In vivo profiling of the Zucchini proximal proteome in the Drosophila ovary.

PIWI-interacting RNAs (piRNAs) are small RNAs that play a conserved role in genome defense. The piRNA processing pathway is dependent on the sequestration of RNA precursors and protein factors in specific subcellular compartments. Therefore, a highly resolved spatial proteomics approach can help identify the local interactions and elucidate the unknown aspects of piRNA biogenesis. Herein, we performed TurboID proximity labeling to investigate the interactome of Zucchini (Zuc), a key factor of piRNA biogenesis in germline cells and somatic follicle cells of the Drosophila ovary. Quantitative mass spectrometry analysis of biotinylated proteins defined the Zuc-proximal proteome, including the well-known partners of Zuc. Many of these were enriched in the outer mitochondrial membrane (OMM), where Zuc was specifically localized. The proximal proteome of Zuc showed a distinct set of proteins compared with that of Tom20, a representative OMM protein, indicating that chaperone function-related and endomembrane system/vesicle transport proteins are previously unreported interacting partners of Zuc. The functional relevance of several candidates in piRNA biogenesis was validated by derepression of transposable elements after knockdown. Our results present potential Zuc-interacting proteins, suggesting unrecognized biological processes.

Zucchini-dependent piRNA processing is triggered by recruitment to the cytoplasmic processing machinery.

The piRNA pathway represses transposable elements in the gonads and thereby plays a vital role in protecting the integrity of germline genomes of animals. Mature piRNAs are processed from longer transcripts, piRNA precursors (pre-piRNAs). In Drosophila, processing of pre-piRNAs is initiated by piRNA-guided Slicer cleavage or the endonuclease Zucchini (Zuc). As Zuc does not have any sequence or structure preferences in vitro, it is not known how piRNA precursors are selected and channeled into the Zuc-dependent processing pathway. We show that a heterologous RNA that lacks complementary piRNAs is processed into piRNAs upon recruitment of several piRNA pathway factors. This processing requires Zuc and the helicase Armitage (Armi). Aubergine (Aub), Argonaute 3 (Ago3), and components of the nuclear RDC complex, which are required for normal piRNA biogenesis in germ cells, are dispensable. Our approach allows discrimination of proteins involved in the transcription and export of piRNA precursors from components required for the cytoplasmic processing steps. piRNA processing correlates with localization of the substrate RNA to nuage, a distinct membraneless cytoplasmic compartment, which surrounds the nucleus of germ cells, suggesting that sequestration of RNA to this subcellular compartment is both necessary and sufficient for selecting piRNA biogenesis substrates.

Identification of ARF genes in Cucurbita pepo L and analysis of expression patterns, and functional analysis of CpARF22 under drought, salt stress.

BACKGROUND: Auxin transcription factor (ARF) is an important transcription factor that transmits auxin signals and is involved in plant growth and development as well as stress response. However, genome-wide identification and responses to abiotic and pathogen stresses of the ARF gene family in Cucurbita pepo L, especially pathogen stresses, have not been reported. RESULTS: Finally, 33 ARF genes (CpARF01 to CpARF33) were identified in C.pepo from the Cucurbitaceae genome database using bioinformatics methods. The putative protein contains 438 to 1071 amino acids, the isoelectric point is 4.99 to 8.54, and the molecular weight is 47759.36 to 117813.27 Da, the instability index ranged from 40.74 to 68.94, and the liposoluble index ranged from 62.56 to 76.18. The 33 genes were mainly localized in the nucleus and cytoplasm, and distributed on 16 chromosomes unevenly. Phylogenetic analysis showed that 33 CpARF proteins were divided into 6 groups. According to the amino acid sequence of CpARF proteins, 10 motifs were identified, and 1,3,6,8,10 motifs were highly conserved in most of the CpARF proteins. At the same time, it was found that genes in the same subfamily have similar gene structures. Cis-elements and protein interaction networks predicted that CpARF may be involved in abiotic factors related to the stress response. QRT-PCR analysis showed that most of the CpARF genes were upregulated under NaCl, PEG, and pathogen treatment compared to the control. Subcellular localization showed that CpARF22 was localized in the nucleus. The transgenic Arabidopsis thaliana lines with the CpARF22 gene enhanced their tolerance to salt and drought stress. CONCLUSION: In this study, we systematically analyzed the CpARF gene family and its expression patterns under drought, salt, and pathogen stress, which improved our understanding of the ARF protein of zucchini, and laid a solid foundation for functional analysis of the CpARF gene.

Structural and functional characterization of genes PYL-PP2C-SnRK2s in the ABA signalling pathway of Cucurbita pepo.

BACKGROUND: The core regulation of the abscisic acid (ABA) signalling pathway comprises the multigenic families PYL, PP2C, and SnRK2. In this work, we conducted a genome-wide study of the components of these families in Cucurbita pepo. RESULTS: The bioinformatic analysis of the C. pepo genome resulted in the identification of 19 CpPYL, 102 CpPP2C and 10 CpSnRK2 genes. The investigation of gene structure and protein motifs allowed to define 4 PYL, 13 PP2C and 3 SnRK2 subfamilies. RNA-seq analysis was used to determine the expression of these gene families in different plant organs, as well as to detect their differential gene expression during germination, and in response to ABA and cold stress in leaves. The specific tissue expression of some gene members indicated the relevant role of some ABA signalling genes in plant development. Moreover, their differential expression under ABA treatment or cold stress revealed those ABA signalling genes that responded to ABA, and those that were up- or down-regulated in response to cold stress. A reduced number of genes responded to both treatments. Specific PYL-PP2C-SnRK2 genes that had potential roles in germination were also detected, including those regulated early during the imbibition phase, those regulated later during the embryo extension and radicle emergence phase, and those induced or repressed during the whole germination process. CONCLUSIONS: The outcomes of this research open new research lines for agriculture and for assessing gene function in future studies.

As assessment of shelf life increasing competence of pectin (Zucchini) based edible coating on tomatoes.

The most recent advancement in food packaging research involves improving the shelf life of perishable foods by utilising bio-based resources that are edible, eco-friendly, and biodegradable. The current study investigated the effect of edible pectin coating on mature green tomatoes to improve shelf life and storage properties. Zucchini pectin was used to make edible coating. The antimicrobial and antioxidant properties of extracted pectin were investigated. The findings indicated that the extracted pectin had antimicrobial (Staphylococcus aureus, Escherichia coli, and Aspergillus niger) and antioxidant (34.32% at 1 mg/mL) properties.Tomatoes were immersed in pectin solutions of varying concentrations, 1, 3, and 5% (w/v). Physiological evaluations of weight loss, total sugar content, titratable acidity pH, and ascorbic acid were performed on tomatoes during their maturing stages of mature green, light red, pure red, and breaking. Coating the tomatoes with pectin (5%) resulted in minimal weight loss while increasing the retention of total sugar, ascorbic acid, and titratable acidity. The shelf life of the pectin-coated tomatoes was extended to 11 days, while the uncoated control tomatoes lasted 9 days. Thus, a 5% edible pectin solution was found to be effective in coating tomatoes. The current study suggests that using 5% pectin as an edible coating on tomatoes can delay/slow the ripening/maturing process while also extending the shelf-life of tomatoes without affecting their physiochemical properties, which is scalable on a large scale for commercial purposes.

Green synthesis of cerium oxide nanoparticles using zucchini peel extract for cytotoxic and photocatalytic properties.

The aim of this study is the green synthesis of cerium oxide nanoparticles (CeO2-NPs) using a natural capping agent and its application in water and wastewater treatment. This study presents the biosynthesis of CeO2-NPs by the exertion of a green method using zucchini (Cucurbita pepo) extract as a capping agent. Synthesized CeO2-NPs were distinguished through TGA/DTA, FT-IR, XRD, FESEM/TEM and EDX/PSA, and DRS procedures. According to the XRD pattern of NPs, the crystallinity structure was a face-centered cubic (fcc) with an Fm3m space group and the size was estimated at 30 nm. The spherical morphology of NPs was confirmed through FESEM/TEM images. In the following, the photocatalytic property of NPs was investigated by the decolorization of methylene blue (MB) dye within UV-A light. Also, the cytotoxicity of NPs on the CT26 cell line was evaluated through the MTT test, and no toxicity was observed in the results, which indicates their biocompatibility.

The influence of different cooking treatments on vegetables' bolus properties.

The influence of boiling, steaming, grilling and sous-vide treatments on bolus properties of vegetables was investigated. Cooking produced potato boluses with large particles or pasty boluses unsuitable for analysis. Celeriac preserved its brittleness and produced more small particles as mastication prolonged. Eggplant and zucchini were similar and both produced relatively large particles throughout the mastication. Saliva incorporation results showed an uncommon trend since boluses from the moment of swallowing did not have the highest moisture content. It was inferred that boiling had similar effects as steaming on one side, and grilling had similar effects as sous-vide on the other.

First report of zucchini yellow mosaic virus infecting bitter melon (Momordica charantia) in South Korea.

Bitter melon (Momordica charantia L., family Cucurbitaceae) is used in traditional medicine for diabetes, cancer, and inflammation-associated diseases due to bioactive compounds in Asia and tropical Africa (Bortolotti et al. 2019). In July 2021, approximately 10% of bitter melon plants in the field showed symptoms such as mosaic, yellowing, and leaf deformation on the leaves, in Samchcuk, South Korea. Cucumber and zucchini plants growing in the same field exhibited symptoms like those of bitter melon plants (Ali et al. 2012). To investigate the causative virus, leaf dip preparations from three symptomatic bitter melon leaf samples with symptoms were analyzed by transmission electron microscopy (TEM). Potyvirus-like particles (approximately 680-730 nm in length and 11-13 nm in diameter) were observed in all samples. To further identify the causal viral pathogens, leaf extracts from five symptomatic bitter melon plants were tested by DAS-ELISA using specific antibodies (Agdia, Elkhart, IN, USA) against cucumber mosaic virus, zucchini yellow mosaic virus (ZYMV), watermelon mosaic virus, and papaya ring spot virus. Positive controls from commercial kits and negative controls from healthy bitter melon plants were included in ELISA assay. The serological assay revealed that all five symptomatic samples positively reacted with the antiserum against ZYMV, but not for other viruses. Total RNA extracted from the five ELISA-positive samples and two healthy bitter melon plants (as negative controls), using Clear-S Total RNA extraction kit (InVirusTech Co., Gwangju, Korea), was tested by RT-PCR with ZYMV-specific primers as previously described (Cho et al. 2011). All amplicons of the expected size (~822 bp) were individually cloned into the pGEM-T Easy Vector (Promega, Madison, WI), and sequenced in both orientations. Thereafter, all the sequenced clones shared 100% nucleotide identity. The sequence of ZYMV-MC1 isolated from bitter melon was deposited in the GenBank (accession no. LC652434). Pairwise comparison of the nucleotide sequence with that of ZYMV isolates in the GenBank revealed 99% sequence identity with ZYMV-chk (MG020559) from Korea, 98% with ZYMV-14-HY-SCS (KU743321) from China, 97% with ZYMV-Y21 (MW345249) from Turkey, 96% with ZYMV-AUIKTPK (KR261951) from Pakstan. Leaf saps from the ZYMV-positive bitter melon samples, prepared in 10 mM phosphate buffer (pH 7.0), were mechanically inoculated in five young, healthy bitter melon plants to fulfil Koch's postulates. ZYMV-MC1 isolate caused mosaic and leaf deformation on bitter melon plants 10 days post-inoculation. The presence of ZYMV in the symptomatic leaves was confirmed by RT-PCR using the mentioned above primers mentioned above followed by nucleotide sequencing of the amplicons. Several cotton aphids (Aphis gossypii) were observed in the bitter melon field, which indicated that they might transmit the virus from ZYMV-infected cucumber or zucchini plants. ZYMV is one of the economically important viruses of cucurbits worldwide and has been recently reported from various crops as natural hosts, including Chayote (Yoon et al. 2018) and balloon flowers (Kim et al. 2021). To the best of our knowledge, this is the first report of ZYMV naturally infecting bitter melon in South Korea. Further large -scale surveys are required to determine its incidence, yield losses, and management in bitter melon in Korea.

First report of mosaic disease associated with zucchini tigre mosaic virus in wax gourd (Benincasa hispida) in Taiwan.

Wax gourd (Benincasa hispida) is a popular cucurbitaceous summer crop used for culinary purposes, confectionery, and refreshing drinks in Asia. During a field survey for cucurbit viral diseases at Beidou, Changhua in May 2017, B. hispida plants with symptoms of mosaic, chlorosis and leaf curl were found. Because more than 90% of plants showed leaf curl symptoms, samples were first examined by PCR with degenerate primers for begomoviruses (Forward: 5'-TGTGAAGGNCCDTGTAARGT-3' and Reverse: 5'-GCRTDGGTACADGCCATATA-3'). Sequence analysis of amplicons revealed that all of tested plants were infected by squash leaf curl Philippine virus (SqLCPhV) (Liao et al., 2007). However, electron microscopic examination revealed potyvirus-like particles and pinwheel-like inclusion bodies on some of begomovirus-infected leaves showing mild curling and mosaic symptoms. RT-PCR with potyvirusdegenerate primers (PNIbF1: 5'-GGBAAYAATAGTGGNCAACC-3' and PCPR1: 5'-GGGGAGGTGCCGTTCTCDATRCACC-3') were used to detect the potyvirus. Among 8 samples examined, all were infected by SqLCPhV (100%) and 5 samples were mixed infection of SqLCPhV and a potyvirus (62.5%). The filamentous virus was isolated from field samples by sap inoculating plants of B. hispida to separate from nonmechanically-transmissible SqLCPhV. Inoculated B. hispida plants showed severe mosaic and deformation. Total RNA was extracted from inoculated B. hispida and used for RT-PCR with a degenerate potyvirus primer PNIbF1 (5'- GGBAAYAATAGTGGNCAACC) pairing with oligo-dT. A 1.8-kb DNA fragment containing partial NIb gene, full-length coat protein (CP) gene, and 3'-UTR was amplified, cloned, and sequenced. The CP gene (293 aa) shared 82.1% nucleotide and 81% amino acid sequence identity to that of zucchini tigre mosaic virus (ZTMV) isolate E11045 (KC345608). Two degenerate primers (fZTMV-F: 5'- AGCRTGTGGYAHCC-3' and gourd-R: 5'-TCCCACCAYTTYTCRAAHGT-3') target the flanking region between the middle of P3 and 5' of CIP genes of ZTMV strains were designed and used to detect ZTMV. An expected 0.7 kb fragment could be amplified from 2 of 5 field-collected samples and all 6 mechanically inoculated B. hispida plants. Upon sap inoculation, ZTMV was also able to infect pumpkin (Cucurbita pepo var. pepo), zucchini (C. pepo var. cylindrica), squash (C. moschata), bottle gourd (Lagenaria siceraria) and Cucumis metuliferus, but not bitter gourd (Momordica charantia), cucumber (Cucumis sativus), watermelon (Citrullus lanatus), luffa (Luffa cylindrica), tobacco (including Nicotiana benthamiana, N. tabacum, and N. glutinoa), Chenopdium quinoa and C. amaranticolor. The infection of ZTMV to all mechanically inoculated plants were tested by RT-PCR using the fZTMV-F and gourd-R primers and/or TEM examinations. To the best of our knowledge, this is the first report of ZTMV infecting B. hispida in Taiwan. ZTMV was originally reported as a T strain of papaya ringspot virus (PRSV-T) (Quiot-Doine et al., 1986), and was later reclassified as a distinct species in the genus Potyvirus (Romay et al., 2014).

First report of zucchini tigre mosaic virus infecting four cucurbit crops in China.

Cucurbits including ridge gourd (Luffa acutangula), Chieh-qua (Benincasa hispida Cogn. var. Chieh-qua How), Trichosanthes anguina, and sponge gourd (L. cylindrica) are important vegetables in most Asian countries. The 90 viruses known to infect cucurbits include 15 species in genus Potyvirus. In October 2020, nine cucurbit samples with leaf distortion, blister and mottle were collected from the same field of Foshan City, Guangdong Province, China. All samples were tested by western blot with potyvirus-specific antibody (Agdia lnc., Elkhart, IN) and RT-PCR with potyvirus degenerate primers Sprimer/M4T (Chen et al. 2001). Seven out of nine samples were positive for potyvirus in both tests, including one ridge gourd, one Chieh-qua, one sponge gourd, two bottle gourd (Lagenaria siceraria) and two T. anguina. All PCR products (~700-bp) were cloned and sequenced. Sequences of seven amplicons (OM522614 to OM522618, OP090158 to OP090159) containing partial nib and cp genes shared 80.3-100% nucleotide (nt) identity among themselves, and 81.2-97.7% nt identity with ZTMV isolates from China (MN267689, LC371337, MK988416). Except for one Chieh-qua sample, papaya ringspot virus (PRSV) was detected in the same samples where ZTMV was found through sequencing of the amplicons mentioned above. The obtained sequences (OM808942 to OM808945, OP090170 to OP090171) were 95.4-100% identical with PRSV isolates from China. Further RT-PCR was conducted with ZTMV-specific primers ZTMVdF/ZTMVdR targeting partial P3 and 6K1 genes, and PRSV-specific primers PRSV3778F/PRSV4630R targeting partial P3, full-length 6K1 and partial CI genes for all nine samples. Consistently, seven samples were positive for ZTMV, among which one Chieh-qua sample was infected with only ZTMV and six samples were co-infected with ZTMV and PRSV. Interspecific recombination event has been reported for ZTMV (Peng et al. 2021), to detect the recombinants, RT-PCR was conducted for all nine samples with primers ZTMV600F/ZTMV2400R covering the interspecies recombination site (Peng et al. 2021). A fragment (~1.8 kb) was amplified from one T. anguina sample and sequenced (OP090172), which had 97.0% nt identity with the reported recombinant ZTMV-KF17 (MK988415). To fulfill Koch's postulates, a Chieh-qua sample detected with ZTMV but not PRSV, was used for mechanical inoculation on Chieh-qua seedlings. Blister and leaf distortion similar to the field symptoms were observed 21 days post-inoculation. ZTMV infection was verified by RT-PCR with primer pairs Sprimer/M4T and ZTMVdF/ZTMVdR, respectively, followed by sequencing. No amplicon was detected with primer pairs PRSV3778F/PRSV4630R and ZTMV600F/ZTMV2400R. To study the incidence of ZTMV and PRSV, 33 samples including T. anguina, ridge gourd and Chieh-qua were collected from three different fields in Foshan City in May 2022, and were tested by RT-PCR with ZTMV and PRSV primers aforementioned. 30.3% (10/33) of the samples were positive for ZTMV, 39.4% (13/33) tested positive for PRSV, and 21.2% (7/33) were co-infected with the two viruses. Amplicons of ZTMV (600 bp) from all positive samples were sequenced (OP090160 to OP090169), and were 84.8-85.5% identical with ZTMV-TW (LC371337). Recombinant of ZTMV was detected in one T. anguina with primers mentioned above and was sequenced (OP090173), which had 96.2% nt identity with ZTMV-KF17. To our knowledge, this is the first report of ZTMV infecting ridge gourd, Chieh-qua, T. anguina and sponge gourd. The results implied that ZTMV had a potential risk to more cucurbit crops in the field.

Effects of Heat Treatments on Various Characteristics of Ready-to-Eat Zucchini Purees Enriched with Anise or Fennel.

Galactagogue herbs, also known as natural lactation adjuvants, are frequently used to stimulate breast milk production. Due to their antioxidant activity and phenolic content, anise (Pimpinella anisum L.) and fennel (Foeniculum vulgare L.) were chosen to increase the added value of zucchini (Cucurbita pepo L.) purees. At the same time, this work aimed to determine the influence of heat treatment on various characteristics of the final product. The phytochemical content, color parameters, and rheological and textural parameters of zucchini purees enriched with herbal aqueous extracts were determined after processing and after one week of storage (4 °C). In the case of antioxidant activity, samples registered a variation between 6.62 ± 1.71 and 38.32 ± 3.85 µM Trolox/g DW for the samples processed by steam convection. The total difference color parameter (ΔE) increased seven times after one week of storage compared to samples at T0. Fennel and anise aqueous extracts helped improve the rheological behavior of zucchini samples both by steam and hot air convection. This study may serve as a springboard for future investigations and clinical trials into the scientific validity and safety of ready-to-eat foods with special destinations.

An unusual case of food protein-induced enterocolitis syndrome due to zucchini.

BACKGROUND: Food protein-induced enterocolitis syndrome (FPIES) is a non-IgE-mediated food allergy characterized by gastrointestinal symptoms, mainly protracted and delayed vomiting. Diagnosis is based on clinical history, and it can be challenging as symptoms are delayed and the causative food is often not very suspicious. OBJECTIVE: This case report highlights the importance of having a high degree of suspicion to reach a correct diagnosis. MATERIALS AND METHODS: We report an unusual case of FPIES due to zucchini. During the follow-up. Two oral food challenges (OFC) were carried out to evaluate tolerance to the food involved. RESULTS: The first OFC was positive and in the second the child tolerated the food without problems. CONCLUSIONS: In this case, the OFC was essential to identify the offending food and to verify that the child had overcome the disease.

First RNA-seq approach to study fruit set and parthenocarpy in zucchini (Cucurbita pepo L.).

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.

Kc167, a widely used Drosophila cell line, contains an active primary piRNA pathway.

PIWI family proteins bind to small RNAs known as PIWI-interacting RNAs (piRNAs) and play essential roles in the germline by silencing transposons and by promoting germ cell specification and function. Here we report that the widely used Kc167 cell line, derived from Drosophila melanogaster embryos, expresses piRNAs that are loaded to Aub and Piwi. Kc167 piRNAs are produced by a canonical, primary piRNA biogenesis pathway, from phased processing of precursor transcripts by the Zuc endonuclease, Armi helicase, and dGasz mitochondrial scaffold protein. Kc167 piRNAs derive from cytoplasmic transcripts, notably tRNAs and mRNAs, and their abundance correlates with that of parent transcripts. The expression of Aub is robust in Kc167, that of Piwi is modest, while Ago3 is undetectable, explaining the lack of transposon-related piRNA amplification by the Aub-Ago3, ping-pong mechanism. We propose that the default state of the primary piRNA biogenesis machinery is random transcript sampling to allow generation of piRNAs from any transcript, including newly acquired retrotransposons. This state is unmasked in Kc167, likely because they do not express piRNA cluster transcripts in sufficient amounts and do not amplify transposon piRNAs. We use Kc167 to characterize an inactive isoform of Aub protein. Since most Kc167 piRNAs are genic, they can be mapped uniquely to the genome, facilitating computational analyses. Furthermore, because Kc167 is a widely used and well-characterized cell line that is easily amenable to experimental manipulations, we expect that it will serve as an excellent system to study piRNA biogenesis and piRNA-related factors.

Transcriptomic changes in Cucurbita pepo fruit after cold storage: differential response between two cultivars contrasting in chilling sensitivity.

BACKGROUND: Zucchini fruit is susceptible to chilling injury (CI), but the response to low storage temperature is cultivar dependent. Previous reports about the response of zucchini fruit to chilling storage have been focused on the physiology and biochemistry of this process, with little information about the molecular mechanisms underlying it. In this work, we present a comprehensive analysis of transcriptomic changes that take place after cold storage in zucchini fruit of two commercial cultivars with contrasting response to chilling stress. RESULTS: RNA-Seq analysis was conducted in exocarp of fruit at harvest and after 14 days of storage at 4 and 20 °C. Differential expressed genes (DEGs) were obtained comparing fruit stored at 4 °C with their control at 20 °C, and then specific and common up and down-regulated DEGs of each cultivar were identified. Functional analysis of these DEGs identified similarities between the response of zucchini fruit to low temperature and other stresses, with an important number of GO terms related to biotic and abiotic stresses overrepresented in both cultivars. This study also revealed several molecular mechanisms that could be related to chilling tolerance, since they were up-regulated in cv. Natura (CI tolerant) or down-regulated in cv. Sinatra (CI sensitive). These mechanisms were mainly those related to carbohydrate and energy metabolism, transcription, signal transduction, and protein transport and degradation. Among DEGs belonging to these pathways, we selected candidate genes that could regulate or promote chilling tolerance in zucchini fruit including the transcription factors MYB76-like, ZAT10-like, DELLA protein GAIP, and AP2/ERF domain-containing protein. CONCLUSIONS: This study provides a broader understanding of the important mechanisms and processes related to coping with low temperature stress in zucchini fruit and allowed the identification of some candidate genes that may be involved in the acquisition of chilling tolerance in this crop. These genes will be the basis of future studies aimed to identify markers involved in cold tolerance and aid in zucchini breeding programs.

De novo assembly of the zucchini genome reveals a whole-genome duplication associated with the origin of the Cucurbita genus.

The Cucurbita genus (squashes, pumpkins and gourds) includes important domesticated species such as C. pepo, C. maxima and C. moschata. In this study, we present a high-quality draft of the zucchini (C. pepo) genome. The assembly has a size of 263 Mb, a scaffold N50 of 1.8 Mb and 34 240 gene models. It includes 92% of the conserved BUSCO core gene set, and it is estimated to cover 93.0% of the genome. The genome is organized in 20 pseudomolecules that represent 81.4% of the assembly, and it is integrated with a genetic map of 7718 SNPs. Despite the small genome size, three independent lines of evidence support that the C. pepo genome is the result of a whole-genome duplication: the topology of the gene family phylogenies, the karyotype organization and the distribution of 4DTv distances. Additionally, 40 transcriptomes of 12 species of the genus were assembled and analysed together with all the other published genomes of the Cucurbitaceae family. The duplication was detected in all the Cucurbita species analysed, including C. maxima and C. moschata, but not in the more distant cucurbits belonging to the Cucumis and Citrullus genera, and it is likely to have occurred 30 ± 4 Mya in the ancestral species that gave rise to the genus.

DDTs-induced antioxidant responses in plants and their influence on phytoremediation process.

Phytoremediation is a low cost technology based on the use of plants to remove a wide range of pollutants from the environment, including the insecticide DDT. However, some pollutants are known to enhance generation of reactive oxygen species (ROS), which can generate toxic effects on plants affecting the phytoremediation efficiency. This study aims to analyze the potential use of antioxidant responses as a measure of tolerance to select plants for phytoremediation purposes. Tomato and zucchini plants were grown for 15 days in soils contaminated with DDTs (DDT + DDE + DDD). Protein content, glutathione-S-transferase (GST), glutathione reductase (GR), glutathione peroxidase (GPx) and catalase (CAT) activities were measured in plant tissues. Exposure to DDTs did not affect protein content or CAT activity in any of the species. GST, GR and GPx activity showed different responses in exposed and control tomato plants. After DDTs exposure, tomato showed increased GR and GPX activity in stems and leaves, respectively, and a decrease in the GST activity in roots. As no effects were observed in zucchini, results suggest different susceptibility and/or defense mechanisms involved after pesticide exposure. Finally, both species differed also in terms of DDTs uptake and translocation. The knowledge about antioxidant responses induced by pesticides exposure could be helpful for planning phytoremediation strategies and for the selection of tolerant species according to particular scenarios.

Molecular characterization of mitochondrial Zucchini and its relation to nuage-piRNA pathway components in Bombyx mori ovary-derived BmN4 cells.

Piwi-interacting RNAs (piRNAs) are a class of small non-coding RNAs that associate with PIWI subfamily proteins, which play an important role in transposon silencing in animal germ cell. The piRNAs biogenesis is divided into two major pathways: primary and secondary, and both pathways are independent of double-stranded RNA-processing enzyme Dicer, which processes the single-stranded RNA transcripts in microRNA (miRNA) and siRNA (small interfering RNA) pathway. Primary piRNAs are processed from long non-coding RNA precursors transcribed from piRNA clusters. Zucchini (Zuc), a mitochondrial phospholipase D (PLD) superfamily protein is conserved among the animals and involved in piRNA biogenesis. Recent studies showed that the Zucchini is an endoribonuclease essential for primary piRNA maturation and production of phased piRNA in secondary piRNA biogenesis of drosophila germ cell. Based on these reports, here we identified and studied the silkworm Zucchini (BmZuc) at subcellular level in ovary-derived BmN4 cell. The silkworm Zuc specifically expressed in germ-related tissues and localized on mitochondria and partially co-localized with perinuclear nuage-piRNA pathway components and nuage marker protein BmVasa. Molecular dissection analyses revealed that the conserved mitochondrial localization sequence, RGV motif, PLDc 2 domain and HKD motif are important for the BmZuc mitochondrial localization. Moreover, the knockdown analyses showed that the piRNA pathway components are independent on BmZuc for their nuage localization, whereas BmZuc depend on piRNA pathway components for the proper localization. Our data provides vital information on mitochondrial BmZuc and its relationship to "nuage" in ovary-derived BmN4 cell.

The Influence of the Osmotic Dehydration Process on Physicochemical Properties of Osmotic Solution.

The osmotic dehydration (OD) process consists of the removal of water from a material during which the solids from the osmotic solution are transported to the material by osmosis. This process is commonly performed in sucrose and salt solutions. Taking into account that a relatively high consumption of those substances might have a negative effect on human health, attempts have been made to search for alternatives that can be used for osmotic dehydration. One of these is an application of chokeberry juice with proven beneficial properties to human health. This study aimed to evaluate the physicochemical properties of the OD solution (chokeberry juice concentrate) before and after the osmotic dehydration of carrot and zucchini. The total polyphenolics content, antioxidant capacity (ABTS, FRAP), dynamic viscosity, density, and water activity were examined in relation to the juice concentration used for the osmotic solution before and after the OD process. During the osmotic dehydration process, the concentration of the chokeberry juice decreased. Compounds with lower molecular weight and lower antioxidant capacity present in concentrated chokeberry juice had a stronger influence on the exchange of compounds during the OD process in carrot and zucchini. The water activity of the osmotic solution increased after the osmotic dehydration process. It was concluded that the osmotic solution after the OD process might be successfully re-used as a product with high quality for i.e. juice production.

The complete genome of the tospovirus Zucchini lethal chlorosis virus.

BACKGROUND: Zucchini lethal chlorosis virus (ZLCV) causes significant losses in the production of cucurbits in Brazil. This virus belongs to the genus Tospovirus (family Bunyaviridae) and seems to be exclusively transmitted by Frankliniella zucchini (Thysanoptera). Tospoviruses have a tripartite and single-stranded RNA genome classified as S (Small), M (Medium) and L (Large) RNAS. Although ZLCV was identified as a member of the genus Tospovirus in 1999, its complete genome had not been sequenced until now. FINDINGS: We sequenced the full-length genome of two ZLCV isolates named ZLCV-SP and ZLCV-DF. The phylogenetic analysis showed that ZLCV-SP and ZLCV-DF clustered with the previously reported isolate ZLCV-BR09. Their proteins were closely related, except the non-structural protein (NSm), which was highly divergent (approximately 90 % identity). All viral proteins clustered similarly in our phylogenetic analysis, excluding that these ZLCV isolates have originated from reassortment events of different tospovirus species. CONCLUSION: Here we report for the first time the complete genome of two ZLCV isolates that were found in the field infecting zucchini and cucumber.