ABSTRACT
Circular RNAs (circRNAs) are a subclass of non-coding RNAs (ncRNAs) formed through a process known as back-splicing. They play a crucial role in the genetic regulation of various biological processes. Currently, circRNAs have been identified as participants in the antiviral response within mammalian cells. However, circRNAs in shrimp infected with the yellow head virus (YHV) remain largely unexplored. Therefore, this study aims to identify circRNAs in the hemocytes of Litopenaeus vannamei during YHV infection. We discovered 358 differentially expressed circRNAs (DECs), with 177 of them being up-regulated and 181 down-regulated. Subsequently, eight DECs, including circ_alpha-1-inhibitor 3, circ_CDC42 small effector protein 2, circ_hemicentin 2, circ_integrin alpha V, circ_kazal-type proteinase inhibitor, circ_phenoloxidase 3, circ_related protein rab-8B, and circ_protein toll-like, were randomly selected for analysis of their expression patterns during YHV infection using qRT-PCR. Furthermore, the circRNAs' characteristics were confirmed through PCR, RNase R treatment, and Sanger sequencing, all of which were consistent with the features of circRNAs. These findings contribute to a better understanding of circRNAs' involvement in the antiviral response in shrimp.
Subject(s)
MicroRNAs , Penaeidae , Roniviridae , Animals , Antiviral Agents , Gene Expression Regulation , MicroRNAs/genetics , RNA, Circular/genetics , Penaeidae/virologyABSTRACT
Microalgae have emerged as a promising, next-generation sustainable resource with versatile applications, particularly as expression platforms and green cell factories. They possess the ability to overcome the limitations of terrestrial plants, such non-arable land, water scarcity, time-intensive growth, and seasonal changes. However, the heterologous expression of interested genes in microalgae under heterotrophic cultivation (dark mode) remains a niche area within the field of engineering technologies. In this study, the green microalga, Chlorella sorokiniana AARL G015 was chosen as a potential candidate due to its remarkable capacity for rapid growth in complete darkness, its ability to utilize diverse carbon sources, and its potential for wastewater treatment in a circular bioeconomy model. The aims of this study were to advance microalgal genetic engineering via dark cultivation, thereby positioning the strain as promising dark-host for expressing heterologous genes to produce high-value phytochemicals and ingredients for food and feed. To facilitate comprehensive screening based on resistance, eleven common antibiotics were tested under heterotrophic condition. As the most effective selectable markers for this strain, G418, hygromycin, and streptomycin exhibited growth inhibition rates of 98%, 93%, and 92%, respectively, ensuring robust long-term transgenic growth. Successful transformation was achieved through microalgal cell cocultivation with Agrobacterium under complete darkness verified through the expression of green fluorescence protein and ß-glucuronidase. In summary, this study pioneers an alternative dark-host microalgal platform, using, Chlorella, under dark mode, presenting an easy protocol for heterologous gene transformation for microalgal host, devoid of the need for expensive equipment and light for industrial production. Furthermore, the developed genetic transformation methodology presents a sustainable way for production of high-value nutrients, dietary supplements, nutraceuticals, proteins and pharmaceuticals using heterotrophic microalgae as an innovative host system.
ABSTRACT
Melanin is a functional pigment that is used in various products. It can be produced by Streptomyces antibioticus NRRL B-1701 when supplemented with L-tyrosine. Arthrospira (Spirulina) platensis is a cyanobacterium with high protein content, including the protein phycocyanin (PC). During PC's extraction, biomass residues are generated, and these residues still contain various amino acids, especially L-tyrosine, which can be used as a low-cost supplement for melanin production. Thus, this study employed a hydrolysate of A. platensis biomass residue for L-tyrosine substitution. The effects of two drying methods, namely, lyophilization and dying via a hot air oven, on the proximate composition and content of L-tyrosine in the biomass residue were evaluated. The highest L-tyrosine (0.268 g L-tyrosine/100 g dried biomass) concentration was obtained from a hot-air-oven-dried biomass residue hydrolysate (HAO-DBRH). The HAO-DBRH was then used as a low-cost L-tyrosine supplement for maximizing melanin production, which was optimized by the response surface methodology (RSM) through central composite design (CCD). Using the RSM-CCD, the maximum level of melanin production achieved was 0.24 g/L, which is approximately four times higher than it was before optimization. This result suggests that A. platensis residue hydrolysate could be an economically feasible and low-cost alternative source of L-tyrosine for the production of melanin.
