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.

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。

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.

水分胁迫对沙棘(Hippophae rhamnoides)和中间锦鸡儿(Caragana intermedia)蒸腾作用影响的比较

多年生灌木沙棘和中间锦鸡儿是黄土高原生态重建的重要物种,设计人工模拟水分胁迫实验,测量沙棘和中间锦鸡儿蒸腾作用的各种指标,研究其蒸腾特性对水分胁迫的适应方式。结果表明,同等水分处理条件下,中间锦鸡儿单叶水平上的蒸腾速率高于沙棘。沙棘和中间锦鸡儿的蒸腾速率日进程在晴天、阴雨天和生长发育的不同阶段明显不同。夜间蒸腾占全天蒸腾的比例相当大,夜间蒸腾在不同物种之间、不同供水量之间存在明显差异,而且越干旱的环境比例越大。两种植物气孔阻力的季节变化格局在不同水分处理间大体相似。沙棘的昼夜蒸腾节律在各种水分处理条件下都表现出明显的气孔振荡现象,而中间锦鸡儿没有。叶片温度、光合有效辐射和气孔阻力是各种水分条件下沙棘和中间锦鸡儿蒸腾作用的共同的限制因子,相对于沙棘,中间锦鸡儿还更多地受到空气相对湿度的影响。

中国沙棘(Hippophae rhamnoides ssp. Sinensis)根瘤内Frankia菌的遗传多样性

为了研究中国沙棘亚种共生菌Frankia的遗传多样性,利用PCR-RFLP分子标记方法,对从青海西宁到内蒙古库伦17个地点采集的106个中国沙棘根瘤样品进行遗传多样性分析。供试样品nifD-nifK基因间隔区（IGS）扩增产物分别用3种内切酶（HinfⅠ、HaeⅢ和MboⅠ）酶切,共产生21条酶切谱带,其中17条为多态性条带,多态位点百分比（PPL）为80.99%,所有样品可被划分为9个基因型。结果表明中国沙棘根瘤内的Frankia菌有丰富的遗传多样性,土壤质量较好地点的丰富度高于土壤质量较差地点,海拔较高地区的丰富度高于海拔较低地区,多数地点至少有2种不同基因型的Frankia菌。聚类分析显示Frankia菌不同基因型间的遗传距离在4.88%～55.96%之间,它们在不同地点的分布是不均匀的,没有发现不同基因型菌间的亲缘关系与地点有相关性。中国沙棘根瘤中Frankia菌可分为两个基因型组,组内基因型分布比较一致,而组间有明显差异。

俄罗斯大果沙棘(Hippophae rhamnoides L.)种子萌发特性

俄罗斯丘依斯克大果沙棘(Hippophae rhamnoides L.)为优良的沙棘引进种。经测定,俄罗斯大果沙棘丘依斯克种子的千粒重为17.75g,与其它沙棘属的种子相比,其千粒重较大,为其2倍左右。四唑(TTC,1.0%)染色测种子的生活力的结果表明:有生活力的种子占97.75%,说明俄罗斯大果沙棘种子饱满度好;染色结果与对比发芽实验的结果很接近,说明用四唑染色来测定俄罗斯大果沙棘种子的发芽能力是较准确的方法。在水、气适宜的条件下,分别研究了温度分别为15、20、25、30℃,光照强度分别为25%、50%、75%、100%的不同处理对丘依斯克种子萌发的影响,并采用胚根生长量和全株生物量对沙棘种子发芽效果进行了评价,提出了沙棘种子发芽的最适温度和光强。结果表明,在25℃条件下,种子萌发最早,发芽势高达33.0%±4.76%,发芽率高达95.5%±1.5%,且主根生长量和生物量最大,分别为(4.5±0.09)cm和(0.137±0.002)g;75%光强最适合沙棘种子萌发,种子萌发最早,发芽势高达61.5%±1.7%,发芽率高达86.0%±1.1%,且主根生长量和生物量最大,分别为(3.7±0.2)cm和(0.108±0.004)g。因此,在直播营造大果沙棘林时,应首选25℃的温度条件,同时,建议进行适度遮荫处理。

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.

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.

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.

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.

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.

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.

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.

The Effects of Total Flavones of Hippophae Rhamnoides L. on Cardiac function and Hemodynamics of Normal Subjects

Healthy volunteers were randomly divided into TFH group(22 subjects) and placebo control group (20 subjects). We used double-blind method. The cardiac performance and hemodynamics of the subjects before and after administration of TFH or placebo were determined by Admittance and STI methods, Oral TFH in a dose of 10 mg could significantly decrease the PEP/LVET, ICT/LVET and ICT of the mormal subjects(P<0.05). Oral TFH could increase SV(P<0.05). CO, CI, SWI, CWI and LI also tended to increase. Oral TFH also markedly reduced TPR (P<0.05) and increasedc (P<0.05). After administration of TFH, HR and HP did not significantly change(P>0.05). After administration of placebo, the parameters observed did not significantly change(P>0.05). The above results show that TFH can strengthen myocardial contractility and pump function of the heart, reduce total peripheral vascular resistance and increase vascular clasticity in normal subjects.

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.

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.