Insights into the differential molecular response of non-germinated and germinated spores of Ameson portunus in vitro by comparative transcriptome analysis.

Ameson portunus, the recently discovered causative agent of "toothpaste disease" of pond-cultured swimming crabs in China has caused enormous economic losses in aquaculture. Understanding the process of spore germination is helpful to elucidate the molecular mechanism of its invasion of host cells. Here, we obtained mature and germinating spores by isolation and purification and in vitro stimulation, respectively. Then, non-germinated and germinated spores were subjected to the comparative transcriptomic analysis to disclose differential molecular responses of these two stages. The highest germination rate, i.e., 71.45 %, was achieved in 0.01 mol/L KOH germination solution. There were 9,609 significantly differentially expressed genes (DEGs), with 685 up-regulated and 8,924 down-regulated DEGs. The up-regulated genes were significantly enriched in ribosome pathway, and the down-regulated genes were significantly enriched in various metabolic pathways, including carbohydrate metabolism, amino acid metabolism and other metabolism. The results suggested that spores require various carbohydrates and amino acids as energy to support their life activities during germination and synthesize large amounts of ribosomal proteins to provide sites for DNA replication, transcription, translation and protein synthesis of the spores of A. portunus within the host cells. Functional genes related to spore germination, such as protein phosphatase CheZ and aquaporin, were also analyzed. The analysis of transcriptome data and identification of functional genes will help to understand the process of spore germination and invasion.

New records of three commercial fish hosts for two Unicapsula spp. and Kudoa megacapsula (Myxozoa: Myxosporea: Multivalvulida).

Infections caused by multivalvulid myxosporeans belonging to genera Unicapsula and Kudoa (Cnidaria: Myxozoa) occasionally affect commercial marine fish species. Postmortem myoliquefaction caused by a variety of Kudoa spp., including K. thyrsites, and unsightly cyst or pseudocyst formation, caused by K. amamiensis, U. muscularis, and other kudoid species, negatively affect commercial values of fillets. However, multivalvulid infections are often latent and imperceptible in the market. Biodiversity, host range, and epidemiology remain to be explored. Here, myxosporean infection was detected in four commercial fish species from southern China, using morphological and molecular analyses. Three Unicapsula spp. (U. pyramidata in Nemipterus japonicus; U. pflugfelderi in Dentex angolensis transported from the Eastern Central Atlantic Ocean, off West African coast; and U. aequilobata in Decapterus macarellus) and Kudoa megacapsula in Nemipterus virgatus were observed to form pseudocysts in the myofibers of the host trunk muscles. All fish hosts identified here, except for U. pyramidata, are new records. Kudoa megacapsula was morphologically characterized by gigantic, cruciform myxospores with four wing-like shell valves morphologically comparable to previous Japanese records of the same species in aquaculture facilities, acquiring fly from China or Korea (Sphyraena pinguis and Seriola quinqueradiata, respectively). Molecular analyses established the conspecificity of the present Chinese isolate with previously recorded Japanese isolates. To our knowledge, for the first time, a partial large subunit ribosomal RNA gene sequence of K. megacapsula was obtained, showing close phylogenetic relationships with Kudoa spp. harboring cruciform myxospores, such as K. thyrsites, K. gunterae, K. whippsi, and K. lateolabracis.

Efficient sugar utilization and high tolerance to inhibitors enable Rhodotorula toruloides C23 to robustly produce lipid and carotenoid from lignocellulosic feedstock.

The utilization of lignocellulosic substrates for microbial oil production by oleaginous yeasts has been evidenced as an economically viable process for industrial-scale biodiesel preparation. Efficient sugar utilization and tolerance to inhibitors are critical for lipid production from lignocellulosic substrates. This study investigated the lignocellulosic sugar utilization and inhibitor tolerance characteristics of Rhodotorula toruloides C23. The results demonstrated that C23 exhibited robust glucose and xylose assimilation irrespective of their ratios, yielding over 21 g/L of lipids and 11 mg/L of carotenoids. Furthermore, C23 exhibited high resistance and efficiently degradation towards toxic inhibitors commonly found in lignocellulosic hydrolysates. The potential molecular mechanism underlying xylose metabolism in C23 was explored, with several key enzymes and signal regulation pathways identified as potentially contributing to its superior lipid synthesis performance. The study highlights R. toruloides C23 as a promising candidate for robust biofuel and carotenoid production through direct utilization of non-detoxified lignocellulosic hydrolysates.