Untargeted metabolomics reveal rhizosphere metabolites mechanisms on continuous ramie cropping

Ramie is an important fiber feed dual-purpose crop in China and plays an important role in the national economy. However, ramie yield and quality can be reduced after many years of continuous cultivation. Currently, relatively little research has been conducted on rhizosphere metabolites and their pathways in continuous ramie cropping. Therefore, a healthy group (CK) and obstacle groups (XZQG, JZ, DJY, and GXD) with 8 years of continuous cultivation were selected for the study. LC-MS and GC-MS untargeted metabolomics were used to explore and analyze ramie rhizosphere metabolites and pathways. The results revealed that significant differences in the agronomic traits of ramie occurred after 8 years of continuous cultivation, with dwarfed plants and decreased yields in the obstacle groups. Metabolomic analysis identified 49 and 19 rhizosphere metabolites, including lipids, organic acids, phenols, and amino acids. In addition, four differential metabolic pathways (phenylpropanoid biosynthesis, fatty acid metabolism, amino acid metabolism, and ascorbate and aldarate metabolism) were elucidated. It was also clarified that sinapic acid, jasmonic acid, glutamine, and inositol might be the main metabolites affecting ramie continuous-cropping obstacle groups, and they were significantly correlated with ramie agronomic traits and physiological indicators. This provided important insights into the mechanisms affecting continuous ramie cropping. Accordingly, it is expected that the increase or decrease of sinapic acid, jasmonic acid, glutamine, and inositol in the soil will alleviate obstacles to continuous ramie cropping and promote the healthy development of the ramie industry in the future

Antitumor Effects of Berberine on Gliomas via Inactivation of Caspase-1-Mediated IL-1β and IL-18 Release

Gliomas arise in the glial cells of the brain or spine and are the most prevalent and devastating type of brain tumors. Studies of tumor immunology have established the importance of the tumor micro-environment as a driver of oncogenesis. Inflammatory mediators such as IL-1β and IL-18 released by monocytes regulate transcriptional networks that are required for malignant cell growth. Berberine is a natural botanical alkaloid that is widely found in the Berberis species. Although it has been widely used as an anti-diarrheal treatment in North America for several decades, our study is the first to investigate berberine as an anti-tumor agent in glioma cells. In this study, we demonstrate that berberine significantly inhibits inflammatory cytokine Caspase-1 activation via ERK1/2 signaling and subsequent production of IL-1β and IL-18 by glioma cells. Moreover, we found that berberine treatment led to decreased motility and subsequently cell death in U251 and U87 cells. In addition, our study is the first to indicate that berberine can reverse the process of epithelial-mesenchymal transition, a marker of tumor invasion. Taken together, our work supports berberine as a putative anti-tumor agent targeting glioma cells

Response of fermentation quality and microbial community of oat silage to homofermentative lactic acid bacteria inoculation

Oat (Avena sativa L.) is one of the important forage crops in the world. However, oat grown in Southwest China has higher moisture content and their preservation face significant challenges. In addition, existing commercial lactic acid bacteria (LAB) have poor fermentation effects in hot and humid regions. Consequently, the current study investigated the response of oat fermentation quality and microbial community to self-selected LAB inoculation. The treatments were: CK, sterilized water; LP694, Lactobacillus plantarum 694; LR753, Lactobacillus rhamnosus 753; and LPLR, LP694 combined with LR753, followed by 1, 3, 7, 14, and 60 days (d) of fermentation. The results showed that LAB inoculation significantly raised the lactic acid content, and decreased the level of pH value, acetic acid, and ammonia-N in oat silage. The LR753 group had a significantly higher (p < 0.05) lactic acid content (60.95 g kg–1 DM), and lower pH value (3.95) and ammonia-N content (10.1 g kg–1 DM) followed by the LPLR group. The LR753 showed lower NDF (54.60% DM) and ADF (39.73% DM) contents than other groups. The Lactobacillus was a prevalent genus in LAB-treated groups, and its relative abundance reached maximum in LP694 (69%) on day 3, while in the LR753 group (72%) on 60 days. The Lactobacillus rhamnosus, Lactobacillus plantarum, and Lactobacillus fermentum became the dominant species in LAB-treated groups with fermentation time. The Lactobacillus genus was positively correlated with WSC (R = 0.6, p < 0.05), while negatively correlated with pH (R = −0.5, p < 0.05), and BA (R = −0.5, p < 0.01). Overall, the LR753 group had better fermentation quality and preservation of nutritional components providing theoretical support and guidance for future oat silage production in Southwest China

Effect of storage time on the silage quality and microbial community of mixed maize and faba bean in the Qinghai-Tibet Plateau

Tibetan Plateau is facing serious shortage of forage in winter and spring season due to its special geographical location. Utilization of forages is useful to alleviate the forage shortage in winter and spring season. Consequently, the current study was aimed to evaluate the influence of storage time on the silage quality and microbial community of the maize (Zea mays L.) and faba bean (Vicia faba L.) mixed silage at Qinghai-Tibet Plateau. Maize and faba bean were ensiled with a fresh weight ratio of 7:3, followed by 30, 60, 90, and 120 days of ensiling. The results showed the pH value of mixed silage was below 4.2 at all fermentation days. The LA (lactic acid) content slightly fluctuated with the extension of fermentation time, with 33.76 g/kg DM at 90 days of ensiling. The AA (acetic acid) and NH3-N/TN (ammonium nitrogen/total nitrogen) contents increased with the extension of fermentation time and no significantly different between 90 and 120 days. The CP (crude protein) and WSC (water soluble carbohydrate) contents of mixed silage decreased significantly (P < 0.05) with ensiling time, but the WSC content remained stable at 90 days. The Proteobacteria was the predominant phyla in fresh maize and faba bean, and Pseudomonas and Sphingomonas were the predominant genera. After ensiling, Lactobacillus was the prevalent genus at all ensiling days. The relative abundance of Lactococcus increased rapidly at 90 days of ensiling until 120 days of fermentation. Overall, the storage time significant influenced the silage fermentation quality, nutrient content, and microbial environment, and it remained stable for 90 days of ensiling at Qinghai-Tibet Plateau. Therefore, the recommended storage time of forage is 90 days in Qinghai-Tibet Plateau and other cool areas

Transcriptomic and Physiological Responses of Qingye Ramie to Drought Stress

Qingye ramie is a multi-purpose crop, used as a feed ingredient in southern China, that is susceptible to drought. Qingye ramie was studied to investigate the effects of high temperatures and drought on its growthh. The results show that, after drought, ramie leaves turn yellow and that the height of the plant, the number of tillers, and its antioxidant activity decreased. To elucidate the molecular mechanism of drought tolerance, we performed RNA sequencing (RNA-seq) on drought-stressed samples and found that 3893 differentially expressed genes showed significant changes; 1497 genes were upregulated, and 2796 genes were downregulated. These genes were categorized into four metabolic pathways and were mainly enriched in plant hormone signal transcription, phenylpropanoid biosynthesis, amino sugar and nucleotide sugar metabolism, and starch and sucrose metabolism. Among these, we mapped the regulatory mechanism of Qingye ramie under drought and adversity. Of these, the expression of MAPK-related genes in the plant hormone pathway was the most significant. The expression of three MAPK serine/threonine protein kinase genes was upregulated by 2.62- to 3.45-fold and the expression of PP2C-related genes increased by 3.34- to 14.12-fold. The expression of PYR/PYL genes decreased significantly by 2.92–7.09-fold. Furthermore, in addition to NAC, ERF, MYB, bHLH, bZIP, C2H2, GeBP, and WRKY transcription factors that have been shown to regulate drought. Some other transcription factors, such as CCL, ASD, SAU, and SPS, were also up- or downregulated in Qingye ramie. Then, the samples were analyzed by qRT-PCR and the variations were consistent with the sequencing results. Consequently, we suggest that the changes after drought stress in green-leaf ramie may be regulated by these transcription factors. Further studies can be carried out in the future, which will provide valuable and important information on the plant’s drought resistance mechanism and deepen our understanding of the mechanisms of drought resistance in Qingye ramie

<i>Bnt05G007257</i>, a Novel NAC Transcription Factor, Predicts Developmental and Synthesis Capabilities of Fiber Cells in Ramie (<i>Boehmeria</i> nivea L.)

NAC transcription factors are one of the largest transcription factor families in plants, and they play a key role in the growth and development of a secondary cell wall. Despite the fact that ramie is well-known for its high fiber yield, the role of NAC transcription factors in ramie secondary cell wall synthesis and fiber development remains unknown. In this study, based on our previous study, we describe the characterization, physicochemical property analysis, protein structure and function prediction, subcellular localization, and functional validation of Bnt05G007257, which encodes an NAC transcription factor from ramie, in transgenic A. thaliana. Our findings show that the open reading frame of Bnt05G007257 was 1035 bp long and encodes for a protein comprising 344 amino acids, having a relative molecular mass of 39.0945 kDa and a theoretical isoelectric point of 6.55. The secondary structure of the encoded protein mainly consisted of random coiling, with a typical conserved structural domain of NAC. The phylogenetic tree revealed that Bnt05G007257 is a homolog of the NAC transcription factor SND2, which regulates secondary wall biosynthesis in A. thaliana. Subcellular localization showed that Bnt05G007257 was tentatively predicted to be localized in the cytoplasm. Furthermore, in stem sections, the secondary wall fiber cells’ thickness in Bnt05G007257 transgenic plants was 31.50% thicker than that in wild-type plants, and the radial width was significantly increased by approximately 21.75%. This indicates that the NAC family homolog Bnt05G007257 may have the potential function of promoting fiber cell development and secondary cell wall synthesis, providing a theoretical basis for the selection of high-fiber-yielding ramie varieties in the future

Mixture Composition Influenced the Biomass Yield and Nutritional Quality of Legume–Grass Pastures

A two-year field experiment was conducted to address the effects of mixture composition and legume-grass seeding ratio on the biomass yield and nutritional quality of legume–grass mixtures. Alfalfa (Medicago sativa L.), white clover (Trifolium repens L.), red clover (Trifolium pratense L.), orchardgrass (Dactylis glomerata L.), perennial ryegrass (Lolium perenne L.), and tall fescue (Festuca arundinacea Schreb.) were selected as plant materials. A total of seven legume–grass mixtures (A1: white clover, orchardgrass, and tall fescue; A2: alfalfa, orchardgrass, and tall fescue; B1: alfalfa, white clover, orchardgrass, and tall fescue; B2: red clover, white clover, orchardgrass, and tall fescue; C1: alfalfa, white clover, orchardgrass, tall fescue, and perennial ryegrass; C2: red clover, white clover, orchardgrass, tall fescue, and perennial ryegrass; and D: alfalfa, red clover, white clover, orchardgrass, tall fescue, and perennial ryegrass) were sown in two legume-grass seeding ratios (L:G) of 4:6 and 5:5. The results showed that A2 produced a higher two-year average biomass yield (14.20 t/ha) in L:G of 4:6 than that of other mixtures. The grasses biomass yield proportion decreased while legume biomass yield proportion increased with prolonged establishment time. A2 showed a higher crude protein yield (2.5 t/ha) in L:G of 4:6. C2 and A1 showed lower neutral detergent fiber (4.6 t/ha) and acid detergent fiber (2.8 t/ha) yields in L:G 5:5, while diverse mixtures showed higher water-soluble carbohydrate yields. Overall, A2 showed a relative feed value of 146.50 in L:G of 4:6, indicating that it has not only produced the higher biomass yield but also had a better nutritional quality

Effects of Maize Varieties on Biomass Yield and Silage Quality of Maize&ndash;Soybean Intercropping in the Qinghai&ndash;Tibet Plateau

Forage deficiency is the bottleneck that restricts the development of plateau animal husbandry. Maize (Zea mays L.)&ndash;soybean (Glycine max L.) intercropping can improve the forage biomass yield and silage quality. This experiment was conducted in Ganzi Tibetan Autonomous Prefecture to explore the effects of four maize varieties (M1, Rongyu Silage No. 1; M2, Yayu 04889; M3, Demeiya No. 1; M4, Zhenghong 505) on biomass yield, nutritional composition, and silage quality in maize&ndash;soybean intercropping. The results showed that M1S had the highest total dry matter yield (18.03 t ha&minus;1), M3S had the highest crude protein (CP) content (8.46% DM), and soybeans had the highest water-soluble carbohydrate (WSC) content (8.55% DM). After silage, the CP content (13.44% DM) of mixed silage in M3S was higher, and the contents of neutral detergent fiber (39.42% DM) and acid detergent fiber (25.42% DM) were lower than those in maize silage alone. The WSC content (4.45% DM) of mixed silage in M3S was higher and the pH value (4.46) and ammonia&ndash;nitrogen to total nitrogen (3.97%) were lower than those of soybean silage alone. The results of membership function analysis showed that M3S was the best in fresh feeding and silage utilization, followed by M1S. Therefore, M3S (Demeiya No 1. intercropped with soybeans) is recommended in high-altitude areas

Genome-Wide Association Study of Six Forage Traits in Ramie (Boehmeria nivea L. Gaud)

Genome-wide association study (GWAS) of six forage traits using whole-genome sequencing data generated from 301 ramie accessions found that traits were continuously distributed; the maximum variant coefficient was fresh weight per clump (FWPC) (2019) and individual plant height (IPH) (2019) minimum. Correlation analysis demonstrated that 2019 and 2020 results were similar; all traits were correlated. GWAS analysis demonstrated that six traits exhibited consistent and precise association signals. Of the latter, 104 were significant and detected in 43 genomic regions. By screening forage trait-associated single nucleotide polymorphisms and combining Manhattan map with genome annotation, signals were categorized according to functional annotations. One loci associated with fresh weight per plant (FWP) (chromosome 5; Bnt05G007759), two associated with FWPC (chromosome 13; Bnt13G018582, and Bnt13G018583), and two associated with leaf dry weight per plant (LDWP) and dry weight per plant (DWP) (chromosome 4; Bnt04G005779 and Bnt04G005780), were identified. We describe forage trait candidate genes that are highly correlated with FWP and FWPC; Bnt05G007759 may be involved in nitrogen metabolism, while Bnt13G018582 and Bnt13G018583 may encode TEOSINTE branch 1/CYCLOIDEA/proliferating cytokine 1 (TCP) domains. Bnt04G005779 and Bnt04G005780, which may regulate growth and development, are highly related to LDWP and DWP. These genomic resources will provide a basis for breeding varieties