Evaluation on Quality and Utilization Value of Bupleurum falcatum Linne

[Objectives]This study aimed to evaluate the quality of Bupleurum falcatum Linne interplanted with walnut in Ganluo and study the development and utilization value of other parts.[Methods]The contents of saikosaponins a and d were determined with RP-HPLC,and the contents of 7 kinds of harmful elements were determined by ICP-MS.The quality of B.falcatum Linne was compared with that of other Bupleurum species.[Results]The total saikosaponin content in Radix Bupleurum Falcatum(15.894 mg/g)was higher than that in Radix Bupleuri of other origins(8.748 mg/g).The total saikosaponin content in leaves of B.falcatum Linne(7.518 mg/g)is more than twice the limit promulgated by Chinese Pharmacopoeia.The contents of Al,Cr,Cu,As,Cd,Pb and Hg in B.falcatum Linne were all lower than the limits promulgated by the pharmacopoeia.In short,the quality of Radix Bupleurum Falcatum was better than that of Radix Bupleurum Marginatum(S3,S4)and Radix Bupleurum Sachalinense(S5).The leaves of B.falcatum Linne contained more saikosaponins and less harmful elements.[Conclusions]The method and technology of interplanting B.falcatum Linne with walnut in Ganluo are mature.The quality of the medicinal materials produced is superior,and the leaves are also rich in saikosaponins a and d,and can be used as the raw material for extracting saikosaponins a and d.This study provides a basis for further in-depth research on the cultivation of B.falcatum Linne in the domestic market.

柴胡属(Bupleurum L)植物研究进展

参阅了国内外有关文献 ,概述了近年来柴胡属植物的研究进展概况 ,着重介绍了柴胡的生物学、化学成分。

蒙古柴胡属Bupleurum L.(伞形科)植物的分类和分布(英文)

根据作者近年来在蒙古的野外采集,以及在俄罗斯圣彼德堡科码洛夫植物研究所标本馆LE和北京中国科学院植物研究所标本馆PE等的研究,确认蒙古产7种柴胡属(Bupleurum L.)植物。作出了它们的检索表。按照蒙古植物区系区划,分析和描述了这7种植物的分布。

Effects of Different Treatments on Seed Germination of Bupleurum chinensis

In order to solve the problem of low emergence rate of Bupleurum chinense seeds, and screen out the best germination conditions, the seeds of Bupleurum chinense L. were treated with different soaking time, germination temperature and exogenous regulating substances to improve the germination rate. The results showed that the germination rate of fresh water at 12 h was significantly higher than that at 8 h and 36 h; The germination rate of seeds treated with constant temperature 15℃ and variable temperature 25 ℃/15 ℃ （day/night） was significantly higher than that of other temperature treatments. Seed soaking time and culture temperature also influenced seed germination start day and peak day. Different concentrations of regulatory substances could promote seeds germination. The germination rate of seeds induced by 0.50 mg/L 6-BA was the highest, the start day and peak day were 4 days and 5 days earlier than control check （CK） respectively.

Analysis of codon usage patterns in Bupleurum falcatum chloroplast genome

Objective In order to distinguish the traditional Chinese medicine Bupleurum falcatum and its adulterants effectively and develop a better understanding of the factors affecting synonymous codon usage,codon usage patterns of chloroplast genome,we determine the complete chloroplast(cp)genome of B.falcatum and clarify the main factors that influence codon usage patterns of 78 genes in B.falcatum chloroplast genome.Methods The total genomic DNA of fresh leaves from a single individual of B.falcatum was extracted with EASYspin plus Total DNA Isolation Kit and 2μg genome DNA was sequenced using Illumina Hiseq 2500 Sequencing Platform.The cp genome of B.falcatum was reconstructed with MITObim v1.8 and annotated in the program CPGAVAS2 with default parameters.Python script and Codon W were used to calculate the codon usage bias parameters.Results The full length of B.falcatum cp genome was 155851 bp,132 different genes were annotated in this cp genome containing 80 protein-coding genes,30 tRNA genes,and four rRNA genes.The codon usage models tended to use A/T-ending codons.The neutrality plot,ENC plot,PR2-Bias plot and correspondence analysis showed that both compositional constraint under selection and mutation could affect the codon usage models in B.falcatum cp genome.Furthermore,three optimal codons were identified and most of these three optimal codons ended with G/U.Conclusion The cp genome of B.falcatum has been characterized and the codon usage bias in B.falcatum cp genome is influenced by natural selection,mutation pressure and nucleotide composition.The results will provide much more barcode information for species discrimination and lay a foundation for future research on codon optimization of exogenous genes,genetic engineering and molecular evolution in B.falcatum.

宜昌市幼龄皱皮木瓜与三岛柴胡间作栽培技术

皱皮木瓜是宜昌市道地药材大单品。为解决皱皮木瓜新建园从定植到丰产,其间(4~5年)园田空间利用率低、杂草防控难、种植收益低的问题,结合三岛柴胡(Bupleurum falcatum L.)生长前期需遮阴、后期需打顶等的田间管理要求,探索出幼龄皱皮木瓜与三岛柴胡的药-药间作栽培模式。结合生产实际,对皱皮木瓜和三岛柴胡生长习性、生长发育特性、间作模式结构、栽培管理技术、间作模式综合效益等进行介绍,以期为宜昌市及其他皱皮木瓜产区发展提供参考。

反相HPLC法测定四种柴胡的柴胡皂式a,c,d含量

The saikosaponin a, c, d in Chinese Thorowax were determined by RP-HPLC with UV detecter at the wavelength of 205 nm. The optimal experiment conditions were ODS (5 micron) colum, mobile phase of acenitrile-water (4: 6) and quinoestrol as internal standard. This method is simple, rapid and the reproducibility is good.

HPLC法测定柴胡属植物地上部分黄酮类含量

目的:建立柴胡属植物地上部分中槲皮素、异鼠李素、山奈素HPLC的含量测定方法。方法:伊利特-C18柱(250 mm×4．6 mm,5μm),以甲醇-水(60:40),磷酸调节pH至3．0为流动相,流速:1．0 ml·min-1,检测波长:360 nm。结果:槲皮素在0．70-5．57μg(r槲=0．999 7),异鼠李素在0．16～1．30μg(r异=0．999 2),山奈素在0．09-0．71μg(r山=0．999 4)范围内呈良好的线性关系。平均回收率分别为97．0％,96．8％,97．4％,RSD分别为2．2％,2．2％,1．1％。结论:本法专属性强,灵敏度高,重现性好,适于黄酮类成分的分离与测定。

加味大柴胡汤联合L-精氨酸对阻塞性黄疸大鼠肾功能的影响

目的研究阻塞性黄疸时,加味大柴胡汤联合L-精氨酸(L-Arg)对肾功能的保护作用。方法SD大鼠40只随机分为假手术组、单纯胆总管结扎组、胆总管结扎+L-Arg组(L-Arg组)和胆总管结扎+大柴胡汤+L-Arg组(中西医结合组),每组l0只。造模成功后第2天起,假手术组及单纯胆总管结扎组每天分别腹腔注射生理盐水,L-Arg组腹腔注射L-Arg,中西医结合组既注射L-Arg又喂加味大柴胡汤,连用9 d,观察各组肾功能的变化,同时测定血和肾组织内皮素(ET)、一氧化氮(NO)水平。结果中西医结合组肾功能明显得到改善,且血和肾组织ET下降,NO升高。结论精氨酸和加味大柴胡汤联合使用较单用精氨酸更能减轻阻塞性黄疸时的肾功能损伤,其机制可能与降低体内ET水平、升高NO水平有关。

海拔、播种期和播种量对三岛柴胡生长的影响

比较不同处理对三岛柴胡(Bupleurum falcatum L.)生长发育的影响,为其规范化栽培提供依据。以三岛柴胡为试验材料,设置不同海拔、播期、播种量处理,观察物候期,测定相关生长指标。结果表明,随着海拔升高,三岛柴胡苗期、抽茎拔节期、孕蕾期、开花期和果期相应推迟,海拔600 m株高最高,不同海拔处理间主根长、根粗、单根重、小区根产量和折合产量无显著差异;2月23日播种的三岛柴胡苗期、抽茎拔节期、孕蕾期、开花期和果期相应推迟,两个播期处理的主根长、根粗、单根重、小区根产量和折合产量无显著差异;随着播种量的加大,主根长、单根重均表现为逐渐减小趋势,根粗和小区根产量无显著差异。海拔1100 m以下适宜三岛柴胡生长;11月和2月播种均可;播种量以15.0 kg/hm^(2)为宜,综合经济效益较好。

聚左旋乳酸非溶剂挥发诱导成孔机制与纳米多孔纤维膜制备

为实现聚左旋乳酸纳米纤维孔隙结构可控,以氯仿和丙酮为混合溶剂,通过在聚左旋乳酸中加入柴胡,采用静电纺丝技术制备纳米多孔纤维膜。探索了非溶剂挥发诱导相分离成孔机制,研究了柴胡质量分数、溶剂配比、聚左旋乳酸质量分数对纤维膜孔隙结构的影响。结果表明:在非溶剂溶液体系中添加柴胡能够提升纤维膜的孔隙率,随着柴胡质量分数的增加,纳米多孔纤维膜的孔隙率逐渐增加,当柴胡质量分数为2%时孔隙率最大,为70.98%;改变氯仿与丙酮配比会对纤维膜孔隙结构造成影响,当氯仿和丙酮质量比为8∶1时,纤维膜的孔隙率达到最大,为82.09%;聚左旋乳酸质量分数为9%时纤维膜的孔隙率达到最大。

Auxin induces lateral root formation in Bupleurum: A heme oxygenase dependent approach

Objective: The content of saikosaponins in genus Bupleurum is increased with numbers of lateral root, but the genetic mechanisms are largely unknown. This study aims to identify the heme oxygenase(HO) gene family members of B. chinense and B. scorzonerifolium, and assess their role in the root development in Bupleurum.Methods: The gene sequences of HO family were selected from iso-seq full-length transcriptome data of B. chinense and B. scorzonerifolium, and were analyzed in physicochemical properties, conserved domains,motifs and phylogenetic relationship. In addition, the expression patterns of HO gene in different parts of roots were compared via transcriptome sequencing and qRT-PCR in the two species.Results: Five Bupleurum HO genes(BcHO1-BcHO5) belonging to the HO1 subfamily were identified from the transcriptome data, whereas the HO_(2) subfamily member was not identified. The expression levels of BcHO1 and BcHO_(2) were significantly higher than those of other three HO members in the transcriptome analysis. In addition, the expression profile of BcHO1 showed consistency with lateral root development in B. chinense and B. scorzonerifolium.Conclusion: Hos might participate in the auxin-induced morphogenesis of lateral roots. The yield of saikosaponin may be improved by manipulating expression of these genes.

响应面法优化超声提取柴胡皂苷a、c、d的工艺研究

目的:应用响应面法优化3种柴胡皂苷a、c、d的最佳提取工艺。方法:采用超声提取法,在单因素试验基础上,根据中心组合实验设计原理采用3因素3水平的响应面分析法进行工艺优化,HPLC法测定柴胡皂苷a、c、d的提取率,色谱柱为symmetry C18柱(150 mm×4.6 mm,5μm),流动相为乙腈-水梯度洗脱,流速1.0 mL/min,检测波长200 nm,检测温度25℃。结果:柴胡皂苷a、c、d的最佳条件为:提取时间93 min、超声功率150 W、液固比60,3种总皂苷含量预测值为13.66 mg/g,验证值为13.64 mg/g,与预测值的相对误差为0.15%。结论:该工艺合理、可靠、有效成分提取率高。

不同密度下三岛柴胡干物质积累和柴胡皂苷含量的动态变化

本实验研究不同密度下的三岛柴胡干物质积累和柴胡皂苷含量动态变化。三岛柴胡个体干物质积累随着密度的增大而减少,而群体密度在生长后期随着密度的增大而增加。在结实期三岛柴胡单株根干物质分配比例随密度的增大而减少。高密度1250062株/公顷柴胡皂苷含量较低,而在生长前期密度714285株/公顷柴胡皂苷含量较高,生长后期两个低密度处理柴胡皂苷含量较高。结实期时三岛柴胡根总皂苷产量以500000株/公顷最高。适宜的密度对三岛柴胡干物质积累与分配、根中柴胡皂苷含量及群体皂苷产量极为重要。

三岛柴胡种子化学成分分析

应用ＧＣ／ＭＳ技术对云南省文山县引种的日本三岛柴胡种子的挥发油和脂肪酸成分 进行定性定量分析，检出５３个挥发油成分和２８个脂肪酸成分。挥发油以４，４，５－三甲基－２－乙 烯、２，２，４三甲基－３－戊烯－１－醇、２，３－二甲基丁烯－３－醇和２，３－二甲基戊烷为主；脂肪酸以十八碳 烯酸，十八烷酸，壬二酸和辛二酸为主。从种子的正了醇提取部分还分离出８个皂甙。其中 ３个鉴定为柴胡皂甙ｄ，ｃ和６”－Ｏ－乙酰基柴胡皂甙ｄ。

种子成熟度及种皮对北柴胡和三岛柴胡种子萌发的影响

目的:了解影响柴胡种子萌发的主要因素。方法:取不同成熟度柴胡种子浸泡、挑破或去除种皮后,在光照培养箱中做发芽试验。结果:柴胡不同级别花序发育时期差异较大,生产上收获的种子成熟度参差不齐,果Ⅰ～果Ⅲ期种子基本没有发芽力,果Ⅳ～Ⅶ期种子发芽率有显著差异;柴胡种皮对种子水分和气体吸收没有障碍,但去除种皮显著提高了种子发芽率,而去皮后浸泡48 h不能进一步提高发芽率。结论:种子成熟度及种皮内含有的发芽抑制物是影响柴胡种子萌发的重要因素。

三岛柴胡种子中的柴胡皂甙(英文)

利用溶剂萃取、正相硅胶柱层析及RP -8硅胶柱层析 ,从伞形科柴胡属植物三岛柴胡的种子中分离出三种柴胡皂甙 ,即 :柴胡皂甙c ,柴胡皂甙d ,6″-乙酰基柴胡皂甙d。它们的结构通过IR、UV、MS、1HNMR、13CNMR等现代波谱技术以及与标准样品对照确定。

柴胡属5种植物RAPD分析与分类鉴定

目的 探讨正品柴胡与同属近似种包括柴胡 Bupleurum chinense、狭叶柴胡 B.scorzonerifolium、竹叶柴胡B.marginatum、小叶黑柴胡 B.smithii var.parvifolium、大叶柴胡 B.longiradiatum 5种柴胡属植物鉴定的分子证据。方法 CTAB法提取柴胡属原植物总 DNA,以随机引物进行非特异扩增。结果 用随机引物 OPA- 1(5’-CAGGCCCTTC- 3’) ,OPD- 8(5’- GTGTGCCCCA- 3’)和 OPD- 11(5’- AGCGCCATTG- 3’)能有效地鉴定这 5种柴胡属植物。结论 RAPD法可准确鉴定正品柴胡及其近似种。

北柴胡与三岛柴胡种子萌发特性研究

本文做了不同温度、种子提取物、不同温度水浸种及药剂对北柴胡与三岛柴胡种子萌发的影响研究,结果表明:两种柴胡种子均以20℃发芽率最高,种子内都存在着抑制种子萌发的物质,3％的双氧水能促进这两种柴胡种子的萌发。

不同激素配比对三岛柴胡悬浮细胞生长发育的影响

[目的]研究不同配比激素对三岛柴胡悬浮细胞生长发育的影响。[方法]采用均匀设计试验方法,对三岛柴胡悬浮细胞培养基的激素进行优化,并以最优激素配方为基础拟合三岛柴胡悬浮细胞培养中鲜重、干重与培养时间的关系曲线。[结果]三岛柴胡悬浮细胞培养的最优激素配方为:N6+0.5 mg/L NAA+0.75 mg/L 6-BA+1.5 mg/L KT;细胞鲜重与培养时间的关系拟合方程为Y=5.703 6X2-41.429X+136.44;细胞干重与培养时间的关系拟合方程为Y=2.021 8X2-22.595X+57.19。[结论]试验为研究三岛柴胡细胞和实现其大规模培养奠定了基础。