荸荠营养器官的发育与解剖学研究

荸荠同化茎起源于肉质主茎倒２或倒３叶的叶腋内。同化茎基部着生二鞘状叶,鞘状叶对早期同化茎穿出土面具保护作用。匍匐茎大多起源于同化茎基部鞘状叶的叶腋内。当植株开始抽生花茎时,地下匍匐茎顶端开始膨大。球茎的膨大是匍匐茎顶端５－６节的基本组织经细胞有丝分裂,增加细胞数目,然后由细胞体积的扩大来实现的。球茎具足够的营养物质供来年顶芽萌发的需要,故属水生植物冬芽的性质。     

直立百部的一个新异名

根据对直立百部Stemona sessilifolia（Miq．）Miq．大量标本的研究和野外调查及引种栽培观察,结合对山东百部Stemona shandongensis D．K．Zang模式标本产地的调查,发现直立百部的形态性状是多变的。研究表明它的习性受生境影响而多变,从直立半灌木到蔓生都有;叶型多变;花通常出自叶腋,而花梗有时与叶柄,甚至主脉合生;内轮花被片大多明显大于外轮。山东百部的形态性状处于直立百部的变异范围内,因此作为独立的种是难以成立的,应处理为直立百部的新异名。     

直立百部遗传多样性的ISSR分析

利用ISSR分子标记,对直立百部4个居群的84个样本进行遗传多样性研究,探讨山东百部种的地位。结果表明:(1)15个引物共检测到184条清晰的谱带,其中多样性条带153条;直立百部的遗传多样性水平较高,其物种水平多样性条带百分率(PPF)为83.15%,有效等位基因数(Ae)为1.425,Shannon多样性指数(I)为0.388,Nei’s多样性指数(Hpop)为0.254。(2)居群平均水平上多样性条带百分率为63.04%,有效等位基因数为1.351,Shannon多样性指数为0.310,Nei’s多样性指数为0.206。(3)AMOVA分析显示直立百部居群间遗传分化水平(ΦST)为0.126 3,POPGENE分析显示居群间的遗传分化系数(GST)为0.191 1,二者均表明居群内变异大于居群间变异;直立百部居群间的基因流(Nm)为2.116 6,说明居群间的基因交流较频繁。(4)聚类及主成分分析显示,直立百部4个居群聚为两支,其中泰山居群与其他居群关系较远,以蔓生百部和大百部为外类群分析的结果支持将山东百部作为直立百部的异名,Mental检测显示居群间遗传距离与地理距离间没有显著相关性。     

Effect of Stemona curtisii root extract on P-glycoprotein and MRP-1 function in multidrug-resistant cancer cells

Multidrug resistance (MDR) is the result of overexpression of membrane bound proteins that efflux chemotherapeutic drugs from the cells. Two proteins, P-glycoprotein (P-gp) and multidrug-resistance associated protein-1 (MRP-1) efflux chemotherapeutic agents out of the cancer cell that decrease intracellular drug accumulation, thereby decreasing the effectiveness of many chemotherapeutic agents. In the present study, the ethanolic extract of the roots of Stemona curtisii Hook. was tested for the potential ability to modulate the MDR phenotype and function of P-gp and MRP-1. The S. curtisii extract reversed the resistance to putative chemotherapeutic agents, including vinblastine, paclitaxel and colchicine of KB-V1 cells (MDR human cervical carcinoma with high P-gp expression) in a dose-dependent manner, but not in KB-3-1 cells (drug sensitive human cervical carcinoma, which lack P-gp expression). The root extract also increased the intracellular uptake and retention of (3)[H]-vinblastine in KB-V1 cells dose dependently. The extract did not influence MDR phenotype-mediated MRP-1 in MRP1-HEK293 (human embryonic kidney cells stably transfected with pcDNA3.1-MRP1-H10 which show high MRP-1 expression) and pcDNA3.1-HEK293 (wild type). In summary, the S. curtisii root extract modulated P-gp activity but not MRP-1 activity. The result obtained from this study strongly indicated that S. curtisii extract may play an important role as a P-gp modulator as used in vitro and may be effective in the treatment of multidrug-resistant cancers. The purified form of the active components of S. curtisii extract should be investigated in more details in order to explain the molecular mechanisms involved in P-gp modulation. This is the first report of new biological activity in this plant, which could be a potential source of a new chemosensitizer.     

Morphological and chemical variation of Stemona tuberosa from southern China – Evidence for heterogeneity of this medicinal plant species

• The occurrence of bioactive alkaloids and tocopherols was studied in 15 different provenances of Stemona tuberosa Lour. collected in southern China, to examine chemical variation of individuals that show notable differences in flower characteristics. Morphological variations stimulated examination of chemical characteristics of these individuals.
• Methanolic root extracts of 15 individuals of S. tuberosa were comparatively assessed with HPLC‐UV‐DAD/ELSD. Five of seven compounds were co‐chromatographically identified. Two compounds were isolated and their structure elucidated using NMR and MS. Amounts of alkaloids and tocopherols were determined using HPLC‐UV‐DAD/ELSD with the external standard method.
• Five alkaloids, tuberostemonine ( 1 ), tuberostemonine A ( 2 ), neotuberostemonine ( 3 ), tuberostemonine N ( 4 ), stemoninine ( 5 ) and two 3,4‐dehydrotocopherol derivatives were identified. Within S. tuberosa alkaloid accumulation tends either towards tuberostemonine ( 1 ) or stemoninine ( 5 ). All individuals show a notable co‐occurrence of compounds 1 or 5 and 3,4‐dehydro‐δ‐tocopherol ( 6 ). These results coincide with differences in flower morphology of S. tuberosa.
• Stemona tuberosa, as defined in the Flora of China, shows a remarkable variation in flower morphology and additionally in the accumulation of alkaloids. The obtained data show the need for future species delimitation to either species or subspecies level.

     

百部内生放线菌的分离、分类及次级代谢潜力

【目的】以对叶百部块根为材料分离内生放线菌,并对分离菌株进行分类、抗菌活性和次级代谢产物合成基因研究。【方法】样品经过严格的表面消毒,选用4种培养基分离百部内生放线菌;分离菌株通过形态观察和16S rRNA序列分析进行分类鉴定;采用琼脂移块法测试分离菌株的抗菌活性;通过PCR检测分离菌株的PKS/NPRS和卤化酶基因;使用HPLC-UV/VIS-ESI-MS/MS分析发酵产物。【结果】从6个样品中获得18株内生放线菌,分属链霉菌属(Streptomyces)、小单孢菌属(Micromonospora)、假诺卡氏菌属(Pseudonocardia)和甲基杆菌属(Methylobacterium)。分离菌株绝大部分具有抗菌活性和次级代谢产物合成基因,其中13株对耐药金黄色葡萄球菌和/或绿脓杆菌有拮抗活性,17株具有PKS/NRPS基因,8株菌具有卤化酶基因,且卤化酶阳性代表菌株的发酵产物具有抗细菌活性和卤代化合物特征。【结论】百部作为一种传统中药,其内生放线菌以链霉菌和小单孢菌为主,在次级代谢产物合成方面具有很好的潜力,可作为一类重要微生物资源进行活性产物开发。     

Physiological responses to salt stress in young umbu plants

Soil salinity affects plant growth and development due to harmful ion effects and water stress caused by reduced osmotic potential in the soil solution. In order to evaluate the effects of salt stress in young umbu plants, research was performed in green house conditions at the Laboratory of Plant Physiology at Federal Rural University of Pernambuco, Brazil. Growth, stomatal behaviour, water relations, and both inorganic and organic solutes were studied aiming for a better understanding of the responses of umbu plants to increasing salinity. Plants were grown in washed sand with Hoagland and Arnon nutrient solution with 0, 25, 50, 75, and 100 mM NaCl. Growth, leaf water potential, transpiration, and diffusive resistance were evaluated. Na⁺, K⁺, Cl⁻, soluble carbohydrates, and free amino acid contents were measured in several plant organs. Most variables were affected with salinity above 50 mM NaCl showing decreases in: number of leaves, plant height, stems diameter, and dry masses, and increases in root-to-shoot ratio. Reductions in ψ_pd were observed in plants grown under 75 and 100 mM NaCl. All salt levels above zero increased Na⁺ and Cl⁻ contents in leaves. However, K⁺ content was not affected. Na⁺ and Cl⁻ in stems and roots reached saturation in treatments above 50 mM NaCl. Organic solute accumulation in response to salt stress was not observed in umbu plants. These results suggest that umbu plants tolerate salt levels up to 50 mM NaCl without showing significant physio-morphological alterations.     

New stemona alkaloids from the roots of Stemona sessilifolia

From the roots of Stemona sessilifolia, three new stemona-type alkaloids, namely stemosessifoine (1), isooxymaistemonine (2), and isomaistemonine (3), along with eight known alkaloids (bisdehydrostemoninine, isobisdehydrostemoninine, tuberostemonine, bisdehydrotuberostemonine, bisdehydrostemoninine, isobisdehydrostemoninine, stemoninine, and protostemonine), were isolated. Their structures were determined on the basis of extensive 2D-NMR spectroscopic-data analysis and by comparison with reported values in the literature. Compound 1 is a structurally unprecedented alkaloid, and it is depicted to be bioconverted from tuberostemonine as the precursor. Isooxymaistemonine (2) showed a positive effect on the human high-density lipoprotein (HDL) receptor gene CD36 and LIMP II analogous-1 (CLA-1) at the dosage of 10 microg/ml.     

蔓生百部的化学成分研究

从蔓生百部(Stemona japonica)根中分离得到13个化合物,通过波谱数据,它们鉴定为β-谷甾醇(1)、豆甾醇(2)、5,11-豆甾二烯-3β-醇(3)、苯甲酸(4)、4-甲氧基苯甲酸(5)、1,8-二羟基-3-甲基蒽醌(6)、1,8-二羟基-6-甲氧基-3-甲基蒽醌(7)、氧代狭叶百部碱(8)、百部定碱(9)、异狭叶百部碱(10)、绿原酸(11)、栀子苷(12)和藏红花素A(13).所有化合物均为首次从该植物中分离得到.