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The present work describes the in vitro biological activities of the crude extracts (petroleum ether, ethyl acetate and n-butanol) prepared from the species Ephedra altissima Desf. The estimation of total phenolic, flavonoid and tannin contents were carried out using the Folin-Ciocalteu, trichloroaluminum and acidified vanillin methods, respectively. The evaluation of the in vitro antioxidant activities were performed by three different methods namely: scavenging of the free radical ABTS, permanganate reducing antioxidant capacity, and potentiometric assay. In addition, the antibacterial activity was assessed by the agar disk diffusion assay against seven bacterial strains. The results of the phytochemical screening revealed the presence of several types of secondary metabolites. The EtOAc extract exhibited the highest content of phenols (125.62 ± 1.51 μg EGA mg-1 of extract). The greatest flavonoid and tannin contents were observed for n-BuOH extract (19.18 ± 0.39 µg EQ mg-1 of extract and 8.95 ± 1.70 μg EC mg-1 of extract, respectively). Moreover, the EtOAc extract revealed potent antioxidant activity in all the tested methods. Furthermore, the aqueous extract from the species E. altissima showed a good ability to reduce iron III to iron II with a value of 0.68 ± 0.3 moL eq L-1 in potentiometric assay. All the crude extracts (PE, EtOAc and n-BuOH) displayed inhibition of bacterial growth against at least three strains with values of MIC ranging from 3.125 to 50 µg mL-1. Therefore, these results suggest that Ephedra altissima could be used as an important source of natural bioactive compounds with antioxidant and antibacterial properties.

Although gymnosperms are well known for their large size, being the largest, tallest and most massive living organisms on earth, an exceptionally diminutive gymnosperm, Ephedra monosperma Gmelin ex C. A. Meyer (Ephedraceae), from southwestern China is compared with the much more massive gymnosperms that tend to receive far greater attention and publicity. Photographs of E. monosperma and its natural habitat are provided and voucher specimens are cited.

Ephedra foeminea is a traditional medicinal plant used in the Eastern Mediterranean region. This study aims to investigate the chemical profiles of different solvent extracts of E. foeminea via an untargeted metabolomics approach, alongside determining their antioxidant capacities. E. foeminea samples collected from Jordan were macerated in solvents of varying polarities; dichloromethane/methanol, methanol, ethanol, ethyl acetate, and acetone. The crude extracts were subjected to comprehensive chemical profiling and metabolomics study using Gas chromatography–Mass spectrometry (GC–MS), Liquid chromatography–Mass spectrometry (LC–MS), and Nuclear Magnetic Resonance (NMR). The obtained data were analyzed using Venn diagrams, Principle Component Analysis (PCA), and Metabolite Enrichment Set Analysis (MESA). ABTS assay was performed to measure the crude extracts’ antioxidant activity. MESA revealed the dominant chemical groups as amino acids, fatty acids, carboxylic acids, and carbohydrates. Results indicated that dichloromethane/methanol and methanolic extracts had the most distinct composition as well as the most unique compounds. The methanolic extract had the most potency (IC50 249.6 µg/mL) in the ABTS assay. However, no significant differences were found. In conclusion, solvents influenced the recovery of metabolites in E. foeminea and the antioxidant activity of the E. foeminea methanolic extract could be correlated to the abundant presence of diverse bioactive compounds.

Background: The genus Ephedra (Ephedraceae) is a dioecious gymnosperm, where female individuals produce the pharmacologically active ephedrine alkaloids. Identifying the sex of specimens without reproductive cones is challenging due to their xeromorphic and morphological similarity. The challenges in sex identification complicate conservation and propagation efforts. Methods: Sequence-Related Amplified Polymorphism (SRAP) markers were applied to distinguish genders in five Ephedra species, particularly the vegetative branches, as well as powdered and fragmented specimens. The fresh material for the five studied Ephedra species and two sexes per species (totaling 10 samples; 5 females & 5 males) was collected from Sinai, Egypt. Results: The SRAP marker results revealed an exclusively male-specific band, and this is not applicable in females in the studied species. The applied SRAP markers grouped males and females in different UPGMA clusters and proved their efficiency in distinguishing between males and females in the five studied species. The Polymorphic Information Content (PIC) values are low (0.16–0.38); this suggests moderate genetic diversity between the females of the studied species, reflecting slow evolutionary rates. Conclusions: The SRAP markers are efficient for identifying Ephedra species at the species and gender levels, even in the absence of sex organs and molecular sequences. Recommendation: This study recommends the use of SRAP markers for conserving and propagating female plants for ephedrine production and suggests sequencing a 95 bp male-specific band to determine if it corresponds to a known sex-linked gene.

Background The evolution of the Jehol Biota of western Liaoning in China includes three phases, initiation in the Dabeigou phase, radiation in the Yixian phase, and decline in the Jiufotang phase. Numerous ephedroid macrofossils were reported from the Lower Cretaceous Yixian Formation. However, so far none has been found in the younger Jiufotang Formation (ca. 120.3 Ma) of western Liaoning. Results Here we report a new species Jianchangia verticillata gen. et sp. nov. with unusual morphology from the Lower Cretaceous of the Jiufotang Formation, Lamadong Village, Jianchang County, Liaoning. This species is the first record of gnetophytes from the Jiufotang Formation. It is similar to other ephedroid species from the Yixian Formation in possessing linear leaves with parallel veins, jointed shoots with swollen nodes and longitudinally furrowed internodes, and ovulate cones possessing two whorls of bracts enclosing two chlamydosperms, but differs from all known species by the ovulate cone having multiple fine linear verticillate bracts. Conclusions This study expands our knowledge about the diversity of early gnetophytes in the Lower Cretaceous, and demonstrates the lineage continuity of gnetophytes from the Yixian Formation to the younger Jiufotang Formation.

The genus Ephedra Tourn. ex L. (Ephedraceae) is an important source with pharmacological and environmental potential. Conversely, Ephedra spp. still exhibit taxonomic complexity, especially for the specimens lacking reproductive cones. This complexity is attributed to its xeromorphic features, notably the reduced leaves and analogous assimilating branches, which make the species identification a real challenge. The current study provides a pioneering approach to distinguish fragments of Ephedra species at the gender level. This study was based on the stem anatomy and stem epidermal features using a light microscope for five species (E. alata Decne., E. aphylla Forssk., E. ciliata Fisch. & C.A.Mey., E. foeminea Forssk. and E. Ephedra pachyclada Boiss.) represented by ten genders collected from S. Sinai, Egypt. Anomocytic and brachyparacytic stomata, tanniniferous idioblasts, annual rings, a terete and furrowed outline, the number and width of tracheids, patches of cortical fibers, unicellular trichomes, druses, solitary crystals, and the activity of interfascicular cambium were among the distinguishing features that were found. Different statistical analyses were applied to explore the diversity at interspecific and intra-generic levels. This study revealed that the stem anatomy was not only an efficient tool for identifying the investigated five Ephedra species at the species level but also presented a differential key to distinguish between genders and species. In addition, our results indicated that the epidermal features played a critical role in differentiating the studied Ephedra species at the gender level. This study confirms the efficacy of stem anatomy as an identification approach for the Ephedra species at the gender level and recommends this approach to identify the fragmented Ephedra for taxonomical, pharmaceutical, and medical applications.

belongs to the genus Ephedra (Ephedraceae) and plays a significant role in desertification control. In this study, we assembled the complete chloroplast genome of for the first time, revealing a typical quadripartite structure. The chloroplast genome was 109,545 base pair (bp) long with a GC content of 36.8%, encoding 118 complete genes. Phylogenetic reconstruction resolved , , , , , , and formed a monophyletic group, indicating their close relationship. The chloroplast genome of establishes a good foundation for further genetic and genomic studies of the .

Bracts of female cones of extant gymnosperm Ephedra (Joint fir) are either colorful and fleshy (section Ephedra), or dry-winged and membranous (section Alatae), or dry and coriaceous (section Asarca), which have played a crucial role in long-distance seed dispersal that is responsible for a wide distribution of the genus in semiarid and arid areas of Eurasia, North Africa, North America, and South America. Recent molecular systematic studies on Ephedra have suggested that the fleshy bracts in character evolution may be plesiomorphic relative to the dry, membranous and coriaceous bracts. However, little is known about when the fleshy bracts of Ephedra have made their debut in the geological past. Herein, we describe a novel, fleshy bract-bearing female cone macrofossil from the Early Cretaceous (ca. 120-125 Ma) Yixian Formation in Liaoning, northeastern China. This cone bears three ellipsoid seeds subtended by only one whorl of fleshy bracts. Each seed has a thin outer envelope and an inner integument that extends upward and passes through the opening of the outer envelope, forming a thin and straight micropylar tube. Such a syndrome shows the closest similarity to an extant triovulate species Ephedra intermedia in the section Ephedra, but the latter bears a whorl of terminal fertile bracts and more than one whorl of inferior sterile bracts, and a thick outer envelope. Hence, we establish a new fossil species Ephedra carnosa. Our discovery provides the first direct macrofossil evidence for the previous molecular systematics of Ephedra, implying that the origin of fleshy bracts in Ephedra should not have been later than that of the membranous and coriaceous bracts by at least the Early Cretaceous.

Phytochemical investigation of the n-BuOH-soluble fraction of the EtOH extract of the herbaceous stems of Ephedra sinica, which is known as Ephedrae Herba in Traditional Chinese Medicine, led to the isolation and identification of 12 A-type proanthocyanidins, containing five dimers, two trimers and five tetramers [i.e., (+)-epigallocatechin-(2α→O→7,4α→8)-(-)-catechin, named ephedrannin D1, a dimer; epigallocatechin-(2α→O→7,4α→8)-epigallocatechin-(4α→8)-catechin (ephedrannin Tr1), a trimer; and epigallocatechin-(2α→O→7,4α→8)-epigallocatechin-(4α→8)-epigallocatechin-(2α→O→7,4α→8)-gallocatechin, named ephedrannin Te1, a tetramer). Tetramers composed of gallocatechin are reported for the first time in Ephedraceae. Catechin, epicatechin, gallocatechin, epigallocatechin and four known dimers were also isolated. The structures were elucidated by extensive spectroscopic analysis. The absolute configurations of the 4α linkages, which were confirmed by NOESY and CD experiments, are the outstanding characteristic of most of these isolated A-type proanthocyanidins. The antimicrobial activities of these compounds were tested by measuring the minimum inhibitory concentrations (MIC) against bacteria (both Gram positive and Gram negative) and fungi, and were found to be in the range of 0.00515–1.38 mM. Compounds 6, 8, 10 and 11 exhibited moderate antimicrobial activities against Canidia albicans.

Gymnosperms show a significantly higher mean (1C=18.16, 1Cx=16.80) and a narrow range (16.89-fold) of genome sizes as compared with angiosperms. Among the 12 families the largest ranges of 1C values is shown by Ephedraceae (4.73-fold) and Cupressaceae (4.45-fold) which are partly due to polyploidy as 1Cx values vary 2.41 and 1.37-fold respectively. In rest of the families which have only diploid taxa the range of 1C values is from 1.18-fold (Cycadaeae) to 4.36-fold (Podocarpaceae). The question is how gymnosperms acquired such big genome sizes despite the rarity of recent instances of polyploidy. A general survey of different families and genera shows that gymnosperms have experienced both increase and decrease in their genome size during evolution. Various genomic components which have accounted for these large genomes have been discussed. The major contributors are the transposable elements particularly LTR-retrotransposons comprising of , and superfamilies which are most widespread. The genomes of gymnosperms have been acquiring diverse LTR-RTs in their long evolution in the absence of any efficient mechanism of their elimination. The epigenetic machinery which silences these large tracts of repeat sequences into the stretches of heterochromatin and the adaptive value of these silenced repeat sequences need further investigation.