Aging-Induced Reduction in Safflower Seed Germination via Impaired Energy Metabolism and Genetic Integrity Is Partially Restored by Sucrose and DA-6 Treatment.

Seed storage underpins global agriculture and the seed trade and revealing the mechanisms of seed aging is essential for enhancing seed longevity management. Safflower is a multipurpose oil crop, rich in unsaturated fatty acids that are at high risk of peroxidation as a contributory factor to seed aging. However, the molecular mechanisms responsible for safflower seed viability loss are not yet elucidated. We used controlled deterioration (CDT) conditions of 60% relative humidity and 50 °C to reduce germination in freshly harvested safflower seeds and analyzed aged seeds using biochemical and molecular techniques. While seed malondialdehyde (MDA) and fatty acid content increased significantly during CDT, catalase activity and soluble sugar content decreased. KEGG analysis of gene function and qPCR validation indicated that aging severely impaired several key functional and biosynthetic pathways including glycolysis, fatty acid metabolism, antioxidant activity, and DNA replication and repair. Furthermore, exogenous sucrose and diethyl aminoethyl hexanoate (DA-6) treatment partially promoted germination in aged seeds, further demonstrating the vital role of impaired sugar and fatty acid metabolism during the aging and recovery processes. We concluded that energy metabolism and genetic integrity are impaired during aging, which contributes to the loss of seed vigor. Such energy metabolic pathways as glycolysis, fatty acid degradation, and the tricarboxylic acid cycle (TCA) are impaired, especially fatty acids produced by the hydrolysis of triacylglycerols during aging, as they are not efficiently converted to sucrose via the glyoxylate cycle to provide energy supply for safflower seed germination and seedling growth. At the same time, the reduced capacity for nucleotide synthesis capacity and the deterioration of DNA repair ability further aggravate the damage to DNA, reducing seed vitality.

Comparative changes in sugars and lipids show evidence of a critical node for regeneration in safflower seeds during aging.

During seed aging, there is a critical node (CN) where the population viability drops sharply. Exploring the specific locations of the CN in different species of plants is crucial for understanding the biological storage properties of seeds and refining seed life span management. Safflower, a bulk oil crop that relies on seeds for propagation, has a short seed life. However, at present, its biological characteristics during storage are not clear, especially the changes in metabolic capability and cell structures. Such knowledge is needed to improve the management of safflower seed life span and effective preservation in gene banks. Here, the seed survival curve of oilseed safflower under the controlled deterioration conditions of 60% relative humidity and 50°C was detected. The seed population showed an inverted S shape for the fall in germination. In the first 12 days of aging, germination remained above 86%. Prior to the CN at approximately day 10 (C10), when viability was in the "plateau" interval, seed vigor reduced at the same imbibition time point. Further analysis of the changes in sugar concentration found that the sucrose content decreased slowly with aging and the content of raffinose and two monosaccharides decreased abruptly at C10. Differentially metabolized lipids, namely lysophospholipids [lyso-phosphatidylcholine (LPC) and lyso-phosphatidylethanolamines (LPE)] and PMeOH, increased at day 3 of aging (C3). Fatty acid content increased by C6, and the content of phospholipids [phosphatidylcholines (PC), phosphatidylethanolamines (PE), and phosphatidylinositols (PI) and glycolipids [digalactosyl diacylglycerol, monogalactosyl diacylglycerol, and sulphoquinovosyl diglycerides (SQDG)] decreased significantly from C10. In addition, the activities of raffinose hydrolase alpha-galactosidase and the glyoxylate key enzyme isocitrate lyase decreased with seed aging. Confocal microscopy and transmission electron microscopy revealed shrinkage of the seed plasma membrane at C10 and the later fragmentation. Seedling phenotypic indicators and 2,3,5-triphenyltetrazolium chloride activity assays also verified that there were significant changes in seeds quality at the CN. In summary, the time point C10 is a CN during seed population aging. Before the CN, sugar and lipid metabolism, especially fatty acid metabolism into sugar, can make up for the energy consumed by aging. After this point, the seeds were irreversibly damaged, and their viability was greatly and rapidly reduced as the cell structure became increasingly destroyed.

Study on the potential mechanism, therapeutic drugs and prescriptions of insomnia based on bioinformatics and molecular docking.

Insomnia is a very common disease worldwide. It seriously affects the quality of human life and even endangers health. Traditional Chinese medicine (TCM) has unique advantages in the intervention and treatment of insomnia. However, its underlying mechanism has yet to be elucidated. This study was performed to explore the potential biomarkers and mechanisms of insomnia, and treatment TCM and classical prescriptions. The gene microarray data of insomnia is downloaded and preprocessed. Differentially expressed genes (DEGs) and GO and KEGG enrichment analyses were performed. The protein-protein interaction network (PPI) was constructed. Small molecule drugs for curing insomnia were identified using cMap and CTD databases. We searched the TCM corresponding to small molecule drugs and the classic prescriptions corresponding to TCM by the TCMSP database. We constructed a network of "ingredient-TCM-classic prescriptions". The molecular docking was performed to validate the screening results. We obtained a total of 124 DEGs, including 78 up-regulated genes, 46 down-regulated genes, 10 Hub genes and 3 key modules. A total of 125 significant GO entries and 15 significant KEGG were enriched (P < 0.05). The main biological processes involve neuronal apoptosis, autophagy, cell growth and apoptosis, etc. These signaling pathways may be involved in molecular regulatory mechanisms of insomnia, such as autophagy regulation, Alzheimer's disease, pathways to neurodegenerative diseases and neurotrophic factor signaling pathways. We identified 10 traditional Chinese medicines and 2 classical prescriptions of potential value. In addition, the molecular docking results indicated that small molecule ligands were nicely bound to the Hub gene, and the binding affinity ranged from -7.6 to -9.7 kcal/mol. This study provides a foundation for the clinical treatment of insomnia, explains the molecular mechanisms, and efficiently develops TCM and classical prescriptions.

Three Previously Undescribed Chlorophenyl Glycosides from the Bulbs of Lilium regale.

Three previously undescribed chlorophenyl glycosides, (2,4,6-trichloro-3-hydroxy-5-methoxyphenyl)methyl ß-D-glucopyranoside (1), (2,4-dichloro-3,5-dimethoxyphenyl)methyl 6-O-ß-D-glucopyranosyl-ß-D-glucopyranoside (2) and 4-chloro-3-methoxy-5-methylphenyl 6-O-(6-deoxy-ß-L-mannopyranosyl)-ß-D-glucopyranoside (3) were obtained from Lilium regale. The absolute configurations of these new finds were elucidated by comprehensive analyses of spectroscopic data combined with acid hydrolysis derivatization. (2,4-dichloro-3,5-dimethoxyphenyl)methyl 6-O-ß-D-glucopyranosyl-ß-D-glucopyranoside (2) can inhibit the proliferation of lung carcinoma A549 cells with an IC50 value of 29 µΜ.