人工栽培唐古特大黄中蒽醌含量水平的分析

采用醋酸镁比色法测定青海省不同海拔种植基地中栽培唐古特大黄的总蒽醌含量,并和野生大黄药材进行了比较。结果表明,在海拔较高地区栽培的二、三年生唐古特大黄中,其总蒽醌含量均明显高于较低海拔地区栽培的同龄唐古特大黄,3年生人工栽培大黄总蒽醌含量只有野生大黄药材的一半左右。     

甘肃省药用植物资源及其开发利用现状

甘肃省有着丰富的药用植物资源和悠久的药材种植历史,中药材种植面积及产量位居全国之首。笔者分别对该省近年来已经实施了人工种植的药用植物和尚处于野生状态的药用植物资源的开发利用现状进行了分析,为该省药用植物资源利用及中药产业的进一步发展提供参考。     

野生阿魏与栽培阿魏植物不同部位的挥发油及阿魏酸含量测定

目的:为了提高新疆阿魏资源储量分布和种植面积,保护野生资源,对阿魏药材的人工栽培品与野生品进行质量对比分析,为人工栽培环境采收的阿魏药材的质量评价提供参考依据。方法:采用挥发油测定法以及高效液相色谱法,对新疆不同产地野生阿魏和栽培品阿魏的挥发油和阿魏酸进行了含量测定。结果:人工压榨阿魏树脂,测得的挥发油含量中,各地野生品均符合药典标准(挥发油含量≥10%),栽培品中,阜康阿魏和伊宁产新疆阿魏达到药典标准。阿魏酸含量测定结果中,阿勒泰栽培品,阜康栽培品和伊宁栽培品的根和叶的阿魏酸含量,均大于野生品阿魏酸含量。结论:人工栽培环境采收的阿魏药材前景可观,不同部位均可以作为提取物的制备原料。     

药用植物植物工厂

<正>我国是世界上使用和出口中草药最多的国家,而中草药的80%来自药用植物,所以,药用植物产业在我国有着特殊的意义。面对全球药用植物市场的需求不断攀升,药用植物材料遭到过渡采挖,野生资源日渐枯竭,人工栽培早已成了满足市场需求的重要手段,但传统的人工种植存在诸如病虫害发生严重导致的农药泛用而药材农残严重、不同区域不同药农种植的药用植物外形特征各异、产量和质量稳定性差等诸多缺陷。     

神农架药用植物

国家对野生药材资源实行保护、采猎相结合的原则,并创造条件开展人工种养。国家重点保护的野生药材物种分为三级:一级:濒临灭绝状态的稀有珍贵野生药材物种;二级:分布区域缩小、资源处于衰竭状态的重要野生药材物种;三级:资源严重减少的主要常用野生药材物种。禁止采猎一级保护野生药材物种。破坏野生药材资源情节严重,构成犯罪的,由司法机关依法追究刑事责任。摘编自《野生药材资源保护管理条例》神农架拥有独特的地理位置和优越的气候条件,是当今北半球中纬度内陆地区保存较为完好的亚热带森林生态系统。她的植物成分丰富多彩,也为药用植物的孕育和繁衍提供了良好的生态条件。据考察,神农架有高等维管束植物3200多种,其中的药用植物达1 800多种。因此,神农架也有了"植物王国、天然药库"的美誉。     

药用植物淫羊藿资源可持续利用现状与展望

药用植物淫羊藿主要分布于我国, 在中药、功能性食品和园林观赏等领域具有重要用途。目前淫羊藿药材主要依靠野外采摘, 过度的采挖已经对淫羊藿野生资源造成严重威胁和破坏, 必须采取有效策略, 推进淫羊藿资源的保育和可持续利用。人工栽培、野生抚育与人工半抚育是保护淫羊藿野生资源的重要途径。此外还可采用植物代谢工程、细胞工程、组织培养与微生物发酵工程技术生产淫羊藿有效成分黄酮类化合物, 减少野生药材的采挖与消耗。全面调查不同种淫羊藿中药理活性成分的种类和含量以及提高有效成分的提取收率对于野生药材保护也具有一定的作用。对于一些珍稀濒危的淫羊藿种质资源, 还应采取必要的行政监管手段加以保护。     

山茱萸资源的开发与综合利用

祖国浩瀚的林海,不但蕴育着极为丰富的野生药材资源,而且有些树种本身也可作为名贵药源。如此树种很多,而山茱萸则是其中的佼佼者。近几年来,由于山茱萸的药用价值愈来愈高,所以种植面积也在逐渐扩大,但产量还是有限,在国内外药材贸易中仍是供不应求。可见充分利用现有的资源优势,大量推广种植山茱萸具有广阔前景。     

药用植物淫羊藿资源可持续利用现状与展望

药用植物淫羊藿主要分布于我国,在中药、功能性食品和园林观赏等领域具有重要用途.目前淫羊藿药材主要依靠野外采摘,过度的采挖已经对淫羊藿野生资源造成严重威胁和破坏,必须采取有效策略,推进淫羊藿资源的保育和可持续利用.人工栽培、野生抚育与人工半抚育是保护淫羊藿野生资源的重要途径.此外还可采用植物代酣工程、细胞工程、组织培养与微生物发酵工程技术生产淫羊藿有效成分黄酮类化合物,减少野生药材的采挖与消耗.全面调查不同种淫羊藿中药理活性成分的种类和含量以及提高有效成分的提取收率对于野生药材保护也具有一定的作用.对于一些珍稀濒危的淫羊藿种质资源,还应采取必要的行政监管手段加以保护.     

湘潭市岳塘区中药资源调查

目的：对湘潭市岳塘区的药用植物资源进行调查，为当地中药资源保护与开发提供参考。方法：对野生中药资源的调查采用代表区域-样地-样方套-样方的调查方式，选取草丛、灌丛、阔叶林和针叶林4种代表区域进行调查。同时，对栽培药材、中医药传统知识及中药材市场进行调查。结果：共调查47个样地，发现药用植物115科439种。当地药用植物以野生为主，无人工栽培药用植物。调查了3家中药材企业，掌握了园区中药饮片企业所利用中药材的品种、来源、产地、价格信息、年需求量、供应量、销售去向等信息资料。访问民间医师、药农10人，共收录民间传统验方15个，对现代中医药研究具有一定的价值。结论：岳塘区野生药用植物资源丰富、种类繁多，但野生资源利用程度低，无栽培药用植物种植，可为岳塘区野生中药资源的保护、利用、开发提供科学参考。     

甘肃省款冬资源调查与保护策略

目的：通过对甘肃省款冬资源分布及变化情况进行调查,制定款冬资源的利用和保护策略。方法：通过走访调查款冬野生资源分布区域、蕴藏量、采挖量及人工种植面积、产量、交易量、市场价格等。结果：甘肃省款冬资源丰富,野生款冬年蕴藏量约300～500 t,采挖做药用约占蕴藏量的10%。人工种植2020年全省款冬种植面积约1600 hm2,年产量约1300 t,种植面积受价格下跌影响呈下降趋势。结论：加强款冬野生资源的保护,开展款冬生产的创新研究,对于甘肃省款冬产业可持续发展具有十分重要的作用。     

Ri质粒介导的毛状根体系建立及其在植物次生代谢产物合成中的研究进展

发根农杆菌Ri质粒可诱导植物产生毛状根体系,该体系具有遗传性状稳定且增殖速度快的特点,可用于药用植物次生代谢产物的生产研究,为利用生物反应器技术进行药用植物有效成分工业化水平的发酵培养开辟了新途径。本文主要综述了发根农杆菌Ri质粒介导的植物毛状根体系遗传转化机理,并对毛状根体系在药用植物次生代谢产物生产中的研究现状进行了深入分析,为从基因水平上调控植物次生代谢产物的合成提供新思路。     

Hairy root research: recent scenario and exciting prospects

High stability of the production of secondary metabolites is an interesting characteristic of hairy root cultures. For 25 years, hairy roots have been investigated as a biological system for the production of valuable compounds from medicinal plants. A better understanding of the molecular mechanism of hairy root development, which is based on the transfer of Agrobacterium rhizogenes T-DNA into the plant genome, has facilitated its increasing use in metabolic engineering. Hairy roots can also produce recombinant proteins from transgenic roots, and thereby hold immense potential for the pharmaceutical industry. In addition, hairy roots offer promise for phytoremediation because of their abundant neoplastic root proliferation. Recent progress in the scaling-up of hairy root cultures is making this system an attractive tool for industrial processes.     

Transformed root cultures for biotechnology

This review is concerned with the application of hairy roots, i.e. plant roots formed from plant cells after transformation by Agrobacterium rhizogenes for the production of bioactive compounds. Transformed root cultures have been established from numerous species of dicotyledonous plants. The plants, as well as the main products accumulated in hairy root cultures derived from these plants, are listed in this paper. Data are presented on novel compounds, hitherto detected only in transformed roots but not occurring in the corresponding intact plants. The possible use of hairy root cultures for the over-production of secondary metabolites and biotransformation of chemicals is discussed. In order to enhance the productivity of hairy root cultures, various methods have been derived, and optimized procedures are proposed. They include selection of high-producing clones, elicitation, composition of growth media, culture conditions and genetic approach. Hairy roots usually store secondary metabolites in vacuoles inside the cells. Therefore, several methods have been used to increase the amount of products released into the medium. Unfortunately, no general procedure is known that works in all cases, and the excretion behaviour of hairy root cultures varies from one species to another and even within one species from one clone to another. Special attention is given to the cultivation methods and bioreactor systems for hairy root cultures. Hairy roots are cultivated usually in shake flasks; however, shake flask culture is not suitable for the complex optimization and continuous control of the culture conditions. In this paper, we are going to present bioreactors proposed for the cultivation of hairy roots under more or less controlled conditions. Modifications of typical bacterial bioreactors, i.e. stirred tanks, airlift loop reactors and other constructions, are presented. A very special type of bioreactor providing good conditions for loose root mass multiplication without oxygen or substrate limitations, is the mist bioreactor. Nowadays, it is practically impossible to select the one best bioreactor type for hairy root culture.     

三裂叶野葛毛状根的培养及其葛根素的产生

发根农杆菌ATCC15834感染三裂叶野葛叶片外植体20天后,从其切口叶脉处产生的愈伤组织上产生毛状根。感染35天后约85％的叶片外植体产生毛状根。毛状根能在无外源生长调节剂的MS固体和液体培养基上自主生长,但在液体培养基中培养的毛状根生长更迅速,也不会形成愈伤组织。毛状根线粒体膜电势的荧光染色结果表明,液体培养的毛状根细胞线粒体的膜电势比固体培养的毛状根高11．8倍。PCR结果证实,发根农杆菌Ri质粒的rolB和rolC基因已在三裂叶野葛毛状根基因组中整合并得到表达。HPLC测定结果表明,三裂叶野葛毛状根中的葛根素含量约为对照根(种子萌发产生的幼苗根)的2．5倍,达1．190mg/g．dry．wt;并比多年生葛根生药片的葛根素含量高6．7％。     

Production of transgenic Aralia elata regenerated from Agrobacterium rhizogenes-mediated transformed roots

Transgenic hairy roots were induced from petiole and root segments of in vitro plant Aralia elata, a medicinal woody shrub, after co-cultivation with A. rhizogenes ATCC 15834. The percentage of putative hairy root induction from root segments was higher (26.7%) than petiole explants (10.0%). Hairy roots showed active production of lateral roots with vigorous elongation. Transgenic plants were regenerated from hairy roots via somatic embryogenesis. These plants had wrinkled leaves, short petioles and numerous lateral hairy roots. The RT-PCR analysis showed the expression of rol A, B, C, D, aux 1 and 2 genes differed between the transgenic lines. Endogenous IAA level was higher in transgenic than non-transgenic plants. Conclusively, transgenic hairy roots were developed for first time in A. elata and the transgenic hairy root lines showed distinct morphological growth pattern and gene expression.     

三裂叶野葛毛状根的培养及其葛根素的产生

发根农杆菌ATCC15834感染三裂叶野葛叶片外植体20天后,从其切口叶脉处产生的愈伤组织上产生毛状根。感染35天后约85%的叶片外植体产生毛状根。毛状根能在无外源生长调节剂的MS固体和液体培养基上自主生长,但在液体培养基中培养的毛状根生长更迅速,也不会形成愈伤组织。毛状根线粒体膜电势的荧光染色结果表明,液体培养的毛状根细胞线粒体的膜电势比固体培养的毛状根高11.8倍。PCR结果证实,发根农杆菌Ri质粒的rolB和rolC基因已在三裂叶野葛毛状根基因组中整合并得到表达。HPLC测定结果表明,三裂叶野葛毛状根中的葛根素含量约为对照根(种子萌发产生的幼苗根)的2.5倍,达1.190 mg/g.dry.wt;并比多年生葛根生药片的葛根素含量高6.7%。     

Expression of transferred genes during hairy root development in pea

Root border cell development and expression of reporter genes were evaluated in transgenic pea hairy roots. Successful induction of hairy roots in pea is conditioned by bacterial strain and plant genotype, as well as by developmental and environmental factors. Morphological changes sometimes occur when hairy roots are transferred from infected plants to tissue culture media, but such changes are confined to specific clones. Expression of reporter genes under the control of promoters from bean (Phaseolus vulgaris L.) stress genes encoding phenylalanine ammonia lyase and chalcone synthase were evaluated. Expression patterns vary between hairy roots taken directly from infected plants, and those grown in culture; most hairy roots taken from infected plants exhibit expression throughout all tissues, whereas expression in cultured hairy roots is most often localized to specific tissues. Patterns of expression that occur during different stages of hairy root development are very similar to those observed in transgenic plants expressing the same fusion genes. Border cell separation and release in hairy roots is normal, and expression of glucuronidase in border cells of some transgenic roots resulted in development of bright blue single cells. Cultured hairy roots should provide a very useful model for studying the effect of defined changes in root border cells on microbial associations with roots of this important legume.Abbreviations YEM yeast extract-mannitol - GUS glucuronidase - PAL phenylalanine ammonium lyase - CHS chalcone syntase     

Application of cell technologies for production of plant-derived bioactive substances of plant origin

Bioactive substances (BAS) of plant origin are known to play a very important role in modern medicine. Their use, however, is often limited by availability of plant resources and may jeopardize rare species of medicinal plants. Plant cell cultures can serve as a renewable source of valuable secondary metabolites. To the date, however, only few examples of their commercial use are known. The main reasons for such a situation are the insufficient production of secondary metabolites and high cultivation costs. It is possible to increase the performance of plant cell cultures by one or two orders of magnitude using traditional methods, such as selection of highly productive strains, optimization of the medium composition, elicitation, and addition of precursors of secondary metabolite biosynthesis. The progress in molecular biology methods brought about the advent of new means for increasing of the productivity of cell cultures based on the methods of metabolic engineering. Thus, overexpression of genes encoding the enzymes involved in the synthesis of the target product or, by contrast, repression of these genes significantly influences the cell biosynthetic capacity in vitro. Nevertheless, the attempts of the production of many secondary metabolites in plant cell culture were unsuccessful so far, probably due to the peculiarities of the cell culture as an artificial population of plant somatic cells. The use of plant organ culture or transformed roots (hairy root) could turn to be a considerably more efficient solution for this problem. The production of plant-derived secondary metabolites in yeast or bacteria transformed with plant genes is being studied currently. Although the attempts to use metabolic engineering methods were not particularly successful so far, new insights in biochemistry and physiology of secondary metabolism, particularly in regulation and compartmentation of secondary metabolite synthesis as well as mechanisms of their transport and storage make these approaches promising.     

人参发根的诱导及其适宜培养条件的研究

利用发根农杆菌A₄菌株在人参根外植体上直接诱导产生发根。在1/2MS固体培养基上建立起发根离体培养系,经连续多代的培养,发根仍保持旺盛生长状态。PCR扩增结果表明,发根农杆菌R_I质粒的rolC基因已在人参发根基因组中整合并得到表达。液体培养基中发根生长速度约为固体培养的2倍。经对发根中人参皂苷含量及比生长速率的测定,筛选出高产发根系R9923。利用HPLC法测定了R9923发根系中单体皂苷Rg1、Re、Rf、Rb1、Rc、Rb2和Rd的含量,人参总皂苷含量达15.2mg/g。确定1/2MS培养液（30g/L蔗糖）、摇床转速110r/min、每２周更换一次培养液、继代培养时间4周,为人参发根生长适宜条件。探讨了培养容积、发根初始接种量以及分级放大培养工艺对发根大规模生产过程中生物产量和皂苷含量的影响。