甘肃棘豆草中苦马豆素的提取分离工艺比较研究

以甘肃棘豆草为试验材料,通过工业酒精热回流提取、水提取和酸水提取3种工艺提取其中的总生物碱,总生物碱经硅胶柱层析分离后减压升华得到,并比较了3种工艺对甘肃棘豆中总生物碱得率和苦马豆素得率的差异。结果表明,3种工艺总生物碱平均得率和苦马豆素平均得率依次分别为7.91、6.79、6.22 mg/g和18.61、12.22、6.23μg/g,由此确定工业酒精热回流为提取苦马豆素的最佳提取分离工艺。     

苦马豆叶绿体基因组结构及其特征分析

利用Illumina高通量测序平台对苦马豆（Sphaerophysa salsula）进行测序和组装,获得完整的苦马豆叶绿体基因组序列。组装结果表明,苦马豆叶绿体基因组全长123 327 bp,存在IR区丢失,不具有四分体结构;注释结果显示,苦马豆叶绿体基因组共编码108个基因,其中包含74个蛋白编码基因、4个rRNA基因和30个tRNA基因;叶绿体基因组序列上共检测到99个SSR位点,包含75个单核苷酸、17个二核苷酸和7个三核苷酸重复单元;系统发育分析显示,苦马豆和骆驼刺为姊妹群,亲缘关系最近。这为今后苦马豆遗传多样性、群体遗传结构和物种形成机制研究奠定了基础。     

新疆黑粉菌目一新种

本文报道新疆黑粉菌目一新种,苦马豆楔孢黑粉菌(Thecaphora sphaerophysae ZY Zhao et Xi)。新种有中文和拉丁文描述,并附有冬孢子光学显徽镜和扫描电镜照片。     

新疆荒漠植物的锈菌Ⅰ.豆科和藜科植物上的单胞锈菌

报道了采自新疆塔里木盆地和准格尔盆地荒漠植物豆科和藜科植物上寄生的单胞锈菌20个种。寄生在疏花骆驼刺Alhagi sparsifolia上的骆驼刺单胞锈菌Uromyces alhaginis、寄生在木地肤Kochia prostrata上的地肤单胞锈菌Uromyces kochiae和寄生在苦马豆Swainsonia salsula上的苦马豆单胞锈菌Uromyces sphaerophysae为中国新记录;寄生在新疆沙冬青Ammopiptanthus nanus上的欧黄华单胞锈菌Uromyces anagyridis和寄生在长豇豆Vigna sesquipedalis上的豇豆单胞锈菌Uromyces vignae为新疆新记录;盐穗木Halostachys caspica和小蓬Nanophyton erinaceum为藜单胞锈菌Uromyces chenopodii的新寄主。所有研究标本保存在新疆农业大学真菌标本室(HMACC)。     

新疆荒漠植物的锈菌Ⅰ.豆科和藜科植物上的单胞锈菌

报道了采自新疆塔里木盆地和准格尔盆地荒漠植物豆科和藜科植物上寄生的单胞锈菌20个种.寄生在疏花骆驼刺 Alhagi sparsifolia 上的骆驼刺单胞锈菌 Uromyces alhaginis、寄生在木地肤 Kochia prostrata 上的地肤单胞锈菌 Uromyces kochiae 和寄生在苦马豆 Swainsonia salsula 上的苦马豆单胞锈菌Uromyces sphaerophysae 为中国新记录;寄生在新疆沙冬青 Ammopiptanthus nanus 上的欧黄华单胞锈菌Uromyces anagyridis 和寄生在长豇豆 Vigna sesquipedalis 上的豇豆单胞锈菌 Uromyces vignae 为新疆新记录;盐穗木Halostachyscaspica和小蓬Nanophyton erinaceum为藜单胞锈菌Uromyces chenopodii的新寄主.所有研究标本保存在新疆农业大学真菌标本室(HMACC).     

青藏高原冰川棘豆(Oxytropis glacialis)内生菌核心微生物组的界定及其互作网络分析

【背景】植物内生菌(endophyte)对寄主植物的益生作用有利于植物的生存与扩散,而菌群互作网络为内生菌生态功能的实现提供了基础保障。【目的】了解影响西藏畜牧业发展的主要疯草冰川棘豆(Oxytropis glacialis)内生菌核心微生物组及其菌群互作网络,为青藏高原疯草类有毒植物的科学治理和利用提供基础参考依据。【方法】利用高通量测序技术分析冰川棘豆中内生菌核心微生物组,构建内生菌相关性网络,并以与苦马豆素代谢相关内生菌为例分析冰川棘豆内生菌的互作模式。【结果】内生细菌测序共得到175791条有效序列,注释到428个细菌OTU,分属于19个门和267个属;内生真菌测序共得到757 113条有效序列,注释到391个真菌OTU,分属于7个门和149个属。Venn图分析表明,根、叶组织的核心内生细菌菌属数目(54、62)大于核心内生真菌(22、13),根组织中的核心内生菌种类与叶组织相当(76、75)。系统发育树分析表明,冰川棘豆中存在产生苦马豆素内生真菌链格孢属(Alternaria)和降解苦马豆素的内生细菌短波单胞杆菌属(Brevundimonas)。相关性网络分析表明,内生菌菌群间以正向反馈的互作网络关系为主,核心菌群可能主要通过间接性的互作方式将影响传递到微生物群落,其中链格孢属与短波单胞杆菌属作为核心菌属通过间接性的显著相关关系(|ρ|0.6,P0.01)参与菌群间互作网络。【结论】冰川棘豆核心菌群以间接的方式参与内生菌菌群间的互作网络,高度的菌属连接性为内生菌生态功能的实现提供可能。     

西北部分地区苦马豆根瘤菌的表型多样性研究

将采集自甘肃、新疆、宁夏等地区的苦马豆根瘤,经分离、纯化获得48株未知菌株,并选取7株参比菌株,进行唯一碳、氮源利用、对抗生素及染料抗性、耐盐性、初始pH生长、生长温度范围及酶活性等共113项生理生化测定;采用数值分类方法对未知根瘤菌进行表型多样性分析。结果表明：供试菌株在碳氮源利用、抗生素敏感性、抗逆性等方面存在着差异。该地区苦马豆根瘤菌具有较强的耐盐、耐碱能力,所有菌株均能在初始pH9~12的YMA培养基上生长,35%菌株可耐受6%的NaCl。从数值分类树状图可见,未知供试菌株在56%的相似水平上聚在一起,在72%的相似水平上分为5个表观群。群Ⅰ有39株菌,在74.6%相似水平聚合,中心菌株为CCNWGS0215;群Ⅱ有5株菌,在76%的相似水平聚合,中心菌株为CCNWGS0228;群Ⅳ有2株菌,在81.5%的相似水平聚合,群Ⅲ和群Ⅴ分别只有1株菌,它们与模式株分离,可能为潜在的新种。     

黄花棘豆的喹诺里西定生物碱

豆科棘豆属有毒植物含多种有毒生物碱,有关棘豆属中生物碱成分的研究,早在1929年Couch由兰伯氏棘豆(Oxytropis lambertii DC.)中分离到一种多羟基化的含氮有机化合物,这是最早报道由棘豆属植物中分离出的生物碱成分,由于当时条件限制未能确定其结构。1982年美国农业部西部研究中心的植物化学家Molyneux从绢毛棘豆(Oxytropis sericea)中分离到苦马豆碱(swainsonine)等吲哚里西定生物碱(indolizidinealkaloids);1989年沈阳药学院于荣敏等由小花棘豆(Oxytropis glabra DC.)中分离出多种喹诺里西定生物碱。喹诺里西定生物碱具有多种生理活性,对试验动物中枢神经系统产生抑制,呼吸抑制或兴奋,致幻、流产和致畸等作用。本文报道由黄花棘豆(Oxytropis ochrocephala Bunge)中测得4种喹诺里西定生物碱。     

苦马豆和披针叶黄华种子硬实特性与活力关系

将苦马豆和披针叶黄华种子在恒温25℃下吸胀,每24 h取出吸胀种子,16 d后未吸胀的种子为硬实种子（H）,硬实种子用硫酸处理后恒温吸胀24 h,与非硬实种子进行发芽试验和各项活力指标测定。结果显示每日内吸胀的种子数量随时间推移以一定比列下降,苦马豆非硬实种子第3天后吸胀率下降到1%,第13～16天突然上升后又下降到1%,披针叶黄华非硬实种子第3天后下降到1%,第9、10天突然上升后又下降到1%。两种豆类都显示出硬实种子发芽率、发芽指数、活力指数、脱氢酶活性、呼吸速率和超氧化物歧化酶（SOD）活性均高于非硬实种子,而电导率、浸出液可溶性糖和丙二醛（MDA）含量低于非硬实种子,缓慢吸胀的硬实种子活力指标高于快速吸胀的硬实种子,这表明硬实种子活力高于非硬实种子,硬实种子吸胀过程中存在吸胀损伤。而在非硬实种子中,根据以上活力指标判断,晚吸胀的种子比早吸胀的种子活力高。     

土壤含水量和土壤类型对豆大蓟马蛹期发育和羽化的影响

豆大蓟马Megalurothrips usitatus(Bagrall)是近年来严重制约海南豇豆生产的重要害虫。针对该蓟马在土壤中化蛹的习性,本文研究了在室内条件下不同土壤含水量和土壤类型对豆大蓟马化蛹的影响。结果表明,土壤含水量和土壤类型显著影响豆大蓟马蛹的发育历期和羽化率,土壤含水量为15%且土壤类型为砂壤土时,豆大蓟马的发育历期最短为3.62±0.15 d,羽化率最高为52.08%±0.07%,低含水量(5%)和高含水量(25%)均不利于豆大蓟马化蛹。土壤类型以砂壤土最适宜豆大蓟马化蛹,粘土最不适合豆大蓟马化蛹,其羽化率最低为19.17%±0.02%。     

<Emphasis Type="Italic">Rhizobium helanshanense</Emphasis> sp. nov., a bacterium that nodulates <Emphasis Type="Italic">Sphaerophysa salsula</Emphasis> (Pall.) DC. in China

Studying rhizobia in the root nodules of Sphaerophysa salsula (Pall.) DC in the northwest of China, we obtained five strains classified as genus Rhizobium on the basis of their 16S rRNA gene sequences. The sequence similarity of strain CCNWQTX14^T with the most related species was 99.0%. Further phylogenetic analysis of housekeeping genes (recA and atpD) suggested the five strains comprised a novel lineage within Rhizobium. The nifH and nodD gene sequences of CCNWQTX14^T were phylogenetically closely related with those of Sinorhizobium kummerowiae and R. sphaerophysae, respectively. The five strains isolated from different places were also distinct from related Rhizobium species using ERIC fingerprint profiles. The DNA–DNA hybridization value was 41.8% between CCNWQTX14^T and Rhizobium sphaerophysae CCNWGS0238^T. Our novel strains were only able to form effective nodules on its original host Sphaerophysa salsula. Our data showed that the five Rhizobium strains formed a unique genomic species, for which a novel species Rhizobium helanshanense sp. nov. is proposed. The type strain is CCNWQTX14^T (=ACCC 16237^T =HAMBI 3083^T).     

Synthesis and antioxidant activity of 4-[2-(3,5-dimethoxyphenyl)ethenyl]-1,2-benzenediol, a metabolite of Sphaerophysa salsula

4-[2-(3,5-Dimethoxyphenyl)ethenyl]-1,2-benzenediol (7), a stilbene isolated from Sphaerophysa salsula, was synthesized from 3,4-dihydroxybenzaldehyde (1) in five steps in an overall yield of 33%. The spectral data for synthetic 7 are in good agreement with those of the natural product. Hydroxystilbene 7 showed potent antioxidative activity.     

二倍体蔷薇FT基因序列多样性研究

以3个类群73个二倍体蔷薇属(Rosa)植物为材料,克隆获得其FLOWERING LOCUS T(FT)同源基因,并对该基因的编码区序列进行多态性分析以及多维尺度(MDS)聚类分析。结果显示,73个二倍体蔷薇植物的FT基因共检测到215个核苷酸多态性位点,其中包括214个SNP和1个缺失突变,平均185个碱基发生1次突变;氨基酸多态性分析结果显示共有35个氨基酸发生变异,平均379.6个氨基酸残基发生1次突变;突变位点统计分析结果发现39、258、426 bp位点是高频突变位点,其碱基由A或C突变为T。MDS聚类分析结果表明,3个类群FT基因编码区序列的碱基组内差异依次排序为:野生种月季组中国古老月季,氨基酸组内差异依次排序为:中国古老月季月季组野生种,推测中国古老月季在长期栽培驯化过程中,其FT基因可能经历了较强的人工选择压力,月季组的种和变种可能是古老月季的重要亲本来源。     

A study on the medicinal plants of the genus Atractylodes

Genus Atractylodes (Fam. Compositae) is the main source of two important Chinese traditional drugs, “Baizhu”and “Cangzhu”, both being long used as a stomachic. After a general survey and taxonomical study, it has been found that “Baizhu” was only derived from A. macrocephala Koidz. (A. ovata auct. Fl. Orient. Asiat. non A. P. DC.), while “Cangzhu” were mainly from A. lancea (Thunb.) DC. and A. lancea DC. var. chinensis Kitam. Comparison of the components in the rhizomes of Chinese Atractylodes has been made by TLC and GLC. The results have shown to be in accordance with their morphological features and pharmaceutical merits. “Baizhu”, A. macrocephala, with its leave pinnately incised, is characterized by the presence of rich atractylon and absence or lack of atractylodin. As for “Cangzhu”, A. lancea and A. lancea var. chinensis with their leave not incised or only lobed, are characterized by high contents of atractylodin, β-eudesmol and hinesol, but poor in atractylon. The above conclusion may be of value to both the classification and utilization of this group of Chinese medicinal plants.     

Histoplasma capsulatum cyclophilin A mediates attachment to dendritic cell VLA-5

Histoplasma capsulatum (Hc) is a pathogenic fungus that replicates in macrophages (Mphi). In dendritic cells (DC), Hc is killed and fungal Ags are processed and presented to T cells. DC recognize Hc yeasts via the VLA-5 receptor, whereas Mphi recognize yeasts via CD18. To identify ligand(s) on Hc recognized by DC, VLA-5 was used to probe a Far Western blot of a yeast freeze/thaw extract (F/TE) that inhibited Hc binding to DC. VLA-5 recognized a 20-kDa protein, identified as cyclophilin A (CypA), and CypA was present on the surface of Hc yeasts. rCypA inhibited the attachment of Hc to DC, but not to Mphi. Silencing of Hc CypA by RNA interference reduced yeast binding to DC by 65-85%, but had no effect on binding to Mphi. However, F/TE from CypA-silenced yeasts still inhibited binding of wild-type Hc to DC, and F/TE from wild-type yeasts depleted of CypA also inhibited yeast binding to DC. rCypA did not further inhibit the binding of CypA-silenced yeasts to DC. Polystyrene beads coated with rCypA or fibronectin bound to DC and Mphi and to Chinese hamster ovary cells transfected with VLA-5. Binding of rCypA-coated beads, but not fibronectin-coated beads, was inhibited by rCypA. These data demonstrate that CypA serves as a ligand for DC VLA-5, that binding of CypA to VLA-5 is at a site different from FN, and that there is at least one other ligand on the surface of Hc yeasts that mediates binding of Hc to DC.     

中国裸齿角石蛾属三新种(毛翅目,齿角石蛾科)

描述了裸齿角石蛾属3新种,即脊状裸齿角石蛾Psilotreta vertebrata sp.nov.(广东)、方背裸齿角石蛾Psilotreta cuboides sp.nov.(云南)和凹入裸齿角石蛾Psilotreta excavata sp.nov.(江西)。模式标本保存于南京农业大学昆虫标本馆。     

Antimutagenic activity of extracts from anticancer drugs in Chinese medicine

The antimutagenic activities of extracts of 36 commonly used anticancer crude drugs from Chinese herbs were studied by using the Salmonella/microsomal system in the presence of picrolonic acid or benzo[a]pyrene to test whether they contain direct or indirect antimutagens. Each crude drug was extracted with boiling water for 2 h, the method which is commonly used by Chinese people to prepare the drug for oral intake. The extracts of Pteris multifida P. showed the highest antimutagenic activity against picrolonic acid-induced mutation. The extracts of 6 other different kinds of Chinese herbs were shown to have a moderate antimutagenic activity against picrolonic acid-induced mutation, and they are: Actinidia chinensis P., Artemisia lavendulaefolia DC. and Crotalaria sessiflora L., Prunella vulgaris L., Paris polyphylla S. and Ampelopsis brevipedunculata T. The extracts of Smilax china L., Prunella vulgaris L. and Actinidia chinensis P. were demonstrated to inhibit the mutagenicity of benzo[a]pyrene completely. The 12 other kinds of extracts of Chinese herbs which had a moderate antimutagenic activity against benzo[a]pyrene were: Pteris polyphylla S., Ampelopsis brevipedunculata T., Duchesnea indica F., Gossypium herbaceum L., Lithospermum erythrorrhizon SZ., Artemisia lavendulaefolia DC., Selaginella doederleinii H., Dianthus superbus L., Centipeda minima ABA., Curcuma zedoaria R., Marsdenia tenacissima WA. and Kalopanax septemlobus K. Among them, there were 5 kinds of crude drugs, Actinidia chinensis P., Artemisia lavendulaefolia DC., Prunella vulgaris L., Paris polyphylla S. and Ampelopsis brevipedunculata T., containing antimutagenic factors against both picrolonic acid- and benzo[a]pyrene-induced mutation.     

不同产地苍术药材化学成分的比较

江苏、安徽及湖北５个产地苍术（Ａｔｒａｃｔｙｌｏｄｅｓｌａｎｃｅａ（Ｔｈｕｎｂ．）ＤＣ．）商品药材的化学分析结果表明：它们在某些药材性状、一些常规化学成分、挥发油成分和含量上存在显著差异。江苏茅苍术根茎挥发油的化学成分复杂,由此探讨了它作为地道药材的特殊性。同时对苍术的“断面暴露稍久可析出结晶”的性状和《药典》（１９９０年版）规定的“层析谱上应显有苍术素斑点”的鉴别实验进行讨论。     

Sister chromatid exchanges induced by cyclophosphamide in V79 cells cultured in diffusion chambers in mice

Chinese hamster cells V79 were cultured in diffusion chambers (DC) and implanted into mice. An exponential growth was observed from the 2nd to 4th day after implantation. The maximum growth was reached on the 6th day. After that, cell growth and viable cell counts decreased. Three days after implantation of DC with V79 cells, the hosts received 6 hourly injections of 0.2 ml of 5-bromodeoxyuridine (BUdR) solution at concentrations of 0.125 to 1.0 x 10(-2) M. DC were removed for chromosome and sister-chromatid exchanges (SCE) analyses 24 h after the first BUdR injection. The frequency of metaphases with differentially stained chromatids, with aberrations, and the number of SCE per cell increased with BUdR dose. The frequency of metaphases with differentially stained chromatids was also positively correlated with the duration of BUdR exposure or the number of hourly injections of BUdR-solution. The effects of cyclophosphamide (CY) in V79 cells in DC in mice were studied. Injections of CY at 2.5, 5, 10 and 15 microgram per gram of body weight to the hosts caused an increase in the number of SCE per cell in a linear manner. The results from this study indicate that V79 cells cultured in DC in mice may provide a potential test system for mutagenicity.