Integrated Morphological, Comparative Transcriptomic, and Metabolomic Analyses Reveal Mechanisms Underlying Seasonal Patterns of Variation in Spines of the Giant Spiny Frog (Quasipaa spinosa).

Quasipaa spinosa, commonly known as the spiny frog, is an economically valued amphibian in China prized for its tender meat and nutritional value. This species exhibits marked sexual dimorphism, most notably the prominent spiny structures on males that are pivotal for mating success and species identification. The spines of Q. spinosa exhibit strong seasonal variation, changing significantly with the reproductive cycle, which typically spans from April to October. Sexually mature males develop densely packed, irregularly arranged round papillae with black spines on their chests during the breeding season, which may then reduce or disappear afterward, while females have smooth chest skin. Despite their ecological importance, the developmental mechanisms and biological functions of these spines have been inadequately explored. This study integrates morphological, transcriptomic, and metabolomic analyses to elucidate the mechanisms underlying the seasonal variation in spine characteristics of Q. spinosa. Our results demonstrate that spine density inversely correlates with body size and that spine development is accompanied by significant changes in epidermal thickness and keratinization during the breeding season. Comparative transcriptomic analysis across different breeding stages revealed significant gene expression alterations in pathways related to extracellular matrix interactions, tyrosine metabolism, Wnt signaling, and melanogenesis. Metabolomic analysis further identified significant seasonal shifts in metabolites essential for energy metabolism and melanin synthesis, including notable increases in citric acid and ß-alanine. These molecular changes are consistent with the observed morphological adaptations, suggesting a complex regulatory mechanism supporting spine development and functionality. This study provides novel insights into the molecular basis of spine morphogenesis and its seasonal dynamics in Q. spinosa, contributing valuable information for the species' conservation and aquaculture.

Comprehensive genomic and metabolomic analysis revealed the physiological characteristics and pickle like odor compounds metabolic pathways of Bacillus amyloliquefaciens ZZ7 isolated from fermented grains of Maotai-flavor baijiu.

Pickle like odor (PLO) is one of the main defective flavors of Maotai flavor baijiu (MFB). Understanding and controlling the PLO compounds producing strains not only solves the problem of PLO from the source, but also ensures the high-quality production of MFB. However, the relevant research on PLO compounds producing strains has not been reported in MFB. In this study, we identified a Bacillus amyloliquefaciens ZZ7 with high yield of PLO compounds in the fermented grains of MFB, and measured its physiological characteristics. It produces 627 volatile compounds and 1,507 non-volatile compounds. There are 7 volatile sulfur compounds that cause the PLO, the content of dimethyl disulfide, dimethyl trisulfide, and dimethyl sulfur is relatively high, accounting for 89.43% of the total volatile sulfur compounds. The genome size of B. amyloliquefaciens ZZ7 is 3,902,720 bp with a GC content of 46.09%, and a total of 3,948 protein coding genes were predicted. Moreover, the functional annotation of coding genes and an assessment of the metabolic pathways were performed by genome annotation, showing it has strong ability to transport and metabolize amino acids and carbohydrates. Comprehensive genomic and metabolomic analysis, the metabolic pathway of PLO compounds of B. amyloliquefaciens ZZ7 was revealed, which mainly involves 12 enzymes including sulfate adenylyltransferase, cysteine synthase, cystathionine γ-synthase, etc. This work provides biological information support at both genetic and metabolic levels for the mechanism of B. amyloliquefaciens ZZ7 to synthesize PLO compounds, and provides a direction for the subsequent genetic modification of ZZ7 to solve PLO from the source in the MFB.

Recent advances in the metabolic engineering and physiological opportunities for microbial synthesis of L-aspartic acid family amino acids: A review.

L-aspartic acid, L-threonine, L-isoleucine, l-lysine, and L-methionine constitute the l-aspartate amino acids (AFAAs). Except for L-aspartic acid, these are essential amino acids that cannot be synthesized by humans or animals themselves. E. coli and C. glutamicum are the main model organisms for AFAA production. It is necessary to reconstitute microbial cell factories and the physiological state of industrial fermentation cells for in-depth research into strains with higher AFAA production levels and optimal growth states. Considering that the anabolic pathways of the AFAAs and engineering modifications have rarely been reviewed in the latest progress, this work reviews the central metabolic pathways of two strains and strategies for the metabolic engineering of AFAA synthetic pathways. The challenges posed by microbial physiology in AFAA production and possible strategies to address them, as well as future research directions for constructing strains with high AFAA production levels, are discussed in this review article.

Assessing the impact of green tax reforms on corporate environmental performance and economic growth: do green reforms promote the environmental performance in heavily polluted enterprises?

Based on Porter's hypothesis and Pollution Shelter's hypothesis, this paper firstly constructs a mechanism of environmental protection tax law and corporate environmental performance. Secondly, it empirically examines the impact of green tax reform on corporate environmental performance and its internal mechanism through the difference in difference (DID) method. The study findings firstly reveal that environmental protection tax law significantly and progressively promotes the improvement of corporate environmental performance. Secondly, the heterogeneity analysis results show that the promotion effect of environmental protection tax law on corporate environmental performance is significant in enterprises with strong financial constraints and stronger internal transparency. The environmental performance improvement effect of the state-owned enterprises is stronger, which indicates that state-owned enterprises can set an example in the face of the formal implementation of the environmental protection tax law. In addition, the heterogeneity of corporate governance shows that the background of senior executives is an important factor affecting the effect of environmental performance improvement. Thirdly, the mechanism analysis indicates that the environmental protection tax law mainly promotes the improvement of enterprise environmental performance by strengthening the local government's enforcement rigidity, improving the local government's environmental concern, promoting enterprise green innovation, and resolving the collusion between government and enterprise. Further analysis shows that the environmental protection tax law based on the empirical results of this paper did not significantly trigger the phenomenon of cross-regional negative pollution transfer of enterprises. The findings of the study have important enlightenment on improving the green governance of enterprises and promoting the high-quality development of national economy.

Transcriptome analysis of grass carp (Ctenopharyngodon idella) and Holland's spinibarbel (Spinibarbus hollandi) infected with Ichthyophthirius multifiliis.

Ichthyophthirius multifiliis is a protozoan ciliate that causes white spot disease (also known as ichthyophthiriasis) in freshwater fish. Holland's spinibarbel (Spinibarbus hollandi) was less susceptible to white spot disease than grass carp (Ctenopharyngodon Idella). In this study, grass carp and Holland's spinibarbel are infected by I. multifiliis and the amount of infection is 10,000 theronts per fish. All grass carp died within 12 days after infection, and the survival rate of Holland's spinibarbel was more than 80%. In order to study the difference in sensitivity of these two fish species to I. multifiliis, transcriptome analysis was conducted using gill, skin, liver, spleen and head kidney of Holland's spinibarbel and grass carp at 48 h post-infection with I. multifiliis. A total of 489,296,696 clean reads were obtained by sequencing. A total of 105 significantly up-regulated immune-related genes were obtained by Gene Ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis in grass carp. Cluster of differentiation 40 (CD40), cluster of differentiation 80 (CD 80), tumor necrosis factor-alpha (TNF-α), toll-like receptor 4 (TLR-4), interleukin 1 beta (IL-1ß) and other inflammatory-related genes in grass carp were enriched in the cytokine-cytokine receptor interaction pathway and toll-like receptor pathway. In Holland's spinibarbel, a total of 46 significantly up-regulated immune-related genes were obtained by GO classification and KEGG pathway enrichment analysis. Immune-related genes, such as Immunoglobin heavy chain (IgH), cathepsin S (CTSS), complement C1q A chain (C1qA), complement component 3 (C3) and complement component (C9) were enriched in phagosome pathway, lysosome pathway and complement and coagulation concatenation pathway. C3 was significantly up-regulated in gill and head kidney. Fluorescence in situ hybridization (FISH) showed that the C3 gene was highly expressed in gill tissue of Holland's spinibarbel infected with I. multifiliis. A small amount of C3 gene was expressed in the gill arch of grass carp after infected with I. multifiliis. In conclusion, the severe inflammatory response in vivo after infecting grass carp with I. multifiliis might be the main cause of the death of grass carp. The extrahepatic expression of the gene of Holland's spinibarbel might play an important role in the immune defense against I. multifiliis.

A chromosomal level genome sequence for Quasipaa spinosa (Dicroglossidae) reveals chromosomal evolution and population diversity.

Quasipaa spinosa is an Asian commercial Dicroglossidae species noted for its spiny chest found in adult males. Here, we report the first chromosomal level Q. spinosa genome employing PacBio long read sequencing and high-resolution chromosome conformation capture (Hi-C) technology. The total length of the final assembled genome was 2,839,292,578 bp, with contig N50 of 3.79 Mb and scaffold N50 of 327.44 Mb. Approximately 99.30% of the length of the assembled genome sequences were anchored to 13 chromosomes with the assistance of Hi-C reads. A total of 26,173 protein-coding genes were predicted, and 95.98% of the genes were functionally annotated. The annotated genes covered a total of 92.10% of the complete vertebrate core gene set according to the BUSCO pipeline evaluation. Approximately 41 million years ago, Q. spinosa began to diverge from its dicroglossid sister taxon Nanorana parkeri. The Q. spinosa genome revealed obvious chromosomal fissions compared with Xenopus tropicalis, which probably represented a specific chromosome evolutionary history within frogs. Population analysis showed that Chinese Q. spinosa could be divided into eastern and western genetic clusters, with the western population showing higher diversity than the eastern population. The effective population size of Q. spinosa showed a continuously decreasing trend from one million years ago to 10,000 years ago. In summary, this study sheds light on Q. spinosa evolution and population differentiation, providing a valuable genomic resource for further biological and genetic studies on this species, and other closely related frog taxa.

HYD3, a conidial hydrophobin of the fungal entomopathogen Metarhizium acridum induces the immunity of its specialist host locust.

Conidial hydrophobins in fungal pathogens of plants, insects, and humans are required for fungal attachment and are associated with high virulence. They are believed to contribute to the pathogenesis of infection by preventing immune recognition. Here, we refute this generalisation offering a more nuanced analysis. We show that MacHYD3, a hydrophobin located on the conidial surface of the specialist entomopathogenic fungus Metarhizium acridum (narrow host range, kills only locusts and grasshoppers), activates specifically the humoral and cellular immunity of its own host insect, Locusta migratoria manilensis (Meyen) but not that of other non-host insects. When topically applied to the cuticle, purified MacHYD3 improved the resistance of locusts to both specialist and generalist fungal pathogens (wide host range) but had no effect on the fungal resistance of other insects, including Spodoptera frugiperda and Galleria mellonella. Hydrophobins extracted from the generalist fungal pathogens M. anisopliae and Beauveria bassiana had no effect on the resistance of locusts to fungal infection. Thus, the host locust has evolved to recognize the conidial hydrophobin of its specialist fungal pathogen, whereas conidial hydrophobins from generalist fungi are able to evade recognition. Our results distinguish the immunogenic potential of conidial hydrophobins between specialist and generalist fungi.