Uso de las proteínas de reserva de megagametofito como marcador de la diversidad genética en Abies pinsapo

El pinsapo (Abies pinsapo Boiss.) se localiza exclusivamente en el sur de España, en las comarcas de Cádiz y Málaga, donde ocupa unas 5.000 ha. Diversos factores como la presencia de plagas y el decaimiento de las masas, de origen aún no determinado, hacen temer por el futuro de la especie. A todo ello hay que sumar la circunstancia de que es posible que se esté produciendo un estrechamiento de la base genética de la especie, debido a problemas de consanguinidad y deriva genética. Por esta razón, el desarrollo de una estrategia de conservación de la especie exige la evaluación de su diversidad genética. En este estudio, se han analizado la posibilidad de utilizar las proteínas del megagametofito de pinsapo como marcador de la diversidad genética de las masas de esta especie. La fracción albúminas ha mostrado la existencia de polimorfismo. Un análisis preliminar efectuado en 27 árboles, permitió identificar más de 41 bandas de las que, al menos 12 son polimórficas entre árboles y 8 dentro de árboles. Este resultado permite ser optimista sobre la posibilidad de emplear este marcador con la finalidad señalad

Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis

[EN] The oppA2 gene encodes an oligopeptide-binding protein similar to the periplasmic substrate-binding proteins of the ABC transport systems. However, oppA2 is an orphan gene, not included in an ABC operon. This gene is located in the clavulanic acid (CA) gene cluster of Streptomyces clavuligerus and is essential for CA production. A transcriptomic study of the oppA2-null mutant S. clavuligerus ΔoppA2::aac showed changes in the expression levels of 233 genes from those in the parental strain. These include genes for ABC transport systems, secreted proteins, peptidases, and proteases. Expression of the clavulanic acid, clavam, and cephamycin C biosynthesis gene clusters was not significantly affected in the oppA2 deletion mutant. The genes for holomycin biosynthesis were upregulated 2-fold on average, and the level of upregulation increased to 43-fold in a double mutant lacking oppA2 and the pSCL4 plasmid. Strains in which oppA2 was mutated secreted into the culture the compound N-acetylglycyl-clavaminic acid (AGCA), a putative intermediate of CA biosynthesis. A culture broth containing AGCA, or AGCA purified by liquid chromatography-mass spectrometry (LC-MS), was added to the cultures of various non-CA-producing mutants. Mutants blocked in the early steps of the pathway restored CA production, whereas mutants altered in late steps did not, establishing that AGCA is a late intermediate of the biosynthetic pathway, which is released from the cells when the oligopeptide-binding protein OppA2 is not availableSIThis work was supported by grant BIO2013-34723 from the Spanish Ministry of Economy and Competitiveness. R. Álvarez-Álvarez and Y. Martínez-Burgo received a PFU fellowship from the Spanish Ministry of Science and Innovatio

Esterification of Free Fatty Acids with Glycerol within the Biodiesel Production Framework

Companies in the field of the collection and treatment of waste cooking oils (WCO) for subsequent biodiesel production usually have to cope with high acidity oils, which cannot be directly transformed into fatty acid methyl esters due to soap production. Since glycerine is the main byproduct of biodiesel production, these high acidity oils could be esterified with the glycerine surplus to transform the free fatty acids (FFA) into triglycerides before performing the transesterification. In this work, commercial glycerol was esterified with commercial fatty acids and commercial fatty acid/lampante olive oil mixtures over tin (II) chloride. In the first set of experiments, the esterification of linoleic acid with glycerol excess from 20 to 80% molar over the stoichiometric was performed. From 20% glycerol excess, there was no improvement in FFA reduction. Using 20% glycerol excess, the performance of a biochar obtained from heavy metal-contaminated plant roots was compared to that of SnCl2. Then, the effect of the initial FFA content was assessed using different oleic acid/lampante olive oil mixtures. The results illustrated that glycerolysis was impeded at initial FFA contents lower than 10%. Finally, the glycerolysis of a WCO with 9.94% FFA was assayed, without success

Molecular genetics of naringenin biosynthesis, a typical plant secondary metabolite produced by Streptomyces clavuligerus

Background: Some types of flavonoid intermediates seemed to be restricted to plants. Naringenin is a typical plant metabolite, that has never been reported to be produced in prokariotes. Naringenin is formed by the action of a chal cone synthase using as starter 4-coumaroyl-CoA, which in dicotyledonous plants derives from phenylalanine by the action of a phenylalanine ammonia lyase. Results: A compound produced by Streptomyces clavuligerus has been identified by LC–MS and NMR as naringenin and coelutes in HPLC with a naringenin standard. Genome mining of S. clavuligerus revealed the presence of a gene for a chalcone synthase (ncs), side by side to a gene encoding a P450 cytochrome (ncyP) and separated from a gene encoding a Pal/Tal ammonia lyase (tal). Deletion of any of these genes results in naringenin non producer mutants. Complementation with the deleted gene restores naringenin production in the transformants. Furthermore, narin genin production increases in cultures supplemented with phenylalanine or tyrosine. Conclusion: This is the first time that naringenin is reported to be produced naturally in a prokariote. Interestingly three non-clustered genes are involved in naringenin production, which is unusual for secondary metabolites. A ten tative pathway for naringenin biosynthesis has been proposed

Effect of mycoviruses on growth, spore germination and pathogenicity of the fungus Fusarium circinatum

Producción CientíficaAim of the study: To assess the impact on two mycoviruses recently described in F. circinatum mitovirus 1, and 2-2 (FcMV1 and FcMV2-2) on i) mycelial growth, ii) spore germination and iii) relative necrosis. Material and methods: Fourteen monosporic strains of F. circinatum (one of each pair infected with mycoviruses and the other without them) of the pathogen with and without viruses were selected for the assay. The statistical analysis, were a linear mixed model of analysis of variance considering one between-subjects factor (isolate) and one within-subjects factor with four levels (1=without viruses, 2=only virus FcMV1, 3=only virus FcMV2-2 and 4=both viruses). Main results: Colony growth rates of F. circinatum isolates were significantly reduced in presence of mycoviruses (p=0.002). The spore germination was also reduced in the F. circinatum isolates containing mycovirus as compared to mycovirus-free isolates (p<0.001). No significant differences in lesion lengths caused by F. circinatum were found in relation to the presence/absence of mycovirus (p<0.61). Research highlights: Reduction of the percentage of spore germination in the isolates of F. circinatum with mycovirus infections, as compared to free isolates, provides indications of reduction of metabolic activity and plant physiology are discussed. The lack of significant differences found in the length of the lesions caused by F. circinatum with respect to the presence/absence of mycovirus, indicates that further studies with a larger number of variables are required.Ministerio de Economía, Industria y Competitividad (projects AGL2012-39912 and AGL2015- 69370-R)European Cooperation in Science and Technology (COST Action FP1406 PINESTRENGTH)Portuguese Foundation for Science and Technology (grant SFRH/BPD/122928/2016