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。

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.

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.

Comparative Analysis of Antioxidants in Hippophae rhamnoides L. Leaves by UPLC

[Objectives]The research aimed to compare antioxidation abilities of different extraction phases of Hippophae rhamnoides L. leaves extract and find its active marker. [Methods] H. rhamnoides L. leaves were extracted firstly by 75% of ethanol,and then were extracted by petroleum ether,chloroform,ethyl acetate and n-butanol in turn after decompression concentration. Via DPPH and ABTS^+ radical scavenging experiments and Fe^(3+) reduction experiment,antioxidation activity of each phase was evaluated. And chemical compositions related to its antioxidation activity were determined by using UPLC for qualitative identification. [Results] Antioxidation activity sequence of each phase was ethyl acetate phase≈n-butanol phase > alcohol extract of raw material > water phase > chloroform phase > petroleum ether phase. 33 kinds of chemical compositions were isolated from alcohol extract and extracts by using UPLC. [Conclusions] UPLC and antioxidation activity test were used simultaneously to analyze alcohol extract and extracts of H. rhamnoides L. leaves,which could fast and accurately determine extraction phase with the strongest antioxidation ability. The research is significant to study active ingredients and mechanism of antioxidation from H. rhamnoides L. leaves.

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.

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.

Relationships between morphological descriptors and RAPD markers for assessing genetic variability in Hippophae rhamnoides L.

Knowledge of genetic relatedness among accessions of germplasm is necessary for the development of breeding strategies to produce improved cultivars. The present investigation on Hippophae rhamnoides was carried out to assess its genetic variability in Himachal Pradesh, India, by employing morphological and RAPD markers. Different areas of Himachal Pradesh were surveyed and eight sites were finally selected Twenty four genotypes were selected for further studies, i.e., three genotypes from each site. On the basis of morphological studies, the genotypes of the Ropa site （Kinnaur） were considered elite genotypes. To assess the variability at the molecular level, RAPD patterns were studied by random primers. The total number of bands amplified was 607, out of which 487 bands were identified as polymorphic, depicting 80.23 per cent variability. Six unique bands were produced from three primers （OPA- 05, OPA-10 and OPD-08） specific for five genotypes, including three genotypes of the Ropa site, Kinnaur District.

Protective Effects of Polyphenol Extracts from Sea Buckthorn (Hippophaë rhamnoides L.) on Rat Hearts

The objective of present study was to investigate the antioxidative and cardioprotective effects of polyphenol extracts from sea buckthorn (PESB) against myocardial ischemia reperfusion injury (MIRI). PESB was analyzed using HPLC-DAD and their antioxidant activities were evaluated by measuring the intrinsic free radicals cavenging capacity and inhibiting the formation of hydroxyl radical capacity. Experiments on the isolated rat hearts were carried out according to the improved Langendorff retrograde perfusion technology. Coronary effluent of lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CK-MB) activities were analyzed to determine the extent of myocardial injury. Protein expression levels of Beclin-1, microtubule-associated protein 1 light chain 3 (LC3) I and II were measured to evaluate the degree of autophagy. In conclusion, coupled results from both in vivo and in vitro experiments have confirmed that PESB has potential health implications in both prevention and amelioration of MIRI, which may be attributed to the inhibition of autophagy.

Comparative quantitative analysis of fruit oil from Hippophae rhamnoides(seabuckthorn) by qNMR, FTIR and GC–MS

Objective: To develop a qNMR method for quantitative analysis of triacylglycerols in fruit oil of Hippophae rhamnoides(seabuckthorn, SBT) and analyze commercial samples of SBT oils using GC–MS and FTIR.Methods: SBT fruit oil(IPHRFH) was extracted with hexane and the triglyceride(TAG) was isolated by vacuum liquid chromatography. Six different branded SBT oils purchased from e-commerce suppliers(Amazon) and in-house prepared SBT oil was analyzed by qNMR and fatty acyl composition of TAGs determined by using NMR. In-house oil was also analysed by GC–MS and FTIR spectroscopy.Results: The qNMR results showed that the oil contained 80.3% of triacylglycerol(TAG). The SBT oil TAGs comprised of linolenate 6.6%, palmitoleate/oleate 65.4%, and total saturated fatty acyl chain including palmitate 28% as determined by qNMR. GC–MS analysis revealed that the major acyl functionalities present in the TAG were palmitoleic acid 36.5%, oleic acid 12.9%, palmitic acid 21.2%, and linoleic acid 18%. Of the six commercial samples analyzed, samples from only one supplier(SW) were fruit oil;All others were the seed oils or mix of fruit oil and seed oil. The labels for samples except for the SW did not indicate whether it was fruit oil or seed oil.Conclusion: The results suggest that SBT oil should be analyzed by combination of GC–MS, FTIR and qNMR for factual content of free fatty acid or TAGs, which are chemically different in nature and affect the quality of oil. GC–MS showed the content of omega free fatty acids after hydrolysis, while qNMR and FTIR showed the content of TAGs. The major acyl functionalities found in SBT fruit oil TAGs are palmitoleate/palmitate/oleate, while linoleate and linonelate make up a minor fraction. Furthermore,analysis of commercial samples showed discrepancies between label claims and actual content.

沙棘(Hippophae rhamnoides)种植对鄂尔多斯砒砂岩地区土壤容重、孔隙度与贮水能力的影响

在砒砂岩地区营造沙棘(Hippophae rhamnoides)林可显著缓解该区蓄水能力差和水土流失严重的问题,但其中的机理还不清楚。通过对2~8年生沙棘林下不同深度土壤的容重、孔隙度、持水量等指标观测,探讨了人工栽植沙棘对砒砂岩地区土壤物理性质动态的影响。结果表明:沙棘人工林显著降低了土壤容重,增加了土壤孔隙数量和土壤持水量,而且这种作用随林龄的增加而增加,但随土层深度增加而降低,主要表现在0~10cm土层。可见,在砒砂岩地区营造沙棘人工林有利于改善土壤水分物理状况,原因可能是由于枯落物和根系的作用。

沙棘(Hippophae rhamnoides)对经客土改良石漠化土壤的适应性

石漠化地区基岩裸露、砾石堆积,水土流失严重,土壤与植被覆盖率低,因此引发一系列的生态恶化问题,而客土恢复植被是一项快速有效的治理途径。本试验在贵州毕节地区实施,以当地黄壤客土(T1,黄壤与砾石比为1∶1;T2,黄壤与砾石比为2∶1;T3,黄壤与砾石比为3∶1;T4,黄壤与砾石比为1∶0)改良石漠化,从耐瘠薄的灌木——沙棘(Hippophae rhamnoides)为试材,探究其幼苗在客土改良后的石漠化土壤中的适应性,栽种期为2年。结果表明:客土处理对沙棘幼苗的冠幅、粗根的根长密度、粗根根重密度均无显著影响;百分之百客土即T4会显著抑制沙棘株高、根系生物量,显著促进细根比根长的增长;当黄壤与砾石比例1/2—2/3时,2年后沙棘细根的根重密度与土壤含水量均会显著增加;当黄壤与砾石比大于2/3时会显著抑制沙棘根系长度总和的增加,且2年后对土壤水分改善作用不显著;沙棘根系指标与2年后的土壤含水量呈现出显著的相关关系。在贵州毕节地区客土改良石漠化时,并非客土越多越好,当黄壤与砾石的比例在1/2—2/3时,栽种沙棘2年可显著改善土壤含水量。

Preparation of a Hippophae rhamnoides Linn oil nanoemulsion and the visualization of its transdermal permeation

A Hippophae rhamnoides Linn oil nanoemulsion was developed, and its physicochemical properties and in vitro transdermal characteristics were investigated. Then its skin irritation was investigated. The optimum Hippophae rhamnoides Linn oil nanoemulsion prescription was determined using Cremophor EL as an emulsifier, Transcutol P as a co-emulsifier, and isopropyl myristate(IPM) as the oil phase(IPM%: Cremophor EL%: Hippophae rhamnoides Linn oil%= 5:16:20, Km = 4:1). The prepared Hippophae rhamnoides Linn oil nanoemulsion was pale yellow and transparent. Globular droplets were observed under a transmission electron microscope(TEM). The average particle size was 52.2±4.8 nm. The loading capacity was 10.68%, and the oil-in-water(O/W) type nanoemulsion was stable after centrifugation. The CLSM results showed that the fluorescence intensity of the OB Hippophae rhamnoides Linn oil nanoemulsion group was stronger than that of the Hippophae rhamnoides Linn oil cream and Hippophae rhamnoides Linn oil groups in each layer of skin. The study also showed that skin surface treated with the Hippophae rhamnoides Linn oil nanoemulsion had strong fluorescence. The skin depth had weak fluorescence, while the areas near the hair follicle and its appendages had the strongest fluorescence. The skin irritation test showed that the prepared Hippophae rhamnoides Linn oil nanoemulsion was applied to the normal and damaged skin of the rabbit, and there was no apparent redness and swelling. These results suggested that the nanoemulsion coated with Hippophae rhamnoides Linn oil was a potential delivery system for skin disorders.