Induction of Isochromanones by Co-Cultivation of the Marine Fungus Cosmospora sp. and the Phytopathogen Magnaporthe oryzae

Microbial co-cultivation is a promising approach for the activation of biosynthetic gene clusters (BGCs) that remain transcriptionally silent under artificial culture conditions. As part of our project aiming at the discovery of marine-derived fungal agrochemicals, we previously used four phytopathogens as model competitors in the co-cultivation of 21 marine fungal strains. Based on comparative untargeted metabolomics analyses and anti-phytopathogenic activities of the co-cultures, we selected the co-culture of marine Cosmospora sp. with the phytopathogen Magnaporthe oryzae for in-depth chemical studies. UPLC-MS/MS-based molecular networking (MN) of the co-culture extract revealed an enhanced diversity of compounds in several molecular families, including isochromanones, specifically induced in the co-culture. Large scale co-cultivation of Cosmospora sp. and M. oryzae resulted in the isolation of five isochromanones from the whole co-culture extract, namely the known soudanones A, E, D (1-3) and their two new derivatives, soudanones H-I (4-5), the known isochromans, pseudoanguillosporins A and B (6, 7), naphtho-γ-pyrones, cephalochromin and ustilaginoidin G (8, 9), and ergosterol (10). Their structures were established by NMR, HR-ESIMS, FT-IR, electronic circular dichroism (ECD) spectroscopy, polarimetry ([α]D), and Mosher’s ester reaction. Bioactivity assays revealed antimicrobial activity of compounds 2 and 3 against the phytopathogens M. oryzae and Phytophthora infestans, while pseudoanguillosporin A (6) showed the broadest and strongest anti-phytopathogenic activity against Pseudomonas syringae, Xanthomonas campestris, M. oryzae and P. infestans. This is the first study assessing the anti-phytopathogenic activities of soudanones.

Experiments in artificial culture: from noisy imitation to storytelling robots

This paper presents a series of experiments in collective social robotics, spanning more than 10 years, with the long-term aim of building embodied models of (aspects of) cultural evolution. Initial experiments demonstrated the emergence of behavioural traditions in a group of social robots programmed to imitate each other’s behaviours (we call these Copybots). These experiments show that the noisy (i.e. less than perfect fidelity) imitation that comes for free with real physical robots gives rise naturally to variation in social learning. More recent experimental work extends the robots’ cognitive capabilities with simulation-based internal models, equipping them with a simple artificial theory of mind. With this extended capability we explore, in our current work, social learning not via imitation but robot–robot storytelling, in an effort to model this very human mode of cultural transmission. In this paper, we give an account of the methods and inspiration for these experiments, the experiments and their results, and an outline of possible directions for this programme of research. It is our hope that this paper stimulates not only discussion but suggestions for hypotheses to test with the Storybots. This article is part of a discussion meeting issue ‘The emergence of collective knowledge and cumulative culture in animals, humans and machines’.

Genetic identity of mint cultivars after in vitro conservation, assessed by ISSR primers

The species of the genus Mentha have been known since ancient times and have significant value in the pharmaceutical, cosmetic and food industries, as well as in medicine. For the widespread use of mint, including in a variety of breeding programs, and the preservation of genetic diversity, effective methods of maintaining cultivars and collection samples are required. Thanks to the development of biotechnological methods, in particular, the creation of slow-growing collections, are now actively used as an effective alternative to field collections. It is known that the cultivation of tissues and organs on artificial culture media can cause somaclonal variability. The purpose of this work is to study the effect of in vitro storage at 4-6°C without illumination after 3 and 4 in vitro conservation cycles on the genetic stability of three cultivars of mint Azhurnaya, Bergamotnaya and Zagrava using ISSR primers. 1 cycle: 1 year of in vitro conservation, microcutting and 2 subcultures of regrowth in a culture room. After conservation, the number of viable explants was 70.0-82.1%. Callus formation at the base of the shoots was not observed in any of the cultivars. After 3 and 4 cycles of in vitro conservation, genetic stability was assessed using 11 ISSR primers. It was found that all three mint genotypes showed full compliance (length and number of amplicons) with the profiles of control samples for all studied markers. It was also found that the markers used by ISSR are highly informative for mint cultivars.

A Comparative Analytical Study of Fresh Honey Collected from Honey Comb of Apis Mellifera and Market Samples of Honey

Honey is a natural product which was known to ancient seers of Ayurveda since Veda period. The nutritional and therapeutic benefits of honey were well known and also documented in authentic texts of Ayurveda. However the abundant availability of honey has gone decreasing since industrialization and deforestation. It has resulted in artificial culture and marketing of honey. The quality of such honey is matter of concern as it is easy to adulterate honey but is very hard to find out the difference between natural and adulterated honey. Considering this issue, present work has been planned to compare freshly collected natural honey and marketed samples of honey. For the comparative analytical study four different samples were collected. Three samples of three different pharmacies were procured from market (honey of branded Ayurvedic companies) and compared with collected honey sample from honeycomb of Apis mellifera. The collected four samples of honey have almost similar organoleptic characters i.e. colour, odour, taste and touch. Considerable differences are detected in physico-chemical analysis, nutritional value and HMF value. The observations are within normal range however the differences are suggestive of need of establishing quality control parameters based on source of honey, collection season, temperature and storage condition and also providing these details while labelling the samples. Finding of present study revealed that freshly collected natural honey have best desired attributes compared all three marketed samples.

Cultivation of Artificial Algal Crust and its Effect on Soil Improvement in Sandy Area

Algae are the pioneer species of biological soil crusts. Cyanobacteria, microschwannophyta and pseudocladophyta can form fixed quicksand algae crusts on the surface of sand surface. Through artificial culture, soil crusts can be formed in a short time. The development and succession of algeal-sand crust promoted the enrichment of nutrients in the sand surface layer, and created conditions for the reproduction of micro-soil organisms and the colonization of herbaceous plants, thus promoting the desert ecosystem to enter a virtuous cycle. This chapter will focus on the cultivation process of artificial soil crust and its effect on soil improvement (soil organic matter and nitrogen) in sandy areas. In conclusion, the application of algal solution can rapidly form algal crusts, and according to the research results, the formation of algal crusts can significantly improve the chemical and biological properties of soil.

Dimorphism in Neopseudocercosporella capsellae, an Emerging Pathogen Causing White Leaf Spot Disease of Brassicas

White leaf spot pathogen: Neopseudocercosporella capsellae causes significant damage to many economically important Brassicaceae crops, including oilseed rape through foliar, stem, and pod lesions under cool and wet conditions. A lack of information on critical aspects of the pathogen’s life cycle limits the development of effective control measures. The presence of single-celled spores along with multi-celled conidia on cotyledons inoculated with multi-celled conidia suggested that the multi-celled conidia were able to form single-celled spores on the host surface. This study was designed to demonstrate N. capsellae morphological plasticity, which allows the shift between a yeast-like single-celled phase and the multi-celled hyphal phase. Separate experiments were designed to illustrate the pathogen’s morphological transformation to single-celled yeast phase from multi-celled hyphae or multi-celled macroconidia in-vitro and in-planta. Results confirmed the ability of N. capsellae to switch between two morphologies (septate hyphae and single-celled yeast phase) on a range of artificial culture media (in-vitro) or in-planta on the host surface before infection occurs. The hyphae-to-yeast transformation occurred through the production of two morphologically distinguishable blastospore (blastoconidia) types (meso-blastospores and micro-blastospores), and arthrospores (arthroconidia).

Analysis of the possible cytogenetic mechanism for overcoming hybrid lethality in an interspecific cross between Nicotiana suaveolens and Nicotiana tabacum

Hybrid lethality is a type of reproductive isolation in which hybrids die before maturation, due to the interaction between the two causative genes derived from each of the hybrid parents. The interspecific hybrid of Nicotiana suaveolens × Nicotiana tabacum is a model plant used in studies on hybrid lethality. While most of the progeny produced from such a cross die, some individuals grow normally and mature. Separately, a technique for producing mature hybrids by artificial culture has been developed. However, the mechanism by which hybrids overcome lethality, either spontaneously or by artificial culture, remains unclear. In the present study, we found that some hybrids that overcome lethality, either spontaneously or by artificial culture, lack the distal part of the Q chromosome, a region that includes the gene responsible for lethality. Quantitative polymerase chain reaction results suggested that the distal deletion of the Q chromosome, detected in some hybrid seedlings that overcome lethality, is caused by reciprocal translocations between homoeologous chromosomes. The results showed that chromosomal instability during meiosis in amphidiploid N. tabacum as well as during artificial culturing of hybrid seedlings is involved in overcoming hybrid lethality in interspecific crosses of the genus Nicotiana.

Artificial Cultivation Anti-tumor Activity of Ganoderma mbrekobenum

Different varieties of Gandoerma have different functional effects. In this study, identification, cultivation, anti-tumor activity characterization and active constituent determination of Ganoderma specimen collected in Tanzania were carried out. The fungi specimen was identified as Ganoderma mbrekobenum by morphological and molecular methods, which was found in Ghana and was firstly reported in 2016. The fruit body was formed in artificial culture using sawdust as the main substrate at the suitable fruiting temperature of 27-30 °C, relative humidity 85-90%, and daily processed lighting for 10 h (300-500 Lux). The fruit body of G.mbrekobenum is hard and has ordinary-yield production, uniform shapes, and high stability. The in vitro experiments showed that the extract from G. mbrekobenum fruit body inhibited cancer cell proliferation of various cells, such as HepG2, MDA-MB-231 and U87. The water extract of the fruit body was tested, and the content of crude polysaccharides was 1.12%. The data showed that G. mbrekobenum was a potential valuable variety. This is the first study that reports the cultivation method and the anti-tumor activity of G. mbrekobenum.

Proteomic Analysis of Mycelial Exudates of Ustilaginoidea virens

Rice false smut (RFS) disease, which is caused by Ustilaginoidea virens, has been widespread all over the world in recent years, causing irreversible losses. Under artificial culture conditions, exudates will appear on colonies of U. virens during the growth of the hyphae. Exudation of droplets is a common feature in many fungi, but the functions of exudates are undetermined. As the executors of life functions, proteins can intuitively reflect the functions of exudates. Shotgun proteomics were used in this study. A total of 650 proteins were identified in the exudate of U. virens, and the raw data were made available via ProteomeXchange with the identifier PXD019861. There were 57 subcategories and 167 pathways annotated with Gene Ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, respectively. Through protein–protein interaction (PPI) network analysis, it was found that 20 proteins participated in the biosynthesis of secondary metabolites. Two separate PPI analyses were performed for carbon metabolism and microbial metabolism in diverse environments. After comparing and annotating the functions of proteins of the exudate, it was speculated that the exudate was involved in the construction and remodeling of the fungal cell wall. Pathogenicity, sporulation, and antioxidant effects might all be affected by the exudate.

Effect of incubation temperature and substrate moisture on embryonic development, hatchling phenotypes and post-hatching growth in the Reeves’ Turtle, Mauremys reevesii

Background Reeves’ Turtles (Mauremys reevesii) are economically important in aquaculture in China. Understanding the effects of incubation temperature and substrate moisture on embryos and hatchlings is of great significance for improving the artificial culture of M. reevesii. However, available studies have not yet determined the thermal and hydric optima for M. reevesii eggs, and the potential interaction between the two factors. Methods In this study, eggs of M. reevesii were incubated at five temperature levels (23, 26, 29, 32 and 35 °C, fluctuation range ± 0.5 °C). In each temperature level, there were three substrate moisture levels (1:0.5, 1:0.9 and 1:1.2, weight ratio of vermiculite to water). Thus, a total of 15 combinations of temperature and moisture were used to examine the effects of incubation temperature and substrate moisture on incubation duration, hatching success, hatchling phenotypes, post-hatching growth and hatchling survival. Results Substrate moisture did not significantly affect most development parameters (except incubation duration and carapace width of hatchlings). Eggs incubated at low moisture level (1:0.5) had a longer incubation duration and produced hatchlings with smaller carapace widths than those incubated at medium (1:0.9) or high (1:1.2) moisture levels. Incubation temperature had a significant effect on incubation duration, hatching success, hatchling phenotypes and hatchling survival. Incubation duration decreased as incubation temperature increased. Eggs incubated at 23, 26 and 29 °C showed higher hatching success than those incubated at 32 and 35 °C. Hatchlings incubated at 32 °C were smaller in body size and mass than those incubated at 23, 26 and 29 °C. At 12 months of age, incubation temperature had no long-lasting effect on body mass, but hatchlings incubated at 23 and 35 °C had lower survival rates than those incubated at 26, 29 and 32 °C. For the development of embryos and hatchlings, the interaction between incubation temperature and substrate moisture was not significant. Conclusions Our results indicate that incubation temperature has a significant influence on the development of embryos and hatchlings of M. reevesii, while substrate moisture only significantly affects the incubation duration and carapace width of hatchlings. The combination of an incubation temperature of 29 ± 0.5 °C and a substrate moisture level of 1:1.2 represented optimal incubation conditions in this experiment. Such incubation conditions are helpful in obtaining higher hatching success, shorter incubation duration and higher survival rates for this aquaculture species.