<Emphasis Type="Italic">OsLAP6/OsPKS1</Emphasis>, an orthologue of <Emphasis Type="Italic">Arabidopsis PKSA/LAP6</Emphasis>, is critical for proper pollen exine formation

Background

Male fertility is crucial for rice yield, and the improvement of rice yield requires hybrid production that depends on male sterile lines. Although recent studies have revealed several important genes in male reproductive development, our understanding of the mechanisms of rice pollen development remains unclear.

Results

We identified a rice mutant oslap6 with complete male sterile phenotype caused by defects in pollen exine formation. By using the MutMap method, we found that a single nucleotide polymorphism (SNP) variation located in the second exon of OsLAP6/OsPKS1 was responsible for the mutant phenotype. OsLAP6/OsPKS1 is an orthologous gene of Arabidopsis PKSA/LAP6, which functions in sporopollenin metabolism. Several other loss-of-function mutants of OsLAP6/OsPKS1 generated by the CRISPR/Cas9 genomic editing tool also exhibited the same phenotype of male sterility. Our cellular analysis suggested that OsLAP6/OsPKS1 might regulate pollen exine formation by affecting bacula elongation. Expression examination indicated that OsLAP6/OsPKS1 is specifically expressed in tapetum, and its product is localized to the endoplasmic reticulum (ER). Protein sequence analysis indicated that OsLAP6/OsPKS1 is conserved in land plants.

Conclusions

OsLAP6/OsPKS1 is a critical molecular switch for rice male fertility by participating in a conserved sporopollenin precursor biosynthetic pathway in land plants. Manipulation of OsLAP6/OsPKS1 has potential for application in hybrid rice breeding.

     

Nitrogen removal characteristics of <Emphasis Type="Italic">Pseudomonas stutzeri</Emphasis> F11 and its application in grass carp culture

This study was designed to investigate the denitrification characteristics of Pseudomonas stutzeri F11 under different environmental conditions and to evaluate the effect of these characteristics on nitrogen removal and the water microbiome in an experimental grass carp aquaculture system. The results showed that the optimal growth conditions of strain F11 were (1) ammonia-nitrogen (N), nitrite-N, or nitrate-N as sole N source; (2) initial ammonia-N concentration of 10–30 mg N/l; (3), initial nitrite-N concentration of 200 mg N/l; (4) sodium citrate as carbon source; (5) rotation (r) speed of 200 r/min; a C:N ratio of between 2 and 10; (6) culture at 32–37 °C. The addition of P. stutzeri F11 to the experimental grass carp aquaculture system reduced the levels of ammonia-N, nitrite-N, and total N in the water over an extended range, but had no effect on nitrate-N level. Results of the 454 pyrosequencing analysis indicated that the structure of the microbial community in the aquaculture water changed significantly after the addition of P. stutzeri F11 preparations. The addition of P. stutzeri F11 to the aquaculture system also altered the microbiome metabolism in the water, especially the bacteria involved in nitrogen metabolism. These results suggest that the addition of P. stutzeri F11 to an experimental grass carp aquaculture system decrease nitrogen levels and alter the microbial community structure of the water; as such, this bacterial strain could be a potential candidate for the regulation of water quality in aquaculture systems.     

Fishmeal alternative from renewable CO2 for rainbow trout feed

The rapid global growth in fish farming and limited supply of fish meal (FM) has consequently reduced FM inclusion levels in compound feeds leading to a higher reliance on alternative protein sources. Sasya is a single cell protein (SCP) product that has a similar amino acid profile as FM. The aim of this study was to evaluate the effects of replacing FM with SCP on in vivo digestibility, growth, feed efficiency, whole‐body proximate/amino acid composition and gene expression levels of various hepatic enzymes in rainbow trout (Oncorhynchus mykiss). Three isonitrogenous (470 g/kg crude protein) and isolipidic (18 g/kg crude lipid) diets were formulated as follows: Diet 1: control (30% FM); Diet 2:24% FM + 6% SCP and Diet 3:18% FM + 12% SCP. Each diet was hand‐fed to triplicate tanks containing 30 rainbow trout fingerlings (4.99 ± 0.20 g) for 9 weeks. Apparent digestibility coefficients of SCP for dry matter, crude protein, lipid and energy were 60, 80, 93 and 74% respectively. Growth performance (final weight: 69–71 g), feed conversion ratios (0.91–0.94) as well as whole‐body protein and amino acid composition were unaffected by diets. However, Diet 3 significantly increased whole‐body crude fat and energy. Fish fed the SCP‐based diets had significantly higher expression for carnitine palmitoyltransferase‐1b (CPT1b), fatty acid delta 6 desaturase (FADS6) and fatty acid elongase 5 compared to the control. Overall, the quality of the SCP was similar as FM. Therefore, this product could enlarge the portfolio of alternative protein sources that can be used in fish diets and thus open a new market opportunity for use of a new feed resource in the feed industry.     

Potential Anti-Aging Substances Derived from Seaweeds

Aging is a major risk factor for many chronic diseases, such as cancer, cardiovascular disease, and diabetes. The exact mechanisms underlying the aging process are not fully elucidated. However, a growing body of evidence suggests that several pathways, such as sirtuin, AMP-activated protein kinase, insulin-like growth factor, autophagy, and nuclear factor erythroid 2-related factor 2 play critical roles in regulating aging. Furthermore, genetic or dietary interventions of these pathways can extend lifespan by delaying the aging process. Seaweeds are a food source rich in many nutrients, including fibers, polyunsaturated fatty acids, vitamins, minerals, and other bioactive compounds. The health benefits of seaweeds include, but are not limited to, antioxidant, anti-inflammatory, and anti-obese activities. Interestingly, a body of studies shows that some seaweed-derived extracts or isolated compounds, can modulate these aging-regulating pathways or even extend lifespans of various animal models. However, few such studies have been conducted on higher animals or even humans. In this review, we focused on potential anti-aging bioactive substances in seaweeds that have been studied in cells and animals mainly based on their anti-aging cellular and molecular mechanisms.     

Dyeing performance of the azo dyes containing trifluoromethyl group for polyester fabrics and its single crystal structure

The structures of disperse dyes and their intermolecular interactions have important impacts on dyeing and printing performances for polyester fabrics. The fluorine dyes show some unique molecular stability and photochemical properties. The dyeing property of the azo dye containing trifluoromethyl group for polyester fabrics, 4'-(N-acetoxyethyl-Nethyl)- amino-2-bromine-4-nitro-6-trifluoromethylazo- benzene (D1), was investigated and compared with the similar structure disperse dye. The results show that the high color yield and good exhaustion of the dyed PET fabrics could be obtained. The polyester fabrics dyed with D1 had excellent light fastness. Its single crystal was prepared and the supramolecular interactions were solved by X-ray diffraction. Dye D1 formed triclinic crystals in a trimeric packing mode. The C-F bond distances of CF3 are 1.2730 Å, 1.2240 Å and 1.2900 Å, respectively. The two benzene rings linked azo unit (-N=N-) are obviously twist. The dihedral angle of the two benzene rings is 50.23 o. There are six weak hydrogen bonds around trifluoromethyl group in the intramolecule and intermolecule. The excellent light stability of the dye should be attributed to its unique supramolecular structure.     

Enzymatic treatment on cotton fibers: degradation kinetics of pectin and influence of shape change on adsorption

Alkaline pectinase was one of the most effective enzymes to treat cotton as alternative agent to replace the conventional alkaline method. Removal of pectin and cutin was considered the explanation for improvement of wettability as well as water adsorption on cotton fiber. However, degradation kinetics of pectin is unclear, and the influence of fiber shape on property changes after enzymatic treatment was ignored. The main objective of this work was to reveal interactions between pectinase and cotton fiber for mechanism study. A heterogeneous catalysis kinetic equation, which is associated with Langmuir adsorption model and enzyme deactivation, was used to describe the heterogeneous catalysis. The enzymatic process conditions were optimized. Raw cotton fibers, pectinase-treated and alkali-treated fibers were characterized by impurities content determination, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and scanning electron microscope (SEM). Mechanism of water adsorption enhancement on treated fibers was discussed. In addition to elimination of the outer impurities, flat fibers with less twist and shape changes of lumen were also obtained to ensure better accessibility and water adsorption after enzymatic treatment.     

Complete nuclear ribosomal DNA sequence analysis of Pacific abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis>

Pacific abalone Haliotis discus hannai (Haliotidae, Gastropoda) is an economically important shellfish species in northern China. The complete nuclear ribosomal DNA (nrDNA) of Pacific abalone was amplified, sequenced, and analyzed. The length of the nrDNA was determined to be around 10.7 kb, and to contain, in order, small subunit ribosomal RNA (nrSSU) genes (1871 bp), internal transcribed spacer (ITS, 759–762 bp), large subunit ribosomal RNA (nrLSU) gene (3411 bp), and an intergenic spacer (IGS, 4624–4654 bp). The SSU and LSU regions were almost identical in different individuals, and show little variation from those of other abalone species. The two different variations of the ITS2 region were presented, and this phenomenon also existed in other species. A phylogeny tree was constructed, based on ITS region sequence datasets, to determine the evolutionary relationships of abalones. Abalones have two major subclades, mainly distributed in the North Pacific, Europe and Australia. The IGS region of the nrDNA was sequenced and analyzed for the first time. Several repeat fragments were present upstream of the sequence, and were significantly different between individuals (93.86% sequence identity). The complete nrDNA sequence will be useful for the classification, identification, phylogeny, germplasm management, and breeding of this shellfish.     

Silymarin inhibits adipogenesis in the adipocytes in grass carp <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis> in vitro and in vivo

In this study, two experiments were performed to explore the function of silymarin in adipogenesis in grass carp (Ctenopharyngodon idellus) using in vitro and in vivo models. In experiment 1, differentiated grass carp pre-adipocytes were treated with silymarin for 6 days. Treatment with 100 μg mL^?1silymarin (SM100 group) significantly reduced triglyceride accumulation at day 6. The adipogenic gene expression levels of PPARγ, C/EBPα, SREBP1c, FAS, SCD1, and LPL, and the protein expression level of PPARγ were significantly down-regulated in the SM100 group. Additionally, the SM100 group had significantly lower reactive oxygen species production and reduced glutathione contents compared with the control in vitro. In experiment 2, the juvenile grass carp (mean body weight= 27.4 ± 0.17 g) were fed six isonitrogenous and isocaloric diets in a factorial design containing 0, 100, or 200 mg kg^?1 silymarin (SM0, SM100, SM200) associated with either 4 or 8% lipid levels (low lipid, LL, and high lipid, HL, respectively) for 82 days. The results demonstrated that dietary silymarin supplementation significantly reduced the elevated intraperitoneal fat index in grass carp fed with high-lipid diets, and the gene expression of adipogenesis (PPARγ, FAS) when supplemented with dietary silymarin was notably lower than when no silymarin was supplemented under the high-lipid diets. Thus, our data suggest that silymarin suppressed lipid accumulation in grass carp both in vitro and in vivo, and the effect might be due to an influence on the expression of adipogenesis factors and ROS production partly associated with effects on antioxidant capability.     

Coordinated expression and regulation of deiodinases and thyroid hormone receptors during metamorphosis in the Japanese flounder (<Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>)

In vertebrates, thyroid hormone receptors (TRs) and deiodinases are essential for developmental events driven by the thyroid hormones (THs). However, the significance of deiodinases during the metamorphosis of the Japanese flounder (Paralichthys olivaceus) remains unclear. Moreover, regulation and response of the TRs and deiodinases to THs in this fish are poorly understood. Therefore, we detected the expression patterns of THs, deiodinases, and TRs in drug-treated larvae and untreated larvae of P. olivaceus by using enzyme-linked immunosorbent assay and quantitative real-time PCR during P. olivaceus metamorphosis. To further understand the roles of these elements, a rescue assay was performed. Our results show the importance of THs, TRs, and deiodinases in flatfish metamorphosis. Our results also confirm that D1 and D2 activate THs and D3 plays the opposite and complementary role. Moreover, we demonstrated that both TRα and TRβ have important but different roles during P. olivaceus metamorphosis.