Molecular identification and functional verification of SPL9 and SPL15 of Lilium.

The transformation of plants from juveniles to adults is a key process in plant growth and development, and the main regulatory factors are miR156 and SQUAMOSA promoter binding protein-like (SPL) transcription factors. Lilium is an ornamental bulb, but it has a long maturation time. In this experiment, Lilium bulbs were subjected to a temperature treatment of 15 °C for 4 weeks to initiate vegetative phase change. Transmission electron microscopy indicated the cell wall of bud core tissue undergoing vegetative phase change became thinner, the starch grains were reduced, and the growth of the juvenile stage was accelerated. The key transcription factors LbrSPL9 and LbrSPL15 were cloned, and the phylogenetic analysis showed they possessed high homology with other plant SPLs. Subcellular localization and transcription activation experiments confirmed LbrSPL9 and LbrSPL15 were mainly located in the nucleus and exhibited transcriptional activity. The results of in situ hybridization showed the expression levels of LbrSPL9 and LbrSPL15 were increased after temperature change treatment. The functional verification experiment of the transgenic plants confirmed that the overexpression of LbrSPL9 and LbrSPL15 could shorten maturation time. These findings help elucidate the regulatory mechanisms of phase transition in Lilium and provide a reference for breeding research in other bulbous flowers.

Caveolin-1 is involved in encephalomyocarditis virus replication in BHK-21 cells.

BACKGROUND: Encephalomyocarditis virus, member of Cardiovirus genus within Picornaviridae family, is an important pathogen that infects different domestic and wild animals. However, the molecular mechanism of its entry remains unclear. In this study, we investigated the mechanism of EMCV infectivity in relation to endocytic pathway using BHK-21 cells. METHODS: The function of numerous cellular key factors implicated in the various endocytic mechanisms were systematically explored using chemical inhibitors. Furthermore, RNA interference (RNAi) as well as the overexpression of dominant protein combined to virus infectivity assays, and confocal microscopy was used to examine EMCV infection in details. RESULTS: The results indicated that the EMCV entry into BHK-21 cells depends on caveolin, dynamin, and actin but not clathrin nor macropinocytosis pathways. The effects of overexpression and knockdown of caveolin-1, one components of the caveolae, was examined on EMCV infection. The results showed that EMCV infection was positive correlation with caveolin-1 expression. Confocal microscopy analysis and internalization assay showed that caveolin-1 is required at the early stage of EMCV infection. CONCLUSIONS: Caveolin-1, dynamin, and actin-dependent endocytosis pathways are necessary for EMCV infection in vitro.

[Study on geographical variation of medicinal ingredients contents in Scutellaria baicalensis from different provenances].

In order to investigate the genetic difference on medicinal components of Scutellaria baicalensis from different provenances on the genetic difference, the S. baicalensis provenance tests were arranged by randomized block design.Excavating the crude drugs that have been growing for three years, with the same drying process, the content of baicalin, baicalein, wogonoside, wogonin and laminarin A in S. baicalensis were detected by HPLC, and then the data were analyzed. The results indicated that the content of baicalin in different provenances of S. baicalensis was significantly different (P<0.05), while the variation of baicalein reached extremely significant level (P<0.01). Cluster analysis showed that if the distance was divided by 5.0, the provenances in Chengde, Hebei province were divided into two independent populations, while the other two populations had large geographic spans. The results show that the significant geographical variations exist in the content of medicinal components in S. baicalensis. The study laid a theoretical foundation of provenance selection of S. baicalensis.

[Influence of cutting seedling on growth, quality and yield of both aerial and underground part by cutting seedling in Scutellaria baicalensis].

By measuring the growth data of Scutellaria baicalensis in different cutting-seedling and determined active ingredient contents by HPLC and ultraviolet spectrophotometric determination. such as flavonoids. baicalin. wogonoside. baicalein. wogonin. oroxylin A. scutellarin. luteolin. and apigenin in the whole plant. Under circumstances of guaranteeing the quality and yield of medicinal materials. the yield of medicinal materials. and stems and leaves reached 193.60,63.21 kg/mu after twice cutting seedling. Not only yield but also active ingredient contents have been improved to some extent. the contents of flavonoids. baicalin. wogonoside. baicalein. wogonin. oroxylin A reached 18.52%. 15.13%. 4.03%. 1.04%. 1.04%. 0.12%. respectively in roots. Luteolin was not detected in young stems and leaves of S. baicalensis,the contents of other active ingredients such as scutellarin. luteolin and apigenin reached 7.00%. 0.96%. 0.04% respectively under twice cutting seedling. Therefore. regular cutting seedling could be regard as a new cultivation technique for wider range of promotion. And gaining high quality and yield of medicinal materials and tea with the purpose of rational utilization of natural resources and promoting the development of integration of herbal combination.

Fluoride induces pyroptosis via IL-17A-mediated caspase-1/11-dependent pathways and Bifidobacterium intervention in testis.

Fluoride, a ubiquitous environmental pollutant, poses a significant public health threat. Our previous study revealed a correlation between fluoride-induced testicular pyroptosis and male reproductive dysfunction. However, the underlying mechanism remains unclear. Wild-type and interleukin 17A knockout mice were exposed to sodium fluoride (100 mg/L) in deionized drinking water for 18 weeks. Bifidobacterium intervention (1 × 109 CFU/mL, 0.2 mL/day, administered via gavage) commenced in the 10th week. Sperm quality, testicular morphology, key pyroptosis markers, spermatogenesis key genes, IL-17A signaling pathway, and pyroptosis pathway related genes were determined. The results showed that fluoride reduced sperm quality, damaged testicular morphology, affected spermatogenesis, elevated IL-17A levels, and induced testicular pyroptosis. Bifidobacterium intervention alleviated adverse reproductive outcomes. Fluoride-activated testicular pyroptosis through both typical and atypical pathways, with IL-17A involvement. Bifidobacterium supplementation attenuated pyroptosis by downregulating IL-17A, inhibiting NLRP3 and PYRIN-mediated caspase-1 and caspase-11 dependent pathways in testis, thereby alleviating fluoride-induced male reproductive damage. In summary, this study uncovers the mechanism underlying fluorine-induced testicular pyroptosis and illustrates the novel protecting feature of Bifidobacterium against fluoride-induced harm to male reproduction, along with its potential regulatory mechanism. These results provide fresh perspectives on treating male reproductive dysfunction resulting from fluoride or other environmental toxins.

Melatonin alleviated fluoride-induced impairment of spermatogenesis and sperm maturation process via Interleukin-17A.

Fluoride-induced male reproductive failure is a major environmental and human health concern, but interventions are still lacking. Melatonin (MLT) has potential functions in regulating testicular damage and interleukin-17 (IL-17) production. This study aims to explore whether MLT can mitigate fluoride-induced male reproductive toxicity through IL-17A, and screen the potential targets. So the wild type and IL-17A knockout mice were employed and treated with sodium fluoride (100 mg/L) by drinking water and MLT (10 mg/kg.BW, intraperitoneal injection per two days starting from week 16) for 18 weeks. Bone F- concentrations, grade of dental damage, sperm quality, spermatogenic cells counts, histological morphology of testis and epididymis, and the mRNA expression of spermatogenesis and maturation, classical pyroptosis related and immune factor genes were detected respectively. The results revealed that MLT supplementations alleviated fluoride-induced impairment of spermatogenesis and maturation process, protecting the morphology of testis and epididymis through IL-17A pathway, and Tesk1 and Pten were identified as candidate targets from 29 regulation genes. Taken together, this study demonstrated a new physiological role for MLT in the protection against fluoride-induced reproductive injury and possible regulation mechanisms, which providing a useful therapeutic strategy for male reproductive function failure caused by fluoride or other environmental pollutants.

Genome-wide identification and evolution of WNK kinases in Bambusoideae and transcriptional profiling during abiotic stress in Phyllostachys edulis.

With-no-lysine (WNK) kinases play vital roles in abiotic stress response, circadian rhythms, and regulation of flowering time in rice, Arabidopsis, and Glycine max. However, there are no previous reports of WNKs in the Bambusoideae, although genome sequences are available for diploid, tetraploid, and hexaploid bamboo species. In the present study, we identified 41 WNK genes in five bamboo species and analysed gene evolution, phylogenetic relationship, physical and chemical properties, cis-elements, and conserved motifs. We predicted the structure of PeWNK proteins of moso bamboo and determined the exposed, buried, structural and functional amino acids. Real-time qPCR analysis revealed that PeWNK5, PeWNK7, PeWNK8, and PeWNK11 genes are involved in circadian rhythms. Analysis of gene expression of different organs at different developmental stages revealed that PeWNK genes are tissue-specific. Analysis of various abiotic stress transcriptome data (drought, salt, SA, and ABA) revealed significant gene expression levels in all PeWNKs except PeWNK11. In particular, PeWNK8 and PeWNK9 were significantly down- and up-regulated, respectively, after abiotic stress treatment. A co-expression network of PeWNK genes also showed that PeWNK2, PeWNK4, PeWNK7, and PeWNK8 were co-expressed with transcriptional regulators related to abiotic stress. In conclusion, our study identified the PeWNKs of moso bamboo involved in circadian rhythms and abiotic stress response. In addition, this study serves as a guide for future functional genomic studies of the WNK genes of the Bambusoideae.

Distribution patterns of the contents of five biologically activate ingredients in the root of Scutellaria baicalensis.

As an important herbaceous plant, Scutellaria baicalensis Georgi (Chinese skullcap) is geographically widespread and commonly used throughout the world. In the Chinese medicine market, S. baicalensis has been divided into two primary types, "Ku Qin" (WXR) and "Tiao Qin" (TST). Moreover, TST is also divided into different grades according to the diameter of roots. To explore the distribution patterns of the contents of five biologically activate ingredients (FBAI), we used six-year-old cultivated S. baicalensis and analyzed its growth characteristics as well as the quality difference among different types and diameters in roots. Throughout the entire root, we discovered that contents of the FBAI all initially increased and subsequently decreased from the top to the bottom of the roots. The baicalin content of WXR was less than that of TST. On the contrary, the contents of baicalein, wogonin, and oroxylin A in WXR were up to about two times higher than that in TST. We also found that the 0 to 40 cm part of the S. baicalensis root possessed about 87% of the root biomass and about 92% of the contents of the active ingredients.

[Observation in situ of differentiation from PGC to hematopoietic system cells in chicken embryo].

To study the relationship between hematopoiesis and primordial germ cells, chick embryos at different developing stages were flatbed and located. After fixed by glutaral, the embryos were PAS and HE stained respectively, dehydrated serially, transparent, mounted, and were observed in situ or in cut sheet condition. The results showed: (1) the cellule amorphous and the disposition in chick embryo of PGCs were coincident no matter stained by PAS or HE staining, and HE staining could disclose the morphologic characteristics more clearly, exactly and completely; (2) genesis of blood island could be observed at the boundary of light and dark region of the extraembryonic blastoderm at about 26 hours; (3) both the blood vessel endothelium cells and free cells of the blood island were differentiated from PGCs. The generating of genuine yolk sac was at about 44 - 48 hours. It is concluded that the initial anatomic site of blood island genesis may be is mesoblast of extraembryonic blastoderm rather than the yolk sac; the blood vessel endothelium cells and the blood cells are generated parallel; the PGCs are the common ancestry of angioblast and HSC.