Response Surface Methodology (RSM) for Optimizing Protein Extraction from Housefly (Musca domestica) Larvae Fed with Toad and Its Structural Characterization.

With the global population on the rise, an escalating interest exists in environmentally sustainable and friendly protein sources. Insects have emerged as multifaceted resources, viewed not only as potential food items, but also as sources of traditional medicines and proteins. This study utilized response surface methodology (RSM) to ascertain the optimal extraction conditions for proteins from Musca domestica used in toad feeding, denoted as MDPs-T. The yield of MDPs-T was elevated to 18.3% ± 0.2% under these optimized conditions. Subsequently, the particle size, ζ-potentials, and structures of MDPs-T were analyzed and compared with the proteins derived from Musca domestica fed on a normal diet (MDPs-ND). This comparative analysis utilized a range of advanced techniques, involving UV spectroscopy, Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), high-performance gel permeation chromatography (HPGPC), and scanning electron microscopy (SEM). The outcomes have revealed a marginal disparity in the physical and chemical properties between MDPs-T and MDPs-ND. Derosination led to a reduction in the particle size of the MDPs by 10.98% to 62.81%. MDPs-T exhibited a higher proportion of low-molecular-weight components relative to MDPs-ND. Additionally, in a comparative analysis of amino acids, MDPs-T displayed a greater abundance of essential and total amino acids relative to MDPs-ND. Consequently, MDPs-T holds potential as a valuable food supplement for human consumption or as a nutrient-rich feed supplement for animals.

Mapping of the novel powdery mildew resistance gene Pm2Mb from Aegilops biuncialis based on ph1b-induced homoeologous recombination.

KEY MESSAGE: A novel powdery mildew resistance gene Pm2Mb from Aegilops biuncialis was transferred into common wheat and mapped to chromosome 2MbL bin FL 0.49-0.66 by molecular cytogenetic analysis of 2Mb recombinants. Aegilops biuncialis, a wild relative of common wheat, is highly resistant to powdery mildew. Previous studies identified that chromosome 2Mb in Chinese Spring (CS)-Ae. biuncialis 2Mb disomic addition line TA7733 conferred high resistance to powdery mildew, and the resistance gene was temporarily designated as Pm2Mb. In this study, a total of 65 CS-Ae. biuncialis 2Mb recombinants were developed by ph1b-induced homoeologous recombination and they were grouped into 12 different types based on the presence of different markers of 2Mb-specificity. Segment sizes and breakpoints of each 2Mb recombinant type were further characterized using in situ hybridization and molecular marker analyses. Powdery mildew responses of each type were assessed by inoculation of each 2Mb recombinant-derived F2 progenies using the isolate E05. Combined analyses of in situ hybridization, molecular markers and powdery mildew resistance data of the 2Mb recombinants, the gene Pm2Mb was cytologically located to an interval of FL 0.49-0.66 in the long arm of 2Mb, where 19 2Mb-specific markers were located. Among the 65 2Mb recombinants, T-11 (T2DS.2DL-2MbL) and T-12 (Ti2DS.2DL-2MbL-2DL) contained a small 2MbL segment harboring Pm2Mb. Besides, a physical map of chromosome 2Mb was constructed with 70 2Mb-specific markers in 10 chromosomal bins and the map showed that submetacentric chromosome 2Mb of Ae. biuncialis was rearranged by a terminal intrachromosomal translocation. The newly developed 2Mb recombinants with powdery mildew resistance, the 2Mb-specific molecular markers and the physical map of chromosome 2Mb will benefit wheat disease breeding as well as fine mapping and cloning of Pm2Mb.

Characterization of the Wheat Heat Shock Factor TaHsfA2e-5D Conferring Heat and Drought Tolerance in Arabidopsis.

Environmental stresses, especially heat and drought, severely limit plant growth and negatively affect wheat yield and quality worldwide. Heat shock factors (Hsfs) play a central role in regulating plant responses to various stresses. In this study, the wheat heat shock factor gene TaHsfA2e-5D on chromosome 5D was isolated and functionally characterized, with the goal of investigating its role in responses to heat and drought stresses. Gene expression profiling showed that TaHsfA2e-5D was expressed constitutively in various wheat tissues, most highly in roots at the reproductive stage. The expression of TaHsfA2e-5D was highly up-regulated in wheat seedlings by heat, cold, drought, high salinity, and multiple phytohormones. The TaHsfA2e-5D protein was localized in the nucleus and showed a transcriptional activation activity. Ectopic expression of the TaHsfA2e-5D in yeast exhibited improved thermotolerance. Overexpression of the TaHsfA2e-5D in Arabidopsis results in enhanced tolerance to heat and drought stresses. Furthermore, RT-qPCR analyses revealed that TaHsfA2e-5D functions through increasing the expression of Hsp genes and other stress-related genes, including APX2 and GolS1. Collectively, these results suggest that TaHsfA2e-5D functions as a positive regulator of plants' responses to heat and drought stresses, which may be of great significance for understanding and improving environmental stress tolerance in crops.

The roles of microRNA in human cervical cancer.

Although a potentially preventable disease, cervical cancer (CC) is the second most commonly diagnosed gynaecological cancer with at least 530,000 new cases annually, and the prognosis with CC is still poor. Studies suggest that aberrant expression of microRNA (miRNA) contributes to the progression of CC. As a group of small non-coding RNA with 18-25 nucleotides, miRNA regulate about one-third of all human genes. They function by repressing translation or inducing mRNA cleavage or degradation, including genes involved in diverse and important cellular processes, including cell cycling, proliferation, differentiation, and apoptosis. Results showed that misexpression of miRNA is closely related to the onset and progression of CC. This review will provide an overview of the function of miRNA in CC and the mechanisms involved in cervical carcinogenesis.

The SPL transcription factor TaSPL6 negatively regulates drought stress response in wheat.

Environmental stresses, such as heat and drought, severely affect plant growth and development, and reduce wheat yield and quality globally. Squamosa promoter binding protein-like (SPL) proteins are plant-specific transcription factors that play a critical role in regulating plant responses to diverse stresses. In this study, we cloned and characterized TaSPL6, a wheat orthologous gene of rice OsSPL6. Three TaSPL6 homoeologs are located on the long arms of chromosomes 4A, 5B, and 5D, respectively, and share more than 98% sequence identity with each other. The TaSPL6 genes were preferentially expressed in roots, and their expression levels were downregulated in wheat seedlings subjected to heat, dehydration, and abscisic acid treatments. Subcellular localization experiments showed that TaSPL6 was localized in the nucleus. Overexpression of TaSPL6-A in wheat resulted in enhanced sensitivity to drought stress. The transgenic lines exhibited higher leaf water loss, malondialdehyde and reactive oxygen species (ROS) content, and lower antioxidant enzyme activities after drought treatment than wild-type plants. Gene silencing of TaSPL6 enhanced the drought tolerance of wheat, as reflected by better growth status. Additionally, RNA-seq and qRT-PCR analyses revealed that TaSPL6-A functions by decreasing the expression of a number of genes involved in stress responses. These findings suggest that TaSPL6 acts as a negative regulator of drought stress responses in plants, which may have major implications for understanding and enhancing crop tolerance to environmental stresses.

Pm57 from Aegilops searsii encodes a tandem kinase protein and confers wheat powdery mildew resistance.

Powdery mildew is a devastating disease that affects wheat yield and quality. Wheat wild relatives represent valuable sources of disease resistance genes. Cloning and characterization of these genes will facilitate their incorporation into wheat breeding programs. Here, we report the cloning of Pm57, a wheat powdery mildew resistance gene from Aegilops searsii. It encodes a tandem kinase protein with putative kinase-pseudokinase domains followed by a von Willebrand factor A domain (WTK-vWA), being ortholog of Lr9 that mediates wheat leaf rust resistance. The resistance function of Pm57 is validated via independent mutants, gene silencing, and transgenic assays. Stable Pm57 transgenic wheat lines and introgression lines exhibit high levels of all-stage resistance to diverse isolates of the Bgt fungus, and no negative impacts on agronomic parameters are observed in our experimental set-up. Our findings highlight the emerging role of kinase fusion proteins in plant disease resistance and provide a valuable gene for wheat breeding.

Extraction, Isolation, Screening, and Preliminary Characterization of Polysaccharides with Anti-Oxidant Activities from Oudemansiella raphanipies.

Response surface methodology (RSM) was used to find the optimal extraction process of Oudemansiella raphanipies polysaccharides (ORPs). The results showed that the optimal extraction parameters were an alkali concentration of 0.02 mol/L, a ratio of material to liquid of 1:112.7 g/mL, an extraction temperature of 66.0 °C, and an extraction time of 4.0 h. Under the optimal conditions, the yield of ORPs was raised to 16.2 ± 0.1%. The antioxidant activities of ORPs-I~V were determined and compared, and ORPs-V was further purified by chromatography, with an average molecular weight (Mw) of 18.86 kDa. The structure of ORPs-V was determined by Fourier transform-infrared spectroscopy (FT-IR), monosaccharide analysis, and nuclear magnetic resonance (NMR) spectroscopy. The ORPs-V comprised fucose, rhamnose, arabinose, glucose, galactose, mannose, xylose, fructose, galacturonic acid, and glucuronic acid at a ratio of 1.73:1.20:1.13:2.87:8.71:2.89:1.42:0.81. Compared to other ORPs, ORPs-V showed the strongest antioxidant activities (ABTS radical cation, hydroxyl radical and DPPH scavenging activities, and reducing power), and were able to significantly increase the activities of superoxide dismutase, catalase, lactate dehydrogenase, and glutathione peroxidase. However, they reduced the malondialdehyde content in mice fed a high-fat diet. These results indicate that ORPs-V may be good anti-oxidant agents to be applied in functional foods.

Wheat heat shock factor TaHsfA2d contributes to plant responses to phosphate deficiency.

Phosphate (Pi) availability has become a major constraint limiting crop growth and production. Heat shock factors (Hsfs) play important roles in mediating plant resistance to various environmental stresses, including heat, drought and salinity. However, whether members of the Hsf family are involved in the transcriptional regulation of plant responses to Pi insufficiency has not been reported. Here, we identified that TaHsfA2d, a member of the heat shock factor family, was strongly repressed by Pi deficiency. Overexpressing TaHsfA2d-4A in Arabidopsis results in significantly enhanced sensitivity to Pi deficiency, evidenced by increased anthocyanin content, decreased proliferation and elongation of lateral roots, and reduced Pi uptake. Furthermore, RNA-seq analyses showed that TaHsfA2d-4A functions through up-regulation of a number of genes involved in stress responses and flavonoid biosynthesis. Collectively, these results provide evidence that TaHsfA2d participates in the regulation of Pi deficiency stress, and that TaHsfA2d could serve as a valuable gene for genetic modification of crop tolerance to Pi starvation.

Development and Characterization of Triticum aestivum-Aegilops longissima 6Sl Recombinants Harboring a Novel Powdery Mildew Resistance Gene Pm6Sl.

Powdery mildew of wheat is a foliar disease that is spread worldwide. Cultivation of resistant varieties is the most effective, economical, and environmentally friendly strategy to curb this disease. Powdery mildew resistance genes (Pm) are the primary resources for resistance breeding, and new Pm genes are in constant demand. Previously, we identified Aegilops longissima chromosome 6Sl#3 as a carrier of powdery mildew resistance and designated the resistance gene as Pm6Sl. Here, we reported the design of 24 markers specific to 6Sl#3 on the basis of the full-length cDNA sequences of 6Sl#3 donor Ae. longissma accession TA1910, and the development of wheat-Ae. longissima 6Sl#3 introgression stocks by ph1b-induced homoeologous recombination. Further, 6Sl#3 introgression lines were identified and characterized by integration analysis of powdery mildew responses, in situ hybridization, and molecular markers and Pm6Sl was mapped to a distal interval of 42.80 Mb between markers Ael58410 and Ael57699 in the long arm of 6Sl#3. Two resistant recombinants, R43 (T6BS.6BL-6Sl#3L) and T27 (Ti6AS.6AL-6Sl#3L-6AL), contained segments harboring Pm6Sl with less than 8% of 6Sl#3 genomic length, and two markers were diagnostic for Pm6Sl. This study broadened powdery mildew resistance gene resources for wheat improvement and provided a fundamental basis for fine mapping and cloning of Pm6Sl to further understand its molecular mechanism of disease resistance.

Response surface methodology for the fermentation of polysaccharides from Auricularia auricula using Trichoderma viride and their antioxidant activities.

Fermentation technology was used to improve the antioxidant activities of Auricularia auricula polysaccharide (AAP). Response surface methodology (RSM) was used to optimize the fermentation conditions. The effects of 4 independent factors: water content (X1: 40-80%), inoculation amount (X2: 2-20%), temperature (X3: 24-32 °C), and time (X4: 4-6 d) on the biological degradation efficiency were evaluated. The RSM results showed that the optimal fermentation conditions were: X1: 61.7%, X2: 12.4%, X3: 31.0 °C, X4: 5.5 d. Verification tests showed no significant differences between the practical and the predictive values for each response. Under the optimal conditions, the degradation rate was 26.89 ± 0.14%, without significant differences with the predicted value (27.03%). The degradation products were classified to different molecular weight (Mw) polysaccharide fragments using membrane separation technology. The FT-IR analysis and monosaccharide composition analysis of degraded AAP (D-AAP-VI) showed that D-AAP-VI was a furan type polysaccharide, which was different from the total AAP (pyran type). In addition, compared to total AAP, the antioxidant activities in vitro of D-AAP-VI were significantly improved (p < 0.05) and D-AAP-VI showed the strongest antioxidant activity. These results indicated that biological degradation may be a suitable way to improve the antioxidant activities of natural polysaccharides.

Isolation, structural characterization and bioactivities of polysaccharides and its derivatives from Auricularia-A review.

Auricularia mushrooms are known for their bioactive compounds, mostly polysaccharides, which have numerous biological activities, such as antioxidant, antitumor, immunomodulatory, hyperlipidemic, antidiabetic, anticoagulant and hepatoprotective effects. Over the past decades, there has been a consistent focus on the isolation, chemical properties and bioactivities of polysaccharides from Auricularia. This review will cover what is known about Auricularia polysaccharides (AP) especially for several common species, including A. auricula-judae, A. auricula, A. polytricha, and A. cornea var. Li. The isolation and purifications, structural characterizations, chemical modifications, and biological activities of these AP and their derivatives will be discussed, thus to provide a foundation for the further investigation, production, and application of AP as functional foods and therapeutic agents.