment of the native enzyme with small amounts of H2O2, followed by dialysis before azide addition. As shown by e.s.r. spectroscopy, Type 2 copper is involved in both types of binding, its signal being converted into that of a species with small hyperfine splitting constant [12 mT (approximately 120 G)] in the case of the low-affinity azide adduct. The spectral similarities of the two types of azide adducts with the corresponding adducts formed by native laccase, which also exhibits Type 3 copper heterogeneity, are discussed.

Ascorbate Oxidase↗

Metabolic Requirement of Cucurbita pepo for Boron.

Lateral roots of intact summer squash seedlings (Cucurbita pepo L.) were used to quantify the effects of boron deficiency on DNA synthesis, protein synthesis, and respiration. The temporal relationship between changes in these metabolic activities and the cessation of root elongation caused by boron deprivation was determined. Transferring 5-day-old squash seedlings to a hydroponic culture medium without boron for 6 hours resulted in a 62% reduction in net root elongation and a 30% decrease in the incorporation of [(3)H]thymidine into DNA by root tips (apical 5-millimeter segments). At this time, root tips from both boron-deficient and boron-sufficient plants exhibited nearly identical rates of incorporation of [(14)C]leucine into protein and respiration as measured by O(2) consumption. After an additional 6 hours of boron deprivation, root elongation had nearly ceased. Concomitantly, DNA synthesis in root apices was 66% less than in the boron-sufficient control plants and protein synthesis was reduced 43%. O(2) consumption remained the same for both treatments. The decline and eventual cessation of root elongation correlated temporally with the decrease in DNA synthesis, but preceded changes in protein synthesis and respiration. These results suggest that boron is required for continued DNA synthesis and cell division in root meristems.

Journal Article↗

The Nature of Light-Induced Inhibition of Photosystem II in Pumpkin (Cucurbita pepo L.) Leaves Depends on Temperature.

Attached leaves of pumpkin (Cucurbita pepo L.) were treated in high or moderate light at room temperature or a 1 degrees C. The symptoms of photoinhibition appearing during light treatments at room temperature could be attributed to a decrease in the primary activity of PSII. However, when the light treatment was given at 1 degrees C, the quantum yield of photosynthetic oxygen evolution decreased much more than would be expected from the decrease in the ratio of variable to maximum fluorescence at 77 degrees K. Also, light treatment at 1 degrees C lowered the chloroplast wholechain electron transfer capacity much more than it affected PSII electron transport (H(2)O to paraphenylbenzoquinone). Light treatments at both room temperature and 1 degrees C led to an increase in B(max), which indicates an increase in the proportion of PSII(beta) centers. PSI was not affected by the light treatments, and the treatments in the dark at 1 degrees C caused only minor changes in the measured properties of the leaves. We conclude that high light always inhibits the primary activity of PSII, but at low temperature there is greater inhibition of electron transfer from primary electron accepting plastoquinone of PSII to the plastoquinone pool, which leads to a drastic decrease in the quantum yield of oxygen evolution in the chilling-sensitive pumpkin.

Journal Article↗