Structural Characterization of Chloroplast Genome in Alpinia japonica(Thunb.)Miq.,a Medicinal Plant of the Genus Alpinia

The analysis of chloroplast gene characteristics in Alpinia japonica(Thunb.)Miq.is of great significance for developing relevant genetic resources.The high-throughput sequencing and bioinformatic research were performed to analyze the chloroplast genome characteristics of A.japonica.The total chloroplast genome length of A.japonica was 161,906 bp,with a typical circular tetrameric structure.And 133 genes were annotated,comprising 87 protein-coding,38 tRNA,and 8 rRNA genes.Furthermore,22 genes contained two copies,and 18 genes owned introns.Repeat sequence analysis showed that it contains 321 simple sequence repeats(SSRs)and 37 long segment repeats.Compared with the chloroplast genomes of eight representative plants in the genus Alpinia,the gene structure,type,and quantity were relatively conservative.Rps12 was the highest variation site in the entire chloroplast gene.A phylogenetic tree showed that the genus Alpinia was the most closely related to the genus Amomum.Meanwhile,A.japonica is the most closely related to Alpinia chinensis belonging to the genus Alpinia.Overall,the chloroplast genome of a new species was reported in the genus Alpinia,and a basis was provided for the utilization of Alpinia plants as a medical resource.

Analysis of Volatile Constituents in Lonicera japonica Thunb. from Different Origins by GC-MS

[Objective] This study aimed to analyze the volatile constituents in Lonicera japonica Thunb. from different origins. [Method] HP-5MS capillary columns were used and column temperature was controlled by a program. MS analysis was performed with EI and quadruple mass analyzer. The volatile constituents in L. japonica Thunb. were identified by NIST02 and Wiley275 libraries, and their relative contents were determined with chromatographic peak area normalization method. [Result] According to GC-MS total ion-current chromatograms, 35 volatile constituents were identified in L. japonica Thunb. from Guangxi Zhuang Autonomous Region, mainly including methyl linolenate, n-hexadecanoic acid and ζ-muurolene; 18 volatile constituents were identified in L. japonica Thunb. from Hunan Province, mainly including n-hexadecanoic acid, linoleic acid and α-curcumene. [Conclusion] Main volatile constituents in L. japonica Thunb. from two different origins varied significantly.

Chemical Constituents of Roots of Ranunculus ternatus Thunb.

Ranunculus ternatus is culus, which is distributed a species of genus Ranunmainly in Henan, Hubei, Guangxi, Sichuan, and Yunnan provinces in China. The root of Ranunculus ternatus has been used for the treatment of faucitis, tuberculosis, neck scrofula, etc in traditional Chinese medicine. Pharmacological experiment indicated that the water extract of Ranunculus ternatus inhibits not only Staphylococcus aureus and pseudomonas aeruginosa, but also S-180, and S37 remarkablely.

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。

Expression of Exogenous Antimicrobial Peptide and Resistance to Stem Rot Disease of Houttuynia cordata Thunb.

[Objective] This study was to identify the expression of exogenous antimicrobial peptide in transgenic Houttuynia cordata Thunb. plants,and analyze their resistance to stem rot disease. [Methods] SDS-PAGE and Western blot analysis were employed to detect expression of exogenous antimicrobial peptide in transgenic H. cordata plants. Both wild type and transgenic H. cordata plants were inoculated with different concentrations of Rhizoctonia solani spores for detecting their resistance. [Results] The exogenous antimicrobial peptide was detected at translation level. The optimal parameters for detecting the resistance of transgenic H. cordata plants to R. solani was inoculation of spores at a concentration of 3×105 ind./ml and cultured for three days. The results showed that resistance of transgenic H. cordata plants to R. solani was enhanced in comparison with CKs. [Conclusion] Expression of exogenous antimicrobial peptide can enhance the resistance of transgenic H. cordata plants to stem rot disease.

忍冬(L.japonica Thunb.)和钻叶紫菀(Aster subulatus Michx.)对较低浓度镉污染的超富集特征比较研究

合适的超富集植物选择对于尾矿重金属污染修复至关重要。针对安丘市石埠子镇废弃矿坑镉(Cd)污染实际,结合忍冬(L.japonica Thunb.)和钻叶紫菀(Aster subulatus Michx.)对重金属Cd的超富集特征比较,分析了30天较低浓度Cd暴露(0.1 mg/kg,0.3 mg/kg,0.9 mg/kg)下两种超富集植物的毒性响应和超富集特征。研究表明,两种植物的株高和干重均受到明显影响(P<0.05),但忍冬对Cd的平均TF和BCF(TF_(叶)=0.494,TF_(茎)=0.357,BCF_(叶)=12.014,BCF_(茎)=8.689,BCF_(根)=24.424)明显高于钻叶紫菀的平均TF和BCF(TF_(茎)=0.127,TF_(叶)=0.205,BCF_(叶)=3.903,BCF_(茎)=2.412,BCF_(根)=19.189)(P<0.05)。这一研究成果对于废弃矿坑较低浓度Cd污染的超富集生态修复植物种类的选择,构建基于超富集植物的尾矿(渣)原位生态修复技术建设具有重要指导意义。

Characteristics of Uptake and Accumulation of Pb and Cd in Osmanthus fragrans(Thunb.) Lour.under Single and Combined Stresses of Pb and Cd

[Objective] This study aimed to investigate the characteristics of Pb and Cd uptake and accumulation in Osmanthus fragrans（Thunb.） Lour.under single and combined stress of Pb and Cd.[Method] Pot experiment was adopted with O.fragrans seedlings as experimental materials to investigate the effect of single and combined stresses of Pb and Cd on the growth of O.fragrans seedlings under 0,50,100,500,1 000 and 2 000 mg/kg of Pb and 0,1,10,50,100 and 200 mg/kg of Cd.[Result] Under the experimental conditions,the growth of O.fragrans seedlings was significantly inhibited under moderate Pb stress（500 mg/kg） and high Cd stress（100 mg/kg）;the decreasing order of Pb uptake rate of different O.fragrans tissues was root stem leaf;the decreasing order of Cd content of different O.fragrans tissues was root stem leaf,while the decreasing order of Cd uptake rate of different O.fragrans tissues was stem root leaf.[Conclusion] Under combined stress,low concentrations of Pb and Cd can mutually promote the uptake and accumulation,while high concentrations of Pb and Cd can mutually inhibit the uptake and accumulation.

Effect of Three Precursors on Accumulation of Principal Volatile Oil Constituents in Tissue Culture Plantlets of Atractylodes lancea (Thunb.) DC.

[Objective] This study was to explore the effect of three precursors on the accumulation of principal volatile oil constituents in tissue culture plantlets of Atractylodes lancea （Thunb.） DC. with the aim to provide references for the improvement of artificial cultivated A. lancea quality. [Method] Three precursors were added into the MS rooting medium for A. lancea tissue culture plantlets and the volatile oil was extracted by ultrasonication after cultured for several days. The content of atractylon, atractylol, β-eucalyptol and atractydin in the volatile oil were determined by using gas chromatography method. [Result] The addition of xylose, isoprene and tetrahydrofuran impacted the growth indicators, yield of volatile oil and relative percentage content of the four constituents of A. lancea tissue culture plantlets. In the 6 g/L xylose optimized medium, the atractylon and β-eucalyptol content reached up to 4.23% and 56.34%, respectively, 1.41% and 1.66% higher than the control; although the addition of isoprene into medium raised the atractylon content, the accumulation of total volatile oil was inhibited that it decreased by 23.67%, 31.06% and 7.10% to the control; for the tetrahydrofuran optimized medium, the content of atractylon, atractylol and atractydin all increased, and the total volatile oil content increased by 49.97% to the control when the concentration of tetrahydrofuran was 0.07 g/L. [Conclusion] The addition of xylose and tetrahydrofuran promoted the accumulation of principal constituents of the volatile oil, whereas the addition of isoprene inhibited the accumulation.

峨眉山不同山峪和海拔高度鱼腥草(Houttuynia cordata Thunb.)居群挥发油成分的变化

以分布于峨眉山不同山峪不同海拔（500-1800m）的17个鱼腥草（Houttuynia cordata Thunb.）居群及生长地土样作为试验材料，按土壤农化分析常规方法对土壤pH值及土壤营养元素进行测定，用水蒸馏法提取植株地上部分的挥发油，并用GCMS对挥发油化学成分进行分离和鉴定，峰面积归一法测定各成分含量。结果显示：峨眉山鱼腥草生长地土壤肥力水平较高，对鱼腥草的生长极为有利。从所有居群中共鉴定出35种化合物，各居群挥发油成分组成及含量存在较大的差异。月桂烯作为四个山峪唯一共有的主要成分在各居群中均能检测到，且在各居群中的平均含量最高。乙酸龙脑酯和trans-β-罗勒烯的含量与海拔高度间分别存在显著和极显著正相关。方差分析及多重比较的结果表明，不同山峪以及海拔高度对各居群挥发油成分和含量有显著影响。通过偏相关分析发现土壤微量元素对鱼腥草挥发油共有成分合成与积累的作用大于大量元素，微量元素中以有效钼的影响最为显著，大量元素中以速效钾的影响最为显著。采用逐步多元回归分析筛选出了对挥发油共有成分含量影响显著的土壤营养元素。

忍冬(Lonicera japonic Thunb.)果实抑菌活性及化学成分研究

【目的】研究忍冬Lonicera japonica Thunb.果实的化学成分,为忍冬果实的开发利用提供依据。【方法】在抑菌试验的基础上,采用硅胶柱色谱、Sephadex LH-20等方法分离纯化化合物,并通过化合物的理化性质和波谱分析鉴定其结构。【结果】从忍冬果实中分离得到7个化合物,分别为熊果酸、齐墩果酸、原儿茶酸、苜蓿素、槲皮素、咖啡酸和木犀草苷。【结论】忍冬果实醇提物和醇提物的不同极性部位具有一定的抑菌活性,且以乙酸乙酯部位的作用最强。从乙酸乙酯部分分离得到的7个化合物均为首次从忍冬果实中分离得到,为研究忍冬果实的药效物质基础及综合开发利用忍冬果实提供了科学依据。

乌饭树(Vaccinium bracteatum Thunb.)的民族植物学

论述中国古代本草关于乌饭树（ＶａｃｉｎｉｕｍｂｒａｃｔｅａｔｕｍＴｈｕｎｂ．）的名称与药效的记载以及现代对乌饭树及同属植物的研究和利用。认为乌饭树自古以来便是黑色保健食品的原料和滋补强壮药，从其枝叶中提取天然黑色素用于保健事业值得进一步开发和利用。

盐和干旱胁迫对燕子掌(Crassula agenten Thunb.)叶片液泡膜H^+-ATPase活性的影响

专性CAM植物燕子掌离体叶片的盐胁迫处理和失水干旱处理后 ,液泡膜H+ ATP酶对低浓度(2 0 0、40 0mmol/L)的盐胁迫 (NaCl胁迫 48h)不敏感 ,而当盐浓度达到 6 0 0mmol/L时 ,ATP水解活性和H+转运活性较对照上升 5 5 %～ 6 5 % ,而干旱胁迫 (4 8h ,失水 12 % )使酶活上升约 30 % ,但是上述各种胁迫均不影响ATP水解与H+转运的耦联比率 ,仍旧维持在12。用Westernblot证实在逆境下 ,H+ ATP酶的 3种主要亚基A、B、c在膜上的蛋白含量均有所增加 ,且以调节亚基 (B)

Extract of Cycas revoluta Thunb. enhances the inhibitory effect of 5-fluorouracil on gastric cancer cells through the AKT-mTOR pathway

BACKGROUND Gastric cancer is one of the most common and deadly malignancies worldwide.Despite recent medical progress, the 5-year survival rate of gastric cancer is still unsatisfactory. 5-fluorouracil(5-Fu) is one of the first-line antineoplastic treatments for gastric cancer, as it can effectively induce cancer cell apoptosis.However, the effect of 5-Fu is limited due to drug resistance of the malignant tumor. Previous studies have reported that Sotetsuflavone from Cycas revoluta Thunb. can markedly suppress lung cancer cell proliferation by apoptosis,though its effect on gastric cancer remains unknown.AIM To investigate the inhibitory effect of Cycas revoluta Thunb. and to determine whether it can overcome gastric cancer cell drug resistance to 5-Fu.METHODS Cell viability was examined to determine whether the natural extract of Cycas revoluta Thunb. induced gastric cancer cell death. The half-maximal effective concentration and the half-maximal lethal concentration were calculatede.Wound-healing and transwell assays were performed to examine gastric cancer cell motility. Clonogenic assays were performed to investigate the synergistic effects of Cycas revoluta Thunb. with 5-Fu, and apoptotic bodies were detected by Hoechst staining. Western blotting was performed to examine the expression of related proteins and to investigate the molecular mechanism of Cycas revoluta Thunb.-induced cancer cell apoptosis. The expressions of proteins, including mammalian target of rapamycin(mTOR) and p-AKT, were detected in different combinations of treatments for 48 h, then analyzed by ECL detection.RESULTS Gastric cancer cells were more sensitive to the natural extract of Cycas revoluta Thunb. compared to normal gastric epithelial cells, and the extract effectively inhibited gastric cancer cell migration and invasion. The extract improved the anti-cancer effect of 5-Fu by enhancing the chemosensitization of gastric cancer cells. Extract plus 5-Fu further reduced the expression of the drug-resistancerelated proteins p-AKT and mTOR after 48 h compared to 5-Fu alone. Compared to 5-Fu treatment alone, mTOR and p-AKT expression was significantly reduced by about 50% and 75%, respectively. We also found that the natural extract of Cycas revoluta Thunb. further increased 5-Fu-induced gastric cancer cell apoptosis. Expression of apoptosis-related protein X-linked inhibitor of apoptosis protein and apoptosis inducing factor were significantly reduced and increased,respectively, in the 5-Fu-resistant gastric cancer line SGC-7901/R treated with extract plus 5-Fu, while the expression of survivin did not change.CONCLUSION The natural extract of Cycas revoluta Thunb. effectively inhibited gastric cancer cell growth and enhanced the anti-cancer effect of 5-Fu through the AKT-mTOR pathway.

忍冬(Lonicerajaponic Thunb.)果实中的环烯醚萜苷类成分

【目的】在前期研究的基础上,进一步研究忍冬Lonicera japonica Thunb.果实的化学成分。【方法】采用硅胶柱色谱、Sephadex LH-20等方法对忍冬果实乙醇提取物的乙酸乙酯部分进行分离纯化,并通过波谱技术和参考文献鉴定其结构。【结果】分离得到了6种环烯醚萜苷类成分,分别鉴定为金吉苷、马钱素、马钱素-7-酮、獐牙菜苷、Demethylsecologanol、裂环氧化马钱素。【结论】化合物1～6为首次从忍冬果实中分离得到。

鸭跖草科杜若(Pollia japonica Thunb.)的减数分裂研究

首次对鸭跖草科杜若(Pollia japonicaThunb.)进行了花粉母细胞减数分裂观察,并重新报道了该种的染色体数目为2n=32。结果显示,减数分裂中期I构型为16Ⅱ,并且观察到次级联会现象。减数分裂后期I和后期Ⅱ存在落后染色体、染色体断片、二次分裂不同步等异常现象,统计各时期畸形率都低于10%。随机统计花粉粒活性,成熟率达到90%以上。这说明杜若的减数分裂过程基本正常,也证明了2n=32的体细胞染色体数目是可信的。

小毛茛(Ranunculus ternatus Thunb.)花粉流潜能初探

在自然居群中对小毛茛传粉机制与距离作了观测，并对开花后花粉萌发能力的变化作了研究，结果是：该种兼具虫媒或风媒传粉机制，以虫媒传粉为主；传粉距离多数局限于２ｍ以内，但不排除远距离传粉的可能性；体外萌发试验表明，该种花粉萌发率在开花第一天甚低，开花后２４ｈ达到最高值。作者推测，小毛茛花粉以近距离散布为主，但具有远距离传播的潜能。

Anti-encystment and amoebicidal activity of Lonicera japonica Thunb. and its major constituent Chlorogenic acid in vitro

Objective:To examine the acanthamoebicidal effects of ethyl acetate,aqueous and butanol fractions of dried flower buds of Lonicera japonica(L.japonica) Thunb.(Flos Lonicerae) in vitro.Methods:Acanthamoeba triangularis isolates were obtained from environmental water samples and identified by PCR.They were exposed to ethyl acetate,water and butanol fractions of L.japonica Thunb.at concentrations ranging from 0.5 mg/m L to 1.5 mg/m L.The extracts were evaluated for growth inhibition at 24,48 and 72 h,respectively.Chlorogenic acid at a concentration of 1 mg/mL was examined for inhibition of encystment.Results:Ethyl acetate fraction at a concentration of 1.5 mg/mL evoked a significant reduction of trophozoite viability by 48.9% after 24 h,49.2% after 48 h and 33.7% after 72 h Chlorogenic acid,the major active constituent of L.japonica Thunb.at the concentration of 1 mg/mL reduced the cysts/trophozoite ratio by 100% after 24 h,84.0% after 48 h and 72.3% after 72 h.This phenolic compound at concentration of 1 mg/mL concurrent with 0.6% hydrogen peroxide inhibited hydrogen peroxide-induced encystment by 92.8% at 72 h.Conclusions:Results obtained from this study show that ethyl acetate fraction at 1.5 mg/m L is the most potent fraction of L.japonica Thunb.and its major constituent chlorogenic acid showed the remarkable inhibition of encystment at a concentration of 1 mg/mL.

Review of Factors Influencing Active Components of Lonicera japonica Thunb.

Lonicera japonica Thunb. is a typical medicinal and edible plant in China. It has antibacterial, anti-oxidant, anti-endotoxin, and anti-inflammatory effects, etc. Its pharmacological action is closely related to its active components. Based on the results of previous studies, this paper summarized the effects of variety, origin, cultivation conditions, pruning, harvesting period and processing methods on the active components of L. japonica , in the hope of providing theoretical guidance for high-yield and high-quality cultivation of L. japonica.

浙江柃木属Eurya Thunb.植物的研究

本文报导了浙江产柃木属Eurya Thunb.12种和1变种,其中金叶微毛柃E.hebeclados Ling var.aureo-punctata为新组合,尖萼毛柃E.acutisepala L.K.Ling及从化柃E.metcalfiana Kobuski为浙江分布新记录。 文中包括分种检索表及每一分类群的列举及其地理分布、生态条件和分类讨论。 本文中所研究和引证的大多数标本藏于杭州植物园植物标本室和杭州大学生物系植物标本室。

Septemloside III: A Nonasaccharide Saponin from the Bark of Kalopanax septemlobus (Thunb.) Koidz.

A nonasaccharide saponin, new hederagenin saponin, had been isolated from the bark of Kalopanax septemlobus (Thunb.) Koidz., and its structure was elucidated by HRESI-MS, NMR experi- ments and chemical analyses as 3-O-β-D-xylopyranosyl-(1→4)-β-D-xylopyranosyl-(1→3)-α-L-rhamno- pyranosyl-(1→2)-α-L-arabinopyranosyl hederagenin 28-O-β-D-xylopyranosyl-(1→3)-β-D–xylopyra- nosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside.