Substrate,Hormone,Winnowing,and Stratification Influence the Seed Germination of Ilex asprella(Hook.et Arn.)Champ.ex Benth

Ilex asprella(Hook.et Arn.)Champ.ex Benth is one of the most important traditional Chinese medicines in southern China.The seeds of Ilex asprella usually have extremely low germination due to their dormancy characteristics,which severely impacts the efficiency of seedling raising and increases labor costs.In this study,to improve the seed germination of I.asprella,the effects of germination substrate,hormone,winnowing,and stratification treatments on the seed germination of I.asprella were investigated.The results of the germination matrix showed that the highest germination percentage of 45.2%was achieved under the 20℃/10℃day/night temperature and vermiculite germination medium conditions.The results of hormone treatments revealed that 100–400 mg/L of gibberellin(GA)and 50–100 mg/L of salicylic acid(SA)were found to be effective in releasing the dormancy of I.asprella seeds.Moreover,winnowing could effectively eliminate unsaturated seeds and impurities,thus improving the seed germination of I.asprella.Furthermore,warm temperature(15℃)stratification could expand the temperature range of I.asprella’s seed germination,which was beneficial for seed germination of I.asprella and for seed nursery at room temperature in production practice.The present study obtained a method to break dormancy and increase seed germination in I.asprella,thereby forming a groundwork for improving the efficiency of large-scale planting of I.asprella.

高效液相色谱-蒸发光散射检测法测定毛冬青药材中Ilexgenin A的含量

目的建立毛冬青药材中Ilexgenin A的高效液相色谱-蒸发光散射检测（HPLC-ELSD）的含量测定方法。方法采用Lichrospher-C18色谱柱（4.6 mm×250 mm，5μm），流动相为乙腈-0.1%醋酸溶液（60∶40），流速1 ml/min，蒸发光散射检测器：漂移管温度96℃，气体流速2.1 L/min。结果Ilexgenin A在0.7-14.5μg浓度范围内线性关系良好，r=0.999 7（n=6）。平均回收率为100.32%，RSD=1.38%（n=6）。结论本法操作快速、准确，可作为毛冬青药材中IlexgeninA的质量控制方法。

不同时期大叶苦丁茶(Ilex latifolia Thunb.)不同器官中咖啡酰奎宁酸类成分的毛细管电泳分析（英文）

大叶苦丁茶(Ilex latifolia Thunb.)含有丰富的咖啡酰奎宁酸。在本研究中,运用胶束电动毛细管色谱法,测定了在不同发育时期的大叶苦丁茶不同器官中的咖啡酰奎宁酸含量。大叶苦丁茶的不同器官在不同发育时期的咖啡酰奎宁酸含量有着显著的变化。在叶片中,10天叶龄的叶的咖啡酰奎宁酸含量是19.78 g/100 g,而当叶龄达到12月时,其中的咖啡酰奎宁酸含量已经降为1.41 g/100 g。随着发育时间的增长,雌花、雄花中的含量变化也有类似的变化。雌花(果)和雄花在发育期为10天时的咖啡酰奎宁酸含量高:分别20.29 g/100 g和18.06 g/100g,随后在雌花发育成的7个月果实中下降到0.16 g/100 g,而在40天期接近脱落的雄花中其含量降为10.36 g/100 g。大叶苦丁茶的器官中,除了嫩叶,雄花由于高含量的咖啡酰奎宁酸和较多的生物量,也是一种好的咖啡酰奎宁酸来源材料。本文首次报道了大叶苦丁茶的花中含有大量的咖啡酰奎宁酸。MEKC方法也是首次被用于苦丁茶中的咖啡酰奎宁酸类成分的测定,使用这种方法,大叶苦丁茶中的6种咖啡酰奎宁酸成分在40分钟内完全分离。相较于以前文献中报道的高效液相色谱法,本文为苦丁茶中的咖啡酰奎宁酸的质量控制提供了一种新的检测方法。

Physiological analysis reveals relatively higher salt tolerance in roots of Ilex integra than in those of Ilex purpurea

Determining the responses of candidate plants to salt stress is a prerequisite for selecting and breeding suitable plants with high salt tolerance to grow in coastal mudfl at areas with high salinity.Here,2-year cutting seedlings of Ilex purpurea Hassk.(local species)and I.integra Thunb.(introduced species)were grown in pots in a glasshouse and irrigated with a Hoagland-NaCl solution at 0,24,and 48 h.Root samples were collected at 0,1,6,24,and 72 h,and concentration of Na^(+)ion;content of proline,soluble carbohydrate,malondialdehyde(MDA),H_(2)O_(2) and ascorbate;and activity of three key antioxidative enzymes were measured.Roots of I.integra accumulated relatively less Na^(+)and had less membrane lipid peroxidation and H_(2)O_(2) during salt stress,thus indicating a relatively higher salt tolerance than roots of I.purpurea.Values for ascorbate content and antioxidant enzymatic activity suggest that the antioxidant ascorbate and antioxidative catalase may play substantial roles for scavenging reactive oxygen species in I.integra roots during salt treatment.Thus,I.integra is apparently more suitable for growing in local highly saline coastal mudfl ats.

An HPLC Fingerprint Identification of Dried Barks of Ilex rotunda and Ilex godajam

[Objectives] A simple and reliable HPLC fingerprint method was developed for the identification of dried barks of Ilex rotunda and I. godajam. [Methods] Nine batches of dried barks of I. rotunda,and seven batches of dried barks of I. godajam collected from different pharmacies and arboretums in different regions of China were used to establish fingerprints. The software Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine( 2004 A Edition) was used to evaluate the fingerprints. [Results]The fingerprints of dried barks of I. rotunda and I. godajam were established. Methodological study met the technical requirements of fingerprints. The similarities of the fingerprints of dried barks of I. rotunda and I. godajam were all more than 0. 8 and 0. 9 respectively. There were 31 and 28 common peaks in I. rotunda and I. godajam,which could be classified into two clusters by principal component analysis( PCA) and hierarchical cluster analysis. [Conclusions] The feasibility and advantages of used HPLC fingerprints were verified,and the results indicated that the HPLC fingerprint as a characteristic distinguishing method combining similarity evaluation,principal component analysis and hierarchical cluster analysis can be successfully used to identify the authenticity of dried barks of I. rotunda and I. godajam.

大鼠肠内菌对毛冬青皂苷ilexsaponin A_1的代谢转化

采用HPLC法研究大鼠肠内菌对毛冬青皂苷ilexsaponin A1的代谢作用。研究采用Kromasil 100-5 C18(250mm×4.6 mm)为色谱柱,流动相为乙腈-0.02%三氟乙酸梯度洗脱,流速:1.0 mL/min,柱温:30℃。将ilexsaponin A1加至大鼠肠内菌培养液中,温孵一定时间后,分析体外代谢产物和代谢模式;大鼠灌胃给予ilexsaponin A1,分析尿液和粪便中的代谢产物,同时测定粪便样品中原型物和代谢产物的含量。结果表明,离体培养大鼠肠内菌可代谢ilexsaponin A1,其主要代谢产物为其苷元ilexgenin A,培养48 h后,约93.12%的ilexsaponin A1被转化成ilexgenin A;大鼠在体实验中,尿液未检测到原型物及代谢物,在粪便中可检测到原型物和代谢物苷元ilexgenin A;ilexsaponin A1经口给予后经原物和代谢产物ilexgenin A排除体外的量高达89.85%。研究结果表明:ilexsaponin A1经口给予后大部分经肠内菌群代谢后被排出体外,主要代谢产物为其苷元。

毛冬青中IlexgeninA和IlexsaponinA1的分离和NMR归属

从岭南地区习用药毛冬青Ilex pubescens Hook．Et．Am．根的乙酸乙酯提取物中分离得到2个化合物，经理化鉴定及波谱技术分析，并结合文献对照，鉴定为乌苏烷型三萜——毛冬青皂苷元A（ilexgenin A，1）和乌苏烷型三萜皂苷——毛冬青皂苷A1（ilexsaponin A1，2）。首次利用2DNMR（^1H-^1H COSY，HMBC和HMQC）技术对此两个化合物的^1H和^13CNMR数据进行了全归属。

毛冬青皂甙Ilexoside O糖链的核磁共振波谱表征及其降解红外光谱分析

为了研究中药毛冬青皂甙Ilexoside O糖链的降解,采取沉淀法和柱色谱法分离纯化Ilexoside O,综合应用一维和二维核磁共振波谱技术,首次对其糖链部分的1H和13C核磁共振信号进行了分析和全归属。采用碱降解法得到次级皂甙,用红外光谱法对降解前后皂甙中主要官能团的结构进行了鉴定。红外光谱分析表明,降解前的皂甙在1733cm-1有较强吸收,降解后该吸收峰消失,并显示在1694cm-1处有很强的振动吸收信号,次级皂甙的红外吸收与毛冬青皂甙Ilexoside J相吻合,初步推断二者为同一化合物。植物成分分离实验表明广西毛冬青中Ilexoside J含量较少而Ilexoside O丰富,而文献表明Ilexoside J具有较好的抗血栓药理活性,因此上述研究为富集毛冬青中Ilexoside J进而使之研发成治疗心血管疾病新药具有重要意义。

Two New Ursane-type Triterpenoid Glycosides from Ilex Cornuta

Ilex cornuta (1.cornuta)is a traditional Chinese medicine (TCM)that has been used in clinical practice for hundreds of years.In order to provide more information about the chemical basis of its pharmacological effects,phytochemical investigation on the roots of L cornuta was conducted in this study.The roots of the plant were firstly extracted with 95% EtOH,and then the crude was partitioned with petroleum ether,EtOAc and n-butyl alcohol.Different chromatographies were employed to isolate the crude step by step and the crude was further purified by semipreparative high performance liquid chromatography (HPLC).As a result,two new triterpenoid saponins (1,2),together with 12 known compounds (3-14),were isolated from the roots of I. cornuta.Their structures were determined based on nuclear magnetic resonance (NMR),mass spectrum (MS)technologies,chemical reactions as well as gas chromatography (GC).Compounds 4,6,8,11,12 and 13 were isolated from this genus for the first time.The structures of compounds 1 and 2 were determined as 3β-O-α-D-xylopyranosyl-(1→3)-α-L-2-O-acetylarabinopyranosyl-(1→2)-β-D glucopyranosyl-23-hydroxyl-20α(H)-urs-12-en-28-oic acid 28-O-β-D-glucopyranosyl ester (1)and 3β-O-α-D-xylopyranosly-(1→3)-α-L-2-O-acetylarabinopyranosyl-(1→2)-β-D-glucopyranosyl-19α,23-dihydroxyl-20α(H)-urs-12-en-28-oic acid 28-O-β-D-glucopyranosyl ester(2).

印度东北地区一种极频危和特有树种(Ilex khasiana Pork.)再生生态学及种群现状

Detailed studies on population structure and regeneration ecology of Ilex khasiana were carried out at five representative natural populations i.e. Mylliem (Population–I), Upper Shillong (Population–II), Shillong Peak (Population–III), Laitkor (Population–IV) and Nongpiyur (Population–V) to probe into the responsible affecting factors for its population structure and regeneration ability in these populations. Experimental observation under three controlled light conditions (full sunlight (100%), intermediate sunlight (50%) and low sunlight (30%)) showed that the growth status and survival rates of seedlings under intermediate and low light were better than those under the high light condition. The density-diameter distribution in population of adult trees (≥5cm dbh) exhibited a typical character of a regenerating population. A higher seedling mortality rate in natural populations of I. khasiana was most probably due to increase in light intensity following vegetation destruction and other anthropogenic disturbances that cause opening of forest canopy. The findings of the present study would be of immense value in formulating appropriate conservation measures for the species.

Effects of Different Processed Products of Ilex hainanensis Merr on Rats with Hyperlipidemia

[Objectives] This study aimed to investigate the hypolipidemic effects of different processed products of Ilex hainanensis Merr to provide experimental basis for the research on the hypolipidemic mechanism of I. hainanensis Merr. [Methods] SD rats were fed high-fat diet to establish hyperlipidemia rat models. The hyperlipidemia rats were administered with different processed products of I. hainanensis Merr by gavage to investigate the preventive and therapeutic effects of the medicinal material. [Results] The levels of total cholesterol(TC), triglyceride(TG), low density lipoprotein cholesterol(LDL-C) in the rats fed high-fat diet were significantly higher than those in of the control group(P<0.01), indicating successful modeling. Among different processing methods, the hypolipidemic effects were in the order as baking=traditional processing=frying=microwaving>drying in shade. Among different baking times, the hypolipidemic effects ranked as 180 ℃>200 ℃=220 ℃. Among different baking times, the hypolipidemic effects ranked as 20 min=40 min>30 min. The hypolipidemic effects of processed product of I. hainanensis Merr with different amounts of active ingredients were different. [Conclusions] The processed products of I. hainanensis Merr all had hypolipidemic effects, but the effects were different. This study will provide a scientific basis for expanding the resources of I. hainanensis Merr.

In Vitro Antibacterial Potential of Plant Extracts and in Vivo Ilex paraguariensis Effect on Oreochromis niloticus Physiology and Resistance to Aeromonas hydrophila

The aim of the study was to investigate the antibacterial potential of garlic(Allium sativum),pinus(Pinus elliottii),eucalyptus(Eucalyptus sp.),yerba mate(Ilex paraguariensis)and guava tree(Psidium guajava Linn.)in vitro and the effect of yerba mate supplementation on Oreochromis niloticus physiology and resistance to Aeromonas hydrophila.The in vitro antibacterial potential of the plants was verified by the minimum inhibitory concentration(MIC)method and by the antimicrobial sensitivity test(TSA).The in vivo assay was performed using diet supplementation with yerba mate and challenge with A.hydrophila.After the supplementation and challenge periods biochemical,immune and physiological parameters were evaluated.The in vitro assays showed that only yerba mate presented antibacterial potential against A.hydrophila and no one of the plant extracts were effective against Streptococcus agalactiae.The use of yerba mate in the diet did not influence the immune and metabolic status of O.niloticus.Also,the supplementation did not act as a growth promoter for the fishes.Considering the challenge of O.niloticus with A.hydrophila,the yerba mate did not improve the resistance of the fishes to the bacteria.The yerba mate extract presented potential to be used as an antibacterial agent for fish diseases.Also,more detailed studies are necessary to stablish effective doses and times required for fish treatments.

Population Ecology of Ilex cornuta in Longgan Lake National Nature Preserve,Hubei Province

The population size class structure, survival curve, height class structure and population distribution patterns of Ilex cornuta in Longgan Lake National Nature Preserve, Hubei Province, were investigated by using the adjacent grid method. The result showed that the population age structure of I. cornuta was of middle-aging type, in the vertical space of population individuals, most of them lived in the shrub layer, and a small number of individuals entered the arborous layer. The distribution pattern of I. cornuta was analyzed by variance /mean ratio method, and the results showed that the distribution pattern of the population was the cluster type. This study can provide references for the further protection, research and development and utilization of I. cornuta .

Primary Research on UVB-irradiation Protective Effect of Total Flavonoids from Large-leaved Kuding Tea (Ilex latifolia Thunb.)

Total flavonoids extracted from large-leaved Kuding tea(Ilex latifolia Thunb.) were investigated to develop anti-UV sun-screening agents. The results showed that total flavonoids of I. latifolia Thunb.(TFILT) showed maximum absorb peak at 325 nm between UVB and UVA by wavelength scanning spectrum. TFILT could inhibit the decrease of cell viability induced by UVB irradiation by CCK-8 method, and 0.25 mg/ml of TFILT showed the most significant protective activity on cells. The dermal thickness and collagen fiber density as skin structural characteristic were analyzed by Histological staining. The results showed that TFILT protected mice skin by significantly attenuating the dermal thickness and inhibiting collagen fiber degradation. The mechanism in vivo needs to be further confirmed in future.

Application of Ilex Plants in Urban Greening

Ilex plants have abundant germplasm resources,with unique aesthetic characteristics endowed by their biological characteristics.Based on the demand of urban greening,this paper introduces the beauty of tree form,leaf form and fruit of Ilex plants,expounds the planting form and application mode of point,line and plane of Ilex plants in urban greening,and puts forward some suggestions for urban greening.

Effects of Canopy Closure on Photosynthetic Characteristics of Ilex latifolia Thunb.in Phyllostachys pubescens Forests

Plantation under the forest is a good way of agroforestry,but the canopy closure has a great influence on understory herbs’growth.In the study,different canopy closures of Phyllostachys pubescens forests were set up to explore its influence on the growth of Ilex latifolia Thunb.The photosynthetic characteristics of Ilex latifolia leaves under different canopy closures were determined by Li-6400 portable photosynthetic system.The results showed that the net photosynthetic rate curve of Ilex latifolia leaves of T1(canopy closure of 0.56)was bimodal with an obvious"midday depression"phenomenon,while the net photosynthetic rate curves of T2(canopy closure of 0.72)and T3(canopy closure of 0.86)were unimodal.The results of light response curve showed that the photosynthetically active radiation and transpiration rate reduced with the increasing of canopy closures.The photosynthetically active radiation,transpiration rate,stomatal conductance,and net photosynthetic rate of Ilex latifolia leaves of T2 were higher than those of T3.Although the net photosynthetic rate of T2 was lower than that of T1,it had no obvious photo-inhibition which affected plant growth.Overall,the canopy closure of 0.72 was more suitable for the growth of Ilex latifolia.The herb plantation in the bamboo forest should be considered with the canopy closure for a better growth.

Study on the Efficient Cutting Propagation Technology for <i>Ilex</i>“China Girl”

In order to realize the efficient industrial production of Ilex “China Girl”, an orthogonal experiment with 4 factors and 3 levels was designed. Firstly, the optimal orthogonal cutting scheme was selected from 9 treatments. Then through the systematic analysis of the effects of cutting position, substrate, exogenous hormones type and concentration on rooting indexes, such as rooting rate, root number, root length and root effect index, the theoretical optimal scheme was predicted and verified. The results showed that the theoretical optimal scheme (3000 mg/L IBA treatment for 15 s, cutting in mixed matrix with peat soil, perlite and vermiculite ratio of 2:1:1) was the optimal cutting rooting scheme of Ilex “China Girl”. After the treatment of this scheme, the rooting rate of Ilex “China Girl” reached 100%, the average root number was 51.67 per plant, and the average root length was 6.13 cm. The rooting time was greatly shortened, the rooting rate and rooting effect were greatly improved. In this study, the efficient cutting propagation technology system of Ilex “China Girl” was established, which laid a foundation for the popularization and application of Ilex “China Girl”, and also provided reference for further improving the cutting propagation efficiency of other evergreen holly. This study laid a foundation for the application of Ilex “China Girl”, and also provided a reference for further improving the cutting propagation efficiency of other evergreen holly.