Genetic improvement of tomato by targeted control of fruit softening

Controlling the rate of softening to extend shelf life was a key target for researchers engineering genetically modified (GM) tomatoes in the 1990s, but only modest improvements were achieved. Hybrids grown nowadays contain 'non-ripening mutations' that slow ripening and improve shelf life, but adversely affect flavor and color. We report substantial, targeted control of tomato softening, without affecting other aspects of ripening, by silencing a gene encoding a pectate lyase

Analyses and localization of pectin-like carbohydrates in cell wall and mucilage of the green alga Netrium digitus

The unicellular, simply shaped desmid Netrium digitus inhabiting acid bog ponds grows in two phases. Prior to division, the cell elongates at its central zone, whereas in a second phase, polar tip growth occurs. Electron microscopy demonstrates that Netrium is surrounded by a morphologically homogeneous cell wall, which lacks pores. Immunocytochemical and biochemical analyses give insight into physical wall properties and, thus, into adaptation to the extreme environment. The monoclonal antibodies JIM5 and JIM7 directed against pectic epitopes with different degrees of esterification label preferentially growing wall zones in Netrium. In contrast, 2F4 marks the cell wall only after experimental de-esterification. Electron energy loss spectroscopy reveals Ca-binding capacities of pectins and gives indirect evidence for the degree of their esterification. An antibody raised against Netrium mucilage is not only specific to mucilage but also recognizes wall components in transmission electron microscopy and dot blots. These results indicate a smooth transition between mucilage and the cell wall in Netrium

Unveiling microbial structures during raw microalgae digestion and co-digestion with primary sludge to produce biogas using semi-continuous AnMBR systems

[EN] Methane production from microalgae can be enhanced through anaerobic co-digestion with carbon-rich substrates and thus mitigate the inhibition risk associated with its low C:N ratio. Acclimated microbial communities for microalgae disruption can be used as a source of natural enzymes in bioenergy production. However, co-substrates with a certain microbial diversity such as primary sludge might shift the microbial structure. Substrates were generated in a Water Resource Recovery Facility (WRRF) and combined as follows: Scenedesmus or Chlorella digestion and microalgae co-digestion with primary sludge. The study was performed using two lab-scale Anaerobic Membrane Bioreactors (AnMBR). During three years, different feedstocks scenarios for methane production were evaluated with a special focus on the microbial diversity of the AnMBR. 57% of the population was shared between the different feedstock scenarios, revealing the importance of Anaerolineaceae members besides Smithella and Methanosaeta genera. The addition of primary sludge enhanced the microbial diversity of the system during both Chlorella and Scenedesmus co-digestion and promoted different microbial structures. Aceticlastic methanogen Methanosaeta was dominant in all the feedstock scenarios. A more remarkable role of syntrophic fatty acid degraders (Smithella, Syntrophobacteraceae) was observed during co-digestion when only microalgae were digested. However, no significant changes were observed in the microbial composition during anaerobic microalgae digestion when feeding only Chlorella or Scenedesmus. This is the first work revealing the composition of complex communities for semi-continuous bioenergy production from WRRF streams. The stability and maintenance of a microbial core over-time in semi-continuous AnMBRs is here shown supporting their future application in full-scale systems for raw microalgae digestion or codigestion.The Ministry of Economy and Competitiveness (MINECO) and the European Regional Development Fund (ERDF) are gratefully acknowledged for their support to this research work through CTM2011-28595-C02-02 and CTM2014-54980-C2-1-R projects. The authors are thankful to Ph.D. Silvia Greses and Ph.D. candidate Rebecca Serna-Garcia (Universitat de Valencia, Spain) for allowing the collection of digestate samples from their bioreactors and providing a brief data characterization of their performance. As well, authors thank the support of Maria Paches (IIAMA, Valencia, Spain) during phytoplankton monitoring in the photobioreactor plant. Finally, the sequencing service from FISABIO (Valencia, Spain) is also gratefully acknowledged for their technical support during the design stage of this work.Zamorano-López, N.; Borrás, L.; Seco, A.; Aguado García, D. (2020). Unveiling microbial structures during raw microalgae digestion and co-digestion with primary sludge to produce biogas using semi-continuous AnMBR systems. The Science of The Total Environment. 699:1-12. https://doi.org/10.1016/j.scitotenv.2019.134365S112699APHA, APHA/AWWA/WEF, 2012. In: Standard Methods for the Examination of Water and Wastewater. Stand. Methods, pp. 541 doi.org/ISBN 9780875532356.Astals, S., Musenze, R. S., Bai, X., Tannock, S., Tait, S., Pratt, S., & Jensen, P. D. (2015). Anaerobic co-digestion of pig manure and algae: Impact of intracellular algal products recovery on co-digestion performance. Bioresource Technology, 181, 97-104. doi:10.1016/j.biortech.2015.01.039Baudelet, P.-H., Ricochon, G., Linder, M., & Muniglia, L. (2017). 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Journal of Environmental Management, 218, 425-434. doi:10.1016/j.jenvman.2018.04.086Herrmann, C., Kalita, N., Wall, D., Xia, A., & Murphy, J. D. (2016). Optimised biogas production from microalgae through co-digestion with carbon-rich co-substrates. Bioresource Technology, 214, 328-337. doi:10.1016/j.biortech.2016.04.119Ju, F., Lau, F., & Zhang, T. (2017). Linking Microbial Community, Environmental Variables, and Methanogenesis in Anaerobic Biogas Digesters of Chemically Enhanced Primary Treatment Sludge. Environmental Science & Technology, 51(7), 3982-3992. doi:10.1021/acs.est.6b06344Kadnikov, V. V., Mardanov, A. V., Beletsky, A. V., Karnachuk, O. V., & Ravin, N. V. (2019). Genome of the candidate phylum Aminicenantes bacterium from a deep subsurface thermal aquifer revealed its fermentative saccharolytic lifestyle. Extremophiles, 23(2), 189-200. doi:10.1007/s00792-018-01073-5Klassen, V., Blifernez-Klassen, O., Wobbe, L., Schlüter, A., Kruse, O., & Mussgnug, J. H. (2016). Efficiency and biotechnological aspects of biogas production from microalgal substrates. Journal of Biotechnology, 234, 7-26. doi:10.1016/j.jbiotec.2016.07.015Klassen, V., Blifernez-Klassen, O., Wibberg, D., Winkler, A., Kalinowski, J., Posten, C., & Kruse, O. (2017). Highly efficient methane generation from untreated microalgae biomass. Biotechnology for Biofuels, 10(1). doi:10.1186/s13068-017-0871-4Leng, L., Yang, P., Singh, S., Zhuang, H., Xu, L., Chen, W.-H., … Lee, P.-H. (2018). A review on the bioenergetics of anaerobic microbial metabolism close to the thermodynamic limits and its implications for digestion applications. Bioresource Technology, 247, 1095-1106. doi:10.1016/j.biortech.2017.09.103Li, R., Duan, N., Zhang, Y., Liu, Z., Li, B., Zhang, D., & Dong, T. (2017). Anaerobic co-digestion of chicken manure and microalgae Chlorella sp.: Methane potential, microbial diversity and synergistic impact evaluation. 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EEF2 Analysis Challenges the Monophyly of Archaeplastida and Chromalveolata

BACKGROUND: Classification of eukaryotes provides a fundamental phylogenetic framework for ecological, medical, and industrial research. In recent years eukaryotes have been classified into six major supergroups: Amoebozoa, Archaeplastida, Chromalveolata, Excavata, Opisthokonta, and Rhizaria. According to this supergroup classification, Archaeplastida and Chromalveolata each arose from a single plastid-generating endosymbiotic event involving a cyanobacterium (Archaeplastida) or red alga (Chromalveolata). Although the plastids within members of the Archaeplastida and Chromalveolata share some features, no nucleocytoplasmic synapomorphies supporting these supergroups are currently known. METHODOLOGY/PRINCIPAL FINDINGS: This study was designed to test the validity of the Archaeplastida and Chromalveolata through the analysis of nucleus-encoded eukaryotic translation elongation factor 2 (EEF2) and cytosolic heat-shock protein of 70 kDa (HSP70) sequences generated from the glaucophyte Cyanophora paradoxa, the cryptophytes Goniomonas truncata and Guillardia theta, the katablepharid Leucocryptos marina, the rhizarian Thaumatomonas sp. and the green alga Mesostigma viride. The HSP70 phylogeny was largely unresolved except for certain well-established groups. In contrast, EEF2 phylogeny recovered many well-established eukaryotic groups and, most interestingly, revealed a well-supported clade composed of cryptophytes, katablepharids, haptophytes, rhodophytes, and Viridiplantae (green algae and land plants). This clade is further supported by the presence of a two amino acid signature within EEF2, which appears to have arisen from amino acid replacement before the common origin of these eukaryotic groups. CONCLUSIONS/SIGNIFICANCE: Our EEF2 analysis strongly refutes the monophyly of the Archaeplastida and the Chromalveolata, adding to a growing body of evidence that limits the utility of these supergroups. In view of EEF2 phylogeny and other morphological evidence, we discuss the possibility of an alternative eukaryotic supergroup

The structure and biochemistry of charophycean cell walls: I. Pectins of Penium margaritaceum

Plant cell walls are essential for proper growth, development, and interaction with the environment. It is generally accepted that land plants arose from aquatic ancestors which are sister groups to the charophycean algae (i.e., Streptophyta), and study of wall evolution during this transition promises insight into structure-function relationships of wall components. In this paper, we explore wall evolutionary history by studying the incorporation of pectin polymers into cell walls of the model organism Penium margaritaceum, a simple single-cell desmid. This organism produces only a primary wall consisting of three fibrillar or fibrous layers, with the outermost stratum terminating in distinct, calcified projections. Extraction of isolated cell walls with trans-1,2-diaminocyclohexane-N,N,N′,N′-tetraacetic acid yielded a homogalacturonan (HGA) that was partially methyl esterified and equivalent to that found in land plants. Other pectins common to land plants were not detected, although selected components of some of these polymers were present. Labeling with specific monoclonal antibodies raised against higher-plant HGA epitopes (e.g., JIM5, JIM7, LM7, 2F4, and PAM1) demonstrated that the wall complex and outer layer projections were composed of the HGA which was significantly calcium complexed. JIM5 and JIM7 labeling suggested that highly methyl esterified HGA was secreted into the isthmus zone of dividing cells, the site of active wall secretion. As the HGA was displaced to more polar regions, de-esterification in a non-blockwise fashion occurred. This, in turn, allowed for calcium binding and the formation of the rigid outer wall layer. The patterning of HGA deposition provides interesting insights into the complex process of pectin involvement in the development of the plant cell wall. © 2006 Springer-Verlag