On Reserves of Hippophae rhamnoides L. Fruit on the Territory of the Zarafshan National Nature Park of Uzbekistan

The article is devoted to the study of stocks of Hippophae rhamnoides fruits on the territory of the Zarafshan National Park. Hippophae rhamnoides is a medicinal plant. This species is a naturally distributed plant. All parts of Hippophae rhamnoides are a source of biologically active substances, especially flavonoids, carotenoids, phytosterols and others. In the course of field studies, 10 monitoring sites were selected in the size of 10 × 10 m, and the productivity of one site in the amount of 10 × 10 averaged 51.33 ± 8.24 kg. According to the results obtained, it was revealed that the biological reserve averages 21.9 tons, respectively, the operational reserve is 13.14 tons and the volume of annual harvesting averages 1.31 tons. When the water level of the Zarafshan River rises, some plants are damaged.

Effects of Different Water Stresses on Eco-physiological Characteristics of Hippophae rhamnoides Seedlings

In order to examine the effects of the decrease of future precipitation on the eco-physiological characteristics of sea buckthorn (Hippophae rhamnoides Linn.) in Huangfuchuan Watershed in Nei Mongol, a water gradient experiment was conducted based on the four specially designed water supply levels, including normal precipitation, slight drought, drought and extreme drought. Results of ANOVE showed that different water gradients had a significant effect on (1) microhabitat factors, such as soil water content and soil temperature; (2) gas exchange, such as net photosynthetic rate, stomatal conductance and transpiration rate; (3) resource use efficiency; and (4) leaf water potential. Water use efficiency of H rhamnoides could increase under moderate water stress, i.e. drought condition, while its net photosynthetic rate and transpiration rate decreased. All kinds of eco-physiological characteristics proved H. rhamnoides seedlings under all water supplies were affected by water stress more or less and that mechanism of intrinsic physiological regulation in seedlings under the extreme drought conditions had the appearance of turbulence to a certain extent. Therefore, H rhamnoides seedlings in Huangfuchuan Watershed could not acclimate to extreme drought conditions.

中国沙棘(Hippophae rhamnoides L．ssp．sinensis Rousi)的开花特性及风媒传粉距离的检测

连续两年对雌雄异株的中国沙棘（Hippophae rhamnoides L．ssp．sinensis Rousi）的花期物候、开花式样进行了观察，检测了柱头可授性、花粉活力和花粉一胚珠比，并进行厂人工授粉、套袋实验，检测结实率。结果显示中国沙棘风媒传粉发生在4月下旬或5月上旬，单花花期约7d；传粉盛期在第3天到第5大，柱头的可授性从第2天到第4天或第5天。中国沙棘花先叶开放、花小、无花冠、花药成熟时萼片从两个侧向的纵缝中裂开，形成有利于化粉散布的对流风洞。花粉活力和花粉．胚珠比都很高，具有典型的适应风媒传粉的花部特征和性状。中国沙棘花粉的传播距离受外界环境影响较大，通过重力玻片法检测可以看出，在风速小于3m／s时，主要集中在15-25m之间，当风速大于3m／s时，顺风向可超过85m以外。传粉效率对结实率的影响较大。

Characterization and comparison of chloroplast genomes from two sympatric Hippophae species(Elaeagnaceae)

The genus Hippophae includes deciduous shrubs or small trees,which provide many ecological,economic,and social benefi ts.We assembled and annotated the chloroplast genomes of sympatric Hippophae gyantsensis(Rousi)Lian and Hippophae rhamnoides Linn subsp.yunnanensis Rousi and comparatively analyzed their sequences.The fulllength chloroplast genomes of H.gyantsensis and H.rhamnoides subsp.yunnanensis were 155,260 and 156,415 bp,respectively;both featured a quadripartite structure with two copies of a large inverted repeat(IR)separated by small(SSC)and large(LSC)single-copy regions.Each Hippophae chloroplast genome contained 131 genes,comprising 85 protein-coding,8 ribosomal RNA,and 38 transfer RNA genes.Of 1302 nucleotide substitutions found between these twogenomes,824(63.29%)occurred in the intergenic region or intron sequences,and 478(36.71%)were located in the coding sequences.The SSC region had the highest mutation rate,followed by the LSC region and IR regions.Among the protein-coding genes,three had a ratio of nonsynonymous to synonymous substitutions(Ka/Ks)>1 yet none were signifi-cant,and 66 had Ka/Ks<1,of which 46 were signifi cant.We found 20 and 16 optimal codons,most of which ended with A or U,for chloroplast protein-coding genes of H.gyantsensis and H.rhamnoides subsp.yunnanensis,respectively.Phylogenetic analysis of fi ve available whole chloroplast genome sequences in the family Elaeagnaceae—using one Ziziphus jujube sequence as the outgroup—revealed that all fi ve plant species formed a monophyletic clade with two subclades:one subclade consisted of three Hippophae species,while the other was formed by two Elaeagnus species,supported by 100%bootstrap values.Together,these results suggest the chloroplast genomes among Hippophae species are conserved,both in structure and gene composition,due to general purifying selection;like many other plants,a signifi cant AT preference was discerned for most proteincoding genes in the Hippophae chloroplast genome.This study provides a valuable reference tool for future research on the general characteristics and evolution of chloroplast genomes in the genus Hippophae.

Research on Rooting Cuttings of Hard Branch of Hippophae rhamnoides

Objective] In order to improve the survival rate of Hippophae rhamnoides hardwood cuttings, cultivate high-quality sea buckthorn seedlings. [Method] This pa-per took hard branches of Russian big fruit H. rhamnoides as material. The days of striking roots, rate of striking roots,root length and number of adventitious root were determined. [Result] The cuttings col ected from upper treetops were obviously inferi-or to the lower ones. Three years cutting is obviously better than one or two years. Cuttings from lower branches were not so good as the cuttings from upper branch-es. Base oblique incisions were clearly superior to paper-cover incision. The optimal length of cuttings was 20-25 cm. The rooting rate of cuttings was the highest by fast dipping with NAA of 50 mg/L. [Conclusion] The study provides theoretical basis for H. rhamnoides artificial cultivation.

Modular growth and clonal propagation of Hippophae rhamnoides subsp, sinensis in response to irrigation intensity

A multipurpose clone plant species, Hippophae rhamniodes has the capacity for indefinite longevity, although under successive drought stress it may often decline or die across large areas. Field trials were con- ducted over 2 years to examine the effects of varied irrigation intensities on modular growth and clonal propagation in a semi-arid area. Irrigation levels included a control, as well as two, four and six times the volume of water that was typically received via local annual average precipitation. Irrigation intensity significantly influenced clonal propagation capacity （number of daughter ramets）, aboveground modular growth （height, base diameter, and crown width）, belowground modular growth included root nodule dry weight, stretching capacity of lateral roots （length of the longest lateral roots, and diameter of first-grade lateral roots）, and branching intensity of lateral roots （number of lateral roots bifurcation grade, number of first- grade lateral roots）. The modular growth and the density of daughter ramets were small under non-irrigation or low irrigation, and became larger with increased irrigation intensity. Beyond a certain threshold, however, further increases in irrigation intensity resulted in a reversion to the development. The optimal irrigation intensities for growth and propagation were 3.48-5.29 times the volume of nominal local annual average precipitation. There were effects of irrigation intensities on the positive significant correlations between aboveground and belowground modular growths, and on clonal propagation capacities. Under various water treatments, H. rhamnoides may adapt to the environment through the regulation of growth and propagation. We concluded that water shortages act to weaken the growth and propagation of H. rhamniodes plantations.

Influence of soil moisture content on pullout properties of Hippophae rhamnoides Linn. roots

Plant root system plays an important role in preventing soil erosion and improving slope stability.However,its performance is significantly affected by soil moisture content,and the role of soil moisture in root reinforcement is not fully understood.In this study,the influence of soil moisture on root pullout properties was studied by experiments.Vertical in-situ pullout tests under four different levels of soil matric suction(12 kPa,18 kPa,24 kPa,30 kPa)were carried out on roots of sea buckthorn plants(Hippophae rhamnoides Linn.)which were artificially cultivated for 7 months.Diameter and length of the root system of sea buckthorn were investigated.The results showed that a very significant correlation was observed between root diameter(D)and root length(L)(P<0.01),and root diameter decreased with soil depth.When soil bulk density was constant,peak pullout force(F)and friction coefficient of root-soil interface(μ)decreased with increasing gravimetric soil moisture content in power functions.Soil moisture content significantly affected root pullout resistance because the increase of soil moisture content decreased the friction coefficient between the roots and soil.Root diameter at breakage point(Db)and length of root segment left in soil(Lb)were increased with soil moisture content.In addition,peak pullout force of the roots increased in a power function with root diameter at the soil surface(D0)and in a linear function with total root length(L).The results provided an experimental basis for quantifying the effects of soil moisture content on soil reinforcement by plant roots.

Variations in seed germination ofHippophae salicifolia with different presoaking treatments

Mature seeds of H. salicifolia, collected from five provenances （i.e. Hanuman Chatti, Helang, Lata, Rambara and Janggal Chatti） in west Himalaya, India, were treated with stratification （at 4℃ for 15, 30 and 60 days） and in different concentrations of GA3 （5, 10, 20 mM）, KNO3 （50, 100, 200 mM） and Thiourea （50, 100, 200 mM） solution to determine the variations in seed germination. Results reveal that the germination rates of seeds from different provenances under different pre-sowing treatments are significantly increased compared to those in control （24%-30%）. The seeds treated with Thiourea （100 mM） have highest germination rate （76%-83% for different seed sources）, followed by those （63%-71% for different seed sources） pretreated with stratification （4℃, 30 days）. GA3 treatment significantly shortens the mean germination time （MGT） and improves seed germination percentage. Considering the practical applicability and cost effectiveness, thiourea （100 mM） and stratification （at 4℃） treatments for seed germination are recommended for mass multiplication through seeds of H. saliciifolia in village/forest nurseries of the west Himalaya, India.

Effects of Total Flavonoids ofHippophae RhamnoidesL.on Intracellular Free Calciumin Cultured Vascular Smooth Muscle Cells of Spontaneously Hypertensive Rats and Wistar-Kyoto Rats

To explore the effects of total flavonoids of Hippophae rhamnoides L. （TFH） quercetin （Que） and isorhamnetin （Isor） on the intracellular free calcium （[Ca^2＋]） in vascular smooth muscle cells （VSMC） of spontaneously hypertensive rats （SHR） and Wistar-Kyoto rats （WKY）. Metheds： Fluo 3-acetoxymethylester（Fluo-3/AM） was used to observe the effects of TFH （100mg/L） and its essential monomers, namely Que （10^-4mol/L） and Isor （10^-4mol/L） on changes of [Ca^2＋]1 in cultured SHR and WKY VSMC （abbr. to Ca-SHR ＆ Ca-WKY） following exposure to high K^＋, norepinephrine （NE） and angiotensin Ⅱ （AngⅡ）, and to compare with the effects of verapamil （Ver）. Results： （1） TFH, Que and Isor had inhibitory effects on resting Ca-SHR （P〈0.05）, but had no significant effects on Ca-WKY （P〉0.05）. （2） High K^＋ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）; TFH, Que and Isor could inhibit the elevation of [Ca^2＋]1 induced by high K^＋ -depolarization, with the effects similar to that of Ver, and the effect on Ca-SHR was more significant than that on Ca-WKY （P〈0.05）. （3） NE and Ang Ⅱ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）, TFH, Que and Isor had remarkably inhibitory effect on the elevation of Ca-SHR and Ca-WKY induced by NE or Ang Ⅱ. （4） In the absence of extracellular Ca^2＋ , TFH, Que and Isor also had certain inhibitory effect on Ca-SHR and Ca-WKY induced by NE, and the effect on the former was more significant than that on the latter（P〈0.05）. Ceaclusiea： TFH, Que and Isor might decrease the levels of [Ca^2＋], in VSMCs by blocking both voltage-dependent calcium channels （VDC） and receptoroperated calcium channels （ROC） in physiological or pathological state, which may be one of the important mechanisms of their hypotensive and protective effects on target organs in patients with hypertension.

Molecular identification of sex in Hippophae rhamnoides L. using isozyme and RAPD markers

In many dioecious plants, gender affects economic value, breeding schemes and opportunities for commercial harvests. Hippophae rhamnoides L. is a dioecious plant species in which female genotypes are commercially preferred over male genotypes. Its berries have rich medicinal, nutritional and pharmaceutical properties because of their large amounts of vitamins, essential oils, proteins, fatty acids, free amino acids and flavanoids. Primary limitation for breeding H. rhamnoides L. is its dioecious nature, since gender cannot be identified by traditional methods. Therefore, some reliable and quick methods need to be developed. This commu- nication deals with the development of isozyme and RAPD markers for early sex identification in this dioecious tree. The isozyme analysis was conducted with four enzyme systems, viz. peroxidase, esterase, malate dehydrogenase and catalase. The peroxidase enzyme system produced a female specific sex marker, which successfully differentiated between the staminate and pistillate geno- types ofH. rhamnoides L. Thirty five random decamer primers were used in our study and one male sex linked marker was identified. OPD-20 （5＇-ACTTCGCCAC-3＇） displayed a band at 911 bp that expressed polymorphism between male and female genotypes. The staminate and pistillate genotypes could be distinguished using RAPD marker OPD-209n. These results revealed the immense poten- tial of peroxidase isozyme patterns and RAPD as genetic markers for sex identification in H. rhamnoides L.

Analysis of Effective Composition of Hippophae Rhamnoides L.

The vitamin E, proteins, amino acids, sugar, trace elements and fatty acids in the oil of Hippophae Rhamnoides L. seeds were quantitatively analyzed. Several methods for analyzing vitamin E were compared with each other and the content of gossypol in the oil was first determined.

Molecular docking studies of anti-cancerous candidates in Hippophae rhamnoides and Hippophae salicifolia

Actinorhizal plants contain numerous antioxidants that may play a crucial role in preventing the formation of tumors.H-Ras p21,a member of the Ras-GTPase family,is a promising target to treat various kinds of cancers.An in silico docking study was carried out to identify the inhibitory potential of compounds of these plants against H-Ras by using Discovery Studio 3.5 and by using Autodock 4.2.Docking studies revealed that four compounds,isorhamnetin-7-rhamnoside,quercetin-3-glucoside-7-rhamnoside（present in H.rhamnoides）,zeaxanthin,and translutein（present in H.salicifolia） significantly bind with binding energies-17.1534,-14.7936,-10.2105 and-17.2217 Kcal/mol,respectively,even though they slightly deviate from Lipinski＇s rule.Absorption,distribution,metabolism,excretion and toxicity（ADME/tox） analyses of these compounds and their stereoisomers showed that they were less toxic and non-mutagenic.Amongst them,isorhamntein-7-rhamnoside showed hepatotoxicity.Hence,these compounds can be further investigated in vivo to optimize their formulation and concentration and to develop potential chemical entities for the prevention and treatment of cancers.

Short-term responses to salinity of seabuckthorn (Hippophae rhamnoides L.) seedlings in the extremely cold and saline Qinghai region of China

The investigation of the mechanisms of plant adaptation to stressor action is one of the leading directions of current biological studies. To understand the mechanism of salt tolerance of seabuckthom （Hippophae rhamnoides L.） and identify its ability to cope with the salinity effect in the saline and extremely cold region of Qinghai, China, a test was conducted with two-year-old seedlings subjected to 0, 200, 400 and 600 mmol·L^-1 NaClsolutions for 30 d. The results show that with an increase in salinity, the biomass of H. rhamnoides seedlings clearly decreased. Leaf water potential （ψw） and relative water content （RWC） were significantly reduced under salinity, with severe water shortages appearing in leaves. At the same time, the total chlorophyll content declined markedly. When salinity increased and stress time prolonged, the net CO2 assimilation rate （A） significantly declined. Intercellular CO2 concentration （Ci） declined at first and was then followed by an increase over the stress time. We conclude that H. rhamnoides grown in the extremely cold and saline region of Qinghai has a certain resistance to salt, which can be planted at appropriate salinity levels.

Direct somatic embryogenesis from leaves,cotyledons and hypocotyls of Hippophae rhamnoides

Plant regeneration via direct somatic embryogenesis from cotyledons, hypocotyls and leaves in Hippophae rhamnoides L. was achieved. The influence of basal media, carbon sources, plant growth regulators (PGRs) with different concentrations and combinations on embryogenesis capacity of explants was studied. The highest frequency of somatic embryo production and germination was obtained on Schenk and Hildebrandt medium (SH) supplemented with 1.0 mg/L kinetin (KIN) and 0.2, 0.5 mg/L indole-3-butyric acid (IAA). Granulated sugar was the optimal carbon source. The embryo-derived plantlets with well-developed roots and shoots were transferred successfully to the greenhouse with a maximum survival rate of 55%. Histological observation revealed that the somatic embryos were similar to those of zygotic embryos.

PIXE ANALYSIS OF TRACE ELEMENTS IN GENUS HIPPOPHAE L

Trace elements in four kinds of the Genus Hippophae L pulps were analysed byPIXE. Optimization of the system performance was done by carefully selecting the absorbers, Analytical sensitivities of the PIXE system were obtained by means of "external standard method".

Studies on the Characteristics of Foliar Surfaces of Hippophae and Related Taxonomy

Trichome forms and stomatal distribution and types were undertaken. The results show that trichome forms are stellate, peltate and stellate-peltate. There is a difference in the position of stomata among the taxa of Hippophae. H. thibetana, H. salici-folia. H. gyantsensis, H. neurocarpa and H. rhamnoides ssp. turkestanica have stomata on both foliar surfaces, while other subspecies have stomata only on the lower surface. Stomata are of the Actinocytic type.

Identify potential treatments of COVID-19 from Tibetan medicine Hippophae Fructus using a network pharmacological approach

OBJECTIVE Since the coronavirus disease 2019(COVID-19)outbreak in December 2019,the search for a potential treatment for COVID-19 has been a constant focus.Therefore,we identified potential treatments for COVID-19 from Hippophae Fructus,a Tibetan medicine that may act on COVID-19,using a network pharmacology approach.METHODS We collected the chemical constituents and corresponding targets of Hippophae Fructus from traditional Chinese medicine system pharmacology(TCMSP).COVID-19 related genes were predicted in pubmed-Gene,OMIM and GeneCards databases.Then,protein-protein interactions(PPIs)of key genes were analyzed by STRING database.Compound-target-diseases network was constructed using Cytoscape software.The potential pathways were determined by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses.Additionally,molecular docking was used to verify the binding effect between the active component and the target.RESULTS A total of 33 components and 192 corresponding targets in Hippophae Fructus were found.50 genes were obtained from the intersection of component targets and disease targets.These genes include IL-6,TNF,MAPK8 and PTGS2,which regulate several pathways associated with COVID-19,involving Hepatitis B,Influenza A,TNF signaling pathway and Tuberculosis.More importantly,high-node compounds such as quercetin and beta-sitosterol can well bind to key targets.CONCLUSION Some components in Hippophae Fructus can act on COVID-19 related genes and regulate multiple pathways.Perhaps Hippophae Fructus has the effect in treating COVID-19.

Safety evaluation of a polyherbal formulation containing hydroalcoholic extracts of Hippophae salicifolia, Nyctanthes arbor-tristis, Ocimum tenuiflorum, and Reinwardtia indica in rodents

Dear Editor, Herbal medicines have been used for prevention and treatment of human diseases and promotion of healthy living. These medicines, often self medicated and consumed as concentrated extracts, are not strictly regulated. Considering their popular use and increasing safety concerns, thorough evaluations of their efficacy and safety are warranted to protect consumers from potential adverse effects. Toxicity tests （acute, sub- acute and chronic） using animals are widely applied to evaulate adverse effects of a drug and thereby determine its ＂No Observed Adverse Effect Level＂ （NOAEL）.

Development of a Genetic Transformation Method for Seabuckthorn (<i>Hippophae rhamnoides</i>L.)

Seabuckthorn (Hippophae rhamnoides L.) is a dioecious plant with berries containing high amounts of several bioactive compounds with nutritional and medicinal traits. It is also planted to control soil erosion. A genetic transformation procedure will facilitate studies of the control of plant development and interactions with symbionts and pathogens, and will provide a tool for plant breeding. Here, we present a particle bombardment method for transforming seabuckthorn. The early stages of induced adventitious shoots from roots were chosen as a novel target tissue for the transformation procedure. The root system was bombarded with gold particles coated with plasmid pRT99gus containing genes for plant kanamycin resistance and for β-glucuronidase expression, and shoots were regenerated under kanamycin selection. PCR analysis of the regenerated transformed lines confirmed the presence of a 603 bp gus (uidA) gene fragment and a 1.5 kb fragment from the 35S promoter in three shoots from independent transformation events.

Essential Oils from Different Parts of the Sea Buckthorn <i>Hippophae rhamnoides</i>L.

The study investigated essential oils of berries and green biomass of the sea buckthorn at various growth stages. Plant material of H. rhamnoídes ssp. mongolica was collected in the experimental fields of the Institute of Cytology and Genetics SB RAS (Siberia, Russia). Volatile compounds and essential oils were detected. Green spring shoots without leaves, first-year seedlings without leaves and berries were analyzed. In all studied exemplars, 103 volatile compounds were defined among which considerable proportion wasn’t found in a sea-buckthorn earlier. Phenylpropanoids prevailed in essential oils composition from seedlings. Alkanes and phenylpropanoids prevailed in shoots. Berries contained mostly carboxylic acids and their esters. These data can be applied to a fingerprint of various cultivars of a sea-buckthorn with relevance to food science and technology.