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.

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。

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 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 communication 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 genotypes of H. 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-20911. These results revealed the immense potential of peroxidase isozyme patterns and RAPD as genetic markers for sex identification in H. rhamnoides L.

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 seabuckthorn (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 NaCl solutions 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.

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.

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.

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。

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 conducted 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 firstgrade lateral roots),and branching intensity of lateral roots(number of lateral roots bifurcation grade,number of firstgrade 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.

基于UPLC-Q-Orbitrap MS技术的沙棘黄酮类成分分析

利用UPLC-Q-Orbitrap MS技术对沙棘(Hippophae rhamnoides L.)果实中黄酮类物质进行系统剖析,并对不同产地沙棘果实中黄酮类化合物进行比较分析,为中药沙棘的化学物质基础和资源利用研究奠定基础。根据保留时间比对,精确分子量匹配和MS/MS碎片信息分析,共鉴定出58个化合物,包括34个黄酮醇、4个黄酮、14个花色素、1个二氢黄烷酮、2个二氢黄烷醇、2个黄烷醇和1个二氢查尔酮,其中14个花色素类化合物为首次在沙棘果实中鉴定。最终筛选得到38种差异积累的黄酮类化合物,其中大部分在青海和西藏产地的沙棘果中呈显著积累。研究丰富了沙棘果中黄酮类化合物化学物质基础研究,揭示了不同产地沙棘果中黄酮类化合物的组成及差异。

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

Objective: To explore the effects of total flavonoids of Hippophae rhamnoides L. (TFH), quercetin (Que) and isorhamnetin (Isor) on the intracellular free calcium (Ca~ 2+ _ i) in vascular smooth muscle cells (VSMC) of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY).Methods: Fluo 3-acetoxymethylester(Fluo-3/AM) was used to observe the effects of TFH (100mg/L) and its essential monomers, namely Que (10~ -4 mol/L) and Isor (10~ -4 mol/L) on changes of Ca~ 2+ _ i 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+ _ i 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 AngⅡcould 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 AngⅡ. (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). Conclusion: TFH, Que and Isor might decrease the levels of Ca~ 2+ _ i in VSMCs by blocking both voltage-dependent calcium channels (VDC) and receptor-operated 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.

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.

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。

沙棘属(Hippophae L.)植物的系统化学成分分析初报

本文主要分析了沙棘果实的特征、水溶性化学成分、油脂及其脂溶性物质 ;沙棘属植物中的黄酮物质、叶片的营养成分 ;柳叶沙棘的迁地保存及引种栽培。

沙棘多糖对脑胶质瘤大鼠的抑瘤作用及对HMGB1、MGMT及TLR4表达的影响

目的:探讨沙棘多糖(HRP)对脑胶质瘤大鼠的抑瘤作用及对高迁移率族蛋白1(HMGB1)、O6-甲基鸟嘌呤-DNA-甲基转移酶(MGMT)和Toll样受体4(TLR4)表达的影响。方法:选取60只SD大鼠,脑部接种C6脑胶质瘤细胞,建立脑胶质瘤模型。造模成功后随机分为模型组(15只)、HRP低剂量组(14只)、HRP高剂量组(14只)、替莫唑胺组(14只)。接种5 d后,HRP低剂量组、HRP高剂量组分别灌胃沙棘多糖100 mg/kg、200 mg/kg,替莫唑胺组灌胃替莫唑胺20 mg/kg,模型组灌胃等量生理盐水,每日1次,连续10 d。磁共振成像(MRI)检查并记录肿瘤大小,测量肿瘤体积,计算抑瘤率;苏木精-伊红(HE)染色观察脑胶质瘤形态学变化情况;流式细胞仪检测肿瘤组织细胞凋亡和细胞周期;实时定量聚合酶链式反应(PCR)检测肿瘤组织中HMGB1、MGMT和TLR4 mRNA水平;蛋白免疫印迹法(Western Blot)检测肿瘤组织HMGB1、MGMT和TLR4蛋白表达。结果:与模型组比较,HRP低剂量组、HRP高剂量组、替莫唑胺组肿瘤体积减小,Sub-G1期、G0/G1期细胞增加,S期、G2/M期细胞减少,凋亡率升高,HMGB1、MGMT和TLR4 mRNA及蛋白相对表达量降低,差异均有统计学意义(P<0.05);与HRP低剂量组比较,HRP高剂量组、替莫唑胺组肿瘤体积减小,抑瘤率升高,Sub-G1期、G0/G1期细胞增加,S期、G2/M期细胞减少,凋亡率升高,HMGB1、MGMT和TLR4 mRNA及蛋白相对表达量降低,差异均有统计学意义(P<0.05);与HRP高剂量组比较,替莫唑胺组抑瘤率降低,Sub-G1期、G0/G1期细胞减少,S期、G2/M期细胞增加,凋亡率降低,HMGB1、MGMT和TLR4 mRNA及蛋白相对表达量升高,差异均有统计学意义(P<0.05)。HE染色显示,模型组脑胶质瘤内肿瘤细胞生长活跃,数量丰富,排列密集,核大深染,肿瘤细胞异型性明显,核浆比例高,核分裂现象多见,呈浸润性生长,无明显细胞萎缩坏死现象;HRP低剂量组、HRP高剂量组和替莫唑胺组细胞数量较模型组减少,细胞生长密度降低,细胞体积减小,出现部分坏死。结论:HRP对脑胶质瘤大鼠具有抑瘤作用,可能与抑制HMGB1、MGMT及TLR4的mRNA转录与蛋白表达有关。