Microorganisms, microbiomes, and their products (e.g., enzymes, metabolites, antibiotics, etc.) are key players in the functioning of both natural and anthropized Earth ecosystems; they can be exploited for both research purposes and biotechnological applications, including fighting the big challenges of our era, such as climate change. Culture collections (CCs) and microbial Biological Resource Centres (mBRCs) are repositories of microorganisms that investigate and safeguard biodiversity and facilitate the scientific and industrial communities’ access to microbial strains and related know-how by providing external users with skills and services. Considering this, CCs and mBRCs are pivotal institutions for the valorisation of microorganisms, the safeguarding of life, and the fostering of excellent bioscience. The aim of this review is to present the state-of-the-art of Italian CCs and mBRCs, highlighting strengths, weaknesses, threats, and opportunities. Italy is, indeed, a hotspot of microbial biodiversity with a high rate of endemism and incredible potential, not only for the food and beverage sector (i.e., “Made in Italy” products), where microorganisms can have a beneficial or a spoiling function, but also to guarantee environmental sustainability and foster the bioeconomy through the design of new bioprocesses and products. However, weaknesses, such as the lack of management rules in accordance with international quality standards, are also analysed and ways of overcoming them are discussed. In this context, an overview is given of the Joint Research Unit MIRRI-IT and the European-funded SUS-MIRRI.IT project, which aims to improve the management and sustainability of Italian microbial collections, and serves as a starting point for an innovative revolution in the context of CCs and mBRCs worldwide.
The concept of fastidious microorganisms currently found in scientific literature is mainly related to the difficulty of isolating/culturing/preserving bacteria. Eukaryotes are investigated much less in this respect, although they represent a fundamental part of the microbial world. Furthermore, not only isolation, but also identification and culturing (in the perspective of long-term preservation) should be considered key aspects often impacting on the study of fastidious microorganisms, especially in terms of preservation in culture collections and biotechnological exploitation. The present review aimed to investigate the current state of the art on fastidious eukaryotes, with special emphasis on the efforts to improve their isolation, identification, culturing and long-term preservation in culture collections practices. A few case studies focused on some fastidious eukaryotic microorganisms (including possible customized solutions to overcome specific issues) are also presented: isolation and preservation of slow-growing fungi, culturing of Haematococcus lacustris, isolation of unialgal strains of Cyanidiophytina (Rhodophyta), identification of Metschnikowia pulcherrima clade yeasts, isolation and preservation of Pyricularia species, preservation of Halophytophtora spp.
The order Lulworthiales, with its sole family Lulworthiaceae, consists of strictly marine genera found on a wide range of substrates such as seagrasses, seaweeds, and seafoam. Twenty-one unidentified Lulworthiales were isolated in previous surveys aimed at broadening our understanding of the biodiversity hosted in the Mediterranean Sea. Here, these organisms, mostly found in association with Posidonia oceanica and with submerged woods, were examined using thorough multi-locus phylogenetic analyses and morphological observations. Maximum-likelihood and Bayesian phylogeny based on nrITS, nrSSU, nrLSU, and four protein-coding genes led to the introduction of three novel species of the genus Paralulworthia: P. candida, P. elbensis, and P. mediterranea. Once again, the marine environment is a confirmed huge reservoir of novel fungal lineages with an under-investigated biotechnological potential waiting to be explored.
The genus Corollospora, typified by the arenicolous fungus Corollospora maritima, consists of twenty-five cosmopolitan species that live and reproduce exclusively in marine environments. Species of this genus are known to produce bioactive compounds and can be potentially exploited as bioremediators of oil spill contaminated beaches; hence their biotechnological importance. In this paper, nine fungal strains isolated in the Mediterranean Sea, from the seagrass Posidonia oceanica (L.) Delile, from driftwood and seawater contaminated by an oil spill, were investigated. The strains, previously identified as Corollospora sp., were examined by deep multi-loci phylogenetic and morphological analyses. Maximum-likelihood and Bayesian phylogeny based on seven genetic markers led to the introduction of a new species complex within the genus Corollospora: Corollospora mediterranea species complex (CMSC). The Mediterranean Sea, once again, proves an extraordinary reservoir of novel fungal species with a still undiscovered biotechnological potential.
The European Culture Collections' Organisation presents two new model documents for Material Deposit Agreement (MDA) and Material Transfer Agreement (MTA) designed to enable microbial culture collection leaders to draft appropriate agreement documents for, respectively, deposit and supply of materials from a public collection. These tools provide guidance to collections seeking to draft an MDA and MTA, and are available in open access to be used, modified, and shared. The MDA model consists of a set of core fields typically included in a 'deposit form' to collect relevant information to facilitate assessment of the status of the material under access and benefit sharing (ABS) legislation. It also includes a set of exemplary clauses to be included in 'terms and conditions of use' for culture collection management and third parties. The MTA model addresses key issues including intellectual property rights, quality, safety, security and traceability. Reference is made to other important tools such as best practices and code of conduct related to ABS issues. Besides public collections, the MDA and MTA model documents can also be useful for individual researchers and microbial laboratories that collect or receive microbial cultures, keep a working collection, and wish to share their material with others.
Microorganisms represent most of the biodiversity of living organisms in every ecological habitat. They have profound effects on the functioning of any ecosystem, and therefore on the health of our planet and of human beings. Moreover, microorganisms are the main protagonists in food, medical and biotech industries, and have several environmental applications. Accordingly, the characterization and preservation of microbial biodiversity are essential not only for the maintenance of natural ecosystems but also for research purposes and biotechnological exploitation. In this context, culture collections (CCs) and microbial biological resource centres (mBRCs) are crucial for the safeguarding and circulation of biological resources, as well as for the progress of life sciences. This review deals with the expertise and services of CCs, in particular concerning preservation and characterization of microbial resources, by pointing to the advanced approaches applied to investigate a huge reservoir of microorganisms. Data sharing and web services as well as the tight interconnection between CCs and the biotechnological industry are highlighted. In addition, guidelines and regulations related to quality management systems (QMSs), biosafety and biosecurity issues are discussed according to the perspectives of CCs and mBRCs.
Grape berry, a nonclimacteric fruit, during ripening turns from green, hard and acidic to coloured, soft and sweet. Many studies have focused on dynamic changes of mRNA levels, metabolites, sugars or individual proteins, but this is the first report of a proteomic approach applied to the screening of the most prominent variations that take place during berry ripening. Vitis vinifera cv. 'Nebbiolo Lampia' berries were collected at 10-day intervals, starting 1 month after flowering to complete ripe stage; total protein extracts from deseeded berries were separated by 2-DE. A total of 730 spots were detected in the 2-DE gels. 118 protein spots, differentially expressed during berry development, were subjected to MALDI-TOF analysis. Ninety-three of them were identified, corresponding to 101 proteins. The majority of proteins were linked to metabolism, energy and protein synthesis and fate. In comparison to published surveys of major berry proteins, fewer proteins related to stress response and more proteins related to cell structure were differentially expressed. Our data confirm a general decrease of glycolysis during ripening, and an increase of PR proteins in the range of 20-35 kDa. They furthermore suggest that oxidative stress decreases during ripening while extensive cytoskeleton rearrangement takes place in this period.
Production of enzymes which degrade plant cell wall macromoleculea has been studied in relatively few ericoid fungal isolates, although these polymers arc a major component of the organic litter and an important source of nutrients for these fungi. Our aims were to investigate whether the ability to degrade the wall pectic component, only reported for one isolate, is a general feature of ericoid fungi. Of about 35 isolates from different geographic regions, all were capable of growing on pectin as the sole carbon source. Polygalacturonase (PG) activity was detected to a different degree in the culture filtrates and independently of the fungal growth rate. Solid and liquid isoelectric focusing allowed separation and identification of several polygalacturonase isoforms. Among the fungal isolates investigated, those from the northern hemisphere produced mostly acidic isoforms, whereas isolates from South Africa secreted more abundantly basic isoforms. However, purification and biochemical characterization of several PG isoforms from the different isolates revealed an optimal activity in the acidic pH range for all the PG enzymes tested. Polygalacturonase enzymes seem to be an important component of the enzymatic arsenal secreted by ericoid fungi during their saprotrophic life. In addition, they could also play a role during root colonization, since penetration across the plant cell wall is a prerequisite for the establishment of endomycorrhizal symbiosis.