Effect of Chitinase on Resistance to Fungal Pathogens in Sea Buckthorn, Hippophae rhamnoides, and Cloning of Class I and III Chitinase Genes

Sea buckthorn (Hippophae rhamnoides L.) is naturally distributed from Asia to Europe. It has been widely planted as an ornamental shrub and is rich in nutritional and medicinal compounds. Fungal pathogens that cause diseases such as dried-shrink disease are threats to the production of this plant. In this study, we isolated the dried-shrink disease pathogen from bark and total chitinase protein from leaves of infected plants. The results of the Oxford Cup experiment suggested that chitinase protein inhibited the growth of this pathogen. To improve pathogen resistance, we cloned chitinase Class I and III genes in H. rhamnoides, designated Hrchi1 and Hrchi3. The full-length cDNA of the open reading frame region of Hrchi1 contained 903 bp encoding 300 amino acids and Hrchi3 contained 894 bp encoding 297 amino acids. Active domain analysis, protein types, and secondary and 3D structures were predicted using online software.     

Molecular Genetic Variation in Chinese Populations of Three Subspecies of Hippophae rhamnoides

To provide a population-level genetic profile for investigation and conservation of genetic diversity of the sea buckthorn (Hippophae rhamnoides L.), 300 individuals of fifteen natural populations of sea buckthorn in China were analyzed by using ISSR (inter-simple sequence repeats) markers. On the basis of Shannon's index and Nei's genetic diversity, the mean genetic diversity detected in the natural populations of Hippophae rhamnoides ssp. yunnanensis, ssp. sinensis, and ssp. gyantsensis was 0.1944, 0.2169, and 0.1372, respectively. The coefficient of gene differentiation (Gst) of seven ssp. yunnanensis populations is 0.2790, and that of seven ssp. sinensis populations is 0.4184. This means that 28% of the total molecular variance of seven ssp. yunnanensis populations existed among populations, and 42% for seven ssp. sinensis populations, suggesting that the subspecies have different genetic structures. No significant correlation between genetic and geographic distances of the populations was found using ISSR markers.     

不同程度的水分胁迫对沙棘幼苗生理生态特征的影响

为探讨未来降水减少对内蒙古皇甫川流域沙棘幼苗生理生态特征的影响,特设计平均降雨水平、偏旱、干旱和极端干旱4种不同的水分梯度处理,开展人工水分梯度实验。方差分析表明,不同的水分梯度显著影响土壤的含水量、土壤温度等微生境因子,并显著影响净光合速率、气孔导度、蒸腾速率等气体交换特征、资源利用效率和叶片水势特征。适度的水分胁迫(干旱环境)能够提高沙棘的水分利用效率,同时却降低净光合速率和蒸腾速率。各种生理生态指标表明,4种水分处理的沙棘幼苗都受到不同程度的水分胁迫的影响,极端干旱环境中沙棘幼苗的内在生理调节机制出现紊乱,皇甫川流域沙棘不适宜在极端干旱环境中生长。     

不同程度的水分胁迫对沙棘幼苗生理生态特征的影响

为探讨未来降水减少对内蒙古皇甫川流域沙棘幼苗生理生态特征的影响,特设计平均降雨水平、偏旱、干旱和极端干旱4种不同的水分梯度处理,开展人工水分梯度实验.方差分析表明,不同的水分梯度显著影响土壤的含水量、土壤温度等微生境因子,并显著影响净光合速率、气孔导度、蒸腾速率等气体交换特征、资源利用效率和叶片水势特征.适度的水分胁迫(干旱环境)能够提高沙棘的水分利用效率,同时却降低净光合速率和蒸腾速率.各种生理生态指标表明,4种水分处理的沙棘幼苗都受到不同程度的水分胁迫的影响,极端干旱环境中沙棘幼苗的内在生理调节机制出现紊乱,皇甫川流域沙棘不适宜在极端干旱环境中生长.     

Chromatographic analysis of carotenol fatty acid esters in Physalis alkekengi and Hippophae rhamnoides

The carotenol fatty acid esters of two potentially valuable sources of plant carotenoids, sepals of Physalis alkekengi (Chinese lantern) and fruits of Hippophae rhamnoides (sea buckthorn), were separated by column chromatography and identified by HPLC-DAD and HPLC-MS. A chemical and an enzymatic hydrolysis were employed to identify the parent carotenoids and to remove the lipid components. Zeaxanthin and beta-cryptoxanthin esters represented the main fraction in P. alkekengi sepals and an important one in H. rhamnoides fruits. Beta-Cryptoxanthin palmitate and zeaxanthin dipalmitate were identified as major compounds in both plants. In P. alkekengi, the carotenoids were mainly (> 90%) esterified with palmitic acid, and a high proportion (> 80%) of saturated medium chain fatty acids was found (by GC-MS) in the total lipid extract. Although the total lipid extract of H. rhamnoides contained significant amounts of unsaturated fatty acids, especially oleic and palmitoleic acids, the xanthophylls were mainly esterified with saturated fatty acids. The oleoresins of both species represent potential sources of carotenoid esters and can be used as food additives, cosmetic ingredients or nutraceuticals.     

Antioxidant capacity of crude extracts containing carotenoids from the berries of various cultivars of Sea buckthorn (Hippophae rhamnoides L.)

Comparative analysis of antioxidant capacity was performed using FRAP and DPPH methods on extracts containing carotenoids acquired from fruits of Sea buckthorn. The examination included nine varieties of Sea buckthorn growing in the comparative cultivation. Conducted analysis allowed to compare the antioxidant capacity with carotenoids content measured with spectrophotometric and HPLC methods. Three of the examined cultivars indicating high antioxidant activity in both, FRAP and DPPH methods, also revealed highest ('Aromatnaya') and high ('Botanicheskaya', 'Arumnyj') total carotenoids content in HPLC analysis.     

Effects of an antioxidant-rich juice (sea buckthorn) on risk factors for coronary heart disease in humans

There is increasing evidence to support the hypothesis that free radical-mediated oxidative processes contribute to atherogenesis. More recently the ability of antioxidant nutrients to affect cell response and gene expression has been reported in vitro, providing a novel mechanistic perspective for the biological activity of antioxidants. Sea buckthorn (Hippopha? rhamnoides L.) is a rich source of antioxidants both aqueous and lipophilic, as well as polyunsaturated fatty acids. The objective of the study was to characterize the antioxidant profile of Sea buckthorn juice (SBJ) and to evaluate its effect on plasma lipids, LDL oxidation, platelet aggregation and plasma soluble cell adhesion protein concentration. Twenty healthy male volunteers were given either a placebo or SBJ for 8 weeks. Additional daily intakes of vitamin C, alpha-tocopherol, beta-carotene and flavonoids through SBJ supplementation were 462, 3.2, 1.0 and 355 mg respectively. There were no significant changes in plasma total cholesterol, LDL-C, platelet aggregation or plasma intercellular cell adhesion molecule 1 (ICAM-1) levels between treatment groups. Although not significant, a 20% and 17% increase in plasma HDL-C and triacylglycerol (TAG) concentrations were observed. SBJ supplementation also resulted in a moderate decrease in the susceptibility of LDL to oxidation.     

Effect of dietary supplementation with sea buckthorn (Hippophaë rhamnoides) seed and pulp oils on the fatty acid composition of skin glycerophospholipids of patients with atopic dermatitis

Sea buckthorn (Hippopha? rhamnoides) seed and pulp oils have traditionally been used for treating skin diseases in China and Russia, but are not widely used in other countries. A placebo-controlled, parallel study was carried out to investigate the effects of these oils on the fatty acid composition of skin glycerophospholipids of patients with atopic dermatitis. Sixteen patients ate 5 g of sea buckthorn seed oil, pulp oil, or paraffin oil daily for 4 months. Skin fatty acids were analyzed with gas chromatography before and after treatment. The seed oil slightly increased the proportion of docosapentaenoic acid (22:5n-3) and decreased the proportion of palmitic acid (16:0) in skin glycerophospholipids (0.05 < P < 0.1). The levels of the other fatty acids remained stable. The results show that the fatty acid composition of skin glycerophospholipids is well buffered against short-term dietary modification.     

不同树种对刺沙棘钉毛蚜的耐害性

在野外自然感染条件下,以新梢卷叶恢复率作为耐虫性指数,分析了20个不同树种对刺沙棘钉毛蚜的抗虫程度及其耐害补偿能力。结果表明:1)除桃树、沙枣和榆树外,其它树种均为刺沙棘钉毛蚜的寄主树种。2)在所有沙棘树种中,沙棘品种和沙棘杂交种最为感虫,而沙棘种和亚种则抗虫性较好。3)不同树种卷叶恢复程度差异较大。在危害高峰期,卷叶率较低的树种,如橙色、棱果沙棘等,在自身耐害补偿功能和自然天敌的影响下,其卷叶可以完全恢复正常,表现出较强的耐害性;而卷叶率高的树种,如优胜、楚伊、乌兰格木等沙棘品种,由于受害程度较重,自身耐害补偿功能力有限,卷叶不能完全恢复正常,恢复率不到50%,表现出较弱的耐害性;苹果和云南沙棘卷叶恢复率分别为55.6%和50%,耐害性居于上述两者之间。     

Growth and physiological responses to drought and elevated ultraviolet-B in two contrasting populations of Hippophae rhamnoides

In the southeast of the Qinghai-Tibetan Plateau of China, sea buckthorn ( Hippophae rhamnoides L.), which is a thorny nitrogen-fixing deciduously perennial shrub, has been widely used in forest restoration as the pioneer species. In our study, two contrasting populations from the low and high altitudinal regions were employed to investigate the effects of drought, ultraviolet-B (UV-B) and their combination on sea buckthorn. The experimental design included two watering regimes (well watered and drought stressed) and two levels of UV-B (with and without UV-B supplementation). Drought significantly decreased total biomass, total leaf area and specific leaf area (SLA), and increased root/shoot ratio, fine root/coarse root ratio and abscisic acid content (ABA) in both populations. However, the high altitudinal population was more responsive to drought than the low altitudinal population. On the other hand, elevated UV-B induced increase in anthocyanins in both populations, whereas the accumulation of UV-absorbing compounds occurred only in the low altitudinal population. The drought-induced enhancement of ABA in the high altitudinal population was significantly suppressed in the combination of drought and elevated UV-B. Moreover, significant drought × UV-B interaction was detected on total biomass in both populations, total leaf area and fine root/coarse root in the low altitudinal population, and SLA in the high altitudinal population. These results demonstrated that there were different adaptive responses between two contrasting populations, the high altitudinal population exhibited higher tolerance to drought and UV-B than the low altitudinal population.     

Inositols and methylinositols in sea buckthorn (Hippophaë rhamnoides) berries

Sea buckthorn (Hippophaë rhamnoides L.) berries, especially of ssp. sinensis, contain significant quantities of an unknown, water-soluble compound, evidently a cyclitol derivative. The compound was isolated by HPLC and analyzed by GC–MS [trimethylsilyl (TMS) derivative, selected ion monitoring (SIM) and total ion chromatogram (TIC) analyses], by ¹H and ¹³C NMR and by optical activity measurements. The results together with analyses of reference compound verified the unambiguous structure (?)-2-O-methyl-l-chiro-inositol (l-quebrachitol). In addition, chiro-inositol and myo-inositol existing in trace amounts were identified based on reference compounds, chromatographic data and mass spectra of the TMS derivatives. Methyl-myo-inositol was tentatively identified based on chromatography and mass spectrometry. Inositols and methyl inositols are bioactive compounds essential for regulating physiological processes of plants and humans. To our knowledge, this is the first report on the presence of chiro-inositol and myo-inositol in sea buckthorn and l-quebrachitol in edible berries. The identification of the inositols and l-quebrachitol in sea buckthorn may bring new insights into the sensory properties and also mechanisms behind the health effects of the berry.     

锰胁迫和性别竞争交互处理下沙棘雌雄幼苗生理响应特征

沙棘(Hippophae rhamnoides)是重要的雌雄异株人工林防护树种,但对其环境胁迫的性别响应差异研究不足,性别竞争与胁迫因子的交互效应响应特征尚不清楚。为了探讨锰胁迫和性别竞争交互处理下沙棘雌雄植株的生理响应特征和耐受能力,旨在为沙棘修复土壤重金属污染提供实践指导,该文研究了锰胁迫(4 000 mg·kg^-1)和3种不同性别组合模式(雌雄、雌雌、雄雄)处理下沙棘的生理响应,分别测定雌雄沙棘叶片中叶绿素、过氧化物酶(peroxidase,POD)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、总酚(total phenols, TP)、游离脯氨酸(free proline,Pro)、可溶性糖(soluble sugar,SS)、甜菜碱(glycine betaine, GB)以及锰含量。结果表明:(1)锰胁迫下,在所有竞争组合中,性间竞争的雄株(M/FM)SOD活性最高,而MDA含量与对照相比未有明显升高,表明雄株的抗氧化能力更好,膜氧化损伤程度更小。(2)锰胁迫时M/FM积累了更多的游离脯氨酸,表现出更好的渗透调节能力和耐受能力。(3)交互效应分析显示性别互作和锰胁迫交互处理显著影响了沙棘雌雄叶片的光合色素、抗氧化酶活性和渗透调节能力; 主成分分析显示SOD、POD、MDA、叶绿素b(chlorophyll b, Chlb)、SS、Pro可作为重要的生理响应指示参数。该研究结果对于利用沙棘修复土壤重金属污染可提供一定的参考。     

Effect of pre-sowing γ-irradiation of sea buckthorn seeds on the content and fatty acid composition of total lipids in the seeds of the first plant generation

Russian Journal of Plant Physiology - Absolute content and FA-composition of sea buckthorn (Hippophaë rhamnoides L.) seed lipids were studied. The seeds of cvs. Vitaminnaya and Zyryanka...     

最优化制备沙棘膏工艺研究

沙棘果为制备沙棘膏的原料,以沙棘膏中总黄酮含量为考察影响工艺因素的指标,通过单因素试验和正交试验,确定最优化制取沙棘膏工艺为:料液比1∶20(g/mL),温度75℃,提取溶剂为80%乙醇,综合考虑成本因素,提取时间4 h。按照该工艺沙棘膏中总黄酮的得率为15.64%。     

Inhibitory effects of sea buckthorn (Hippophae rhamnoides L.) seed on UVB-induced Photoaging in human dermal fibroblasts

Sea buckthorn (Hippophae rhamnoides L.) seed, which has very high nutraceutical, cosmetoceutical and therapeutic properties, has been widely used to treat diseases in Tibetan and Mongolian traditional medicines. In this study, we measured the antioxidant activities of the solvent-dependent SBS (Sea buckthorn seed) extracts using the DPPH and Folin-Ciocalteu assays and we investigated the efficacy of SBS in protecting skin against UVB-induced damage using cultured human dermal fibroblasts. In addition, the inhibitory effects of SBS on UVB-induced skin photoaging were examined by determining the level of Metalloproteinase (MMP)-1 and procollagen after UVB-irradiation. The antioxidant capacity of the SBSE (alcoholic) extract was significantly higher than the SBSH (hexane) and SBSW (water) extracts, as measured based on the free-radical scavenging activity and total phenolic content. SBSE was chosen as the most suitable candidate antioxidant. In human dermal fibroblasts, the cell viability of SBSE extract at doses of 2.5, 5, and 10 ??g/mL was higher than the UVB-treated control. By RT-PCR and Western blot, SBSE treatment inhibited UVB-induced IL-1?? expression in cultured cells. In addition, SBSE restrained UVB-induced IL-6 and COX-2 gene expression in cultured fibroblasts in a dose-dependent manner. Treatment with the SBSE extract after UVB irradiation in human dermal fibroblasts significantly reduced MMP-1 expression and increased procollagen synthesis when compared with UVB-irradiation only. In this study, SBSE was shown to increase the synthesis of procollagen, decrease the expression of MMP-1, and inhibit the production of IL-1??, IL-6, and COX-2 in UVB-irradiated human fibroblasts. These findings suggest that the SBSE extract may be a potential therapeutic agent for preventing and treating skin photoaging.     

祁连山两侧中国沙棘不同居群的遗传多样性研究

应用RAPD技术研究了祁连山中段中国沙棘(Hippophae rhamnoidoes ssp．sinensis)4个居群和1个对照居群的遗传多样性水平,探讨中国沙棘边缘居群的遗传变异以及天然屏障祁连山对中国沙棘居群遗传结构的影响。结果显示,祁连山地区中国沙棘居群水平的Nei’基因多样度(h)和Shannon多态信息指数(I)分别为0．2070和 0．297 4,基因分化系数(Gst)为0．319 3,均高于中国沙棘整个分布区的平均值,表明该地中国沙棘边缘居群遗传多样性水平及居群间的遗传分化都有增加的趋势;居群间每代迁移数(Nm=1．065 9)显著低于异交风媒植物(Nm= 5．380),表明祁连山的隔离对中国沙棘居群间的基因交流有限制作用,造成了该地中国沙棘边缘居群间较高的遗传分化。     

Bioactive Components in Methyl tert-Butyl Ether Extract of Sea Buckthorn (Hippophae rhamnoides L.) Green Waste

Russian Journal of Bioorganic Chemistry - Lipophilic constituents of sea buckthorn leafy shoots, which is a large-scale waste product of sea buckthorn oil production and rejuvenating pruning of...     

Mechanisms underlying host plant selection by Holcocerus hippophaecolus adults

We determined the mechanisms underlying host selection by adults of the seabuckthorn carpenterworm, Holcocerus hippophaecolus Hua, Chou, Fang et Chen. Four sea buckthorn (Hippophae rhamnoides L.) subspecies (varieties) with different degrees of resistance to H. hippophaecolus were chosen for artificial insect infection in cages. The results showed that olfactory and visual cues are very important for the selection of host plants by H. hippophaecolus, but that olfactory stimuli play a more vital role in this process. The relative abundance of branches and leaves had no effect on the likelihood that adults landed on plants from four subspecies (varieties), but did influence landing rates within the same subspecies (varieties). When considering only the most resistant sea buckthorn subspecies (varieties), the presence of luxuriant branches and leaves led to lower landing rates. These results provide a theoretical basis for the understanding of H. hippophaecolus damage to sea buckthorn and the means to implement effective measures of control.     

盐胁迫下沙棘的渗透调节效应

分别用含有0、100、200和300mmol LNaCl的Hoagland培养液处理1年生沙棘(HippophaerhamnoidesL.)苗30d后,测定其鲜重,干重,含水量,可溶性糖、脯氨酸和无机离子(Na 、Cl-)的含量及叶片渗透势和渗透调节能力。结果表明:100mmol LNaCl处理的沙棘地上部和根的鲜重和干重最大,其含水量也最大;NaCl浓度超过100mmol L时,沙棘地上部分和根的鲜重和干重随盐浓度增加而逐步下降,其下降的趋势为地上部大于根部。随NaCl浓度不断升高,沙棘体内Na 和Cl-浓度随之升高,茎叶和根系中Cl-含量明显高于Na ,对Na 的相对吸收量多于Cl-。沙棘对盐胁迫有一定的适应能力,随NaCl浓度的升高,沙棘叶内脯氨酸含量升高,可溶性糖含量增加,渗透势降低,渗透调节能力增强。本结果可为盐碱地营造沙棘林提供依据。