地黄炮制过程氨基酸组分分析

利用氨基酸自动分析仪分别测定加工前后地黄中的总氨基酸和游离氨基酸质量分数。分析、比较炮制前后地黄的氨基酸成分变化。炮制前后地黄的氨基酸种类及含量有明显变化,多数氨基酸的含量在炮制后明显下降甚至消失。结论:炮制前后的地黄中氨基酸质量分数差别明显,其中碱性氨基酸质量分数的差别尤其显著,该差异可能是由于炮制过程中发生美拉德反应造成的。     

Identification of autotoxic compounds in fibrous roots of Rehmannia (Rehmannia glutinosa Libosch.)

Rehmannia is a medicinal plant in China. Autotoxicity has been reported to be one of the major problems hindering the consecutive monoculture of Rehmannia. However, potential autotoxins produced by the fibrous roots are less known. In this study, the autotoxicity of these fibrous roots was investigated. Four groups of autotoxic compounds from the aqueous extracts of the fibrous roots were isolated and characterized. The ethyl acetate extracts of these water-soluble compounds were further analyzed and separated into five fractions. Among them, the most autotoxic fraction (Fr 3) was subjected to GC/MS analysis, resulting in 32 identified compounds. Based on literature, nine compounds were selected for testing their autotoxic effects on radicle growth. Seven out of the nine compounds were phenolic, which significantly reduced radicle growth in a concentration-dependent manner. The other two were aliphatic compounds that showed a moderate inhibition effect at three concentrations. Concentration of these compounds in soil samples was determined by HPLC. Furthermore, the autotoxic compounds were also found in the top soil of the commercially cultivated Rehmannia fields. It appears that a close link exists between the autotoxic effects on the seedlings and the compounds extracted from fibrous roots of Rehmannia.     

地黄种质资源形态及生物学性状的观察与比较

收集我国地黄种质资源30份,并对其叶片形态、株形、开花期、花色、块根形状、块根皮色、块根肉色等进行观察,对叶片长度、叶片宽度、叶片鲜重、株高、叶片数等性状进行测定。结果表明不同地黄品种之间在叶片长度、叶片宽度、叶片鲜重、株高度、叶片数等都存在极显著差异。根据综合鉴定与评价,将地黄种质资源划分为5个类型。     

PP333对怀地黄试管苗形态及生理特性的影响

研究PP333对怀地黄试管苗生长及一些生理指标的影响。通过单因子实验、比色法和愈创木酚法探讨PP333对试管苗的影响。结果表明,不同浓度PP333均促进试管苗芽的萌发,使根系粗壮,根数增加,低浓度PP333(0.01、0.05mg·L-1)促进试管苗茎的伸长生长,高浓度(0.1、2mg·L-1)抑制茎、叶生长,PP333浓度为2mg·L-1时壮苗效果最佳。PP333处理使试管苗生长中期叶片可溶性蛋白含量、POD活力提高。适宜浓度的PP333可以改变试管苗的生理特性,达到培育壮苗的目的。     

地黄源库关系的变化及其与连作障碍的关系

通过正茬剪叶模拟连作地黄光合源不足条件下,测定分析植株源与库的关系,应用同位素示踪技术分析连作地黄光合产物的分配及内源激素的变化,以探讨地黄的连作障碍机制。结果表明:剪叶处理对植株的影响不能达到连作障碍程度,其库容及其活性大于连作地黄;连作地黄出现光合产物滞留于地上部,库的活性影响了植株光合产物向库的运输,在生长前期植株内源激素发生了变化;因此断定,连作地黄库容及其活性过低是限制地黄块根膨大的重要因素,库通过不同激素对源的反馈调节,抑制了地上源的光合能力及光合产物向库的分配,导致连作障碍。     

Alleviatory effect of spent Pleurotus eryngii Quel substrate on replant problem of Rehmannia glutinosa Libosch

Rehmannia glutinosa Libosch. is a medicinal plant cultivated at a commercial scale in China. However, replanting problems result in a severe decline in both the biomass and quality of its roots, which are of greatest medicinal value. This study attempted to remediate the replant soil using spent Pleurotus eryngii Quel substrate for alleviating this issue, and to investigate the underlying mechanisms. Results showed that R. glutinosa grew successfully in fresh soil and remedial replant soil, while no roots were harvested in the unremedied replant soil. Overall, the nutritional status in the remedial soil was higher than that of the unremedied and fresh soil, while the concentration of allelopathic phenolic acids was lower. When planted in unremedied soil, the growth of five plant pathogens was induced and one beneficial fungus was suppressed. When planted in remedied soil, four out of the five pathogens were suppressed, while two beneficial fungi were identified in the remedial soil. This study suggests that the spent P. eryngii substrate significantly alleviates the replant problem of R. glutinosa, and that the alleviatory function reflects a synergetic effect, including the supplementation of soil nutrition, the degradation of allelochemicals, and the remediation of unbalanced microbial community.     

地黄中腺苷含量的影响因素

地黄中腺苷含量的影响因素朱青罗燕燕王瑛龚韬郄春鹏（北京市临床药学研究所,北京１０００３５）ＴｈｅｉｎｆｌｕｅｎｃｅｆａｃｔｏｒｓｏｎａｄｅｎｏｓｉｎｅｃｏｎｔｅｎｔｉｎｔｈｅｒｏｏｔｔｕｂｅｒｏｆＲｅｈｍａｎｎｉａｇｌｕｔｉｎｏｓａＬｉｂｏｓｃｈ．Ｚ...     

怀地黄活性内生菌的分离鉴定及抗菌抗肿瘤活性

利用平板分离法从怀地黄(Rehmannia glutinosa Libosch)中分离出内生菌共130株,包括67株细菌、50株真菌和13株放线菌。利用大肠杆菌、金黄色葡萄球菌和黑曲霉作为供试菌株,通过对峙试验和平板点接法筛选出8株对3种供试菌株均具有较强拮抗作用的内生菌。经菌种形态观察、生理生化实验以及16S rRNA序列测定,这8株活性内生菌均为假单胞菌属的细菌,并分别与Pseudomonas fluorescens、Pseudomonas thivervalensis、Pseudomonas chlororaphis、Pseudomonas koreensis和一个未定种具有最高相似性。对这8株活性内生菌进行液体发酵、不同有机溶剂抽提和抗菌抗肿瘤研究,结果表明它们的乙醇抽提物和乙酸乙酯抽提物均不同程度地对3种供试菌株和食管癌细胞系Ec9706具有抑制作用。其中2-2号菌株(Pseudomonas chlororaphis)抗菌抗肿瘤作用最强,具有重要的开发价值。     

Total phenolics, flavonoids and antioxidant activity of Saptarangi (Salacia chinensis L.) fruit pulp

Salacia chinensis L. has various beneficial properties including antidiabetic and antioxidant activity. The S. chinensis fruit pulp (SCFP) was extracted with four different solvents (Methanol, ethanol, acetone and water) and was screened for total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity (AOA). The AOA was assessed by evaluating the 1, 1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) and metal chelating assay. Methanolic SCFP extract exhibited the highest phenolic content (3.20?±?0.12 mg GAE/g FW) whereas, ethanolic extract showed highest flavonoid content (0.31?±?0.68 mg RE/g FW). The methanolic extract possesses highest antioxidant activity towards DPPH (92.44 %), FRAP (1.939 O.D) and metal chelating activity (74.16 %). AOA (DPPH and FRAP) was significantly correlated with TPC. The results indicated that SCFP is a good natural source of antioxidant compounds for use in food and pharmaceutical industry.     

Hairy root cultures of Rehmannia glutinosa and production of iridoid and phenylethanoid glycosides

Hairy root lines through the infection of Agrobacterium rhizogenes strain (A4) were established from shoot tips and leaves of Rehmannia glutinosa Libosch. Ten lines of hairy roots were selected on the basis of biomass increase in half-strength Gamborg medium (¹/₂ B5). Transgenic status of the roots was confirmed by polymerase chain reaction using rolB and rolC specific primers. Iridoid glycosides (catalposide, loganin, aucubin and catalpol) and phenylethanoid glycosides (verbascoside and isoverbascoside) identified using HPLC?CESI?CMS, and their contents were compared with untransformed root culture and roots of 1-year-old field-grown plants of R. glutinosa by RP-HPLC. The growth and production of secondary metabolites in ten hairy root lines varied considerably as to the media. Woody plant (WP) medium displayed higher growth in terms of fresh (FW) and dry weights (DW) compared to ¹/₂ B5 medium. High-yielding hairy root lines produced higher amounts of loganin, catalposide, verbascoside and isoverbascoside in comparison to the untransformed root culture and roots of 1-year-old field-grown plants. The highest amounts of catalposide and loganin in transformed roots were 4.45?mg?g^?1 DW (RS-2 hairy root line) and 4.66?mg?g^?1 DW (RS-1 hairy root line), respectively. Aucubin and catalpol were detected in some lines in trace amounts. The highest amounts of verbascoside (16.9?mg?g^?1 DW) and isoverbascoside (3.46?mg?g^?1 DW) were achieved in RS-2 root line. The contents of catalposide, verbascoside and isoverbascoside in high-producing lines were several times higher than in untransformed root culture and roots of R. glutinosa plants grown in soil. Loganin and aucubin could not be detected in roots of field-grown plants. However, the levels of catalpol were much lower in the in vitro roots.     

地黄的组织培养和快速繁殖

1　植物名称　地黄 (Ｒｅｈｍａｎｎｉａｇｌｕｔｉｎｏｓａ) ,又名生地。2　材料类别　茎段、茎尖。3　培养条件　 ( 1 )无菌苗获得培养基 :ＭＳ ;( 2 )茎尖分化培养基 :ＭＳ ＮＡＡ 0 .0 1ｍｇ·Ｌ- 1 (单位下同 ) 6 ＢＡ 1 ＧＡ30 .2 ;( 3)增殖培养基 :1 /2ＭＳ 6 ＢＡ     

地黄组织培养及植株再生的研究

以地黄根茎所获无菌苗为材料,对其愈伤组织诱导、分化和再生植株的获取进行了初步研究。结果表明：取叶片、茎段、叶柄进行愈伤组织诱导,筛选出最适培养基为MS附加2,4-D0．5mg／L、BA1．0mg／L,愈伤组织诱导率可达100％。将叶片接种在分化培养基中,诱导不定芽,其最适分化培养基为MS附加BA 3mg／L、NAA 0．1mg／L,分化率为77．5％。试管苗在改良的MS(大量与微量元素、铁盐和有机物质各1／2)附加NAA 0．05mg／L的培养基上,经过15～20d培养,生根率可达100％。     

The activities of antioxidant enzymes in response to oxidative stresses and hormones in paraquat-tolerant Rehmannia glutinosa plants

All members of R. glutinosa show the unique characteristic of intrinsic tolerance to paraquat (PQ). Antioxidant enzymes have been proposed to be the primary mechanism of PQ resistance in several plant species. Therefore, the antioxidant enzyme systems of R. glutinosa were evaluated by comparatively analyzing cellular antioxidant enzyme levels, and their responses of oxidative stresses and hormones. The levels of ascorbate peroxidase (APX), glutathione reductase (GR), non-specific peroxidase (POX), and superoxide dismutase (SOD) were 7.3-, 4.9-, 2.7- and 1.6-fold higher in PQ-tolerant R. glutinosa than in PQ-susceptible soybeans. However, the activity of catalase (CAT) was about 12-fold higher in the soybeans. The activities of antioxidant enzymes reduced after PQ treatment in the two species, with the exception of POX and SOD in R. glutinosa, which increased by about 40 %. Interestingly, the activities of APX, SOD and POX in R. glutinosa, relative to those in soybeans, were further increased by 49, 67 and 93 % after PQ treatment. The considerably higher intrinsic levels, and increases in the relative activities of antioxidant enzymes in R. glutinosa under oxidative stress support the possible role of these enzymes in the PQ tolerance of R. glutinosa. However, the relatively lower levels of SOD versus PQ tolerance, and the mixed responses of antioxidant enzymes to stresses and hormones, suggest a possible alternative mechanism(s) for PQ tolerance in R. glutinosa.     

重茬地黄土壤酚酸的动态积累及其对地黄生长的影响

为了探究酚酸物质与地黄连作障碍之间的关系,检测了重茬地黄土壤中阿魏酸、香草酸、香草醛和对羟基苯甲酸等4种酚酸物质的积累量,并通过水培条件下添加酚酸,研究了4种酚酸对地黄生长的影响。结果表明：阿魏酸、香草酸和香草醛含量在地黄生长期表现为持续增加,而对羟基苯甲酸含量先升高后降低;外加阿魏酸（8 μg·ml^-1）、香草酸（0.8 μg·ml^-1）、香草醛（1.2 μg·ml^-1）和对羟基苯甲酸（3.0 μg·ml^-1）对水培地黄根长、根质量、全株鲜质量和株高有较强的抑制作用;酚酸处理使地黄叶片叶绿素含量降低,除对羟基苯甲酸外差异性极显著;经过酚酸处理的地黄幼苗SOD和POD活性为先升高后下降,丙二醛含量升高,其中阿魏酸的处理使根中酶活性降至最低,根部腐烂死亡。推测4种酚酸可能是造成地黄化感作用的物质。     

怀地黄块根内含梓醇结构的组织化学和超微结构研究

应用半薄切片、组织化学和透射电子显微镜观察相结合的方法,研究了梓醇在地黄块根中的贮存位置及其细胞的超微结构。结果表明,地黄块根的韧皮部和木质部的薄壁细胞是梓醇的贮存场所。     

地黄自毒物质提取及其生物指标测定

连作障碍在药用植物地黄(Rehmannia glutinosa L.)栽培中尤其明显,同一地块上种植一茬地黄后,须经8-10a后方可再种。从怀地黄主产区河南省焦作地区采集种植1a地黄的茬后土壤,采用不同溶剂提取地黄土壤自毒物质并利用生物测试确定抑制率最强的土壤提取液作温室栽培实验以确定对植物生理指标的影响。利用高效液相色谱法(HPLC)对土壤提取液和根系分泌物进行比较,并利用高效液相色谱、傅立叶转换红外光谱(FTIR)和电喷雾离子阱质谱(ESI-MS)技术鉴定地黄连作自毒物质的化学成分。生物测试结果表明水和甲醇提取的地黄茬后土壤自毒物质对地黄生长具有最强的抑制作用;在培养器中加入浓度0.5g/mL的水和甲醇提取液对地黄胚根的抑制率分别达17%和26%,当浓度增至5.0g/mL时,抑制率增加到70%以上。盆栽结果显示甲醇提取的自毒物质导致地黄地上部叶绿素含量减少,根系活力下降,明显抑制了地黄体内超氧化物歧化酶、过氧化物歧化酶、过氧化物酶的活性,加剧了地黄膜质过氧化,以及引起地黄植体内生长素类激素含量下降。高效液相色谱法显示土壤提取液和根系分泌物具有相似图谱。电喷雾离子阱质谱分析共检测到6个特征物质:香草酸、D-甘露醇、二十六烷酸苯羟基乙酯、毛蕊花糖苷、β-谷甾醇和胡萝卜苷。     

Total phenolics,flavonoids and antioxidant properties of three Ceropegia species from Western Ghats of India

The aim of present work was to assess the total phenolic content (TPC), total flavonoid content (TFC), phenolic compounds and antioxidant properties of various extracts of three Ceropegia spp.: Ceropegia spiralis, Ceropegia panchganiensis and Ceropegia evansii from Western Ghats of India. TPC of the samples varied from 0.3 ± 0.2 to 28.5 ± 0.3 mg TAE/g FW, whereas, TFC of the samples ranged between 0.1 ± 0.1 and 15.3 ± 0.3 mg RE/g FW. The major phenolic compounds identified were gallic acid, vanillin, cathechol and ferulic acid. All the extracts possess 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidant power (FRAP) as well as metal chelating ability and this was also supported by significant correlation with TPC and TFC. To the best of our knowledge, this is the first paper presenting comprehensive data on TPC, TFC, phenolic profile and antioxidant properties of the Ceropegia spp.