真菌诱导子对青蒿发根细胞生长和青蒿素积累的影响

３种真菌诱导子（大菌丽花轮枝孢（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｉａｅ Ｋｌｅｂ．）、葡枝根霉（Ｒｈｉｚｏｐｕｓ ｓｔｏｌｏｎｉｆｅｒ（Ｅｈｒｅｎｂ．ｅｘＦｒ．）Ｖｕｉｌｌ）和束状刺盘孢（Ｃｏｌｌｅｔｏｒｉｃｈｕｍ ｄｅｍａｔｉｕｍ（Ｐｅｒｓ．）Ｇｒｏｖｅ）处理青蒿（Ａｒｔｅｍｉｓｉａ ａｎｎｕａＬ．）的发根,均能促进发根中青蒿素的积累,其中以大丽花轮枝孢的诱导效果最好;对细胞生长均没有明显影响,     

红豆杉细胞悬浮培养中不同添加物对细胞生长和紫杉醇合成的影响

采用正交实验检测红豆杉（Ｔａｘｕｓ ｃｈｉｎｅｎｓｉｓ（Ｐｉｌｇｅｒ）Ｒｅｈｄ．）细胞悬浮培养中水杨酸、Ｄ－果糖、甘露醇和硫酸镧对细胞生长和紫杉醇（ｔａｘｏｌ）积累的影响。添加１０ｇ／ＬＤ－果糖,可使细胞的鲜重和干重明显增加;添加６０ｇ／Ｌ甘露醇使细胞的鲜重和干重明显减少;１ｍｇ／Ｌ水杨酸仅使细胞鲜重增加,对干重影响不明显;硫酸镧对细胞生长无明显影响。单独添加这４种物质,紫杉醇含量均下降,同时添加     

高山红景天细胞悬浮培养中红景天甙生物合成代谢调控Ⅱ：真 …

考察了６种真菌诱导物对高山红景天（Ｒｈｏｄｉｏｌａ ｓａｃｈａｌｉｎｅｎｓｉｓ,Ｂｏｒ）细胞生长与红景天甙积累的影响。其中以黑曲霉诱导物效果最好。在细胞培养初期添加浓度为４０ｍｇ（ｃａｒｂｏｈｙｄｒａｔｅ）Ｌ的黑曲霉诱导物能使培养细胞中红景天甙含量提高到０．９９５％。前体与诱导物调控组合运用最终使红景天甙产量达到１６７．４ｍｇ／Ｌ,是对照培养的３．８倍。另外,对真菌诱导物的作用机理也进行了探讨,真     

水母雪莲悬浮培养细胞生长和黄酮类活性成分合成

在ＭＳ培养基上进行水母雪莲（ＳａｕｓｕｒｅａｍｅｄｕｓａＭａｘｉｍ．）细胞悬浮培养,研究了摇床转速、接种量、培养液初始ｐＨ、碳源等的影响。结果表明,摇床转速为９０～１２０ｒ／ｍｉｎ,接种量为５０～８０ｇＦＷ／Ｌ,培养液初始ｐＨ５．５～６．０,对水母雪莲悬浮培养细胞生长和黄酮合成最有利。碳源以蔗糖最适合,蔗糖浓度则以４０ｇ／Ｌ较好,此时细胞生长量为１８～１９ｇＤＷ／Ｌ,总黄酮合成量可达１４２３．２５ｍｇ／Ｌ。用ＨＰＬＣ检测显示４′,５,７三羟基３′,６二甲氧基黄酮（ｊａｃｅｏｓｉｄｉｎ）和４′,５,７三羟基６甲氧基黄酮（ｈｉｓｐｉｄｕｌｉｎ）的含量分别达到总黄酮含量的２２．１１％和０．１５％。     

红豆杉细胞培养的研究

从红豆杉（Ｔａｘｕｓｃｈｉｎｅｎｓｉｓ（Ｐｉｌｇ）Ｒｅｈｄ）的嫩茎及针叶诱导的出愈伤组织,对愈伤组织培养及细胞悬浮培养进行了研究,利用ＨＰＬＣ方法测定它们合成紫杉醇的能力,发现了能够提高培养细胞生长速率及紫杉醇含量的一些因子,红豆杉愈伤组织及悬浮培养细胞的生长速率已分别达到０．２５ｇ／Ｌ．ｄ和０．２８ｇ／Ｌ．ｄ。而他们的紫杉醇含量分别是０．００２６％和０．０１２％。     

新疆的刺球座属、穴壳属及普氏腔孢属

本文报道了新疆腔菌纲座囊菌目刺球座属（Ｌａｓｉｏｂｏｔｒｙｓ）、穴壳属（Ｄｏｔｈｉｏｒａ）和普氏腔孢属（Ｐｌｏｗｒｉｇｈｔｉａ）的六种子囊菌,即：忍冬刺球座菌（Ｌ．ｌｏｎｉｃｅｒａｅ）、花揪穴壳菌（Ｄ．ｓｏｒｂｉ）及其无性阶段花揪疡壳孢（Ｄｏｔｈｉｃｈｉｚａ ｓｏｒｂｉ）、茶Ｂｉａｏ子普氏腔孢菌（Ｐ．ｒｉｂｅｓｉａ）、小檗普氏腔孢菌（Ｐ．ｂｅｒｂｅｒｉｄｉｓ）、沙棘普氏腔孢菌（Ｐ．ｈｉｐｐｏｐｈａ     

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)。     

镧对马尾松苗的根系生长和IAA含量的影响

在０．１～１．０ｍｇ／Ｌ硝酸镧的Ｈｏａｇｌａｎｄ培养液中,马尾松（ＰｉｎｕｓｍａｓｏｎｉａｎａＬａｍｂ．）截根苗的侧根数目明显增加,为对照的１．７５～３．７５倍;根的鲜重、干重显著提高,增幅达３３．７％～４６．４％;根系还原力也提高１３．５％～２８．５％。根长仅在０．０５～０．５ｍｇ／Ｌ的低浓度下比对照有所增加,在浓度高于１．０ｍｇ／Ｌ时,根长只有对照的５３．３％。侧根内的游离态ＩＡＡ含量随溶液中镧的浓度增加而相应增加,ＩＡＡ氧化酶的活性则相应降低。硝酸镧处理对该苗地上部的促进效应不如根部显著。     

辣椒炭疽病菌毒素

辣椒炭疽病菌（辣椒刺盘孢Ｃｏｌｌｅｔｏｔｒｉｃｈｕｍ ｃａｐｓｉｃｉ）在２５－２８℃的Ｃｚａｐｅｋ－Ｄｏｘ培养液中振荡培养时分泌出毒素,这种毒素能引起辣椒叶片形成坏死斑,类似于病原菌侵染形成的症状。辣椒叶煎汁培养液和Ｃｚａｐｅｋ－Ｄｏｘ培养液适于Ｃ．ｃａｐｓｉｃｉ的生长和产毒,生长适宜温度和范围分别为２５℃和ｐＨ６－７,在２５－２８℃,ｐＨ６－７的条件下培养滤液毒性最强;光线和通气条件可促进Ｃ．ｃ     

L-苯丙氨酸与血管平滑肌细胞增殖

本文用氚标胸腺嘧啶核苷掺入ＤＮＡ合成法测定自发性高血压大鼠（ＳＨＲ）与正常对照鼠的培养主动脉血管平滑肌细胞（ＶＳＭＣ）增殖,观察Ｌ－苯丙氨酸对细胞增殖、细胞生长及原癌基因ｃ－ｆｏｓ、ｃ－ｍｙｃ表达的影响。结果显示：（１）Ｌ－苯丙氨酸剂量依赖性地抑制血清、碱性成纤维细胞生长因子及凝血酶诱导的ＤＮＡ合成;（２）Ｌ－苯丙氨酸剂量依赖性地抑制细胞对血清的增殖反应;（３）Ｌ－苯丙氨酸抑制血清诱导的ｃ－ｆｏｓ     

西洋参悬浮细胞发酵工艺研究

探讨了西洋参悬浮细胞分步培养与稀土、D-半乳糖和甘露醇等诱导子对悬浮细胞生长及皂甙产量的影响。发现继代4d后换液一次再继续培养获得的培养物,在皂甙产率和糖利用率等方面优于连续培养;D-半乳糖作为诱导子,对悬浮培养的西洋参细胞生长、皂甙产率及皂甙的分泌等方面都有非常明显的促进作用。     

真菌激发子对人参悬浮培养细胞的生长和人参皂甙生物合成的影响

真菌诱导子处理人参悬浮培养细胞后,人参皂甙的合成有明显增加,诱导处理改变人参皂甙的积累时程,促进人参细胞培养物中次生产物的外泌,同时增强细胞对蔗糖的摄取、吸收并引起细胞H~ 流的变化。     

Effects of elicitation on the production of saponin in cell culture of Panax ginseng

This study was initiated to investigate the impacts of elicitor concentration and elicitor-adding time on the saponin synthesis and the cell growth of Panax ginseng cell suspensions. Both of the elicitors tested, yeast extract and methyl jasmonate, significantly improved saponin production. The highest additive level of the seven ginsenosides tested was 2.07% (dry weight basis), which was 28-fold higher than that in the control. The optimum time to add either elicitor was found to be on the day of inoculation. The addition of either elicitor did not show as significant an influence on cell growth as on saponin production. It was advisable to remove 2,4-dichlorophenoxyacetic acid (2,4-D) from the medium when methyl jasmonate was used as the elicitor as methyl jasmonate interacts antagonistically with 2,4-D. These results suggest that the addition of an elicitor to ginseng cell suspension cultures could stimulate saponin production.     

Regulation of fungal elicitors on the cell growth and biosynthesis of saponin of Panax ginseng cell suspension culture]

Adding fungal elicitors to the Panax ginseng cell suspension cultures, the biosynthesis of saponin was obviously induced, the total productivity of saponin in cultures could increase more than 30% of the control. During elicitation, the accumulation patterns of saponin in suspension cultured cells were changed, the culture time for maximum biosynthesis of saponin was shortened 2-4 days comparing with that of the control, and about 80% of biosynthetic saponin in elicited cells was secreted into medium, meanwhile the uptake for sucrose in medium of cells was enhanced, and the disturbing of pH in medium was observed, which predicated that an ion exchange occurred between elicited cells and medium.     

不同培养基及其元素组成对西洋参愈伤组织悬浮培养物生长和皂苷含量的影响

对MS、67-V和FOX 3种基本基质对西洋参(Panax quinquefolium Linn.)愈伤组织悬浮培养物生长和皂苷含量的影响进行了比较。在3种基本基质中,培养物的鲜重和干重增加量差异不大,而皂苷含量和产量差异较大,其中MS较高,FOX次之,67-V最低。探讨了MS基质中,KNO3、CaCl2和MgO4对培养物生长和皂苷含量的影响。KNO3浓度在237．5mg/L时有利于培养物生长,而浓度在1900mg/L时有利于皂苷合成;CaO2浓度在55．35mg/L时有利于培养物生长,而浓度在332．1mg/L时有利于皂苷合成;MgSO4浓度为92．50mg/L时培养物生长最好,皂苷产量也最高。     

三七.人参和西洋参细胞悬浮培养的比较研究

用薄层层析对三七、人参和西洋参愈伤组织进行的初步鉴定表明,三种愈伤组织都含有皂甙和主要皂甙成分Rb_1、Rg_1,三七愈伤组织还含有一种抗癌皂甙Rh_1。对愈伤组织的生长,三七低于人参高于西洋参;对愈伤组织中总皂甙含量,三七均高于人参和西洋参。三种植物细胞悬浮培养结果类似于他们的愈伤组织培养,但生长又进一步提高。三七细胞悬浮培养中皂甙产生的时间进程几乎与生长平行,合适的收获期为培养30天。寡糖素不仅增强三七培养细胞的皂甙形成而且促进细胞生长,较合适的浓度为1.25 ppm。通过以上研究,使三七悬浮培养细胞的生长(干重增加178毫克)为最初培养愈伤组织的4倍以上,总皂甙产率高达20.6毫克,为最初培养愈伤组织的8.5倍。     

Improved Taxol production by combination of inducing factors in suspension cell culture of Taxus baccata

To date enormous attempts have been devoted to improve Taxol production exploiting various methodologies from bioprocess engineering to biotechnological and synthetic approaches. We have developed a 2-stage suspension cell culture of Taxus baccata L. using modified B5 medium in order to improve cell growth as well as productivity. After callus induction and cell line selection, B5 medium was supplemented with vanadyl sulfate (0.1 mg/l), silver nitrate (0.3 mg/l) and cobalt chloride (0.25 mg/l) at the first day of stage I culture to maximize cell growth. This medium was further supplemented with sucrose (1%) and ammonium citrate (50 mg/l) on day 10 and sucrose (1%) and phenylalanine (0.1 mM) on day 20 (i.e., biomass growth medium). At stage II (day 25), two different concentrations of several elicitors such as methyl jasmonate (10 or 20 mg/l), salicylic acid (50 or 100 mg/l) and fungal elicitor (25 or 50 mg/l) were added to the biomass growth medium with the aim of improving cellular productivity. For morphological analysis, microscopic inspection was carried out during cultivation. Cell-associated and extracellular amount of Taxol were detected and measured using HPLC methodology. At stage I, overall Taxol amount of biomass growth medium was 13.75 mg/l (i.e., 5.6-fold higher than that of untreated B5 control). At stage II, treated cells with methyl jasmonate (10 mg/l), salicylic acid (100 mg/l) and fungal elicitor (25 mg/l) produced the highest amount of Taxol (39.5 mg/l), which is 16-fold higher than that of untreated B5 control (2.45 mg/l). Microscopic analyses of Taxus cells in suspension cultures showed various positional auto-fluorescence showing direct correlation with Taxol production. Our studies revealed that intervallic supplementation of B5 medium with combination of biomass growth factors at stage I and mixture of elicitors at stage II could significantly increase Taxol production. Thus, we suggest that the exploitation of this methodology may improve the production of Taxol since demands for Taxol pharmaceuticals are increasingly growing and resource paucities have limited its direct harvesting from Taxus trees.     

米根霉诱导因子对紫草细胞培养中紫草宁色素分泌的影响

在紫草细胞培养中,加入采根霉粗提物可显著提高紫草宁色素产量,并可加快胞内色素分泌到培养液中的速率和数量。在细胞培养的第6天加入米根霉诱导因子时,其促进紫草宁色素分泌的作用最大,培养液中紫草宁色素含量是对照的2．24倍。此外,同时加入正十六烷和米根霉诱导因子对紫草宁色素的分泌具有协同作用。     

<Emphasis Type="Italic">Fusarium oxysporum</Emphasis> cell elicitor enhances betalain content in the cell suspension culture of <Emphasis Type="Italic">Celosia cristata</Emphasis>

We started a cell suspension culture from magenta coloured calli of cockscomb to study the effect of biotic and abiotic elicitors on the biosynthesis of betalain pigments. The cultures were grown in a flask containing 30 ml MS media fortified with 13.5 μM 2,4-D and 0.44 μM BAP. These cultures were elicited during its log-phase of growth using fungal elicitors (prepared from mycelia of Fusarium oxysporum), yeast extract, copper sulphate and cobalt chloride. The elicitation reduced the cell count, cell viability and percent pigmented cell in the suspension culture. Similarly, it also resulted in reduced betalain content by all the elicitors except 0.125 × 10^?3% fungal elicitor. Rather, fungal elicitor at this concentration significantly enhanced the amaranthin, betanin, betalamic acid and betaxanthin content in the culture. Besides this, copper sulphate doubled the pigment contribution (ratio of particular pigment content to total pigment content) of betaxanthin at all the concentrations. Therefore, we conclude that fungal elicitor can further be investigated to enhance the content of betalain pigments in suspension culture at a larger scale.