Desentrañando los intricados patrones de variación en el complejo diploid-poliploide complex Veronica subsect. Pentasepalae Benth. (Veronica L., Plantaginaceae sensu APG IV). Relación con las variaciones climáticas del periodo cuaternario

Tesis por compendio de publicaciones[ES]La poliploidía y la hibridación serán los leitmotive de los capítulos que componen esta Tesis Doctoral, cuyo objetivo principal es contribuir al entendimiento de la historia evolutiva y las relaciones entre las entidades que forman parte del complejo diploide-poliploide Veronica subsección Pentasepalae Benth

Conservation of genetic diversity in Mediterranean endemic species: Arenaria balearica (Caryophyllaceae)

Background – Biodiversity loss is a problem that needs to be urgently addressed, particularly with the uncertainties of climate change. Current conservation policies principally focus on endangered species but they often give little consideration to the evolutionary processes, genetic diversity, or the rarity of nonendangered species. Endemic species occurring in rocky habitats that are undergoing exceptional habitat loss appear to be one of the most important candidates for conservation. The aim is to establish in situ and ex situ conservation recommendations for the Mediterranean endemic species Arenaria balearica. Material and methods – Arenaria balearica is a species endemic to the Mediterranean with a disjunct distribution range throughout Majorca, Corsica, Sardinia, and other small Tyrrhenian islands. A combination of molecular techniques (AFLPs and plastid DNA) was employed to determine genetic diversity and rarity across populations and to calculate the Relevant Genetic Units for Conservation (RGUCs). Moreover, Species Distribution Models (SDMs) were developed to assess the potential current distribution and the expected situation under future climatic scenarios. Key results – To preserve the genetic diversity and rarity of the species, in situ conservation is proposed for six populations as RGUCs. Moreover, as the RGUCs can only account for a part of the phylogeographic signal, ex situ conservation is also suggested for some additional populations. According to the results, the habitat suitability in the 2050 scenario is limited and suitable areas for A. balearica could have disappeared by 2070. Therefore, the persistence of the species could be in danger in a short period of time and conservation planning becomes necessary.Instituto de Salud Carlos III Spanish Government CGL2010-1635

Ferrocene-Decorated Phenol Derivatives by Trapping ortho-Quinone Methide Intermediates with Ferrocene

The InCl3-catalyzed reaction of ferrocene with ortho-hydroxybenzyl alcohols is reported and represents a convenient route for the synthesis of ferrocenyl phenols. This carbon–carbon bond forming process is believed to proceed through an ortho-quinone methide intermediate that can be intercepted by ferrocene through a Friedel–Crafts-type process. Preliminary cytotoxic screening carried out on several cancer cell lines revealed that some of the compounds exhibit moderate cytotoxicity

Characterization of 12 Polymorphic SSR Markers in Veronica Subsect.Pentasepalae (Plantaginaceae) and Cross-Amplification in 10 Other Subgenera

[EN] • Premise of the study: Microsatellite primers were developed in the perennial herbs of the diploid-polyploid complex Veronica subsect. Pentasepalae (Plantaginaceae) to investigate the role that hybridization has played in the evolution of the group, which includes several endangered species. • Methods and Results: Twelve pairs of primers leading to polymorphic and readable markers were identifi ed and optimized from V. jacquinii and V. orbiculata using a microsatellite-enriched library method and 454 GS-FLX technique. The set of primers amplifi ed dinucleotide to pentanucleotide repeats, and the number of alleles per locus ranged from one to six, one to 11, and one to nine for V. orsiniana , V. javalambrensis , and V. rosea , respectively. Transferability analyses were performed in 20 species corresponding to 10 different subgenera. • Conclusions: These results indicate the utility of the newly developed microsatellites across Veronica subsect. Pentasepalae , which will help in the study of gene fl ow patterns and genetic structure

Trapping para-quinone methide intermediates with ferrocene: Synthesis and preliminary biological evaluation of new phenol-ferrocene conjugates

The reaction of para-hydroxybenzyl alcohols with ferrocene in the presence of a catalytic amount of InCl3 provided ferrocenyl phenol derivatives, an interesting class of organometallic compounds with potential applications in medicinal chemistry. This transformation exhibited a reasonable substrate scope delivering the desired products in synthetically useful yields. Evidence of involvement of a para-quinone methide intermediate in this coupling process was also provided. Preliminary biological evaluation demonstrated that some of the ferrocene derivatives available by this methodology exhibit significant cytotoxicity against several cancer cell lines with IC50 values within the range of 1.07–4.89 μM

IAPT chromosome data 28

[EN] Veronica angustifolia (Vahl) Bernh.; Veronica austriaca subsp. jacquinii (Baumg.) Watzl; Veronica dalmatica N.Pad.Gar., Rojas-Andrés, López-González & M.M.Mart.Ort.; Veronica kindlii Adam.; Veronica orsiniana Ten.; Veronica prostrata L.; Veronica rosea Desf.; Veronica sennenii (Pau) M.M.Mart.Ort. & E.Rico; Veronica tenuifolia subsp. javalambrensis (Pau) Molero & J.Pujadas; Veronic tenuifolia Asso subsp. tenuifolia; Veronica teucrium L.; Veronica thracica Velen.. In: K. Marhold & J. Kucera, IAPT chromosome data 28

Combined application of N-fixing PGPB and rice straw mulch compensates N immobilization by straw, improving crop growth

[EN] Background Rice straw, often treated as waste, provides many benefits to crops when used as a mulch. However, straw degradation promotes nitrogen immobilisation due to its high C:N ratio, causing N competition between soil microorganisms and the crop. Currently, sustainable practices to remedy nitrogen immobilisation are hardly being implemented. In microcosm conditions we assessed whether the inoculation with N-fixers could offset the transient nitrogen deficiency caused by straw mulch, thereby harnessing the benefits of straw while mitigating its negative impact on nitrogen depletion and exerting a synergistic effect on crop growth. Results Inoculation with N-fixers increased the nitrogen content in the soil (the increase ranged from 14% up to 90% for NH4 + and from 20% to 60% for NO3 −) and, in most cases, also the nitrogen content in the plant (ranging from 10% to 15% increase), compared to the non-inoculated control. Therefore, inoculation would compensate for the lack of nitrogen caused by nitrogen immobilisation, and this resulted in an increased biomass production by the crop compared with the uninoculated control (the increase ranged from 25% to 85%). In addition, inoculation with N-fixers did not lead to a permanent change in the bacterial community composition, whereas straw addition increased the biodiversity of the soil microbiome. Conclusions The results obtained in microcosm conditions are a first indication that complementing straw mulching with the inoculation of N-fixers could avoid the transient N immobilisation produced during straw degradation. Thus, the benefits of the combination would be a yield increase, while improving the biodiversity of the soil microbiome, stabilising soil temperatures and increasing water soil content.SIThis work was supported by the Spanish Ministry of Science, Innovation and Universities (project RTC-2017-6249-2

Development of 14 Microsatellite Markers in Odontites vernus s.l. (Orobanchaceae) and Cross-Amplification in Related Taxa

[EN] • Premise of the study: Microsatellite primers were developed for the fi rst time in the root hemiparasite herb Odontites vernus (Orobanchaceae). These markers will be useful to investigate the role of polyploidization in the evolution of this diploid-tetraploid complex, as well as the extent of gene fl ow between different ploidy levels. • Methods and Results: Fourteen polymorphic and reproducible loci were identifi ed and optimized from O. vernus using a microsatellite-enriched library and 454 Junior sequencing. The set of primers amplifi ed di- to pentanucleotide repeats and showed two to 13 alleles per locus. Transferability was tested in 30 taxa (19 belonging to Odontites and 11 from eight other genera of Orobanchaceae tribe Rhinantheae). • Conclusions: The results indicate the utility of the newly developed microsatellites in O. vernus and several other species, which will be useful for taxon delimitation and conservation genetics studies

De novo assembly and characterization of leaf transcriptome for the development of functional molecular markers of the extremophile multipurpose tree species Prosopis alba

Background: Prosopis alba (Fabaceae) is an important native tree adapted to arid and semiarid regions of north-western Argentina which is of great value as multipurpose species. Despite its importance, the genomic resources currently available for the entire Prosopis genus are still limited. Here we describe the development of a leaf transcriptome and the identification of new molecular markers that could support functional genetic studies in natural and domesticated populations of this genus. Results: Next generation DNA pyrosequencing technology applied to P. alba transcripts produced a total of 1,103,231 raw reads with an average length of 421 bp. De novo assembling generated a set of 15,814 isotigs and 71,101 non-assembled sequences (singletons) with an average of 991 bp and 288 bp respectively. A total of 39,000 unique singletons were identified after clustering natural and artificial duplicates from pyrosequencing reads. Regarding the non-redundant sequences or unigenes, 22,095 out of 54,814 were successfully annotated with Gene Ontology terms. Moreover, simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs) were searched, resulting in 5,992 and 6,236 markers, respectively, throughout the genome. For the validation of the the predicted SSR markers, a subset of 87 SSRs selected through functional annotation evidence was successfully amplified from six DNA samples of seedlings. From this analysis, 11 of these 87 SSRs were identified as polymorphic. Additionally, another set of 123 nuclear polymorphic SSRs were determined in silico, of which 50% have the probability of being effectively polymorphic. Conclusions: This study generated a successful global analysis of the P. alba leaf transcriptome after bioinformatic and wet laboratory validations of RNA-Seq data. The limited set of molecular markers currently available will be significantly increased with the thousands of new markers that were identified in this study. This information will strongly contribute to genomics resources for P. alba functional analysis and genetics. Finally, it will also potentially contribute to the development of population-based genome studies in the genera.Fil: Torales, Susana. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Recursos Naturales. Instituto de Recursos Biológicos; ArgentinaFil: Rivarola, Maximo Lisandro. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pomponio, María Florencia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Recursos Naturales. Instituto de Recursos Biológicos; ArgentinaFil: González, Sergio Alberto. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Acuña, Cintia Vanesa. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Fernández, Paula del Carmen. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: López Lauenstein, Diego. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Fisiología y Recursos Genéticos Vegetales; ArgentinaFil: Verga, Aníbal Ramón. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Fisiología y Recursos Geneticos Vegetales; ArgentinaFil: Hopp, Horacio Esteban. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Paniego, Norma Beatriz. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Marcucci Poltri, Susana Noemí. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentin

Low sperm to egg ratio required for successful in vitro fertilization in a pair-spawning teleost, Senegalese sole (Solea senegalensis)

Cultured Senegalese sole (Solea senegalensis) breeders fail to spawn fertilized eggs. The implantation of large-scale in vitro fertilization protocols, to solve this problem, has been frustrated by low production of poor quality sperm. Cultured females were induced to ovulate with a 5 µg kg−1 single injection of gonadotropin-releasing hormone agonist (GnRHa) and viable eggs (82.6 ± 9.2% fertilization) were stripped 41:57 ± 1:46 h after the injection. Sperm was collected from cultured males, diluted in modified Leibovitz and used fresh to fertilize the eggs. Males were not treated with hormones. A nonlinear regression, an exponential rise to a maximum (R = 0.93, p < 0.0001) described the number of motile spermatozoa required to fertilize a viable egg and 1617 motile spermatozoa were sufficient to fertilize 99 ± 12% (±95% CI) of viable eggs. Similar, spermatozoa egg−1 ratios of 592 ± 611 motile spermatozoa egg−1 were used in large-scale in vitro fertilizations (190 512 ± 38 471 eggs). The sperm from a single male (145 ± 50 µl or 8.0 ± 6.8 × 108 spermatozoa) was used to fertilize the eggs. The mean hatching rate was 70 ± 14% to provide 131 540 ± 34 448 larvae per fertilization. The viability of unfertilized eggs stored at room temperature decreased gradually, and the sooner eggs were fertilized after stripping, the higher the viability of the eggs. The collection of sperm directly into a syringe containing modified Leibovitz significantly increased the percentage of motile spermatozoa (33.4 ± 12.2%) compared with other collection methods. The spz egg−1 ratios for Senegalese sole were at the lower end of ratios required for fish. Senegalese sole have a pair-spawning reproductive behaviour characterized by gamete fertilization in close proximity with no sperm competition. The provision of a large-scale in vitro fertilization protocol (200 µl of sperm per 100 ml of eggs) will enable the industry to operate sustainably and implement breeding programmes to improve production.publishedVersio