加工方法对黄花蒿提取青蒿素含量的影响

通过对采收后的黄花蒿植株进行适当的处理及干燥温度和贮藏时间对比试验,采用HPLC法测定,探讨提高青蒿素含量的加工新方法。结果表明:整株立式阴晾一定时间后晒干的处理随着阴晾时间的增加青蒿素含量呈抛物线状变化,4～5d最高,达显著水平,之后逐渐下降;随着干燥温度的升高青蒿素含量呈下降趋势,40℃时叶片青蒿素含量较高;随着贮藏时间的延长青蒿素含量逐渐下降,贮藏100 d后下降明显。采收后整株立式阴晾4～5 d后再晒干方法能提高黄花蒿叶片的青蒿素含量。40℃的干燥温度能使叶片中青蒿素含量损耗较少。黄花蒿叶片的保质贮藏时间约90 d。     

无载体固定化细胞的研究进展

以无载体因三化酵母细胞酒精连续发酵的成功工业化应用为实例,并与通常的载休固定化细胞技术比较,无载体固定化细胞技术的优缺点,系统提出了无载体固定化细胞技术的概念,进而对载体固定化细胞技术在其它微生物发酵和动植物细胞培养过程的应用前景进行了展望。     

广西黄花蒿类型调查研究

本文报道广西黄花蒿类型调查结果。１）生长在石山上的黄花蒿类型植株长势比生长在平地、路边、土坡、房前屋后的类型差,且产量也较低。２）同一分布区内,生长在石山的黄花蒿类型,有效成分（青蒿素）含量明显高于生长在平地、路边的类型,前者比后者含量提高００３％～０２８％。     

黄花蒿培养细胞中青蒿素合成代谢的体外调节

黄花蒿培养细胞通过两步培养积累青蒿素．第１步在含有０．２～０．４ｍｇ／Ｌ６－苄基氨基嘌呤（６－ＢＡ）和３～４ｍｇ／Ｌ吲哚乙酸（ＩＡＡ）的Ｎ６培养基中进行细胞的增殖培养,第２步将培养好的细胞转入含０．２～０．４ｍｇ／Ｌ６－ＢＡ和０．２～０．４ｍｇ／ＬＩＡＡ的改良Ｎ６培养基中进行青蒿素的合成．青蒿素的合成量为１９０μｇ／ｇ干细胞左右．当在第２步培养中加入青蒿素合成前体青蒿酸,青蒿素合成量比仅靠激素诱导提高了３倍多．青蒿素的合成途径是植物固醇合成途径的分支途径,当在青蒿素合成过程即第２步培养中加入固醇生物合成抑制剂双氯苯咪唑和氯化氯胆碱处理,可使代谢向合成青蒿素的方向移动,青蒿素合成量明显提高．经２００ｍｇ／Ｌ氯化氯胆碱处理２ｄ,黄花蒿细胞合成青蒿素量为３７２μｇ／ｇ干细胞;经２０ｍｇ／Ｌ双氯苯咪唑处理４ｄ,黄花蒿细胞合成青蒿素量为１５４０μｇ／ｇ干细胞,比靠激素诱导提高了８倍多,与诱导脱分化细胞的黄花蒿叶中所含的青蒿素（３０００μｇ／ｇ干细胞）处于同一个数量级．以上结果表明：在通过植物激素调节可以合成青蒿素的黄花蒿培养细胞中,缺乏青蒿素合成前体是青蒿素合成量低的重要原因．因此,在青蒿素合成的过程中通过体外调节,     

不同土壤环境对黄花蒿生长和青蒿素含量的影响研究

通过田间小区试验,比较研究了施肥与不施肥条件下,4种土壤环境（沙土、旱地土、水稻土和棕色石灰土）对黄花蒿的生长、生物量分配和青蒿素含量的影响。结果表明：黄花蒿对土壤养分的适应性较强,在沙土、旱地土、水稻土和石灰土上均能生长发育,养分水平低时,分配更多的生物量到根,根生物量分数和根/冠比增大;养分水平高时,分配更多的生物量到叶,叶生物量分数增加。黄花蒿的生长和青蒿素含量显著受土壤养分的影响,不施肥时,石灰土和水稻土栽培黄花蒿的株高、地径、总生物量、叶生物量和青蒿素含量显著大于旱地土,而旱地土又显著大于沙土。但在施肥条件下,以上参数不同土壤间无显著差异,且显著高于不施肥。因此,只要根据土壤养分状况合理施肥,黄花蒿在不同养分土壤栽培均能获得较高的青蒿素产量。     

内生青霉菌对黄花蒿组培苗生长和青蒿素合成的影响

从黄花蒿茎中分离得到了17株内生真菌,其中内生青霉菌(Penicilliumsp.Y2)能有效促进黄花蒿组培苗生长及青蒿素合成。内生青霉菌悬浮培养5d后,分别将培养液与菌丝匀浆后经过高压灭菌处理,或将培养液经过高压灭菌、过滤除菌处理获得3种内生菌诱导子(A、B和C)。结果表明,3种内生菌诱导子对植株生长、抗氧化酶活性及青蒿素合成都有促进作用,诱导子C青蒿素合成诱导效果最好,可促进黄花蒿组培苗的干重增长44.44%、可溶性糖含量提高38.24%,诱导超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)活性,从而提高青蒿素合成达58.86%,黄花蒿组培苗青蒿素含量达4.701mg.g-1(干重)。     

无载体固定化细胞的研究进展

以无载体固定化酵母细胞酒精连续发酵的成功工业化应用为实例,并与通常的载体固定化细胞技术比较,阐述了无载体固定化细胞技术的优缺点,系统提出了无载体固定化细胞技术的概念,进而对无载体固定化细胞技术在其它微生物发酵和动植物细胞培养过程的应用前景进行了展望。     

黄花蒿中青蒿素含量的RP-HPLC法测定

建立黄花蒿中青蒿素的高效液相色谱测定方法,采用柱前衍生RP-HPLC法测定黄花蒿中青蒿素的含量,采用ZORBAXXDB-C18(4.6mm×150mm,5μm)色谱柱,甲醇-0.01mol/L醋酸钠-醋酸缓冲液(pH5.8,体积比62∶38)为流动相;检测波长:260nm;流速:0.8mL/min;柱温:30℃。对广西、沈阳、北京、郑州、苏州和杭州等不同产地的野生黄花蒿样品、以及同一产地不同采集时间黄花蒿样品进行检测,结果表明不同地区青蒿素含量差异很大,同一地区不同采集时间黄花蒿样品的青蒿素含量也有差异。该法准确可靠、重现性好,能准确地反映青蒿素含量的检测。     

青蒿倍半萜合酶(环化酶)研究进展

青蒿素是从中药青蒿中分离得到的抗疟有效单体,是含有过氧基团的新型倍半萜内酯化合物,是目前世界上最有效的疟疾治疗药物。青蒿素的生物合成途径属于类异戊二烯代谢途径中的倍半萜类分支途径,倍半萜合酶是该途径的关键酶之一,目前已从青蒿中克隆了多个倍半萜合酶基因。综述了青蒿中已克隆的几种倍半萜合酶基因的研究进展。     

State of the art of the production of the antimalarial compound artemisinin in plants

For more than three centuries we have relied on the extracts of the bark of Cinchona species to treat malaria. Now, it seems we may be changing to the leaves of a Chinese weed, Artemisia annua, and its active compound artemisinin. Artemisinin-derived drugs have been proved particularly effective treatments for severe malaria, even for multi-drug-resistant malaria. However, this promising antimalarial compound remains expensive and is hardly available on a global scale. Therefore, many research groups have directed their investigations toward the enhancement of artemisinin production in A. annua cell cultures or whole plants in order to overproduce artemisinin or one of its precursors. This article provides a brief review of the state of art of the different aspects in A. annua research.     

适于青蒿芽生长和青蒿素积累的光、温和培养方式探讨

探讨了光照、温度和培养方式对青蒿芽生长和青蒿素合成的影响。适宜芽生长和青蒿素积累的光照强度约为３ ０００ ｌｘ,照光时间为２０ ｈ／ｄ ;芽生长和青蒿素积累的最适温度分别为２５ ℃和３０ ℃,通过先２５ ℃（２５ ｄ）后３０ ℃（５ ｄ） 的温度转变二步培养法可以提高青蒿素的产量;青蒿芽生长和青蒿素积累的最佳培养方式为非浸没低转速摇瓶培养。     

青蒿素生物合成分子调控研究进展

青蒿素是目前世界上最有效的疟疾治疗药物。通过对青蒿素的生物合成途径,青蒿素生物合成途径的关键酶,青蒿素生物合成的分子调控的介绍,综述了青蒿素生物合成分子调控的最新研究进展。     

温度对青蒿毛状根生长和青蒿素生物合成的影响

本实验研究了不同温度(15℃～35℃)对青蒿毛状根生长和青蒿素生物合成的影响,发现25℃有利于毛状根生长,30℃促进了青蒿素生物合成。通过温度改变的二步培养技术(培养前20d温度控制在25℃,后10d温度提高到30℃),青蒿素的产量得到明显提高,高于在恒温培养时(25℃或30℃)的结果。     

Modulation of artemisinin biosynthesis by elicitors,inhibitor, and precursor in hairy root cultures of Artemisia annua L.

Artemisinin is frequently used in the artemisinin-based combination therapy to cure drug-resistant malaria in Asian subcontinent and large swath of Africa. The hairy root system, using the Agrobacterium rhizogenes LBA 9402 strain to enhance the production of artemisinin in Artemisia annua L., is developed in our laboratory. The transgenic nature of hairy root lines and the copy number of transgene (rol B) were confirmed using polymerase chain reaction and Southern Blot analyses, respectively. The effect of different concentrations of methyl jasmonate (MeJA), fungal elicitors (Alternaria alternate, Curvularia limata, Fusarium solani, and Piriformospora indica), farnesyl pyrophosphate, and miconazole on artemisinin production in hairy root cultures were evaluated. Among all the factors used individually for their effect on artemisinin production in hairy root culture system, the maximum enhancement was achieved with P. indica (1.97 times). Increment of 2.44 times in artemisinin concentration by this system was, however, obtained by combined addition of MeJA and cell homogenate of P. indica in the culture medium. The effects of these factors on artemisinin production were positively correlated with regulatory genes of MVA, MEP, and artemisinin biosynthetic pathways, viz. hmgr, ads, cyp71av1, aldh1, dxs, dxr, and dbr2 in hairy root cultures of A. annua L.     

青蒿素的发现与研究进展

1967年北京《5.23抗疟计划》付诸实施,在全国多个研究单位协作下,组织植物化学与药理学等专业200多人,搜寻对抗耐氯喹恶性疟疾的新药.他们追索我国历代抗疟方剂,用约200种草药制成380多种抽提物,再筛查其对小鼠疟疾模型的疗效.最后确定,在60℃用乙醚萃取中药青蒿的黄花蒿所得第191号中性抽提物,对感染鼠疟和猴疟的小鼠与猴以及21例恶性疟、间日疟患者均能奏效.1972年11月,从该抽提物分离出相对分子质量0.282 kD的无色结晶,命名为青蒿素.它属于一种新型的倍半萜内酯.继后青蒿素纯品的胶囊被用于临床数千例疟疾患者.2006年以来,为克服耐药性,世界卫生组织(WHO)宣布改用青蒿素的联合疗法(ACT),挽救了80个国家100多万人的生命.因在发现青蒿素过程中的关键作用,屠呦呦被授予2011年度拉斯克临床医学研究奖.本文对青蒿素发现的争议,如何恰当评价其他科学家的贡献,以及开发植物界以外青蒿素新药源的近况,均作了简要报道.     

Abscisic acid (ABA) treatment increases artemisinin content in <Emphasis Type="Italic">Artemisia annua</Emphasis> by enhancing the expression of genes in artemisinin biosynthetic pathway

Artemisinin, a sesquiterpene lactone endoperoxide derived from Artemisia annua L., is the most effective antimalarial drug. In an effort to increase the artemisinin production, abscisic acid (ABA) with different concentrations (1, 10 and 100 μM) was tested by treating A. annua plants. As a result, the artemisinin content in ABA-treated plants was significantly increased. Especially, artemisinin content in plants treated by 10 μM ABA was 65% higher than that in the control plants, up to an average of 1.84% dry weight. Gene expression analysis showed that in both the ABA-treated plants and cell suspension cultures, HMGR, FPS, CYP71AV1 and CPR, the important genes in the artemisinin biosynthetic pathway, were significantly induced. While only a slight increase of ADS expression was observed in ABA-treated plants, no expression of ADS was detected in cell suspension cultures. This study suggests that there is probably a crosstalk between the ABA signaling pathway and artemisinin biosynthetic pathway and that CYP71AV1, which was induced most significantly, may play a key regulatory role in the artemisinin biosynthetic pathway.