Synthesis and/or accumulation of bioactive molecules in the in vivo and in vitro root

Abstract

Roots of many species are studied because of the presence of high-value bioactive molecules, yet few studies have attempted to determine the biosynthetic pathways of these compounds or the way in which synthesis is regulated. The presence of secondary metabolites in the root does not necessarily mean that this organ is also the site of synthesis. Thus the identification of organ-specific intermediate precursors and key enzymes is important for understanding the biosynthetic pathway and the regulation of bioactive molecules. This knowledge could allow researchers to predict the suitability of in vitro systems, such as regenerated roots and hairy roots, for producing the molecules of interest. In the present review, the production of bioactive molecules in in vivo roots is compared to that in in vitro untransformed and transformed roots, concentrating on recent developments in the study of the biosynthesis of the anti-cancer alkaloid camptothecin in Camptotheca acuminata Decne. The results of a recent study performed in our laboratory on the production of camptothecin and other secondary metabolites in in vivo and in vitro C. acuminata roots are also presented.     

A new species of the wild banana genus,Musa (Musaceae), from Borneo

A new species of wild banana, Musa bauensis Häkkinen & Meekiong, is described and illustrated. It is from the Bau limestone area, Sarawak, East Malaysia.     

中国特有植物喜树的雌性不育形态解剖学研究

为阐明雌性不育与喜树（Camptotheca acuminata Decne.）开花期花序大量掉落的关系,本文采用数码体视显微镜及石蜡切片技术,对各个发育时期的喜树的花进行解剖学观察。喜树花均为两性花,雄蕊发育正常,雌蕊柱头有三种不同形态,即上位、中位和下位;柱头上位的小花,花柱伸长正常,柱头三裂,外翻,子房膨大,胚珠饱满,胚囊发育正常;柱头中位的小花,雌蕊选择性败育,柱头和花柱伸长缓慢,柱头外翻异常,局部子房细胞液泡化,胚珠退化,胚囊发育异常;柱头下位的小花,花柱退化,柱头不露,珠被细胞发育异常,大孢子母细胞未能减数分裂,胚珠萎缩,子房外形瘦长,室内中空。喜树雌性器官的发育与其柱头在子房上的表现形态有着严格的对应关系,雌性不育发生于大孢子母细胞减数分裂前后,而且雌性不育是喜树花序掉落的内部原因之一。     

增强UV-B辐射对喜树幼苗生物量和两种生物碱含量的影响

通过盆栽试验,以自然辐射为对照,研究人工增强UV-B辐射下(5.0 μW/cm2)喜树幼苗生物量和各器官中喜树碱、10-羟基喜树碱含量的变化,试验结果表明:(1 )UV-B辐射处理前20d,处理组幼苗的全株以及各器官的鲜重与对照相差很少.至辐射处理的40d,处理组幼苗的全株以及各器官的生物量均高于正常条件的幼苗.试验后期,处理组单株生物量降低,长时间的UV-B辐射使喜树植株矮化、基茎加粗,同时还改变了喜树生长过程中干物质的分配,较多的干物质分配到喜树的茎和根中,而较少进入叶中;(2)UV-B辐射增强能明显增加喜树地上器官中喜树碱的含量,而对10-羟基喜树碱含量影响不明显.(3)各器官中生物碱含量与生物量的积累速率具有一定的相关性,生物量增长过快时单位质量植物体中的生物碱含量下降.     

喜树花序内性别表达及性别分配

 花序内性别表达和性别分配的差异在被子植物中非常普遍。常见的变化模式是: 在顺次开放的花序内, 从早开的花到晚开的花, 生殖结构大小和数量依次减少, 晚开的花偏向雄性。通常认为资源竞争和位置效应是形成这种差异的主要原因。在喜树(Camptotheca acuminata)的聚伞花序内, 早开的头状花序在花序直径、小花直径、短雄蕊长度、坐果率、果序重量等方面与晚开的头状花序差异显著。通过去除喜树聚伞花序一级或一级和二级头状花序来控制花序内的资源分配, 从而分析影
响喜树花序内花性状、性别分配及生殖能力的主要因子。结果显示, 去除部分头状花序后, 剩余头状花序的花序直径、小花直径明显增加, 短雄蕊长度明显缩短, 位置间差异消失; 而坐果率、果序重量的位置间差异依然显著。表明资源限制对花序直径、小花直径和短雄蕊长度等影响显著, 而坐果率和果序重量受位置效应影响明显。喜树雄全同株的性别分配可能是对特定生殖状况和资源状况适应的结果。     

喜树种子内生放线菌的分离鉴定及抗菌活性物质初分离

【目的】探究含有抗肿瘤活性喜树碱的喜树(Camptotheca acuminata Decne.)种子中内生放线菌的多样性,以及从内生放线菌中寻找新型活性化合物产生菌。【方法】利用放线菌富集培养基分离喜树种子内生放线菌,并根据菌落形态及16S rRNA基因序列同源性分析进行菌种鉴定;以及利用与病原微生物共培养方法检测分离菌株的抗菌活性。【结果】从上海交通大学校园的喜树种子中共分离到33株内生疑似放线菌,经基于16S rRNA基因序列分析的分子鉴定,确定其中21株属于链霉菌,1株属于拟诺卡氏菌,另有3株由于序列相似性低于95%而无法分类;8株因未能克隆16S rRNA基因序列而未确定分类。这11株疑似内生放线菌是否是新放线菌(种)类群,有待后续深入研究。初步的抗细菌、真菌活性检测发现,分离到的内生放线菌中42.42%以上对立枯丝核菌(Rhizoctonia solani Kühn)有很强的抑制菌核形成的作用,54.54%内生菌有较强抑制芽孢杆菌生长的作用。并且挑选其中具有代表性的菌株Streptomyces sp.CXSZ1进行代谢产物的分析,初步发现其代谢产物中含有抗细菌活性成分。【结论】喜树种子内生放线菌的分离、鉴定及生物活性物质分离鉴定的研究工作,一方面可以为揭示喜树等高等植物种子内生菌的起源、演化等提供有价值的信息,同时也为新结构、新活性化合物发掘提供实验材料。     

一株高效DEHP降解菌的分离、鉴定及其降解特性

【目的】分离得到高效的邻苯二甲酸二乙基己基酯(DEHP)降解菌。【方法】采用富集培养法筛选分离菌株,并对菌株进行驯化;通过PCR扩增得到其16S rRNA和gyrB基因序列,进行同源序列分析及分子系统发育树的构建,同时结合形态学观察和生理生化实验对菌株进行初步鉴定;采用高效液相色谱(HPLC)分析菌株对DEHP的降解特性。【结果】分离得到一株能以DEHP为唯一碳源和能源生长的菌株,命名为HS-NH1,初步鉴定其为戈登氏菌(Gordoniasp.)。菌株HS-NH1最适的生长和降解条件为30°C、pH 7.0,在此条件下,该菌株60 h内能够将浓度为500 mg/L的DEHP降解90%以上。高效液相色谱(HPLC)分析表明,菌株HS-NH1在降解DEHP过程中产生了一种重要的中间代谢产物——邻苯二甲酸。底物广谱性试验证明,菌株HS-NH1能够有效地利用多种常见的邻苯二甲酸酯(PAEs)与芳香族衍生物。【结论】筛选得到了一株DEHP降解菌Gordonia sp.HS-NH1,该菌降解效率高,具有良好的底物广谱性,在邻苯二甲酸酯类化合物的污染治理中将会有一定的应用潜力。     

Transformation of p-hydroxybenzonitrile to p-hydroxybenzoic acid using nitrilase activity of Gordonia terrae

Abstract

Resting cells of Gordonia terrae with nitrilase (EC 3.5.5.1) activity were investigated for transformation of p-hydroxybenzonitrile to p-hydroxybenzoic acid. The maximum conversion was observed in 0.1 M potassium phosphate buffer, pH 8.0, using 40 mM substrate and resting cells corresponding to 0.70 Uml^{? 1} nitrilase activity at 35°C. A 500 mL fed batch reaction was designed for synthesis of p-hydroxybenzoic acid with six feedings of substrate at an interval of 1 h. A total of 14.4 g of p-hydroxybenzoic acid (> 98.7%) was obtained in 6 h with a productivity of 0.78 gh^{? 1}g^{? 1}_DCW of G. terrae.     

Inhibition of Biosynthesis of Polyisoprenol Sugars in Chick Embryo Microsomes by Tunicamycin

Changes in the phytoalexin content in unripe fruit of banana, Musa acuminata, were analyzed after various treatments. The results show that level of hydroxyanigorufone started to increase 1-2 day after either wounding or inoculation with conidia of Colletotrichum musae. Inoculation followed by wounding induced the formation of many other phenylphenalenones. The accumulation of hydroxyanigorufone decreased, after its transient maximum, on ripening by exposure of the wounded fruit to ethylene. The level of production of hydroxyanigorufone in ripe fruit treated by wounding and/or by inoculation was much lower than that in unripe fruit. 2-Aminooxyacetic acid, an inhibitor of phenylalanine ammonia-lyase (PAL), inhibited the accumulation of hydroxyanigorufone in wounded fruit, and the PAL activity increased after wounding and ethylene treatment, respectively. Feeding experiments with [1-¹³C] and [2-¹³C]cinnamic acids, and [2-¹³C]malonate show that two molecules of cinnamic acid and one of malonate were incorporated into each molecule of hydroxyanigorufone. The phytoalexins isolated from fruit to which deuterated hydroxyanigorufone and irenolone had been administered revealed that 2-(4′-hydroxyphenyl)-1,8-naphthalic anhydride was biosynthesized from hydroxyanigorufone rather than from irenolone.     

Efficiency of certain miscible oils and chlorpyriphos methyl insecticide against the soft scale insect,Kilifia acuminata Signoret (Homoptera: Coccidae) and their toxicities on rats

Abstract

The toxicity and efficacy of using three miscible oils; cabl-2, citrole and bio-dux and a chemical insecticide (chlorpyrifos-methyl) were studied against a soft scale insect, Kilifia acuminata (Signoret) infesting mango trees. The sub-chronic toxicity of tested chemicals on white albino rats was also studied. Considering the general mean number of K. acuminata stages after 3 months post spraying, chlorpyrifos methyl and capl-2 were the most efficient compounds followed by citrol and bio-dux in descending order. Chlorpyrifos-methyl was the most effective treatment for all months after spraying, the mean reduction percentage being 94.10, 91.63 and 92.00% while it gave the highest toxic effect after three months; 90.27, 87.84 and 89.73% reduction in infestation on pre-adult, adult and gravid female stages, respectively. Results also indicate that the general mean percent reduction in K. acuminata infestation were 82.62, 86.67, 78.87 and 89.28% in treated trees by citrole, cabl-2, bio-dux and chlorpyrifos-methyl, respectively. Citrole, bio-dux, cabl-2 and chlorpyrifos-methyl caused significant increase in WBCs counts, ALT and AST activities of treated rats after 15 and 30 days from treatment comparison with control. Changes occurring in the creatinine concentration showed a significant increase in rats treated with the tested chemicals 30 days from treatment, except in the case of bio-dux oil, while there were no significant changes after 15 days in rats treated with tested oils. On the contrary, tested chemicals caused a significant decrease in RBCs count and hemogobin values after 15 and 30 days form treatment, except in rats treated with bio-dux oil. Hemoglobin content showed no significant changes over the same periods.