Cloning and Functional Characterization of a Novel ß-GRP Gene From Melanotus cribricollis.

In this study, a novel ß-1,3-glucan recognition protein gene (ß-GRP) was identified from Melanotus cribricollis, and its potential role in the immune response was investigated. The full length of ß-GRP cDNA (Accession Number: MT941530) was 1644 bp, encoding a protein composed of 428 amino acids. The theoretical molecular weight and the isoelectric point were 51.53 kDa and 6.17, respectively. The amino acid sequence of ß-GRP from M. cribricollis was closely related to that of. ß-GRP-like from Photinus pyralis, and was predicted to contain conserved GH16 domain without glucanase active site. The results of real-time quantitative PCR showed that fungal infection of Metarhizium pingshaense may significantly upregulated the expression level of ß-GRP gene. The RNAi suppression of ß-GRP gene expression significantly increased the corrected cumulative mortality. Meanwhile, antimicrobial peptide genes defensin and lysozyme were significantly downregulated after interference of ß-GRP. Taken together, these results suggest that ß-GRP of M. cribricollis probably participates in the host immune system by mediating the expression of antimicrobial peptides. This study provides comprehensive insights into the innate immune system of insect larvae.

[Selection and validation of internal reference genes for qPCR in Polygonatum cyrtonema tubers at different development stages and in response to abiotic stress].

In order to analyze the expression of genes involved in steroidal saponin biosynthesis pathway in Polygonatum cyrtonema tubers, it is very important to select internal reference genes that are stably expressed at different development stages and in response to abiotic stress. According to the previously established P. cyrtonema transcriptome database and reported internal reference genes in plant, this study systematically analyzed eight candidate internal reference genes including histone H2 A, glyceraldehyde-3-phosphate dehydrogenase, ACTIN, ß-tubulin, ubiquitin-conjugating enzyme-E2-10, elongation factor 1-alpha isoform, 18 S rRNA and α-tubulin 4 for expression stability in P. cyrtonema tubers at different development stages and in response to methyl jasmonate(MeJA) stress by using Real time fluorescence quantitative PCR(qPCR). Based on the statistical analysis of qPCR results by using GeNorm, NormFinder and BestKeeper softwares, the expression of ubiquitin-conjugating enzyme-E2-10 and elongation factor 1-alpha isoform are the most stable in P. cyrtonema tubes at different development stages and in response to MeJA stress. The two internal reference genes were further validated by analyzing the expression of 4 genes involved in steroidal saponin biosynthesis pathways. In conclusion, ubiquitin-conjugating enzyme-E2-10 and elongation factor 1-alpha isoform can be used as the most appropriate internal reference genes for qPCR analysis in P. cyrtonema. This study also provide a foundation for future investigate the molecular mechanism of steroidal saponin biosynthesis pathways in P. cyrtonema.

Investigation of the Functional Components in Health Beverages Made from Polygonatum cyrtonema Rhizomes Provides Primary Evidence to Support Their Claimed Health Benefits.

This study aims to understand the functional component compositions of traditional herbal health beverages made from Polygonatum cyrtonema rhizomes and to reveal the pharmacodynamic chemical basis for their claimed health benefits. Two traditional methods, rhizome decoction and rhizome infusion, were used to make health herbal beverages, including "Huangjin" tea and "Huangjin" wine, respectively. The secondary metabolites of "Huangjin" beverages were investigated and compared by widely targeted metabolomics. The results clearly showed that the major functional components in "Huangjin" beverages were phenolic acids, flavonoids, and alkaloids. The "Huangjin" wine has a greater variety of flavonoids and alkaloids than "Huangjin" tea, and the functional components in "Huangjin" wine were more abundant than those in "Huangjin" tea. Homoisoflavones and amide alkaloids were the dominating flavonoids and alkaloids in "Huangjin" wine, respectively. Continuous rhizome infusion could not increase the content of functional components in "Huangjin" wine. In conclusion, this study not only provides primary evidence to support the claimed health benefits of "Huangjin" beverages but also suggests that making traditional herbal beverages by rhizome infusion has superior health benefits than making them by rhizome decoction, which is attributed to the higher yields of functional components extracted by Chinese liquor than hot water. Therefore, Chinese liquor shows advantages in its use as a superior binary ethanol-water solvent in making herbal health beverages to enhance the solubility of poorly water-soluble functional components.

Traditional Processing Can Enhance the Medicinal Effects of Polygonatum cyrtonema by Inducing Significant Chemical Changes in the Functional Components in Its Rhizomes.

The aims of this study were to explore the significant chemical changes in functional components induced by the traditional processing method and evaluate whether this method based on nine cycles of steaming and drying can effectively enhance the medicinal effects of Polygonatum cyrtonema rhizome. A global analysis on dynamic changes in secondary metabolites during nine processing cycles was performed, and the significantly differentially accumulated secondary metabolites were initially identified based on the secondary metabolome. Unsupervised principal component analysis (PCA), hierarchical clustering analysis (HCA), and orthogonal partial least squares discriminant analysis (OPLA-DA) on secondary metabolites clearly showed that processing significantly increased the global accumulation of secondary metabolites in processed P. cyrtonema rhizomes compared to unprocessed crude rhizomes. The first six processing cycles induced drastic changes in the accumulation of functional components, while the last three did not induce further changes. The accumulations of most functional components were significantly enhanced after the first three cycles and stabilized after six cycles; meanwhile, the first three cycles also led to numerous new components. However, the enhancing effects were unavoidably reversed or weakened under continued processing lasting 6-9 cycles. Furthermore, continued processing also reduced the contents of a small number of original components to undetectable levels. Processing induced some significantly enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, among which the first three processing cycles enhanced the synthesis of various secondary metabolites and significantly affected the metabolisms of amino acids. In conclusion, this study not only reveals that processing can effectively enhance the medicinal effects, by two main mechanisms including enhancing chemical synthesis and inducing structural transformation of functional components, but also provides theoretical guidance for the optimization of the traditional processing method based on nine cycles of steaming and drying for achieving optimal effects on enhancing the medicinal effects of P. cyrtonema rhizome.

Quality assessment and nutrient uptake and utilization in Luohan pine (Podocarpus macrophyllus) seedlings raised by chitosan spraying in varied LED spectra.

Target seedling cultivation pursues high quality and nutrient utilization instead of increasing growth and size. Exposure to light-emitting diode (LED) spectra is a well-known approach that can accelerate growing speed in tree seedlings, but it is still unknown whether seedling quality and nutrient utilization would be further improved with exogeneous polymer additives. Luohan pine (Podocarpus macrophyllus) seedlings were exposed to red (red-green-blue lights, 71.7%-13.7%-14.6%), green (26.2%-56.4%-17.4%), and blue (17.8%-33.7%-48.5%) LED-light spectra with half receiving leaf spray by chitosan oligosaccharides (Cos) at a rate of 2 ppm (w/w) and the other half receiving only water. The red-light spectrum promoted height, biomass, nutrient utilization, and quality assessment (DQI) in water-sprayed seedlings. The Cos spray enhanced fine-root growth, protein, and chlorophyll-b contents with elevated nutrient utilization and quality in seedlings in the green-light spectrum. DQI was found to have a positive relationship with phosphorus utilization. In conclusion, although the red-light LED spectrum can promote seedling growth, green light combined with Cos spray is recommended with the aim of maintaining seedling quality and increasing P utilization in Luohan pine seedlings.