Three-dimensional reconstruction and multiomics analysis reveal a unique pattern of embryogenesis in Ginkgo biloba.

Ginkgo (Ginkgo biloba L.) is one of the earliest extant species in seed plant phylogeny. Embryo development patterns can provide fundamental evidence for the origin, evolution, and adaptation of seeds. However, the architectural and morphological dynamics during embryogenesis in G. biloba remain elusive. Herein, we obtained over 2,200 visual slices from 3 stages of embryo development using micro-computed tomography imaging with improved staining methods. Based on 3-dimensional (3D) spatiotemporal pattern analysis, we found that a shoot apical meristem with 7 highly differentiated leaf primordia, including apical and axillary leaf buds, is present in mature Ginkgo embryos. 3D rendering from the front, top, and side views showed 2 separate transport systems of tracheids located in the hypocotyl and cotyledon, representing a unique pattern of embryogenesis. Furthermore, the morphological dynamic analysis of secretory cavities indicated their strong association with cotyledons during development. In addition, we identified genes GbLBD25a (lateral organ boundaries domain 25a), GbCESA2a (cellulose synthase 2a), GbMYB74c (myeloblastosis 74c), GbPIN2 (PIN-FORMED 2) associated with vascular development regulation, and GbWRKY1 (WRKYGOK 1), GbbHLH12a (basic helix-loop-helix 12a), and GbJAZ4 (jasmonate zim-domain 4) potentially involved in the formation of secretory cavities. Moreover, we found that flavonoid accumulation in mature embryos could enhance postgerminative growth and seedling establishment in harsh environments. Our 3D spatial reconstruction technique combined with multiomics analysis opens avenues for investigating developmental architecture and molecular mechanisms during embryogenesis and lays the foundation for evolutionary studies of embryo development and maturation.

Hyperspectral Imaging Combined with Deep Transfer Learning to Evaluate Flavonoids Content in Ginkgo biloba Leaves.

Ginkgo biloba is a famous economic tree. Ginkgo leaves have been utilized as raw materials for medicines and health products due to their rich active ingredient composition, especially flavonoids. Since the routine measurement of total flavones is time-consuming and destructive, rapid, non-destructive detection of total flavones in ginkgo leaves is of significant importance to producers and consumers. Hyperspectral imaging technology is a rapid and non-destructive technique for determining the total flavonoid content. In this study, we discuss five modeling methods, and three spectral preprocessing methods are discussed. Bayesian Ridge (BR) and multiplicative scatter correction (MCS) were selected as the best model and the best pretreatment method, respectively. The spectral prediction results based on the BR + MCS treatment were very accurate (RTest2 = 0.87; RMSETest = 1.03 mg/g), showing a high correlation with the analytical measurements. In addition, we also found that the more and deeper the leaf cracks, the higher the flavonoid content, which helps to evaluate leaf quality more quickly and easily. In short, hyperspectral imaging is an effective technique for rapid and accurate determination of total flavonoids in ginkgo leaves and has great potential for developing an online quality detection system for ginkgo leaves.

Restrictive Regulation of Ionic Liquid Quaternary Ammonium Salt in SBA-15 Pore Channel for Efficient Carbon Dioxide Conversion.

Herein, the synthesis of a new type of catalyst, SBA-M (Schiff complex of different metal types grafted on SBA-15) based on a quaternization reaction, is described. Various amounts of ionic liquid were grafted into the pore channels of SBA-15 using the post-grafting method, which allowed the ionic liquid to be grafted into the pore channels restrictively. Notably, over six cycles, SBA-Mn (0.2) has been shown to maintain its catalytic activity and stability. In addition, a reaction mechanism for the cycloaddition of CO2 with epoxides based on density-functional theory is proposed. The cycloaddition reaction of CO2 and epoxides is an efficient way of carbon fixation. It is demonstrated that the metal coordinated with the oxygen atom of the epoxides and that a halogen attacked the carbon of epoxides. Moreover, theoretical calculations and synthesis strategy provide a new approach for CO2 conversion.

Study of the Cycloaddition of CO2 with Styrene Oxide Over Six-Connected spn Topology MOFs (Zr, Hf) at Room Temperature.

A series of MOFs with a 6-connected spn topology were synthesized (MOF-808-(Zr, Hf), PCN-777-(Zr, Hf), MOF-818-(Zr, Hf)). Through the in situ DRIFTS of NH3 adsorption-desorption, we found that the activated catalyst mainly contains Lewis acid sites. The effects of different organic ligands on the Lewis acid of the Zr6 cluster were analyzed by XPS and NH3 -TPD, and the relative Lewis acidity of the same metal was obtained: PCN-777>MOF-808>MOF-818. In the Py-FTIR results, we confirmed that MOF-818 has a higher acid site density. In the activity test, MOFs with mesoporous structure showed better catalytic activity under normal temperature and pressure. Among them, MOF-818 can still maintain a high degree of crystallinity after catalysis. Finally, we use density functional theory to propose the mechanism of the cycloaddition reaction of carbon dioxide and styrene oxide. The results show that the metal is coordinated with styrene oxide and halogens attack the ß carbon of the epoxide.

The Jasmine (Jasminum sambac) Genome Provides Insight into the Biosynthesis of Flower Fragrances and Jasmonates.

Jasminum sambac (jasmine flower), a world-renowned plant appreciated for its exceptional flower fragrance, is of cultural and economic importance. However, the genetic basis of its fragrance is largely unknown. Here, we present the first de novogenome assembly of J. sambac with 550.12 Mb (scaffold N50 = 40.10 Mb) assembled into 13 pseudochromosomes. Terpene synthase (TPS) genes associated with flower fragrance are considerably amplified in the form of gene clusters through tandem duplications in the genome. Gene clusters within the salicylic acid/benzoic acid/theobromine (SABATH) and benzylalcohol O-acetyltransferase/anthocyanin O-hydroxycinnamoyltransferases/anthranilate N-hydroxycinnamoyl/benzoyltransferase/deacetylvindoline 4-O-acetyltransferase (BAHD) superfamilies were identified to be related to the biosynthesis of phenylpropanoid/benzenoid compounds. Several key genes involved in jasmonate biosynthesis were duplicated, causing an increase in copy numbers. In addition, multi-omics analyses identified various aromatic compounds and many genes involved in fragrance biosynthesis pathways. Furthermore, the roles of JsTPS3 in ß-ocimene biosynthesis, as well as JsAOC1 and JsAOS in jasmonic acid biosynthesis, were functionally validated. The genome assembled in this study for J. sambac offers a basic genetic resource for studying floral scent and jasmonate biosynthesis, and provides a foundation for functional genomic research and variety improvements in Jasminum.

Rejuvenation increases leaf biomass and flavonoid accumulation in Ginkgo biloba.

Rejuvenation refers to the transition from an adult state to a juvenile state. Trunk truncation at the base of the tree can result in tree rejuvenation. However, little is known about the association of rejuvenation with leaf biomass and flavonoid accumulation. The results of this study showed that, compared with control leaves, leaves of renewed Ginkgo biloba shoots were larger, thicker, and more lobed and had higher fresh/dry weights and chlorophyll contents. The leaf biomass per hectare of rejuvenated trees was twofold higher than that of the untruncated controls. Moreover, we observed a marked increase in the accumulation of flavonol glycosides via metabolomic analysis and detected upregulated expression of genes involved in flavonoid biosynthesis, including CHS, FLS, F3'H, DFR, and LAR. Overexpression of GbCHS in ginkgo calli confirmed that GbCHS plays an important role in flavonoid biosynthesis. Interestingly, the contents of gibberellins significantly increased in the rejuvenated leaves. Moreover, exogenous gibberellin treatment significantly increased GbCHS expression and flavonoid contents. Our findings show that truncation can stimulate tree rejuvenation by altering hormone levels, representing an effective and feasible approach for enhancing the biomass and flavonoid content of G. biloba leaves.

Ginkgo seed extract promotes longevity and stress resistance of Caenorhabditis elegans.

Ginkgo seeds are a traditional food in China valued for their nutritional and health benefits. However, little is known about the anti-aging and health-promoting effects of ginkgo seed products. Here, we showed that ginkgo seed powder extract (GSP-E) is abundant in alkaloids and flavonoids, and can extend the lifespan of Caenorhabditis elegans. GSP-E improved most physiological indicators related to aging of C. elegans, including locomotor activity, reproductive capacity, and resistance to oxidation and heat. Moreover, GSP-E reduced the accumulation of lipofuscin and reactive oxygen species (ROS) in C. elegans. Further studies demonstrated that GSP-E improved longevity and stress resistance by mediating lipid metabolism and autophagy, as well as by regulating gene expression (e.g., FASN-1, POD-2, GPX-7, FAT-5). GSP-E has an anti-amyloid effect and delayed amyloid-induced paralysis of C. elegans. These findings could support the utilization of ginkgo seed as a potential dietary supplement for the health food industry, and provide a novel health-promoting resource against aging and aging-related diseases.

The genome of Medicago polymorpha provides insights into its edibility and nutritional value as a vegetable and forage legume.

Medicago polymorpha is a nutritious and palatable forage and vegetable plant that also fixes nitrogen. Here, we reveal the chromosome-scale genome sequence of M. polymorpha using an integrated approach including Illumina, PacBio and Hi-C technologies. We combined PacBio full-length RNA-seq, metabolomic analysis, structural anatomy analysis and related physiological indexes to elucidate the important agronomic traits of M. polymorpha for forage and vegetable usage. The assembled M. polymorpha genome consisted of 457.53 Mb with a long scaffold N50 of 57.72 Mb, and 92.92% (441.83 Mb) of the assembly was assigned to seven pseudochromosomes. Comparative genomic analysis revealed that expansion and contraction of the photosynthesis and lignin biosynthetic gene families, respectively, led to enhancement of nutritious compounds and reduced lignin biosynthesis in M. polymorpha. In addition, we found that several positively selected nitrogen metabolism-related genes were responsible for crude protein biosynthesis. Notably, the metabolomic results revealed that a large number of flavonoids, vitamins, alkaloids, and terpenoids were enriched in M. polymorpha. These results imply that the decreased lignin content but relatively high nutrient content of M. polymorpha enhance its edibility and nutritional value as a forage and vegetable. Our genomic data provide a genetic basis that will accelerate functional genomic and breeding research on M. polymorpha as well as other Medicago and legume plants.

Physiological, Transcriptomic, and Metabolic Responses of Ginkgo biloba L. to Drought, Salt, and Heat Stresses.

Ginkgo biloba L. is highly adaptable and resistant to a range of abiotic stressors, allowing its growth in various environments. However, it is unclear how G. biloba responds to common environmental stresses. We explored the physiological, transcriptomic, and metabolic responses of G. biloba to short-term drought, salt, and heat stresses. Proline, H2O2, and ABA contents, along with CAT activity, increased under all three types of stress. SOD activity increased under salt and heat stresses, while soluble protein and IAA contents decreased under drought and salt stresses. With respect to metabolites, D-glyceric acid increased in response to drought and salt stresses, whereas isomaltose 1, oxalamide, and threonine 2 increased under drought. Piceatannol 2,4-hydroxybutyrate and 1,3-diaminopropane increased under salt stress, whereas 4-aminobutyric acid 1 and galactonic acid increased in response to heat stress. Genes regulating nitrogen assimilation were upregulated only under drought, while the GRAS gene was upregulated under all three types of stressors. ARF genes were downregulated under heat stress, whereas genes encoding HSF and SPL were upregulated. Additionally, we predicted that miR156, miR160, miR172, and their target genes participate in stress responses. Our study provides valuable data for studying the multilevel response to drought, salinity, and heat in G. biloba.

Liquid profiling in plants: identification and analysis of extracellular metabolites and miRNAs in pollination drops of Ginkgo biloba.

The pollination drop (PD), also known as an ovular secretion, is a critical feature of most wind-pollinated gymnosperms and function as an essential component of pollination systems. However, the metabolome and small RNAs of gymnosperm PDs are largely unknown. We employed gas chromatography-mass spectrometry to identify a total of 101 metabolites in Ginkgo biloba L. PDs. The most abundant metabolites were sugars (45.70%), followed by organic acids (15.94%) and alcohols (15.39%) involved in carbohydrate metabolism, glycine, serine and threonine metabolism. Through pollen culture of the PDs, we further demonstrated that the metabolic components of PDs are indispensable for pollen germination and growth; in particular, organic acids and fatty acids play defensive roles against microbial activity. In addition, we successfully constructed a small RNA library and detected 45 known and 550 novel miRNAs in G. biloba PDs. Interestingly, in a comparative analysis of miRNA expression between PDs and ovules, we found that most of the known miRNAs identified in PDs were also expressed in the ovules, implying that miRNAs in PDs may originate from ovules. Further, combining with potential target prediction, degradome validation and transcriptome sequencing, we identified that the interactions of several known miRNAs and their targets in PDs are involved in carbohydrate metabolism, hormone signaling and defense response pathways, consistent with the metabolomics results. Our results broaden the knowledge of metabolite profiling and potential functional roles in gymnosperm PDs and provide the first evidence of extracellular miRNA functions in ovular secretions from gymnosperms.