Alternative Splicing in the Anthocyanin Fruit Gene Encoding an R2R3 MYB Transcription Factor Affects Anthocyanin Biosynthesis in Tomato Fruits

Abstract Tomato (Solanum lycopersicum L.) fruits are typically red at ripening, with high levels of carotenoids and a low content in flavonoids. Considerable work has been done to enrich the spectrum of their health-beneficial phytochemicals, and interspecific crosses with wild species have successfully led to purple anthocyanin-colored fruits. The Aft (Anthocyanin fruit) tomato accession inherited from Solanum chilense the ability to accumulate anthocyanins in fruit peel through the introgression of loci controlling anthocyanin pigmentation, including four R2R3 MYB transcription factor encoding genes. Here, we carried out a comparative functional analysis of these transcription factors in wild type and Aft plants, testing their ability to take part in the transcriptional complexes that regulate the biosynthetic pathway and their efficiency in inducing anthocyanin pigmentation. Significant differences emerged for SlAN2like, both in the expression level and protein functionality, with splicing mutations determining a complete loss-of-function of the wild type protein. This transcription factor thus appears to play a key role in the anthocyanin fruit pigmentation. Our data provide new clues to the long-awaited genetic bases of the Aft phenotype and also contribute to clarify why domesticated tomato fruits display a homogeneous red coloration without the typical purple streaks observed in wild tomato species

The cyclic peptide G4CP2 enables the modulation of galactose metabolism in yeast by interfering with GAL4 transcriptional activity

Genetically-encoded combinatorial peptide libraries are convenient tools to identify peptides to be used as therapeutics, antimicrobials and functional synthetic biology modules. Here, we report the identification and characterization of a cyclic peptide, G4CP2, that interferes with the GAL4 protein, a transcription factor responsible for the activation of galactose catabolism in yeast and widely exploited in molecular biology. G4CP2 was identified by screening CYCLIC, a Yeast Two-Hybrid-based combinatorial library of cyclic peptides developed in our laboratory. G4CP2 interferes with GAL4-mediated activation of galactose metabolic enzymes both when expressed intracellularly, as a recombinant peptide, and when provided exogenously, as a chemically-synthesized cyclic peptide. Our results support the application of G4CP2 in microbial biotechnology and, additionally, demonstrate that CYCLIC can be used as a tool for the rapid identification of peptides, virtually without any limitations with respect to the target protein. The possible biotechnological applications of cyclic peptides are also discussed

Update on the diagnosis of tuberculosis

Background Tuberculosis remains a global public health threat, and the development of rapid and precise diagnostic tools is the key to enabling the early start of treatment, monitoring response to treatment, and preventing the spread of the disease. Objective An overview of recent progress in host- and pathogen-based tuberculosis diagnostics. Sources We conducted a PubMed search of recent relevant articles and guidelines on tuberculosis screening and diagnosis. Content An overview of currently used methods and perspectives in the following areas of tuberculosis diagnostics is provided: immune-based diagnostics, X-ray, clinical symptoms and scores, cough detection, culture of Mycobacterium tuberculosis and identifying its resistance profile using phenotypic and genotypic methods, including next generation sequencing, sputum- and non-sputum-based molecular diagnosis of tuberculosis and monitoring of response to treatment. Implications A brief overview of the most relevant advances and changes in international guidelines regarding screening and diagnosing tuberculosis is provided in this review. It aims at reviewing all relevant areas of diagnostics, including both pathogen- and host-based methods.PostprintPeer reviewe

Updating the approaches to define susceptibility and resistance to anti-tuberculosis agents: implications for diagnosis and treatment

11 páginas, 2 figuras, 1 tablaInappropriately high breakpoints have resulted in systematic false-susceptible AST results to anti-TB drugs. MIC, PK/PD and clinical outcome data should be combined when setting breakpoints to minimise the emergence and spread of antimicrobial resistance.I. Comas was supported by PID2019-104477RB-I00 from the Spanish Science Ministry and by ERC (CoG 101001038)Peer reviewe

Exploring legume-rhizobia symbiotic models for waterlogging tolerance

Unexpected and increasingly frequent extreme precipitation events result in soil flooding or waterlogging. Legumes have the capacity to establish a symbiotic relationship with endosymbiotic atmospheric dinitrogen-fixing rhizobia, thus contributing to natural nitrogen soil enrichment and reducing the need for chemical fertilization. The impact of waterlogging on nitrogen fixation and legume productivity needs to be considered for crop improvement. This review focuses on the legumes-rhizobia symbiotic models. We aim to summarize the mechanisms underlying symbiosis establishment, nodule development and functioning under waterlogging. The mechanisms of oxygen sensing of the host plant and symbiotic partner are considered in view of recent scientific advances

Synthetic Protein Circuits and Devices Based on Reversible Protein-Protein Interactions: An Overview

Protein-protein interactions (PPIs) contribute to regulate many aspects of cell physiology and metabolism. Protein domains involved in PPIs are important building blocks for engineering genetic circuits through synthetic biology. These domains can be obtained from known proteins and rationally engineered to produce orthogonal scaffolds, or computationally designed de novo thanks to recent advances in structural biology and molecular dynamics prediction. Such circuits based on PPIs (or protein circuits) appear of particular interest, as they can directly affect transcriptional outputs, as well as induce behavioral/adaptational changes in cell metabolism, without the need for further protein synthesis. This last example was highlighted in recent works to enable the production of fast-responding circuits which can be exploited for biosensing and diagnostics. Notably, PPIs can also be engineered to develop new drugs able to bind specific intra- and extra-cellular targets. In this review, we summarize recent findings in the field of protein circuit design, with particular focus on the use of peptides as scaffolds to engineer these circuits

Auxin is required for the long coleoptile trait in japonica rice under submergence

Rice coleoptile elongation under submergence guarantees fast seedling establishment in the field. We investigated the role of auxin in influencing the capacity of rice to produce a long coleoptile under water. In order to explore the complexity of auxin's role in coleoptile elongation, we used gene expression analysis, confocal microscopy of an auxin-responsive fluorescent reporter, gas chromatography coupled to mass spectrometry (GC-MS/MS), and T-DNA insertional mutants of an auxin transport protein. We show that a higher auxin availability in the coleoptile correlates with the final coleoptile length under submergence. We also identified the auxin influx carrier AUX1 as a component influencing this trait under submergence. The coleoptile tip is involved in the final length of rice varieties harbouring a long coleoptile. Our experimental results indicate that auxin biosynthesis and transport underlies the differential elongation between short and long coleoptile-harbouring japonica rice varieties