Agriculture faces great challenges to overcome global warming and improve system sustainability, requiring access to novel genetic diversity. So far, wild populations and local landraces remain ...poorly explored. This is notably the case for the two diploid species, Brassica oleracea L. (CC, 2n=2x=18) and B. rapa L. (AA, 2n=2x=20). In order to explore the genetic diversity in both species, we have collected populations in their centre of origin, the Mediterranean basin, on a large contrasting climatic and soil gradient from northern Europe to southern sub-Saharan regions. In these areas, we also collected 14 populations belonging to five B. oleracea closely related species. Our objective was to ensure the absence of species misidentification at the seedling stage among the populations collected and to describe thereafter their origins. We combined flow cytometry, sequencing of a species-specific chloroplast genomic region, as well as cytogenetic analyses in case of unexpected results for taxonomic verification. Out of the 112 B. oleracea and 154 B. rapa populations collected, 103 and 146, respectively, presented a good germination rate and eighteen populations were misidentified. The most frequent mistake was the confusion of these diploid species with B. napus. Additionally for B. rapa, two autotetraploid populations were observed. Habitats of the collected and confirmed wild populations and landraces are described in this study. The unique plant material described here will serve to investigate the genomic regions involved in adaptation to climate and microbiota within the framework of the H2020 Prima project ‘BrasExplor’.
The most common form of the Brassica napus L. is oilseed rape. Because of its physical characteristics the seed is very mobile and therefore disposed to spillage. Uncontrolled seed loss represents ...the potential for the appearance of volunteer and feral populations of B. napus inside and outside production areas; B. napus seed remains viable in the soil for several years. The appearance dynamics of these plants is dependent on the soil seed bank potential and complex interactive characteristics of the genotype seeds and soil and agro-climatic factors. The presence of undefined pollinating resources originated from soil seed bank in the nature presents the potential for spontaneous intra- and inter-species pollination of B. napus reflected also in its genetic structure.
Oilseed rape, Brassica napus L., is one of the most important oil plants of the moderate climatic zone and a typical industrial plant. In the past, dynamics of oilseed rape production in Slovenia was ...reflected in flexible agricultural market situation because of introduction of new crops into rotation and due to financial supports from European Union. In 2010, it was cultivated on 5351ha, accounting for about 1.1% of the total production areas in Slovenia. It is mainly grown in the eastern part of Slovenia (Pomurje, Podravje) and in the Spodnje-Posavska region. From 1984 until 2010, 58 different genotypes of oilseed and fodder rape were grown in Slovenia, of which a total of 28 were registered in the National List of Varieties in each year. Average yield of oilseed rape was ranged between 1.8 and 2.9 t/ha. Under Slovenian fragmented property structure the cross-pollination between B. napus and volunteer or feral populations (within and outside the production area) can occur. In addition to that, the presence of some sexually compatible relatives which have a high affinity to cross-pollination with B. napus are found (B. rapa, B. oleracea, B. nigra, Hirchfeldia incana, Raphanus raphanistrum, Sinapis arvensis, Diplotaxis erucoides, D. tenuifolia, D. muralis, S. alba, R. sativus and Rapistrum rugosum). Uncontrolled gene flow between different forms of B. napus or sexually compatible wild relatives in the case of coexistence of different production systems has a direct impact on the varietal purity of seeds and on crop quality.
Oljna ogrščica, Brassica napus L., je najpomembnejša oljnica zmernega klimatskega pasu in tipična industrijska rastlina. V Sloveniji se je v preteklosti obseg njene pridelave zelo spreminjal zaradi prilagajanja razmeram na trgu, uvajanja novih poljščin v kolobar in zaradi neposrednih plačil s strani Evropske unije. V letu 2010 se je pridelovala na 5351ha, kar predstavlja okrog 1,1% vseh pridelovalnih površin. Največ se je pridela v vzhodni Sloveniji, in sicer v Pomurski, Podravski in Spodnje-posavski regiji. Od leta 1984 do 2010 se je na območju Slovenije pridelovalo 58 različnih genotipov oljne in krmne ogrščice, od tega je bilo 28 vpisanih v Sortno listo v posameznem letu. Povprečni pridelki oljne ogrščice se gibljejo med 1,8 in 2,9 t/ha. V slovenskih razmerah razdrobljene posestne strukture obstajajo možnost oprašitve posevkov B. napus s samosevnimi in podivjanimi populacijami (znotraj in zunaj pridelovalnih površin). Poleg tega se v pri nas pojavljajo nekateri spolno kompatibilni sorodniki, ki imajo visoko sposobnost oprašitve z B. napus (B. rapa, B. oleracea, B. nigra, Hirchfeldia incana, Raphanus raphanistrum, Sinapis arvensis, Diplotaxis erucoides, D. tenuifolia, D. muralis, S. alba, R. sativus and Rapistrum rugosum). Nenadzorovan prenos genov med različnimi pojavnimi oblikami B. napus ali spolno kompatibilnimi sorodniki v primeru soobstoja različnih sistemov pridelave neposredno vpliva na sortno čistost semenskih posevkov in na kvaliteto pridelka.
Eleven microsatellite markers were developed for the leafhoppers of the genus Aphrodes using shotgun pyrosequencing and will be used to study the genetic diversity, population structure and gene flow ...within and between species in this genus in order to assess their conservation status. The number of alleles per locus ranged from 3 to 10, while observed and expected heterozygosity values varied from 0.421–1.000 to 0.542–0.876, respectively. Cross-species amplification was successful among the four congeners.
The objective of this study was to investigate the impact of parturition type on vitality in newborn puppies, their weight gains, and survival in the first week postpartum. One hundred and ...twenty-three puppies were divided in three groups: vaginal parturition (VP), emergency (EM-CS), and elective cesarean section (EL-CS). Apgar scores were assessed 5, 15, and 60 min postpartum. Lactate and glucose concentrations were measured in amniotic fluid and umbilical blood; cortisol concentrations were measured in amniotic fluid and puppy urine. Puppies' weight gain was tracked daily for 7 days postpartum. Apgar score at 5 and 15 min was significantly better in the VP group. EL-CS puppies had significantly lower umbilical blood and amniotic fluid lactate concentrations compared to the VP group, which also had higher umbilical blood lactate concentration than EM-CS puppies. The cortisol concentration in the amniotic fluid and in urine differed significantly between the groups, with the highest concentration in the EM-CS, followed by the VP group. Glucose concentration in amniotic fluid was higher in the VP group than EM-CS group. The type of parturition had no impact on puppies' weight gain or their survival at birth; however, supportive treatment was provided for non-vital puppies. Non-invasive analysis of puppies' fluids could help in the assessment of the neonatal vitality.
This paper reports attempts to introduce management accounting in the Italian Ministry of Finance, in the Municipality of Genua and in the Province of Perugia. The report frames the developments as ...construction of actor-networks, where local translations of a general idea lead to local variations in practice.
This framing reveals that the traditional account of implementation is too simple to grasp all the complexity of introducing new technologies and practices. The actor-network perspective shows also why there cannot exist a prescription for a successful implementation. In all cases under reviews, the actors tried to achieve the same results, mobilizing resources locally accessible to them. The results inevitably differed.
Background. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has rapidly spread worldwide from the beginning of 2020. Quantitative reverse transcription-PCR (RT-qPCR) is, to ...this day, the preferred methodology for viral RNA detection, even if not without problems. To overcome some of the limitations still existing for the detection and quantification of nucleic acids in various applications, the use of one-step reverse transcription-droplet digital PCR (RT-ddPCR) has been established. The purpose of this study was, then, to evaluate the efficacy of ddPCR for the detection of SARS-CoV-2 RNA in nasopharyngeal swabs, optimizing the detection of low-viral load-burdened samples. Methods. The RT-ddPCR workflow was validated for sensitivity, specificity, linearity, reproducibility, and precision using samples from 90 COVID-19-infected patients referred to the Department of Laboratory Medicine of the University Hospital of Udine (Italy). Results. The present study shows that RT-ddPCR allows the detection of as low as 10.3 copies of a SARS-COV-2 E-gene per sample with a higher level of accuracy and precision, especially at low concentration. Conclusion. During the postpeak phase of the SARS-CoV-2 pandemic, it is essential to rely on a highly robust molecular biology method to identify infected subjects, whether they have symptoms or not, in order to prepare appropriate containment measures.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has rapidly spread worldwide from the beginning of 2020. The presence of viral RNA in samples by nucleic acid (NA) molecular ...analysis is the only method available to diagnose COVID-19 disease and to assess patients’ viral load. Since the demand for laboratory reagents has increased, there has been a worldwide shortage of RNA extraction kits. We, therefore, developed a fast and cost-effective viral genome isolation method that, combined with quantitative RT-PCR assay, detects SARS-CoV-2 RNA in patient samples. The method relies on the addition of Proteinase K followed by a controlled heat-shock incubation and, then, E gene evaluation by RT-qPCR. It was validated for sensitivity, specificity, linearity, reproducibility, and precision. It detects as low as 10 viral copies/sample, is rapid, and has been characterized in 60 COVID-19-infected patients. Compared to automated extraction methods, our pretreatment guarantees the same positivity rate with the advantage of shortening the time of the analysis and reducing its cost. This is a rapid workflow meant to aid the healthcare system in the rapid identification of infected patients, such as during a pathogen-related outbreak. For its intrinsic characteristics, this workflow is suitable for large-scale screenings.
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