26 results on '"Lucretti, Sergio"'
Search Results
2. Somatic Embryogenesis and Flow Cytometric Assessment of Nuclear Genetic Stability for Sansevieria spp.: An Approach for In Vitro Regeneration of Ornamental Plants.
- Author
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Catalano, Caterina, Carra, Angela, Carimi, Francesco, Motisi, Antonio, Sajeva, Maurizio, Butler, Alan, Lucretti, Sergio, Giorgi, Debora, Farina, Anna, and Abbate, Loredana
- Subjects
SOMATIC embryogenesis ,ORNAMENTAL plants ,REGENERATION (Botany) ,VEGETATIVE propagation ,SUCCULENT plants ,PLANT regulators - Abstract
Sansevieria Thunb. species are traditionally known as succulent ornamental plants worldwide. They are also cultivated for medicinal, fodder, soil conservation and fiber uses, and for their capacity to reduce environmental pollution. Sansevieria sexual propagation is limited by the lack of viable seeds, and reproduction is largely made via vegetative propagation by suckers or cuttings. For these reasons, genetic improvement by conventional breeding is limited. To overcome this problem and to address the increasing demand from customers for novel Sansevieria varieties, many commercial companies regularly use in vitro propagation, as is the case in the breeding process of several ornamental plants. In this paper, for the first time, we report a procedure for in vitro somatic embryogenesis and plant regeneration starting from three flower explants for seven different Sansevieria genotypes. Regeneration was attempted using stigmas/styles, anther/filament, and ovary which were cultured on a Murashige and Skoog solidified medium under three different plant growth regulator combinations. A good regeneration rate was obtained with all genotypes used under all culture conditions tested from every explant type, with percentages ranging from 0 to 73.3%. "Genetic stability" assessment of regenerated plants in respect to their mother plants was verified through flow cytometry analysis showing a high degree of uniformity, with only S. parva exhibiting a different level of DNA fluorescence among in vitro regenerated plants. This is an interesting achievement in the aim to produce true-to-type plants and new variants with desirable characteristics, both of which are desired features in ornamentals improvement. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
3. Perturbation of Polyamine Catabolism Can Strongly Affect Root Development and Xylem Differentiation
- Author
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Tisi, Alessandra, Federico, Rodolfo, Moreno, Sandra, Lucretti, Sergio, Moschou, Panagiotis N., Roubelakis-Angelakis, Kalliopi A., Angelini, Riccardo, and Cona, Alessandra
- Published
- 2011
4. Synergistic Action of Mild Heat and Essential Oil Treatments on Culturability and Viability of Escherichia coli ATCC 25922 Tested In Vitro and in Fruit Juice.
- Author
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Di Gregorio, Luciana, Tchuenchieu, Alex, Poscente, Valeria, Arioli, Stefania, Del Fiore, Antonella, Costanzo, Manuela, Giorgi, Debora, Lucretti, Sergio, and Bevivino, Annamaria
- Subjects
FRUIT juices ,ESCHERICHIA coli ,ESSENTIAL oils ,BACTERIAL inactivation ,MICROBIAL products ,BACTERICIDAL action - Abstract
The strengthening effect of a mild temperature treatment on the antimicrobial efficacy of essential oils has been widely reported, often leading to an underestimation or a misinterpretation of the product's microbial status. In the present study, both a traditional culture-based method and Flow Cytometry (FCM) were applied to monitor the individual or combined effect of Origanum vulgare essential oil (OEO) and mild heat treatment on the culturability and viability of Escherichia coli in a conventional culture medium and in a fruit juice challenge test. The results obtained in the culture medium showed bacterial inactivation with an increasing treatment temperature (55 °C, 60 °C, 65 °C), highlighting an overestimation of the dead population using the culture-based method; in fact, when the FCM method was applied, the prevalence of injured bacterial cells in a viable but non-culturable (VBNC) state was observed. When commercial fruit juice with a pH of 3.8 and buffered at pH 7.0 was inoculated with E. coli ATCC 25922, a bactericidal action of OEO and a higher efficiency of the mild heat at 65 °C for 5′ combined with OEO were found. Overall, the combination of mild heat and OEO treatment represents a promising antimicrobial alternative to improve the safety of fruit juice. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
5. Assessment of the origin of new citrus tetraploid hybrids (2n = 4x) by means of SSR markers and PCR based dosage effects
- Author
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Ferrante, Sergio Pietro, Lucretti, Sergio, Reale, Silvia, De Patrizio, Alessandro, Abbate, Loredana, Tusa, Nicasio, and Scarano, Maria-Teresa
- Published
- 2010
- Full Text
- View/download PDF
6. Chromosome sorting in tetraploid wheat and its potential for genome analysis
- Author
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Kubalakova, Marie, Kovarova, Pavlina, Suchankova, Pavla, Cihalikova, Jarmila, Bartos, Jan, Lucretti, Sergio, Watanabe, Nobuyoshi, Kianian, Shahryar F., and Dolezel, Jaroslav
- Subjects
Genomics -- Comparative analysis ,Genetic research -- Comparative analysis ,Wheat -- Comparative analysis ,Biological sciences - Abstract
This study evaluates the potential of flow cytometry for chromosome sorting in durum wheat (Triticum turgidum Desf. var. durum, 2n = 4x = 28). Histograms of fluorescence intensity (flow karyotypes) obtained after the analysis of DAPI-stained chromosomes consisted of three peaks. Of these, one represented chromosome 3B, a small peak corresponded to chromosomes 1A and 6A, and a large peak represented the remaining 11 chromosomes. Chromosomes sorted onto microscope slides were identified after fluorescence in situ hybridization (FISH) with probes for GAA microsatellite, pSc119.2, and Afa repeats. Genomic distribution of these sequences was determined for the first time in durum wheat and a molecular karyotype has been developed for this crop. Flow karyotyping in double-ditelosomic lines of durum wheat revealed that the lines facilitated sorting of any arm of the wheat A- and B-genome chromosomes. Compared to hexaploid wheat, flow karyotype of durum wheat is less complex. This property results in better discrimination of telosomes and high purities in sorted fractions, ranging from 90 to 98%. We have demonstrated that large insert libraries can be created from DNA purified using flow cytometry. This study considerably expands the potential of flow cytogenetics for use in wheat genomics and opens the possibility of sequencing the genome of this important crop one chromosome arm at a time.
- Published
- 2005
7. Bivariate flow cytometry DNA/BrdUrd analysis of plant cell cycle
- Author
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Lucretti, Sergio, Nardi, Luca, Nisini, Paola Trionfetti, Moretti, Filippo, Gualberti, Giuliana, and Doležel, Jaroslav
- Published
- 1999
- Full Text
- View/download PDF
8. Localization of seed protein genes on flow-sorted field bean chromosomes
- Author
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Macas, Jiří, Doležel, Jaroslav, Lucretti, Sergio, Pich, Uta, Meister, Armin, Fuchs, Jörg, and Schubert, Ingo
- Published
- 1993
- Full Text
- View/download PDF
9. Chromosome analysis and sorting.
- Author
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Doležel, Jaroslav, Lucretti, Sergio, Molnár, István, Cápal, Petr, and Giorgi, Debora
- Abstract
Flow cytometric analysis and sorting of plant mitotic chromosomes has been mastered by only a few laboratories worldwide. Yet, it has been contributing significantly to progress in plant genetics, including the production of genome assemblies and the cloning of important genes. The dissection of complex genomes by flow sorting into the individual chromosomes that represent small parts of the genome reduces DNA sample complexity and streamlines projects relying on molecular and genomic techniques. Whereas flow cytometric analysis, that is, chromosome classification according to fluorescence and light scatter properties, is an integral part of any chromosome sorting project, it has rarely been used on its own due to lower resolution and sensitivity as compared to other cytogenetic methods. To perform chromosome analysis and sorting, commercially available electrostatic droplet sorters are suitable. However, in order to resolve and purify chromosomes of interest the instrument must offer high resolution of optical signals as well as stability during long runs. The challenge is thus not the instrumentation, but the adequate sample preparation. The sample must be a suspension of intact mitotic metaphase chromosomes and the protocol, which includes the induction of cell cycle synchrony, accumulation of dividing cells at metaphase, and release of undamaged chromosomes, is time consuming and laborious and needs to be performed very carefully. Moreover, in addition to fluorescent staining chromosomal DNA, the protocol may include specific labelling of DNA repeats to facilitate discrimination of particular chromosomes. This review introduces the applications of chromosome sorting in plants, and discusses in detail sample preparation, chromosome analysis and sorting to achieve the highest purity in flow‐sorted fractions, and their suitability for downstream applications. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
10. Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures.
- Author
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Lonoce, Chiara, Salem, Reda, Marusic, Carla, Jutras, Philippe V., Scaloni, Andrea, Salzano, Anna Maria, Lucretti, Sergio, Steinkellner, Herta, Benvenuto, Eugenio, and Donini, Marcello
- Published
- 2016
- Full Text
- View/download PDF
11. First detailed karyo-morphological analysis and molecular cytological study of leafy cardoon and globe artichoke, two multi-use Asteraceae crops.
- Author
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Giorgi, Debora, Pandozy, Gianmarco, Farina, Anna, Grosso, Valentina, Lucretti, Sergio, Gennaro, Andrea, Crinò, Paola, and Saccardo, Francesco
- Subjects
CARDOON ,ARTICHOKES ,REPEATED sequence (Genetics) ,FLOW cytometry ,CHROMOSOMES ,KARYOTYPES - Abstract
Traditionally globe artichoke and leafy cardoon have been cultivated for use as vegetables but these crops are now finding multiple new roles in applications ranging from paper production to cheese preparation and biofuel use, with interest in their functional food potential. So far, their chromosome complements have been poorly investigated and a well-defined karyotype was not available. In this paper, a detailed karyo-morphological analysis and molecular cytogenetic studies were conducted on globe artichoke (Cynara cardunculus Linnaeus, 1753 var. scolymus Fiori, 1904) and leafy cardoon (C. cardunculus Linneaus, 1753 var. altilis De Candolle, 1838). Fluorescent In Situ Hybridization In Suspension (FISHIS) was applied to nuclei suspensions as a fast method for screening of labelling probes, before metaphase spread hybridization. Classic Fluorescent In Situ Hybridization (FISH) on slide, using repetitive telomeric and ribosomal sequences and Simple Sequence Repeats (SSRs) oligonucleotide as probes, identified homologous chromosome relationships and allowed development of molecular karyotypes for both varieties. The close phylogenetic relationship between globe artichoke and cardoon was supported by the very similar karyotypes but clear chromosomal structural variation was detected. In the light of the recent release of the globe artichoke genome sequencing, these results are relevant for future anchoring of the pseudomolecule sequence assemblies to specific chromosomes. In addition, the DNA content of the two crops has been determined by flow cytometry and a fast method for standard FISH on slide and methodological improvements for nuclei isolation are described. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
12. FISHIS: A New Way in Chromosome Flow Sorting Makes Complex Genomes More Accessible.
- Author
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Lucretti, Sergio, Giorgi, Debora, Farina, Anna, and Grosso, Valentina
- Published
- 2014
- Full Text
- View/download PDF
13. FISHIS: Fluorescence In Situ Hybridization in Suspension and Chromosome Flow Sorting Made Easy.
- Author
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Giorgi, Debora, Farina, Anna, Grosso, Valentina, Gennaro, Andrea, Ceoloni, Carla, and Lucretti, Sergio
- Subjects
PLANT genetics ,FLUORESCENCE ,PLANT hybridization ,PLANT chromosomes ,FLOW cytometry ,MICROSATELLITE repeats ,DNA probes ,CYTOGENETICS - Abstract
The large size and complex polyploid nature of many genomes has often hampered genomics development, as is the case for several plants of high agronomic value. Isolating single chromosomes or chromosome arms via flow sorting offers a clue to resolve such complexity by focusing sequencing to a discrete and self-consistent part of the whole genome. The occurrence of sufficient differences in the size and or base-pair composition of the individual chromosomes, which is uncommon in plants, is critical for the success of flow sorting. We overcome this limitation by developing a robust method for labeling isolated chromosomes, named Fluorescent In situ Hybridization In suspension (FISHIS). FISHIS employs fluorescently labeled synthetic repetitive DNA probes, which are hybridized, in a wash-less procedure, to chromosomes in suspension following DNA alkaline denaturation. All typical A, B and D genomes of wheat, as well as individual chromosomes from pasta (T. durum L.) and bread (T. aestivum L.) wheat, were flow-sorted, after FISHIS, at high purity. For the first time in eukaryotes, each individual chromosome of a diploid organism, Dasypyrum villosum (L.) Candargy, was flow-sorted regardless of its size or base-pair related content. FISHIS-based chromosome sorting is a powerful and innovative flow cytogenetic tool which can develop new genomic resources from each plant species, where microsatellite DNA probes are available and high quality chromosome suspensions could be produced. The joining of FISHIS labeling and flow sorting with the Next Generation Sequencing methodology will enforce genomics for more species, and by this mightier chromosome approach it will be possible to increase our knowledge about structure, evolution and function of plant genome to be used for crop improvement. It is also anticipated that this technique could contribute to analyze and sort animal chromosomes with peculiar cytogenetic abnormalities, such as copy number variations or cytogenetic aberrations. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
14. Flow Sorting and Molecular Cytogenetic Identification of Individual Chromosomes of Dasypyrum villosum L (H. villosa) by a Single DNA Probe.
- Author
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Grosso, Valentina, Farina, Anna, Gennaro, Andrea, Giorgi, Debora, and Lucretti, Sergio
- Subjects
PRIONS ,PROTEOLYSIS ,PROTEOLYTIC enzymes ,X-ray crystallography ,MAGNETIC resonance imaging ,GLYCOSYLPHOSPHATIDYLINOSITOL - Abstract
Dasypyrum villosum (L.) Candargy (sin. Haynaldia villosa) is an annual wild diploid grass species (2n = 2x=14; genome VV) belonging to the Poaceae family, which is considered to be an important source of biotic and abiotic stress resistance genes for wheat breeding. Enhanced characterization of D. villosum chromosomes can facilitate exploitation of its gene pool and its use in wheat breeding programs. Here we present the cytogenetic identification of D. villosum chromosomes on slide by fluorescent in situ hybridization (FISH), with the GAA simple sequence repeat (SSR) as a probe. We also describe the isolation and the flow cytometric analysis of D. villosum chromosomes in suspension, resulting in a distinguished flow karyotype. Chromosomes were flow sorted into three fractions, according their DNA content, one of which was composed of a single type of chromosome, namely 6 V, sorted with over 85% purity. Chromosome 6 V is known to carry genes to code for important resistance and seed storage characteristics, and its isolation represents a new source of genetic traits and specific markers useful for wheat improvement. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
15. Assessment of the origin of new citrus tetraploid hybrids (2 n = 4x) by means of SSR markers and PCR based dosage effects.
- Author
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Ferrante, Sergio Pietro, Lucretti, Sergio, De Reale, Silvia, Patrizio, Alessandro, Abbate, Loredana, Tusa, Nicasio, and Scarano, Maria-Teresa
- Subjects
CAPILLARY electrophoresis ,POLYMERASE chain reaction ,SOMATIC hybrids ,DNA primers ,GENE frequency ,CELL fusion - Abstract
We report the accurate determination of the allelic configurations of a total of eight new citrus tetraploid hybrids by means of SSR analysis, coupled with capillary electrophoresis, and PCR based dosage effects. Tetraploid hybrids were spontaneously obtained from different interploid crosses (2x × 4x) between diploid ‘Femminello’ lemon and the allotetraploid somatic hybrid (2 n = 4x = 36) ‘Key’ lime + ‘Valencia’ orange, and between diploid ‘Wilking’ and ‘Fortune’ mandarins and an autotetraploid ‘Dancy’ mandarin (2 n = 4x = 36). To understand the opportunity to employ them in further backcross programs, the cytological mechanisms underlying their ploidy level were unambiguously determined using six SSR primers. PCR conditions were optimized and skewness in template/product ratios were verified. Tetraploid allelic configurations were determined from PCR based dosage effects using electropherogram peak heights to estimate the copy number per allele. In all the tetraploid hybrids we found out that diploginy (2 n eggs) has occurred, contributing the extra haploid genome in the tetraploids. According to the marker genotypes, it was further inferred that the 2 n eggs in ‘Femminello’ lemon resulted from first division restitution (FDR), while in ‘Wilking’ and ‘Fortune’ mandarins 2 n eggs occurred in second division restitution (SDR). These new genotypes, with their improved genetic female background, can be therefore considered very valuable in our citrus genetic improvement program as pollen donors in backcrosses suitable to eliminate negative traits. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
16. DNA Markers and FCSS Analyses Shed Light on the Genetic Diversity and Reproductive Strategy of Jatropha curcas L.
- Author
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Ambrosi, Daria Gigliola, Galla, Giulio, Purelli, Marina, Barbi, Tommaso, Fabbri, Andrea, Lucretti, Sergio, Sharbel, Timothy Francis, and Barcaccia, Gianni
- Subjects
PLANT reproduction ,GENETIC markers ,OILSEED plants ,GENOMES ,PLANT embryology ,ENDOSPERM ,JATROPHA ,GENETIC research ,DEVELOPED countries - Abstract
Jatropha curcas L. (2n = 2x = 22) is becoming a popular non-food oleaginous crop in several developed countries due to its proposed value in the biopharmaceutical industry. Despite the potentials of its oil-rich seeds as a renewable source of biodiesel and an interest in large-scale cultivation, relatively little is known with respect to plant reproduction strategies and population dynamics. Here, genomic DNA markers and FCSS analyses were performed to gain insights into ploidy variation and heterozygosity levels of multiple accessions, and genomic relationships among commercial varieties of Jatropha grown in different geographical areas. The determination of ploidy and the differentiation of either pseudogamous or autonomous apomixis from sexuality were based on the seed DNA contents of embryo and endosperm. The presence of only a high 2C embryo peak and a smaller 3C endosperm peak (ratio 2:3) is consistent with an obligate sexual reproductive system. Because of the lack of either 4C or 5C endosperm DNA estimates, the occurrence of gametophytic apomixis seems unlikely in this species but adventitious embryony cannot be ruled out. The investigation of genetic variation within and between cultivated populations was carried out using dominant RAPD and Inter-SSR markers, and codominant SSR markers. Nei's genetic diversity, corresponding to the expected heterozygosity, was equal to H
e = 0.3491 and the fixation index as low as Fst = 0.2042. The main finding is that seeds commercialized worldwide include a few closely related genotypes, which are not representative of the original Mexican gene pool, revealing high degrees of homozygosity for single varieties and very low genetic diversity between varieties. [ABSTRACT FROM AUTHOR]- Published
- 2010
- Full Text
- View/download PDF
17. Primed in situ labelling facilitates flow sorting of similar sized chromosomes.
- Author
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Pich, Uta, Meister, Armin, Macas, Jiří, Doležel, Jaroslav, Lucretti, Sergio, and Schubert, Ingo
- Published
- 1995
- Full Text
- View/download PDF
18. Comparison of three DNA fluorochromes for flow cytometric estimation of nuclear DNA content in plants.
- Author
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Doležel, Jaroslav, Sgorhati, Sergio, and Lucretti, Sergio
- Subjects
DNA ,PLANT species ,PLANT molecular biology ,CYTOMETRY ,DYES & dyeing ,NUCLEIC acids - Abstract
Flow cytometric estimation of nuclear DNA content was performed in six plant species employing three fluorochromes showing different DNA base preferences: propidium iodide (no base preference). 4',6-diamidino-2-phenylindole (DAPI; AT preference), and mithramycin (GC preference). Nuclei isolated from human leukocytes were used as a primary reference standard. While nuclear DNA contents estimated using propidium iodide were in agreement with published data obtained using other techniques, the values obtained using fluorochromes showing base preference were significantly different. It was found that the differences were caused by the differences in overall AT/GC ratios, and by the species-specific differences in binding of these fluorochromes to DNA. It was concluded that nuclear DNA content estimations performed with fluorochromes showing base preference should be interpreted with caution even when AT/GC ratios of the reference and the sample are equal. The use of intercalating, dyes (e.g. propidium iodide) is recommended for this purpose. On the other hand, comparison of the staining behaviour of intercalating dyes with that of dyes showing base preference may give additional information on chromatin structural differences and arrangement of molecule pairs in DNA. [ABSTRACT FROM AUTHOR]
- Published
- 1992
- Full Text
- View/download PDF
19. Plant Chromosome Analysis and Sorting by Flow Cytometry.
- Author
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Doležel, Jaroslav, Lucretti, Sergio, and Schubert, Ingo
- Published
- 1994
- Full Text
- View/download PDF
20. A Cross between Bread Wheat and a 2D(2R) Disomic Substitution Triticale Line Leads to the Formation of a Novel Disomic Addition Line and Provides Information of the Role of Rye Secalins on Breadmaking Characteristics.
- Author
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Sestili, Francesco, Margiotta, Benedetta, Vaccino, Patrizia, Moscaritolo, Salvatore, Giorgi, Debora, Lucretti, Sergio, Palombieri, Samuela, Masci, Stefania, and Lafiandra, Domenico
- Subjects
TRITICALE ,RYE ,WHEAT ,SELECTION (Plant breeding) ,BREAD ,MOLECULAR weights - Abstract
A bread wheat line (N11) and a disomic 2D(2R) substitution triticale line were crossed and backrossed four times. At each step electrophoretic selection for the seeds that possessed, simultaneously, the complete set of high molecular weight glutenin subunits of N11 and the two high molecular weight secalins of rye, present in the 2D(2R) line, was carried out. Molecular cytogenetic analyses of the BC
4 F8 generation revealed that the selection carried out produced a disomic addition line (2n = 44). The pair of additional chromosomes consisted of the long arm of chromosome 1R (1RL) from rye fused with the satellite body of the wheat chromosome 6B. Rheological analyses revealed that the dough obtained by the new addition line had higher quality characteristics when compared with the two parents. The role of the two additional high molecular weight secalins, present in the disomic addition line, in influencing improved dough characteristics is discussed. [ABSTRACT FROM AUTHOR]- Published
- 2020
- Full Text
- View/download PDF
21. Phyllostictine A, a potential natural herbicide produced by Phyllosticta cirsii: In vitro production and toxicity
- Author
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Zonno, Maria Chiara, Vurro, Maurizio, Lucretti, Sergio, Andolfi, Anna, Perrone, Carmen, and Evidente, Antonio
- Subjects
- *
PHYLLOSTICTA , *TOXINS , *PHYTOTOXICITY , *MICROSCOPY - Abstract
Abstract: Phyllostictine A is a powerful toxin produced by Phyllosticta cirsii, a potential mycoherbicide of Cirsium arvense. To support its potential use as a natural herbicide, toxin production has been studied using different media and cultural conditions. The toxin content in the crude extracts has been determined by using a HPLC method set up for this purpose. Furthermore, its phytotoxicity has been evaluated on tobacco protoplasts by flow cytometric analysis, and on C. arvense protoplasts, by fluorescence microscopy. The best cultural conditions found allowed to produce more than 28mgml−1 of toxin in culture filtrate. The pure metabolite proved to have rapid dose-dependant toxic effects on host and no-host plant protoplasts. [Copyright &y& Elsevier]
- Published
- 2008
- Full Text
- View/download PDF
22. Fingerprinting of three typical macrosperma Italian lentil (Lens culinaris Medik.) landraces using fluorescence-based AFLP markers
- Author
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Fiocchetti, Floriana, Laddomada, Barbara, Roselli, Mariaincoronata, Crinò, Paola, and Lucretti, Sergio
- Subjects
- *
DNA fingerprinting of plants , *LENTILS , *GENETIC markers , *AMPLIFIED fragment length polymorphism , *PLANT diversity , *PLANT genetics , *FLUORESCENCE - Abstract
Abstract: Italian lentil landraces are principally cultivated for self or local consumption. Most of them are disappearing, particularly macrosperma types by being less required by the market. A pre-requisite for the conservation and the efficient use of genetic resources is the better understanding of the extent and the distribution of the existing genetic variation, useful for future breeding programmes. Our study was undertaken to analyse and quantify the genetic diversity within and among three macrosperma Italian lentil landraces (Onano, Altamura and Villalba), using fluorescent AFLP markers. AFLP markers generated information to differentiate among closely related genotypes and group within the same cluster individuals belonging to the same landrace. The total genetic diversity (H T), the genetic diversity within population (H S) and the extent of differentiation between populations (D ST) were 0.198, 0.155 and 0.043, respectively. The fixation index (G ST =0.219) showed that about 78% of the observed total genetic variation can be attributed to within population differences and around 22% is due to differences among populations. The gene flow estimate (N m =1.774) and the mean genetic distance value (0.077) suggested narrow genetic base among the analysed populations, confirming the tendency of Italian lentil landraces to group together. The present study showed that fluorescence-based AFLP technique is a biotechnological tool that can provide significant insights for research in genetic diversity of lentil landraces and their subsequent conservation and utilization in breeding programs. [Copyright &y& Elsevier]
- Published
- 2009
- Full Text
- View/download PDF
23. Flow cytometry, SSR and modified AFLP markers for the identification of zygotic plantlets in backcrosses between ‘Femminello’ lemon cybrids (2n and 4n) and a diploid clone of ‘Femminello’ lemon (Citrus limon L. Burm. F.) tolerant to mal secco disease
- Author
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Scarano, Maria-Teresa, Tusa, Nicasio, Abbate, Loredana, Lucretti, Sergio, Nardi, Luca, and Ferrante, Sergio
- Subjects
- *
LIMONOIDS , *FLOW cytometry , *GEL electrophoresis - Abstract
One of the most important goals in the genetic improvement of lemon in the Mediterranean area is certainly the obtainment of new genotypes tolerant or resistant to mal secco disease. Two lemon cybrids (one diploid and one autotetraploid) with an intermediate degree of resistance to mal secco disease, spontaneously obtained by symmetric protoplast fusion between ‘Valencia’ orange and ‘Femminello’ lemon, were used as mother plants in backcrosses with a diploid clone of ‘Femminello’ lemon tolerant to the mal secco disease (LTMS) to improve tolerance and fruit quality. Since ‘Femminello’ lemon normally reproduces apomictically by nucellar embryony, an embryo-rescue technique was applied 105 days after pollination to recover zygotic embryos. Sixty-six plantlets were regenerated as follows: 20 from the 4nX2n cross and 46 from the 2nX2n cross. In order to distinguish zygotic embryos from nucellars, flow cytometry was applied on all the 66 plantlets; and a modified amplified fragment-length polymorphism (AFLP) technique and microsatellite analysis were carried out on a sample of 20 seedlings (six triploids and 14 diploids). Here, we report the usefulness and reliability of the tested techniques and discuss the results obtained. Flow cytometry (FCM) was useful only in inter-ploid crosses, identifying six triploids, four of which were also detected by simple sequence repeat (SSR) analysis and only two by modified AFLP. Among the 14 tested diploids, five zygotic genotypes were identified by SSR and four by modified AFLP. In our hands, the SSR technique appeared to be more suitable than modified AFLP. [Copyright &y& Elsevier]
- Published
- 2003
- Full Text
- View/download PDF
24. Best practices in plant cytometry.
- Author
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Galbraith D, Loureiro J, Antoniadi I, Bainard J, Bureš P, Cápal P, Castro M, Castro S, Čertner M, Čertnerová D, Chumová Z, Doležel J, Giorgi D, Husband BC, Kolář F, Koutecký P, Kron P, Leitch IJ, Ljung K, Lopes S, Lučanová M, Lucretti S, Ma W, Melzer S, Molnár I, Novák O, Poulton N, Skalický V, Sliwinska E, Šmarda P, Smith TW, Sun G, Talhinhas P, Tárnok A, Temsch EM, Trávníček P, and Urfus T
- Subjects
- Flow Cytometry, Protoplasts
- Published
- 2021
- Full Text
- View/download PDF
25. FISHIS: fluorescence in situ hybridization in suspension and chromosome flow sorting made easy.
- Author
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Giorgi D, Farina A, Grosso V, Gennaro A, Ceoloni C, and Lucretti S
- Subjects
- DNA, Plant genetics, Fluorescent Dyes metabolism, Genomics, Nucleic Acid Amplification Techniques, Nucleic Acid Denaturation, Plant Leaves genetics, Suspensions, Triticum genetics, Chromosomes, Plant genetics, Flow Cytometry methods, In Situ Hybridization, Fluorescence methods
- Abstract
The large size and complex polyploid nature of many genomes has often hampered genomics development, as is the case for several plants of high agronomic value. Isolating single chromosomes or chromosome arms via flow sorting offers a clue to resolve such complexity by focusing sequencing to a discrete and self-consistent part of the whole genome. The occurrence of sufficient differences in the size and or base-pair composition of the individual chromosomes, which is uncommon in plants, is critical for the success of flow sorting. We overcome this limitation by developing a robust method for labeling isolated chromosomes, named Fluorescent In situ Hybridization In suspension (FISHIS). FISHIS employs fluorescently labeled synthetic repetitive DNA probes, which are hybridized, in a wash-less procedure, to chromosomes in suspension following DNA alkaline denaturation. All typical A, B and D genomes of wheat, as well as individual chromosomes from pasta (T. durum L.) and bread (T. aestivum L.) wheat, were flow-sorted, after FISHIS, at high purity. For the first time in eukaryotes, each individual chromosome of a diploid organism, Dasypyrum villosum (L.) Candargy, was flow-sorted regardless of its size or base-pair related content. FISHIS-based chromosome sorting is a powerful and innovative flow cytogenetic tool which can develop new genomic resources from each plant species, where microsatellite DNA probes are available and high quality chromosome suspensions could be produced. The joining of FISHIS labeling and flow sorting with the Next Generation Sequencing methodology will enforce genomics for more species, and by this mightier chromosome approach it will be possible to increase our knowledge about structure, evolution and function of plant genome to be used for crop improvement. It is also anticipated that this technique could contribute to analyze and sort animal chromosomes with peculiar cytogenetic abnormalities, such as copy number variations or cytogenetic aberrations.
- Published
- 2013
- Full Text
- View/download PDF
26. Identification of zygotic and nucellar seedlings in citrus interploid crosses by means of isozymes, flow cytometry and ISSR-PCR.
- Author
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Tusa N, Abbate L, Ferrante S, Lucretti S, and Scarano MT
- Subjects
- Citrus enzymology, Crosses, Genetic, Diploidy, Flow Cytometry, Isoenzymes genetics, Minisatellite Repeats, Plant Diseases genetics, Plant Diseases microbiology, Polymerase Chain Reaction methods, Polyploidy, Seedlings genetics, Citrus genetics
- Abstract
'Milam' (a purported hybrid of Citrus jambhiri Lush) + 'Femminello' lemon (Citrus limon L. Burm. f.) allotetraploid somatic hybrids were used as pollen parents in interploid crosses with diploid 'Femminello' lemon to achieve mal secco tolerance in different populations of seedless triploid lemon types with good fruit quality. A total of 137 plantlets were obtained and subjected to screening experiments, in order to distinguish zygotic embryos from nucellars. Here we report on and discuss the results obtained with three techniques: flow cytometry, isozyme analysis and ISSR-PCR (the inter-simple sequence repeats-polymerase chain reaction). ISSR-PCR resulted to be a very efficient and reliable technique for the identification of zygotic plantlets.
- Published
- 2002
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