Publications by authors named "Fabio Palumbo"

87 Publications

Molecular Hallmarks, Agronomic Performances and Seed Nutraceutical Properties to Exploit Neglected Genetic Resources of Common Beans Grown by Organic Farming in Two Contrasting Environments.

Front Plant Sci 2021 25;12:674985. Epub 2021 May 25.

Department of Crop and Soil Sciences, University of Georgia, Athens, GA, United States.

Common bean ( L.) is an essential source of food proteins and an important component of sustainable agriculture systems around the world. Thus, conserving and exploiting the genetic materials of this crop species play an important role in achieving global food safety and security through the preservation of functional and serependic opportunities afforded by plant species diversity. Our research aimed to collect and perform agronomic, morpho-phenological, molecular-genetic, and nutraceutical characterizations of common bean accessions, including lowland and mountain Venetian niche landraces (ancient farmer populations) and Italian elite lineages (old breeder selections). Molecular characterization with SSR and SNP markers grouped these accessions into two well-separated clusters that were linked to the original Andean and Mesoamerican gene pools, which was consistent with the outputs of ancestral analysis. Genetic diversity in the two main clusters was not distributed equally the Andean gene pool was found to be much more uniform than the Mesoamerican pool. Additional subdivision resulted in subclusters, supporting the existence of six varietal groups. Accessions were selected according to preliminary investigations and historical records and cultivated in two contrasting Venetian environments: sea-level and mountain territories. We found that the environment significantly affected some nutraceutical properties of the seeds, mainly protein and starch contents. The antioxidant capacity was found significantly greater at sea level for climbing accessions and in the mountains for dwarf accessions. The seed yield at sea level was halved than mountain due to a seeds reduction in weight, volume, size and density. At sea level, bean landraces tended to have extended flowering periods and shorter fresh pod periods. The seed yield was positively correlated with the length of the period during which plants had fresh pods and negatively correlated with the length of the flowering period. Thus, the agronomic performance of these genetic resources showed their strong connection and adaptation to mountainous environments. On the whole, the genetic-molecular information put together for these univocal bean entries was combined with overall results from plant and seed analyses to select and transform the best accessions into commercial varieties (, pure lines) suitable for wider cultivation.
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http://dx.doi.org/10.3389/fpls.2021.674985DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8185351PMC
May 2021

Physicochemical and Rheological Characterization of Different Low Molecular Weight Gellan Gum Products and Derived Ionotropic Crosslinked Hydrogels.

Gels 2021 May 26;7(2). Epub 2021 May 26.

Department of "Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche" (STEBICEF), University of Palermo, Via Archirafi, 32, 90123 Palermo, Italy.

A series of four different low molecular weight gellan gum products was obtained by alkaline hydrolysis with the aim to investigate the impact of the molecular weight on the rheological properties of the polysaccharide aqueous dispersions and on the physicochemical characteristics of derived ionotropic crosslinked hydrogels. In particular, thermo-rheological analysis was conducted on aqueous dispersions to study the influence of molecular weight on the thermogelation properties typical of the native polysaccharide while strain sweep experiments were conducted to establish if aqueous dispersion shows a viscoelastic behavior. The effect of different Ca on the rheological properties of hydrogels were studied. Furthermore, ionotropic crosslinked hydrogels were analyzed in terms of morphology on the dried state and swelling behavior, while their viscoelastic properties were studied by means of rheological analysis conducted in frequency sweep regime after different time points of incubation in phosphate buffer at pH 7.4. Release experiments conducted using fluorescein isothiocyanate labelled dextran as a model diffusion agent and was performed to investigate the possibility of using the low molecular weight GG-derived hydrogels as an active molecule-releasing device. Finally, the cytocompatibility of hydrolysis products was investigated, as well as the capacity of hydrogels to encapsulate viable MC3T3-E1 preosteoblastic cells.
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http://dx.doi.org/10.3390/gels7020062DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8162344PMC
May 2021

A Review of Unreduced Gametes and Neopolyploids in Alfalfa: How to Fill the Gap between Well-Established Meiotic Mutants and Next-Generation Genomic Resources.

Plants (Basel) 2021 May 17;10(5). Epub 2021 May 17.

Department of Agronomy Food Natural Resources Animals Environment, Campus of Agripolis, University of Padova, 35020 Padova, Italy.

The gene flow mediated by unreduced gametes between diploid and tetraploid plants of the complex is pivotal for alfalfa breeding. Sexually tetraploidized hybrids could represent the best way to exploit progressive heterosis simultaneously derived from gene diversity, heterozygosity, and polyploidy. Moreover, unreduced gametes combined with parthenogenesis (i.e., apomixis) would enable the cloning of plants through seeds, providing a unique opportunity for the selection of superior genotypes with permanently fixed heterosis. This reproductive strategy has never been detected in the genus , but features of apomixis, such as restitutional apomeiosis and haploid parthenogenesis, have been reported. By means of an original case study, we demonstrated that sexually tetraploidized plants maintain apomeiosis, but this trait is developmentally independent from parthenogenesis. Alfalfa meiotic mutants producing unreduced egg cells revealed a null or very low capacity for parthenogenesis. The overall achievements reached so far are reviewed and discussed along with the efforts and strategies made for exploiting reproductive mutants that express apomictic elements in alfalfa breeding programs. Although several studies have investigated the cytological mechanisms responsible for 2 gamete formation and the inheritance of this trait, only a very small number of molecular markers and candidate genes putatively linked to unreduced gamete formation have been identified. Furthermore, this scenario has remained almost unchanged over the last two decades. Here, we propose a reverse genetics approach, by exploiting the genomic and transcriptomic resources available in alfalfa. Through a comparison with 9 proteins belonging to   known for their involvement in 2 gamete production, we identified 47 orthologous genes and evaluated their expression in several tissues, paving the way for novel candidate gene characterization studies. An overall view on strategies suitable to fill the gap between well-established meiotic mutants and next-generation genomic resources is presented and discussed.
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http://dx.doi.org/10.3390/plants10050999DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8156078PMC
May 2021

An asymmetric electrospun membrane for the controlled release of ciprofloxacin and FGF-2: Evaluation of antimicrobial and chemoattractant properties.

Mater Sci Eng C Mater Biol Appl 2021 Apr 27;123:112001. Epub 2021 Feb 27.

Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Via Archirafi 32, 90123 Palermo, Italy; Institute of Biophysics at Palermo, Italian National Research Council, Via Ugo La Malfa 153, 90146 Palermo, Italy.

Here, an asymmetric double-layer membrane has been designed and fabricated by electrospinning as a tool for a potential wound healing application. A hydrophobic layer has been produced by using a polyurethane-polycaprolactone (PU-PCL) copolymer and loaded with the antibacterial ciprofloxacin whereas an ion responsive hydrophilic layer has been produced by using an octyl derivative of gellan gum (GG-C) and polyvinyl alcohol (PVA) and loaded with the growth factor FGF-2. This study investigated how the properties of this asymmetric membrane loaded with actives, were influenced by the ionotropic crosslinking of the hydrophilic layer. In particular, the treatment in DPBS and the crosslinking in CaCl 0.1 or 1 M of the hydrophilic layer affected the release profile of the bioactive molecules allowing to modulate both the antimicrobial effect, as assayed by logarithmic reduction of the Staphylococcus aureus viable count, and the chemoattractant properties on NIH 3 T3 cell line, as assayed by scratch test and cell chemoattraction assay.
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http://dx.doi.org/10.1016/j.msec.2021.112001DOI Listing
April 2021

A multi-kingdom metabarcoding study on cattle grazing Alpine pastures discloses intra-seasonal shifts in plant selection and faecal microbiota.

Sci Rep 2021 Jan 13;11(1):889. Epub 2021 Jan 13.

Department of Agronomy Food Natural Resources Animals and Environment (DAFNAE), University of Padova, Campus of Agripolis, Viale dell'Università 16, 35020, Legnaro, Padova, Italy.

Diet selection by grazing livestock may affect animal performance as well as the biodiversity of grazed areas. Recent DNA barcoding techniques allow to assess dietary plant composition in faecal samples, which may be additionally integrated by the description of gut microbiota. In this high throughput metabarcoding study, we investigated the diversity of plant, fungal and bacterial taxa in faecal samples of lactating cows of two breeds grazing an Alpine semi-natural grassland during summer. The estimated plant composition of the diet comprised 67 genera and 39 species, which varied remarkably during summer, suggesting a decline of the diet forage value with the advancing of the vegetative season. The fungal community included Neocallimastigomycota gut symbionts, but also Ascomycota and Basidiomycota plant parasite and coprophilous taxa, likely ingested during grazing. The proportion of ingested fungi was remarkably higher than in other studies, and varied during summer, although less than that observed for plants. Some variation related to breed was also detected. The gut bacterial taxa remained stable through the summer but displayed a breed-specific composition. The study provided insights in the reciprocal organisms' interactions affecting, and being affected by, the foraging behaviour: plants showed a high temporal variation, fungi a smaller one, while bacteria had practically none; conversely, the same kingdoms showed the opposite gradient of variation as respect to the animal host breed, as bacteria revealed to be the group mostly characterized by host-specificity.
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http://dx.doi.org/10.1038/s41598-020-79474-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7806629PMC
January 2021

Synthesis of antibacterial composite coating containing nanocapsules in an atmospheric pressure plasma.

Mater Sci Eng C Mater Biol Appl 2021 Feb 9;119:111496. Epub 2020 Sep 9.

Department of Applied Physics, Ghent University, Sint-Pietersnieuwstraat 41 B4, 9000 Ghent, Belgium.

Antibacterial coating is an important strategy preventing bacterial colonization and biofilm formation. One-step synthesis of nanocapsule-containing antibacterial coatings with controlled release of Ag ions was achieved in the current work by aerosol-assisted atmospheric pressure plasma deposition. The experimental parameters of deposition including the discharge power, silver nitrate concentration, aerosol flow rate, continuous and pulsed mode of operation were studied in order to analyze their effects on surface morphology and chemical composition of the coating. Formation of nanocapsules embedded in the polymeric coating was observed. A core-shell structure was found for nanocapsule with silver in the core and polymer in the shell. Antibacterial coatings on polyethylene terephthalate film were studied in terms of Ag ion release, antibacterial properties against Escherichia coli and Staphylococcus aureus, and cytotoxicity with murine fibroblasts. Two-phase release kinetics of Ag ions was observed as initially a short-term burst release followed by a long-term slow release. It was revealed that high antibacterial efficiency of the coatings deposited on polyethylene terephthalate films can be coupled with low cytotoxicity. These biocompatible antibacterial coatings are very promising in different fields including biological applications.
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http://dx.doi.org/10.1016/j.msec.2020.111496DOI Listing
February 2021

The Gene Family in : Genome-Wide Identification and Expression Analysis under Drought and Salinity Stresses.

Genes (Basel) 2020 12 7;11(12). Epub 2020 Dec 7.

Faculty of Agriculture, Shahrood University of Technology, Shahrood 3619995161, Iran.

Members of the / transcription factor family play critical roles in plant development, biosynthesis of key metabolites, and stress response. A detailed study was performed to identify ss in the durum wheat ( ssp. ) genome, which resulted in the identification of 271 genes distributed on chromosomes 1A-7B. By carrying 27 genes, chromosome 6A had the highest number of ss. Furthermore, a duplication assay of ss demonstrated that 70 duplicated gene pairs had undergone purifying selection. According to RNA-seq analysis, the highest expression levels in all tissues and in response to stimuli were associated with and subfamily genes. In addition, the results revealed that genes have tissue-specific expression patterns, and most genes were significantly induced in the root tissue. Additionally, 13 genes (six s, three s, two s, one , and one ) were selected for further analysis via qRT-PCR of their potential in coping with drought and salinity stresses. The genes belonging to the DREB subfamily were markedly induced under both drought-stress and salinity-stress conditions. Furthermore, docking simulations revealed several residues in the pocket sites of the proteins associated with the stress response, which may be useful in future site-directed mutagenesis studies to increase the stress tolerance of durum wheat. This study could provide valuable insights for further evolutionary and functional assays of this important gene family in durum wheat.
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http://dx.doi.org/10.3390/genes11121464DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7762271PMC
December 2020

Hyaluronan alkyl derivatives-based electrospun membranes for potential guided bone regeneration: Fabrication, characterization and in vitro osteoinductive properties.

Colloids Surf B Biointerfaces 2021 Jan 4;197:111438. Epub 2020 Nov 4.

Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Via Archirafi 32, 90123, Palermo, Italy; Institute of Biophysics at Palermo, Italian National Research Council, Via Ugo La Malfa 153, 90146, Palermo, Italy.

The aim of the work was to determine the effects of the chemical functionalization of hyaluronic acid (HA) with pendant aliphatic tails at different lengths and free amino groups in terms of chemical reactivity, degradation rate, drug-eluting features, and surface properties when processed as electrospun membranes (EM) evaluating the osteoinductive potential for a possible application as guided bone regeneration (GBR). To this end, a series of HA derivatives with different aliphatic tails (DD-Cx mol% ≈ 12.0 mol%) and decreasing derivatization of free amino groups (DD mol% from 70.0 to 30.0 mol%) were first synthesized, namely Hn. Then dexamethasone-loaded Hn EM, i.e. HnX were prepared from aqueous polymeric solutions with polyvinyl alcohol (PVA), as a non-ionogenic linear flexible polymeric carrier, and the multifunctional 2-hydroxypropyl- cyclodextrin (HPCD) which acted as a rheological modifier, a stabilizer of Taylor's cone, and a solubilizing agent. A comprehensive characterization of the membranes was carried out through ATR-IR, XRD, and WCA measurements. According to the in vitro hydrolytic and enzymatic degradation and drug release in different aqueous media for two months, the insertion of alkyl pendant grafts and the crosslinking process provided tuneable additional resistance to the whole membrane suitably for the final application of the membranes. Cell culture showed the cytocompatibility and cell proliferation until 7 days. Osteogenic differentiation and mineralization of pre-osteoblastic MC3T3 cells occurred for most of membranes after 35 days as valued by measuring ALP activity (50 nmol 4-np/h/nf DNA) and the deposition of calcium (120-140 μg ml).
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http://dx.doi.org/10.1016/j.colsurfb.2020.111438DOI Listing
January 2021

Potentials and Challenges of Genomics for Breeding Cannabis Cultivars.

Front Plant Sci 2020 25;11:573299. Epub 2020 Sep 25.

DAFNAE-Department of Agronomy, Food, Natural Resources, Animals and Environment, University of Padova, Campus of Agripolis, Legnaro, Italy.

( L.) is an influential yet controversial agricultural plant with a very long and prominent history of recreational, medicinal, and industrial usages. Given the importance of this species, we deepened some of the main challenges-along with potential solutions-behind the breeding of new cannabis cultivars. One of the main issues that should be fixed before starting new breeding programs is the uncertain taxonomic classification of the two main taxa (e.g., and a) of the genus. We tried therefore to examine this topic from a molecular perspective through the use of DNA barcoding. Our findings seem to support a unique species system () based on two subspecies: subsp. and subsp. . The second key issue in a breeding program is related to the dioecy behavior of this species and to the comprehension of those molecular mechanisms underlying flower development, the main cannabis product. Given the role of MADS box genes in flower identity, we analyzed and reorganized all the genomic and transcriptomic data available for homeotic genes, trying to decipher the applicability of the ABCDE model in . Finally, reviewing the limits of the conventional breeding methods traditionally applied for developing new varieties, we proposed a new breeding scheme for the constitution of F hybrids, without ignoring the indisputable contribution offered by genomics. In this sense, in parallel, we resumed the main advances in the genomic field of this species and, ascertained the lack of a robust set of SNP markers, provided a discriminant and polymorphic panel of SSR markers as a valuable tool for future marker assisted breeding programs.
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http://dx.doi.org/10.3389/fpls.2020.573299DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7546024PMC
September 2020

NGS-based barcoding with mini-COI gene target is useful for pet food market surveys aimed at mislabelling detection.

Sci Rep 2020 10 20;10(1):17767. Epub 2020 Oct 20.

Laboratory of Genomics, Department of Agronomy Food Natural Resources Animals and Environment (DAFNAE), University of Padova, Campus of Agripolis, Viale dell'Università 16, 35020, Legnaro, PD, Italy.

Pet food industry has grown considerably in the last few years and it is expected to continue with this rate. Despite the economic impact of this sector and the consumer concerns for the increasing number of food and feed adulteration cases, few studies have been published on mislabelling in pet foods. We therefore investigated the capability of a next generation sequencing-based mini-barcoding approach to identify animal species in pet food products. In a preliminary analysis, a 127 bp fragment of the COI gene was tested on both individual specimens and ad hoc mixed fresh samples used as testers, to evaluate its discrimination power and primers effectiveness. Eighteen pet food products of different price categories and forms available on the market (i.e. kibbles, bites, pâté and strips) were analysed through an NGS approach in biological replicates. At least one of the species listed in the ingredients was not detected in half of the products, while seven products showed supplementary species in addition to those stated on the label. Due to the accuracy, sensitivity and specificity demonstrated, this method can be proposed as food genetic traceability system to evaluate both the feed and food quality timely along the supply chain.
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http://dx.doi.org/10.1038/s41598-020-74918-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7575603PMC
October 2020

A hyaluronic acid/cyclodextrin based injectable hydrogel for local doxorubicin delivery to solid tumors.

Int J Pharm 2020 Nov 12;589:119879. Epub 2020 Sep 12.

Università degli Studi di Palermo, Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Via Archirafi 32, 90123 Palermo, Italy; Institute of Biophysics at Palermo, Italian National Research Council, Via Ugo La Malfa 153, 90146 Palermo, Italy.

Localized delivery of anticancer drugs is often the most useful therapeutic approach for the treatment of solid tumors. The use of injectable polymeric systems that maximize drug concentration in the proximal area of the tumor represents an extremely advantageous therapeutic strategy. Here, the development of an injectable in situ forming hydrogel was accomplished by exploiting the azo-type Michael reaction between an amine derivative of hyaluronic and vinylsulfone functionalized β-cyclodextrins complexing doxorubicin. This injectable system can be easily prepared and administered with timelines compatible with normal operating room procedures, as demonstrated by rheological tests. In vitro experiments revealed that the peculiar physicochemical properties of the hydrogel guarantee a sustained release of the anticancer drug that blocks the growth of colorectal carcinoma micromasses cultured in 3D conditions. In vivo studies have confirmed that the medicated hydrogel can drastically reduce the tumor mass in the animal model without causing cytotoxic side effects in other areas of the body such as the heart. Overall, the proposed system has shown promising characteristics that make it an interesting useful device for localized chemotherapy of solid tumors.
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http://dx.doi.org/10.1016/j.ijpharm.2020.119879DOI Listing
November 2020

New gellan gum-graft-poly(d,l-lactide-co-glycolide) copolymers as promising bioinks: Synthesis and characterization.

Int J Biol Macromol 2020 Nov 7;162:1653-1667. Epub 2020 Aug 7.

Università degli Studi di Palermo, Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Via Archirafi 32, 90123 Palermo, Italy; Institute of Biophysics at Palermo, Italian National Research Council, Via Ugo La Malfa 153, 90146 Palermo, Italy.

This research focused on the aim of tackling the urgent demand of printable biomaterials, hence we synthetized and characterized three gellan gum-graft-poly(d,l-lactide-co-glycolide) copolymers (GGm-PLGA a, b and c) which differed in the graft substitution degree. We investigated the effect of the polyester chain grafted onto hydrophilic backbone of gellan gum in terms of physicochemical properties and the ability of the system to print 3D cell laden constructs. In particular, we evaluated thermo-rheological, ionotropic crosslinking, shear thinning, swelling and stability properties of these copolymers and their derived biomaterials and findings related to the degree of functionalization. Moreover, the optimization of the 3D process parameters and the effect of different water/DPBS mixtures was investigated, demonstrating the feasibility of the system to print 3D constructs. Finally, biological tests revealed that fibroblasts and chondrocytes remained viable after printing and over a culture period of seven days into scaffolds.
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http://dx.doi.org/10.1016/j.ijbiomac.2020.07.254DOI Listing
November 2020

A Reappraisal of the Evolutionary and Developmental Pathway of Apomixis and Its Genetic Control in Angiosperms.

Genes (Basel) 2020 07 28;11(8). Epub 2020 Jul 28.

Research Division of Perugia, Institute of Biosciences and Bioresources, National Research Council (CNR), Via Madonna Alta 130, 06128 Perugia, Italy.

Apomixis (agamospermy) is asexual reproduction by seed. In angiosperms it represents an easy byway of life cycle renewal through gamete-like cells that give rise to maternal embryos without ploidy reduction (meiosis) and ploidy restitution (syngamy). The origin of apomixis still represents an unsolved problem, as it may be either evolved from sex or the other way around. This review deals with a reappraisal of the origin of apomixis in order to deepen knowledge on such asexual mode of reproduction which seems mainly lacking in the most basal angiosperm orders (i.e., Amborellales, Nymphaeales and Austrobaileyales, also known as ANA-grade), while it clearly occurs in different forms and variants in many unrelated families of monocots and eudicots. Overall findings strengthen the hypothesis that apomixis as a whole may have evolved multiple times in angiosperm evolution following different developmental pathways deviating to different extents from sexuality. Recent developments on the genetic control of apomixis in model species are also presented and adequately discussed in order to shed additional light on the antagonist theories of gain- and loss-of-function over sexuality.
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http://dx.doi.org/10.3390/genes11080859DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7466056PMC
July 2020

The Effect of Soil on the Biochemical Plasticity of Berry Skin in Two Italian Grapevine ( L.) Cultivars.

Front Plant Sci 2020 26;11:822. Epub 2020 Jun 26.

Department of Agronomy, Food, Natural Resources, Animals and Environment, University of Padova, Legnaro PD, Italy.

Grapevine represents a particularly interesting species as concerns phenotypic plasticity, considering that the , meaning the contribution of the geography, geology, and climate of a certain place, together with the agronomical practices utilized, may deeply influence the berry phenotype at the physiological, molecular, and biochemical levels. This phenomenon leads to the production of wines that, although produced from the same variety, exhibit different enological profiles and represents an issue of increasing interest from both a biological and an economic point of view. The main objective of the present study was to deepen the understanding of phenotypic plasticity in grapevine, trying to dissect the role of one its important components - the soil - by investigating the singular effect that different physico-chemical soil properties can produce in terms of berry plasticity at the phenological, physiological, and biochemical levels in a red and a white variety of great economic importance in Italy and overseas: Corvina and Glera. The results indicated a genotype-dependent response to the soil factor, with higher biochemical plasticity in Corvina with respect to Glera and suggested a key role of specific soil properties, including the skeleton, texture, and mineral composition, on the metabolite profile of berry skin.
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http://dx.doi.org/10.3389/fpls.2020.00822DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7333541PMC
June 2020

The Mitochondrial Genome Assembly of Fennel () Reveals Two Different Gene Sequences in Cytoplasmic Male Sterile Accessions.

Int J Mol Sci 2020 Jun 30;21(13). Epub 2020 Jun 30.

Department of Agronomy Food Natural Resources Animals and Environment (DAFNAE), University of Padova, Campus of Agripolis, Viale dell'Università 16, 35020 Legnaro, PD, Italy.

Cytoplasmic male sterility (CMS) has always aroused interest among researchers and breeders, being a valuable resource widely exploited not only to breed F1 hybrid varieties but also to investigate genes that control stamen and pollen development. With the aim of identifying candidate genes for CMS in fennel, we adopted an effective strategy relying on the comparison between mitochondrial genomes (mtDNA) of both fertile and sterile genotypes. mtDNA raw reads derived from a CMS genotype were assembled in a single molecule (296,483 bp), while a draft mtDNA assembly (166,124 nucleotides, 94 contigs) was performed using male fertile sample (MF) sequences. From their annotation and alignment, two -like sequences were identified. , the putative mutant copy with a 300 bp truncation at the 5'-end, was found only in the mtDNA of CMS samples, while the wild type copy () was detected only in the MF mtDNA. Further analyses (i.e., reads mapping and Sanger sequencing), revealed an copy also in CMS samples, probably in the nuclear DNA. However, qPCRs performed on different tissues proved that, despite its availability, is expressed only in MF samples, while mRNA was always detected in CMS individuals. In the light of these findings, the energy deficiency model could explain the pollen deficiency observed in male sterile flower. could represent a gene whose mRNA is translated into a not-fully functional protein leading to suboptimal ATP production that guarantees essential cellular processes but not a high energy demand process such as pollen development. Our study provides novel insights into the fennel mtDNA genome and its genes, and paves the way for further studies aimed at understanding their functional roles in the determination of male sterility.
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http://dx.doi.org/10.3390/ijms21134664DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7370444PMC
June 2020

Genotyping by RAD Sequencing Analysis Assessed the Genetic Distinctiveness of Experimental Lines and Narrowed Down the Genomic Region Responsible for Leaf Shape in Endive ( L.).

Genes (Basel) 2020 04 23;11(4). Epub 2020 Apr 23.

Department of Agronomy, Food, Natural Resources, Animals and Environment (DAFNAE), University of Padova, 35020 Legnaro PD, Italy.

The characterization of genetic diversity in elite breeding stocks is crucial for the registration and protection of new varieties. Moreover, experimental population structure analysis and information about the genetic distinctiveness of commercial materials are essential for crop breeding programs. The purpose of our research was to assess the genetic relationships of 32 endive ( L.) breeding lines, 18 from var. (escarole) and 14 from var. (curly), using heterologous -derived simple sequence repeats (SSR) markers and single-nucleotide polymorphisms (SNP) markers. We found that 14 out of 29 SSR markers were successfully amplified, but only 8 of them were related to polymorphic loci. To overcome the limitation of the low number of informative SSR marker loci, an alternative SNP-based approach was employed. The 4621 SNPs produced by a restriction site-associated DNA marker sequencing approach were able to fully discriminate the 32 endive accessions; most importantly, as many as 50 marker loci were found to distinguish the curly group from the escarole group. Interestingly, 24 of the marker loci mapped within a peripheral segment of chromosome 8 of lettuce ( L.), spanning a chromosomal region of 49.6 Mb. Following Sanger sequencing-based validation, three genes were determined to carry nonsynonymous SNPs, and one of them matched a putative ortholog of , subunit 1 of the Elongator complex. Considering that several previously characterized Elongator complex subunit mutants exhibited elongated and/or curly leaf phenotypes, this gene should be taken into consideration for a better understanding of the underlying mechanism controlling leaf shape in endive.
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http://dx.doi.org/10.3390/genes11040462DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7231076PMC
April 2020

Sustainable and Tunable Mg/MgO Plasmon-Catalytic Platform for the Grand Challenge of SF Environmental Remediation.

Nano Lett 2020 May 10;20(5):3352-3360. Epub 2020 Apr 10.

Institute of Nanotechnology, CNR-NANOTEC, via Amendola 122/D, 70126 Bari, Italy.

Sulfur hexafluoride (SF) is one of the most harmful greenhouse gases producing environmental risks. Therefore, developing ways of degrading SF without forming hazardous products is increasingly important. Herein, we demonstrate for the first time the plasmon-catalytic heterogeneous degradation of SF into nonhazardous MgF and MgSO products by nontoxic and sustainable plasmonic magnesium/magnesium oxide (Mg/MgO) nanoparticles, which are also effective as a plasmon-enhanced SF chemometric sensor. The main product depends on the excitation wavelength; when the localized surface plasmon resonance (LSPR) is in the ultraviolet, then MgF forms, while visible light LSPR results in MgSO. Furthermore, Mg/MgO platforms can be regenerated in few seconds by hydrogen plasma treatment and can be reused in a new cycle of air purification. Therefore, this research first demonstrates effectiveness of Mg/MgO plasmon-catalysis enabling environmental remediation with the concurrent functionalities of monitoring, degrading, and detecting sulfur and fluorine gases in the atmosphere.
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http://dx.doi.org/10.1021/acs.nanolett.0c00244DOI Listing
May 2020

Production and physicochemical characterization of a new amine derivative of gellan gum and rheological study of derived hydrogels.

Carbohydr Polym 2020 May 18;236:116033. Epub 2020 Feb 18.

Università Degli Studi Di Palermo, Dipartimento Di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Via Archirafi 32, 90123 Palermo, Italy; Institute of Biophysics at Palermo, Italian National Research Council, Via Ugo La Malfa 153, 90146 Palermo, Italy.

The production of an amine derivative of gellan gum, named GG-EDA, was here obtained by functionalizing the polysaccharide backbone with pendant ethylenediamine moieties. The obtained derivative was characterized by spectroscopic, colorimetric, chromatographic and rheological analyses to study the effect of the free amino groups on the physicochemical properties of the macromolecule. A titration experiment was conducted to study the acid-base dissociation constants in aqueous media for the carboxylic and amino groups in the GG-EDA and to shed light on the possibility that the derivative shows a polyampholyte structure under physiological conditions. The rheological analysis conducted on both physical and chemical hydrogels based on GG-EDA revealed that the presence of amino groups plays a fundamental role in influencing the viscoelastic properties and stability of the produced samples.
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http://dx.doi.org/10.1016/j.carbpol.2020.116033DOI Listing
May 2020

Initiated Chemical Vapor Deposition of Crosslinked Organic Coatings for Controlling Gentamicin Delivery.

Pharmaceutics 2020 Mar 1;12(3). Epub 2020 Mar 1.

Institute of Solid State Physics, NAWI Graz, Graz University of Technology, Petersgasse 16, 8010 Graz, Austria.

A coating consisting of a copolymer of methacrylic acid and ethylene glycol dimethacrylate was deposited over a gentamicin film by initiated chemical vapor deposition with the aim of controlling the drug release. Gentamicin release in water was monitored by means of conductance measurements and of UV-vis Fluorescence Spectroscopy. The influence of the polymer chemical composition, specifically of its crosslinking density, has been investigated as a tool to control the swelling behavior of the initiated chemical vapor deposition (iCVD) coating in water, and therefore its ability to release the drug. Agar diffusion test and microbroth dilution assays against Staphylococcus aureus and Pseudomonas aeruginosa on cellulose coated substrates confirmed that the antibacterial activity of the drug released by the coating was retained, though the release of gentamicin was not complete.
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http://dx.doi.org/10.3390/pharmaceutics12030213DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7150873PMC
March 2020

Gellan gum-based delivery systems of therapeutic agents and cells.

Carbohydr Polym 2020 Feb 18;229:115430. Epub 2019 Oct 18.

Università degli Studi di Palermo, Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Via Archirafi 32, 90123 Palermo, Italy; Institute of Biophysics at Palermo, Italian National Research Council, Via Ugo La Malfa 153, 90146 Palermo, Italy.

The purpose of this review is to make a summary of high quality research trends using gellan gum (GG) as a polymeric constituent for the design of innovative drug delivery systems and devices for biomedical applications, such as cell therapy and regenerative medicine. The use of gellan gum is described both in its native form and as chemically functionalized derivatives or physically mixed with natural or synthetic materials. Starting from a systematic study of recent research works, the main properties of the native polysaccharide have been highlighted and therefore some improvements have been focused thanks to the design of chemically functionalized derivatives and the use of composite materials.
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http://dx.doi.org/10.1016/j.carbpol.2019.115430DOI Listing
February 2020

Hyaluronic acid based nanohydrogels fabricated by microfluidics for the potential targeted release of Imatinib: Characterization and preliminary evaluation of the antiangiogenic effect.

Int J Pharm 2020 Jan 20;573:118851. Epub 2019 Nov 20.

Università degli Studi di Palermo, Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Via Archirafi 32, 90123 Palermo, Italy; Institute of Biophysics at Palermo, Italian National Research Council, Via Ugo La Malfa 153, 90146 Palermo, Italy.

Microfluidics is emerging as an innovative technique for the "on chip" fabrication of nanoparticles for drug delivery applications. Here, by using an amphiphilic derivative of hyaluronic acid as a starting macromolecule, nanohydrogels loaded with Imatinib were produced by the microfluidic procedure in order to develop an innovative therapeutic tool for the treatment of retinal neovascularization. Both cyRGDC functionalized and non-functionalized nanohydrogels were designed and fabricated by using the same technique. The targeting efficiency of the obtained nanosystems was studied in vitro on human retinal pigment epithelial cells (HRPEpiC) and human umbilical vein endothelial cells (HUVEC), the latter chosen as generic cellular model to assay inhibiting effect on cellular sprouting of Imatinib loaded nanohydrogels. The suitability of microfluidic approach for nanohydrogel production and drug loading was demonstrated. The cyRGDC functionalized nanosystems loaded with Imatinib, showed in vitro an enhanced ability to inhibit HUVEC organization into a capillary like structure.
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http://dx.doi.org/10.1016/j.ijpharm.2019.118851DOI Listing
January 2020

The Molecular Determination of Hybridity and Homozygosity Estimates in Breeding Populations of Lettuce ( L.).

Genes (Basel) 2019 11 9;10(11). Epub 2019 Nov 9.

Department of Agronomy, Food, Natural Resources, Animals and Environment, University of Padova, 35020 Legnaro PD, Italy.

The development of new varieties of horticultural crops benefits from the integration of conventional and molecular marker-assisted breeding schemes in order to combine phenotyping and genotyping information. In this study, a selected panel of 16 microsatellite markers were used in different steps of a breeding programme of lettuce ( L., 2 = 18). Molecular markers were first used to genotype 71 putative parental lines and to plan 89 controlled crosses designed to maximise recombination potentials. The resulting 871 progeny plants were then molecularly screened, and their marker allele profiles were compared with the profiles expected based on the parental lines. The average cross-pollination success rate was 68 ± 33%, so 602 F1 hybrids were completely identified. Unexpected genotypes were detected in 5% of cases, consistent with this species' spontaneous out-pollination rate. Finally, in a later step of the breeding programme, 47 different F3 progenies, selected by phenotyping for a number of morphological descriptors, were characterised in terms of their observed homozygosity and within-population genetic uniformity and stability. Ten of these populations had a median homozygosity above 90% and a median genetic similarity above 95% and are, therefore, particularly suitable for pre-commercial trials. In conclusion, this study shows the synergistic effects and advantages of conventional and molecular methods of selection applied in different steps of a breeding programme aimed at developing new varieties of lettuce.
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http://dx.doi.org/10.3390/genes10110916DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6895879PMC
November 2019

Diversity Analysis of Sweet Potato Genetic Resources Using Morphological and Qualitative Traits and Molecular Markers.

Genes (Basel) 2019 10 24;10(11). Epub 2019 Oct 24.

Department of Agronomy, Food, Natural resources, Animals and Environment (DAFNAE) University of Padova, Agripolis Campus, Viale dell'Università, 16-35020 Legnaro, Italy.

The European Union (EU) market for sweet potatoes has increased by 100% over the last five years, and sweet potato cultivation in southern European countries is a new opportunity for the EU to exploit and introduce new genotypes. In view of this demand, the origins of the principal Italian sweet potato clones, compared with a core collection of genotypes from Central and Southern America, were investigated for the first time. This was accomplished by combining a genetic analysis, exploiting 14 hypervariable microsatellite markers, with morphological and chemical measurements based on 16 parameters. From the molecular analyses, Italian accessions were determined to be genetically very similar to the South American germplasm, but they were sub-clustered into two groups. This finding was subsequently confirmed by the morphological and chemical measurements. Moreover, the analysis of the genetic structure of the population suggested that one of the two groups of Italian genotypes may have descended from one of the South American accessions, as predicted on the basis of the shared morphological characteristics and molecular fingerprints. Overall, the combination of two different characterization methods, genetic markers and agronomic traits, was effective in differentiating or clustering the sweet potato genotypes, in agreement with their geographical origin or phenotypic descriptors. This information could be exploited by both breeders and farmers to detect and protect commercial varieties, and hence for traceability purposes.
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http://dx.doi.org/10.3390/genes10110840DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6895877PMC
October 2019

In vivo functionalization of diatom biosilica with sodium alendronate as osteoactive material.

Mater Sci Eng C Mater Biol Appl 2019 Nov 16;104:109897. Epub 2019 Jun 16.

Dipartimento di Chimica, Università degli Studi "Aldo Moro", Via Orabona 4, I-70126 Bari, Italy. Electronic address:

Bisphosphonates are a class of drugs widely used in the clinical treatment of disorders of bone metabolism, such as osteoporosis, fibrous dysplasia, myeloma and bone metastases. Because of the negative side effects caused by oral administration of bisphosphonates, various silica mesoporous materials have been investigated for a confined and controlled release of these drugs. Here, we propose biosilica from diatoms as suitable substrate for alendronate local activation of bone cells. Following a novel strategy, sodium alendronate can be in vivo incorporated into biosilica shells of cultured Thalassiosira weissflogii diatoms, by feeding the algae with an aqueous solution of the drug. After acid/oxidative treatments for removing organic matter, the resulting bisphosphonate-functionalized mesoporous biosilica was characterized and tested as osteoinductive support. Effects on osteoblast growth and anti-osteoclast activity have been examined by evaluating SaOS-2, BMSC, J774 cell viability on the alendronate-"doped" biosilica. The loading percentage of sodium alendronate into biosilica, estimated as 1.45% w/w via TGA, was able to decrease metabolic activity of J774 osteoclasts-like cells till 5% over glass control. We demonstrated a good osteoconductive ability and activation of a tissue regeneration model together with osteoclasts inhibition of the functionalized biosilica, opening the way to interesting applications for diatom microalgae as a bioinspired mesoporous material for tissue engineering.
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http://dx.doi.org/10.1016/j.msec.2019.109897DOI Listing
November 2019

Blend scaffolds with polyaspartamide/polyester structure fabricated via TIPS and their RGDC functionalization to promote osteoblast adhesion and proliferation.

J Biomed Mater Res A 2019 12 3;107(12):2726-2735. Epub 2019 Sep 3.

Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Università di Palermo, Palermo, Italy.

Target of this work was to prepare a RGDC functionalized hybrid biomaterial via TIPS technique to achieve a more efficient control of osteoblast adhesion and diffusion on the three-dimensional (3D) scaffolds. Starting from a crystalline poly(l-lactic acid) (PLLA) and an amorphous α,β-poly(N-2-hydroxyethyl) (2-aminoethylcarbamate)-d,l-aspartamide-graft-polylactic acid (PHEA-EDA-g-PLA) copolymer, blend scaffolds were characterized by an appropriate porosity and pore interconnection. The PHEA-EDA-PLA interpenetration with PLLA improved hydrolytic susceptibility of hybrid scaffolds. The presence of free amino groups on scaffolds allowed to tether the cyclic RGD peptide (RGDC) via Michael addition using the maleimide chemistry. Cell culture test carried out on preosteoblastic cells MC3T3-E1 incubated with scaffolds, has evidenced cell adhesion and proliferation. Furthermore, the presence of distributed bone matrix on all scaffolds was evaluated after 70 days compared to PLLA only samples.
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http://dx.doi.org/10.1002/jbm.a.36776DOI Listing
December 2019

Genetic Structure of Cultivated Varieties of Radicchio ( L.): A Comparison between F1 Hybrids and Synthetics.

Plants (Basel) 2019 Jul 10;8(7). Epub 2019 Jul 10.

Department of Agronomy, Food, Natural Resources, Animals and Environment, University of Padova, 35020 Legnaro PD, Italy.

L., well known in Italy with the common name "Radicchio", is an important leafy vegetable that is prevalently reproduced by allogamy due to very efficient barriers of self-incompatibility. Marker-assisted breeding is widely used by seed firms to develop new hybrid varieties that manifest genetic distinctiveness, uniformity and stability. A total of 29 mapped microsatellite markers were used for genotyping 504 samples of the Red of Chioggia biotype: First, two synthetics, four F1 hybrids and two derived F2 populations were compared to assess the distinctiveness of their gene pool and structure; then, the uniformity and stability of 3 years of production of a commercial F1 variety were also investigated. Genetic similarity and diversity statistics as well as the genetic structure of populations were analysed, including allele and genotype frequencies. The mean estimates and ranges of genetic similarity enabled the molecular discrimination of OP synthetics from F1 varieties and their F2 progenies and the determination of individual plant memberships. Moreover, the genetic structure of F1 hybrids produced in 3 years unexpectedly revealed two main clusters that discriminate the first 2 years from the 3rd, mainly because of the presence of uncommon specific alleles and different allele frequencies. Overall, this molecular information will enable breeders to determine the genetic distinctness, uniformity and stability of commercial and experimental varieties, as well as their genetic relationships and relatedness. Hence, this work provides a useful tool for achieving the molecular characterisation and genetic identification of different radicchio populations.
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http://dx.doi.org/10.3390/plants8070213DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6681312PMC
July 2019

Production of a Double-Layer Scaffold for the "On-Demand" Release of Fibroblast-like Limbal Stem Cells.

ACS Appl Mater Interfaces 2019 Jun 12;11(25):22206-22217. Epub 2019 Jun 12.

Department of "Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche" (STEBICEF) , University of Palermo , Via Archirafi 32 , 90123 Palermo , Italy.

The production and characterization of a double-layer scaffold, to be used as a system for the "on-demand" release of corneal limbal stem cells, are reported here. The devices used in the clinics and proposed so far in the scientific literature, for the release of corneal stem cells in the treatment of limbal stem cell deficiency, cannot control the in vivo space-time release of cells as the biomaterial of which they are composed is devoid of the stimuli-responsive feature. Our approach was to produce a scaffold composed of two different polymeric layers that give the device the appropriate mechanical properties to be placed on the ocular surface and the possibility of releasing the stem cells following a noninvasive and cell-friendly treatment. This device consists of an electrospun microfibrillar scaffold of poly-l-lactic acid coated by a polymeric film based on an amphiphilic derivative of hyaluronic acid sensitive to the ionic strength of the external medium and to the presence of a complexing agent. The latter represents the "sacrificial" cell containing layer of the scaffold that can be dissolved "on demand" by the treatment with a solution of cyclodextrins. The rapid removal of the external polymeric film from the device is exploited to control the space-time release of the cells. In vitro and ex vivo experiments showed that fibroblast-like limbal stem cells cultured on the scaffold without the use of the feeder layer maintained their characteristics of stem cells and can be released "on demand" on the culture well coated with Matrigel or on the decellularized bovine cornea, respectively.
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http://dx.doi.org/10.1021/acsami.9b06757DOI Listing
June 2019

Data from functionalization of diatom mesoporous biosilica with bisphosphonates.

Data Brief 2019 Jun 16;24:103831. Epub 2019 Mar 16.

CNR- ICCOM, Department of Chemistry, University of Bari "Aldo Moro", Via E. Orabona, 4, 70126 Bari, Italy.

Diatoms are unicellular photosynthetic microalgae that produce a sophisticated mesoporous biosilica shell called . Easy to achieve and extract, diatom frustules represent a low-cost source of mesoporous biocompatible biosilica. In this paper, the possibility to functionalize the diatom biosilica with bisphosphonates (BPs) was investigated. In particular, two BPs were tested: the amino-containing sodium alendronate (ALE) and the amino-lacking sodium etidronate (ETI). According to first SEM-EDX analysis, the presence of the amino-moiety in ALE structure allowed a better incorporation of this BP into living diatom biosilica, compared to ETI. Then, diatom growth was deeply investigated in presence of ALE. After extraction of functionalized frustules, ALE-biosilica was further characterized by XPS and microscopy, and ALE release was evaluated by ferrochelation assay. Moreover, the bone regeneration performances of ALE-functionalized frustules were preliminarily investigated on bone osteoblast-like cells, Comassie staining. Data are related to the research article " functionalization of diatom biosilica with sodium alendronate as osteoactive material".
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http://dx.doi.org/10.1016/j.dib.2019.103831DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6451687PMC
June 2019

Genomics of Flower Identity in Grapevine ( L.).

Front Plant Sci 2019 21;10:316. Epub 2019 Mar 21.

Department of Agronomy, Food, Natural Resources, Animals, and Environment, University of Padua, Legnaro, Italy.

The identity of the four characteristic whorls of typical eudicots, namely, sepals, petals, stamens, and carpels, is specified by the overlapping action of homeotic genes, whose single and combined contributions have been described in detail in the so-called ABCDE model. Continuous species-specific refinements and translations resulted in this model providing the basis for understanding the genetic and molecular mechanisms of flower development in model organisms, such as and other main plant species. Although grapevine ( L.) represents an extremely important cultivated fruit crop globally, studies related to the genetic determinism of flower development are still rare, probably because of the limited interest in sexual reproduction in a plant that is predominantly propagated asexually. Nonetheless, several studies have identified and functionally characterized some ABCDE orthologs in grapevine. The present study is intended to provide a comprehensive screenshot of the transcriptional behavior of 18 representative grapevine ABCDE genes encoding MADS-box transcription factors in a developmental kinetic process, from preanthesis to the postfertilization stage and in different flower organs, namely, the calyx, calyptra, anthers, filaments, ovary, and embryos. The transcript levels found were compared with the proposed model for to evaluate their biological consistency. With a few exceptions, the results confirmed the expression pattern expected based on the data.
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http://dx.doi.org/10.3389/fpls.2019.00316DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6437108PMC
March 2019

Construction of the First SNP-Based Linkage Map Using Genotyping-by-Sequencing and Mapping of the Male-Sterility Gene in Leaf Chicory.

Front Plant Sci 2019 11;10:276. Epub 2019 Mar 11.

Laboratory of Genomics for Plant Breeding, Department of Agronomy Food Natural Resources Animals and Environment (DAFNAE), University of Padova, Legnaro, Italy.

We report the first high-density linkage map construction through genotyping-by-sequencing (GBS) in leaf chicory ( subsp. var. , 2 = 2 = 18) and the SNP-based fine mapping of the linkage group region carrying a recessive gene responsible for male-sterility (). An experimental BC population, segregating for the male sterility trait, was specifically generated and 198 progeny plants were preliminary screened through a multiplexed SSR genotyping analysis for the identification of microsatellite markers linked to the locus. Two backbone SSR markers belonging to linkage group 4 of the available consensus map were found genetically associated to the gene at 5.8 and 12.1 cM apart. A GBS strategy was then used to produce a high-density SNP-based linkage map, containing 727 genomic loci organized into 9 linkage groups and spanning a total length of 1,413 cM. 13 SNPs proved to be tightly linked to the locus based on a subset of 44 progeny plants analyzed. The map position of these markers was further validated by sequence-specific PCR experiments using an additional set of 64 progeny plants, enabling to verify that four of them fully co-segregated with male-sterility. A mesosynteny analysis revealed that 10 genomic DNA sequences encompassing the 13 selected SNPs of chicory mapped in a peripheral region of chromosome 5 of lettuce ( L.) spanning about 18 Mbp. Since a MYB103-like gene, encoding for a transcription factor involved in callose dissolution of tetrads and exine development of microspores, was found located in the same chromosomal region, this orthologous was chosen as candidate for male-sterility. The amplification and sequencing of its CDS using accessions with contrasting phenotypes/genotypes (i.e., 4 male sterile mutants, , and 4 male fertile inbreds, ) enabled to detect an INDEL of 4 nucleotides in its second exon, responsible for an anticipated stop codon in the male sterile mutants. This polymorphism was subsequently validated through allele-specific PCR assays and found to fully co-segregate with male-sterility, using 64 progeny plants of the same mapping BC population. Overall, our molecular data could be practically exploited for genotyping plant materials and for marker-assisted breeding schemes in leaf chicory.
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http://dx.doi.org/10.3389/fpls.2019.00276DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6421318PMC
March 2019