Stefano Cassanelli

Bacterial cellulose (BC) is a biopolymer boasting potential applications in food and biomedical fields. Although acetic acid bacteria (AAB) are recognized as high BC-producers, several factors may affect the amount produced, making the BC synthesis variable and strain-dependent. Therefore, strategies to improve BC production are of practical relevance. In this study, 12 synthetic microbial communities (SynComs), consisting of acetic acid bacteria (AAB) and/or lactic acid bacteria (LAB) and/or yeasts (Y) were assembled and tested to boost BC production. Specifically, a sequential chemometric strategy based on two Plackett-Burman designs (PBDs) and a circumscribed central composite design (CCC) was implemented to construct and screen SynComs, as well to optimize the culture conditions for BC production. The design of experiments (DOE) approach used in this study allowed for the identification of a suitable SynCom (including both LAB and Y as helper strains) that, in the optimized conditions (50 g/L glucose, 32 °C, 7 days) reached a BC titre (10.40  ±  0.40 g/L) comparable with that of the best BC-producer (AAB monoculture Δgdh K2G30; BC titre 10.79  ±  0.36 g/L). The optimized level, moreover, was consistent with the BC production predicted by the CCC regression model (10.53 g/L). Although further metabolic and population dynamics analyses are required to elucidate the roles and interactions among the community members, our results offer a new perspective on the use of model microbial consortia for BC synthesis and underlines their potential for the development of future and more versatile processes. Key points: • PBD and CCC sequential designs are useful for SynComs assembly and optimization • The composition of SynComs significantly affects the BC production • Several SynComs are more effective than some AAB monocultures for BC production

Bacterial cellulose synthesis from defined media and waste products has attracted increasing interest in the circular economy context for sustainable productions. In this study, a glucose dehydrogenase-deficient Δgdh K2G30 strain of Komagataeibacter xylinus was obtained from the parental wild type through homologous recombination. Both strains were grown in defined substrates and cheese whey as an agri-food waste to assess the effect of gene silencing on bacterial cellulose synthesis and carbon source metabolism. Wild type K2G30 boasted higher bacterial cellulose yields when grown in ethanol-based medium and cheese whey, although showing an overall higher d-gluconic acid synthesis. Conversely, the mutant Δgdh strain preferred d-fructose, d-mannitol, and glycerol to boost bacterial cellulose production, while displaying higher substrate consumption rates and a lower d-gluconic acid synthesis. This study provides an in-depth investigation of two K. xylinus strains, unravelling their suitability for scale-up BC production.

In this study, twelve strains of acetic acid bacteria (AAB) belonging to five different genera were tested for their ability to produce levan, at 70 and 250 g/L of sucrose concentration, respectively. The fructan produced by the bacterial strains was characterized as levan by NMR spectroscopy. Most of the strains produced levan, highlighting intra- and inter-species variability. High yield was observed for Neoasaia chiangmaiensis NBRC 101099 ^T, Kozakia baliensis DSM 14400 ^T and Gluconobacter cerinus DSM 9533 ^T at 70 g/L of sucrose. A 12-fold increase was observed for N. chiangmaiensis NBRC 101099 ^T at 250 g/L of sucrose concentration. Levan production was found to be affected by glucose accumulation and pH reduction, especially in Ko. baliensis DSM 14400 ^T. All the Gluconobacter strains showed a negative correlation with the increase in sucrose concentration. Among strains of Komagataeibacter genus, no clear effect of sucrose on levan yield was found. Results obtained in this study highlighted the differences in levan yield among AAB strains and showed interdependence between culture conditions, carbon source utilization, and time of incubation. On the contrary, the levan yield was not always related to the sucrose concentration.

The evolution of resistance to insecticides threatens the sustainable control of many of the world’s most damaging insect crop pests and disease vectors. To effectively combat resistance, it is important to understand its underlying genetic architecture, including the type and number of genetic variants affecting resistance and their interactions with each other and the environment. While significant progress has been made in characterizing the individual genes or mutations leading to resistance, our understanding of how genetic variants interact to influence its phenotypic expression remains poor. Here, we uncover a mechanism of insecticide resistance resulting from transposon-mediated insertional mutagenesis of a genetically dominant but insecticide-susceptible allele that enables the adaptive potential of a previously unavailable recessive resistance allele to be unlocked. Specifically, we identify clones of the aphid pest Myzus persicae that carry a resistant allele of the essential voltage-gated sodium channel (VGSC) gene with the recessive M918T and L1014F resistance mutations, in combination with an allele lacking these mutations but carrying a Mutator-like element transposon insertion that disrupts the coding sequence of the VGSC. This results in the down-regulation of the dominant susceptible allele and monoallelic expression of the recessive resistant allele, rendering the clones resistant to the insecticide bifenthrin. These findings are a powerful example of how transposable elements can provide a source of evolutionary potential that can be revealed by environmental and genetic perturbation, with applied implications for the control of highly damaging insect pests. Significance The ability to control damaging plant pests and vectors of animal disease is threatened by the emergence of insecticide resistance. Developing effective strategies to prevent, slow, or overcome resistance requires an understanding of the underlying genetic mechanisms. Here, we present an example of the evolution of insecticide resistance arising from transposable element–mediated disruption of a dominant insecticide-susceptible allele, resulting in expression of a recessive resistance allele and potent resistance. Our findings demonstrate how the adaptive potential of transposable elements can be revealed by environmental and genetic perturbation and that this can have profound and unexpected impacts on resistance. They also illustrate how combinations of mutations that individually confer no fitness benefit can interact to provide strong context-dependent fitness benefits.

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.

Cyantraniliprole is a novel insecticide for control of multiple chewing and sucking insect pest species including the sweetpotato whitefly Bemisia ta

The European grapevine moth, Lobesia botrana (Denis & Schiffermüller) (Lepidoptera: Tortricidae), is the key pest of vineyard, Vitis vinifera L. I

Sexual reproduction and breeding systems are driving forces for genetic diversity. The mating-type (MAT) locus represents a mutation and chromosome rearrangement hotspot in yeasts. Zygosaccharomyces rouxii complex yeasts are naturally faced with hostile low water activity (aw) environments and are characterized by gene copy number variation, genome instability, and aneuploidy/allodiploidy. Here, we investigated sex-determination system in Zygosaccharomyces sapae diploid strain ABT301T, a member of the Z. rouxii complex. We cloned three divergent mating type-like (MTL) α-idiomorph sequences and designated them as ZsMTLα copies 1, 2, and 3. They encode homologs of Z. rouxii CBS 732T MATα2 (amino acid sequence identities spanning from 67.0 to 99.5%) and MATα1 (identity range 81.5–99.5%). ABT301T possesses two divergent HO genes encoding distinct endonucleases 100% and 92.3% identical to Z. rouxii HO. Cloning of MATa-idiomorph resulted in a single ZsMTLa locus encoding two Z. rouxii-like proteins MATa1 and MATa2. To assign the cloned ZsMTLα and ZsMTLa idiomorphs as MAT, HML, and HMR cassettes, we analyzed their flanking regions. Three ZsMTLα loci exhibited the DIC1-MAT-SLA2 gene order canonical for MAT expression loci. Furthermore, four putative HML cassettes were identified, two containing the ZsMTLα copy 1 and the remaining harboring ZsMTLα copies 2 and 3. Finally, the ZsMTLa locus was 3′-flanked by SLA2, suggesting the status of MAT expression locus. In conclusion, Z. sapae ABT301T displays an aααα genotype missing of the HMR silent cassette. Our results demonstrated that mating-type switching is a hypermutagenic process in Z. rouxii complex that generates genetic diversity de novo. This error-prone mechanism could be suitable to generate progenies more rapidly adaptable to hostile environments.

Esterase-based resistance in the peach-potato aphid, Myzus persicae (Sulzer), is generally due to one of two alternative amplified carboxylesterase genes, E4 or FE4 (fast E4). The E4 amplified form is distributed worldwide and it is correlated with a particular translocation between autosomes 1 and 3, whereas the FE4 form, which has hitherto not been found to be associated with chromosomal rearrangements, is typical of the Mediterranean regions. In this study, we present for the first time cytogenetic and molecular data on some M. persicae parthenogenetic lineages, which clearly show a chromosomal A1-3 translocation associated with esterase FE4 genes and unrelated to high levels of esterase-based resistance.

In this study, we present cytogenetic data regarding 66 Myzus persicae strains collected in different regions of Italy. Together with the most common 2n = 12 karyotype, the results showed different chromosomal rearrangements: 2n = 12 with A1–3 reciprocal translocation, 2n = 13 with A1–3 reciprocal translocation and A3 fission, 2n = 13 with A3 fission, 2n = 13 with A4 fission, 2n = 14 with X and A3 fissions. A 2n = 12–13 chromosomal mosaicism has also been observed. Chromosomal aberrations (and in particular all strains showing A1–3 reciprocal translocation) are especially frequent in strains collected on tobacco plants, and we suggest that a clastogenic effect of nicotine, further benefited by the holocentric nature of aphid chromosomes, could be at the basis of the observed phenomenon.