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Background Streptococcus pneumoniae is the leading reason for invasive diseases including pneumonia and meningitis, and also secondary infections following viral respiratory diseases such as flu and COVID-19. Currently, serotype-dependent vaccines, which have several insufficiency and limitations, are the only way to prevent pneumococcal infections. Hence, it is plain to need an alternative effective strategy for prevention of this organism. Protein-based vaccine involving conserved pneumococcal protein antigens with different roles in virulence could provide an eligible alternative to existing vaccines. Methods In this study, PspC, PhtD and PsaA antigens from pneumococcus were taken to account to predict B-cell and helper T-cell epitopes, and epitope-rich regions were chosen to build the construct. To enhance the immunogenicity of the epitope-based vaccine, a truncated N-terminal fragment of pneumococcal endopeptidase O (PepO) was used as a potential TLR2/4 agonist which was identified by molecular docking studies. The ultimate construct was consisted of the chosen epitope-rich regions, along with the adjuvant role (truncated N-PepO) and suitable linkers. Results The epitope-based vaccine was assessed as regards physicochemical properties, allergenicity, antigenicity, and toxicity. The 3D structure of the engineered construct was modeled, refined, and validated. Molecular docking and simulation of molecular dynamics (MD) indicated the proper and stable interactions between the vaccine and TLR2/4 throughout the simulation periods. Conclusions For the first time this work presents a novel vaccine consisting of epitopes of PspC, PhtD, and PsaA antigens which is adjuvanted with a new truncated domain of PepO. The computational outcomes revealed that the suggested vaccine could be deemed an efficient therapeutic vaccine for S. pneumoniae ; nevertheless , in vitro and in vivo examinations should be performed to prove the potency of the candidate vaccine.

The oligopeptidase PepO from Streptococcus thermophilus A was purified to protein homogeneity by a five-step chromatography procedure. It was estimated to be a serine metallopeptidase of 70 kDa, with maximal activity at pH 6.5 and 41°C. PepO has endopeptidase activity on oligopeptides composed of between five and 30 amino acids. PepO was demonstrated to be active and stable at the pH, temperature and salt concentrations found in Swiss-type cheese during ripening. Using a battery of PCR techniques, the pepO gene was amplified, subcloned and sequenced, revealing an open reading frame of 1893 nucleotides. The amino acid sequence analysis of the pepO gene-translation product shows homology with PepO enzymes from other lactic acid bacteria and contains the signature sequence of the metallopeptidase family.

The oligopeptidase PepO from Streptococcus thermophilus A was purified to protein homogeneity by a five-step chromatography procedure. It was estimated to be a serine metallopeptidase of 70 kDa, with maximal activity at pH 6.5 and 41 degrees C. PepO has endopeptidase activity on oligopeptides composed of between five and 30 amino acids. PepO was demonstrated to be active and stable at the pH, temperature and salt concentrations found in Swiss-type cheese during ripening. Using a battery of PCR techniques, the pepO gene was amplified, subcloned and sequenced, revealing an open reading frame of 1893 nucleotides. The amino acid sequence analysis of the pepO gene-translation product shows homology with PepO enzymes from other lactic acid bacteria and contains the signature sequence of the metallopeptidase family.

Fruit peel color is a major factor that influences fruit quality and customers’ demand. However, the molecular mechanisms underlying the green fruit peel color trait of Cucurbita pepo L. remain unknown. Two parental lines, RP16 and RP38, were used to study the fruit peel color trait in C. pepo . The parental line RP16 shows white peel color, whereas RP38 exhibits green peel color. 384 F 2 populations were used to identify the inheritance pattern associated with green fruit and white fruit peel in Cucurbita pepo L. 293 F 2 individuals were white, and 91 F 2 individuals were green, resulting in a ratio of 3:1. Hence, white peel is dominant over the green fruit peel trait, and a single recessive green peel gene ( Cpgp ) controls the green fruit peel. The fruit chlorophyll (Chll) content decreases as fruit matures in the RP16 line. In contrast, Chll increases during the fruit growing periods on fruit peels of the RP38 line. The BSA-sequence analysis revealed the Cpgp locus on Chr5, within a 2.3 Mb region. Subsequent fine-mapping analysis, using 699 F 2 plants, narrowed down this region to 23.90 kb on the same chromosome. Within this region, two annotated genes, namely Cp4.1LG05g02070 and Cp4.1LG05g02060 , are present. These genes are predicted to encode a two-component Arabidopsis Pseudo-Response Regulator 2-like protein (APRR2), which may be involved in green pigmentation processes in plants. Consequently, sequence alignment and gene expression analyses at various fruit development stages supported that Cp4.1LG05g02070 may be the primary candidate gene responsible for regulating the green fruit peel color trait in Cucurbita pepo L. This study may provide a basis for further study on the basic mechanisms that control the fruit peel colors in Cucurbita spp .

Fruit shape index (round shape) and seed size have important effects on pumpkin yield. To investigate these traits and to create the most desirable state, a cross was made between two pumpkin cultivars. This study aimed to estimate the main gene effects (additive, dominant and di-genic epistasis) and to determine the mode of inheritance for fruit shape and seed size by generation mean analysis. Six generations, namely P1, P2, F1, F2, BC1, and BC2 from a cross between Zucchini and hull-less seed Pumpkin, S10 × P25, were constructed and evaluated for fruit length, fruit width, fruit shape index, and some seed-related traits (seed length, seed width, and seed thickness). The experiment was conducted in the research field of the Faculty of Agricultural Science, University of Guilan, Rasht, Iran in 2019. Results showed a significant difference between generations in terms of fruit and seed traits. Scale and joint scale tests showed the presence of epistasis for some traits. According to the results of the average traits of different generations, standard heterosis and hetrobeltiosis were observed. Concerning the fruit shape index and seed width, there was a dominance effect. The broad-sense heritability of the traits was relatively high for all traits and between 52 and 94%. Narrow sense heritability was between 26 and 90% and relatively low for the fruit shape index and seed width. Therefore, selection of elite lines and production of their hybrids are recommended as two methods suitable for breeding to achieve a round shape index and a larger seed size.

A Palmer amaranth biotype (CT-Res) with resistance to glyphosate was recently confirmed in a pumpkin field in Connecticut. However, the underlying mechanisms conferring glyphosate resistance in this biotype is not known. The main objectives of this research were 1) to determine the effect of plant height (10, 20, and 30 cm) on glyphosate resistance levels in CT-Res Palmer amaranth biotype, and 2) to investigate whether the target site–based mechanisms confer glyphosate resistance. To achieve these objectives, progeny seeds of the CT-Res biotype after two generations of recurrent selection with glyphosate (6,720 g ae ha–1) were used. Similarly, known glyphosate-susceptible Palmer amaranth biotypes from Kansas (KS-Sus) and Alabama (AL-Sus) were included. Results from greenhouse dose-response studies revealed that CT-Res Palmer amaranth biotype had 69-, 64-, and 54-fold resistance to glyphosate as compared with the KS-Sus biotype when treated at heights of 10, 20, and 30 cm, respectively. Sequence analysis of the EPSPS gene revealed no point mutations at the Pro106 and Thr102 residues in the CT-Res Palmer amaranth biotype. Quantitative polymerase chain reaction analysis revealed that the CT-Res biotype had 33 to 111 relative copies of the EPSPS gene compared with the AL-Sus biotype. All these results suggest that the EPSPS gene amplification endows a high level of glyphosate resistance in the GR Palmer amaranth biotype from Connecticut. Because of the lack of control with glyphosate, growers should adopt the use of effective alternative preemergence and postemergence herbicides in conjunction with other cultural and mechanical tactics to mitigate the further spread of GR Palmer amaranth in Connecticut. Nomenclature: Glyphosate; Palmer amaranth, Amaranthus palmeri S. Watson; pumpkin (Cucurbita pepo L.)

VQ protein is a plant specific protein, which plays an important role in plant growth and development and biological and abiotic stress response. This study aimed to systematically analyze for the first time the VQ of Cucurbita pepo and understand their expression patterns in response to different stimuli. Herein, 44 VQ genes were identified, which were divided into eight groups (I–VIII) based on phylogenetic analysis. Two genes ( CpVQ1 and CpVQ2 ) could not be located on the chromosome, whereas the remaining CpVQ genes were randomly distributed on the chromosomes, except for chromosomes 15 and 18. Noteworthy, the main event driving the expansion of the VQ gene family was chromosome fragment duplication. Based on qRT-PCR analysis, VQ genes are expressed in different tissues, and VQ genes are differentially regulated under a variety of abiotic stresses and powdery mildew stress, indicating that they play an important role in plant stress response and other aspects. This report presents the first systematic analysis of VQ genes from C. pepo and provides a solid foundation for further research of the specific functions of VQ proteins.

Background Cucurbita pepo L. cultivars display high morphological traits variation. In addition, C . pepo faces numerous threats, such as viral and fungal infections, which significantly influence crop cultivation. Recent genomic advancements improved the understanding of genetic diversity and stress responses in this crop. We investigated genetic variations related to plant morphology and quality traits. Additionally, the inclusion of both powdery mildew (PM) and Zucchini yellow mosaic virus (ZYMV) susceptible and tolerant varieties facilitated the examination of genetic diversity concerning biotic stress. Results The sequencing of eight Cucurbita pepo varieties produced an average of 40 million raw reads with a coverage of reference genome ranging from 22 to 40X. More than 4.7 million genomic variants were identified in all genomes. Based on admixture and PCA analysis, the eight C. pepo genotypes were grouped in two clusters belonging to Cocozelle and Zucchini groups, with “Whitaker” separated from the rest of the accessions. Genes involved in pathways related to gibberellin regulation, leaf development, and pigment accumulation resulted highly affected by variation suggesting that the diversity observed among varieties in plant and fruit morphology could be related to variants identified in such genes. Each variety showed its own set of genetic differences. The genomic comparison of 381e, 968Rb and SPQ allowed the identification of variants in chromosome regions affecting response to Zucchini yellow mosaic virus (ZYMV) and powdery mildew (PM). Variants in key genes associated with resistant traits were identified, suggesting potential pathways and mechanisms involved in biotic stress response and plant immunity. Conclusions Genetic variations affecting morphology and fruit quality in C. pepo emphasize their significance for breeding efforts. Furthermore, the genomic comparison of 381e, 968Rb and SPQ highlighted variants in chromosomal regions influencing zucchini’s response to PM and ZYMV. These findings could pave the way for more targeted and effective genetic improvement strategies, thereby potentially leading to increased agricultural productivity and quality.

Background and aims Plant breeding activities shape the rhizosphere microbiome but less is known about the relationship of both with the seed microbiome. We analyzed the composition of bacterial communities of seeds and rhizospheres of Styrian oil pumpkin genotypes in comparison to bulk soil to elucidate specific microbial signatures to support a concept involving plant-microbe interactions in breeding strategies. Methods The seed and rhizosphere microbiomes of 14 genotypes of oilseed pumpkin and relatives were analyzed using a 16S rRNA gene amplicon sequencing approach, which was assessed by bioinformatics and statistical methods. Results All analyzed microhabitats were characterized by diverse bacterial communities, but the relative proportions of phyla and the overall diversity was different. Seed microbiomes were characterized by the lowest diversity and dominant members of Enterobacteriaceae including potential pathogens (Erwinia, Pectobacterium). Potential plant-beneficial bacteria like Lysobacter, Paenibacillus and Lactococcus contributed to the microbial communities in significant abundances. Interestingly, strong genotype-specific microbiomes were detected for seeds but not for the rhizospheres. Conclusions Our study indicates a strong impact of the Cucurbita pepo genotype on the composition of the seed microbiome. This should be considered in breeding of new cultivars that are more capable of exploiting beneficial indigenous microbial communities.

Background Floral scents play a crucial role in attracting insect pollinators. Among the compounds attractive to pollinators is 1,4-dimethoxybenzene (1,4-DMB). It is a significant contributor to the scent profile of plants from various genera, including economically important Cucurbita species. Despite its importance, the biosynthetic pathway for the formation of 1,4-DMB was not elucidated so far. Results In this study we showed the catalysis of 1,4-DMB in the presence of 4-methoxyphenol (4-MP) by protein extract from Styrian oil pumpkin ( Cucurbita pepo ) flowers. Based on this finding, we identified a novel O -methyltransferase gene, Cp4MP-OMT , whose expression is highly upregulated in the volatile-producing tissue of pumpkin flowers when compared to vegetative tissues. OMT activity was verified by purified recombinant Cp4MP-OMT, illustrating its ability to catalyse the methylation of 4-MP to 1,4-DMB in the presence of cofactor SAM ( S -(5′-adenosyl)- L -methionine). Conclusions Cp4MP-OMT is a novel O -methyltransferase from C. pepo , responsible for the final step in the biosynthesis of the floral scent compound 1,4-DMB. Considering the significance of 1,4-DMB in attracting insects for pollination and in the further course fruit formation, enhanced understanding of its biosynthetic pathways holds great promise for both ecological insights and advancements in plant breeding initiatives.