欧洲红豆杉细胞培养的研究

从欧洲红豆杉（Ｔａｘｕｓｂａｃｃａｔａ）的嫩茎及什叶诱导出愈伤组织，并对之进行了愈伤组织培养及细胞悬浮培养研究；利用ＨＰＬＣ方法测定了培养物合成紫杉醇的能力．探索了提高培养细胞生长率及紫杉醇含量的一些措施．结果表明，欧洲红豆杉愈伤组织及悬浮培养细胞的生长速率已分别达到０．２７ｇ·Ｌ－１·ｄ－１和０．３５ｇ·Ｌ－１·ｄ－１，紫杉醇含量分别为０．００３１％和０．０１７％。     

云南红豆杉的保护和开发利用

云南红豆杉(Taxus yunnanensis W. C. ChengL. K. Fu)是红豆杉属植物中紫杉醇含量最高的树种,同时也是我国红豆杉属中分布最广和资源蕴藏量最丰富的物种.然而,近年来由于人为大量砍伐,云南红豆杉野生资源遭到严重破坏,加上生长缓慢且天然更新困难,目前处于濒危状态.对云南红豆杉天然资源的保护和药用人工林的培育对于云南红豆杉资源的可持续利用具有重要的意义.对云南红豆杉资源分布现状及种群和繁殖生态学等方面的研究进行综述,发现云南红豆杉的濒危是由其自身繁殖机制引起的天然更新困难、对环境的适应能力较低以及人为因素共同导致的.同时从紫杉醇含量的影响因素及获取方式、人工药用林培育方面总结了云南红豆杉的开发利用现状,发现云南红豆杉中紫杉醇的含量受其自身特性及外在因素的综合影响;包含紫杉醇在内的紫杉烷类物质目前主要通过直接提取和半合成两种方式获取;此外在全合成、组织或细胞培养及真菌诱导等方式上也有了新的研究进展.最后,建议从就地保护、迁地保护及引种回归3个方面对云南红豆杉天然资源进行综合保护,同时大力推进紫杉烷工业化合成方式的研究和云南红豆杉人工药用原料林的培育,提高云南红豆杉的资源利用效率,在保护云南红豆杉天然林的同时实现资源的合理利用.(表1参129)     

红豆杉培养细胞紫杉醇的分离及鉴定

红豆杉培养细胞紫杉醇的分离及鉴定甘烦远，郑光植，彭丽萍，沈月毛（中国科学院昆明植物研究所昆明６５０２０４）关键词红豆杉；紫杉醇；细胞培养；分离鉴定ＩＳＯＬＡＴＩＯＮＡＮＤＩＤＥＮＴＩＦＩＣＡＴＩＯＮＯＦＴＡＸＯＬＦＲＯＭＣＵＬＴＵＲＥＤＣＥＬＬＳＯＦ...     

水杨酸诱导黄瓜抗黑星病抗性的部位差异和时效性

寻找能够诱导植物产生抗病性的天然的生物活性物质制剂已成为作物保护研究的重点［１］．水杨酸（ＳＡ）外源处理可诱导一些重要农作物产生针对真菌、细菌、病毒病害的局部或系统性抗性,因而倍受关注［２］．黄瓜黑星病是黄瓜保护地栽培过程中的一种主要病害,寻找能防治此病且不污染环境的生物农药并研究其作用规律已成为当务之急．本文简报以黄瓜植株和黑星病菌的互作系统为材料,观察ＳＡ对黄瓜抗黑星病抗性的诱导作用及其诱导抗性在植株不同位置的差异和时效差异．１　材料与方法１．１　实验材料１．１．１　供试植物　　黄瓜品种为对…     

甘蔗泛素结合酶基因的克隆与表达北大核心CSCD

为克隆鉴定甘蔗泛素结合酶基因（Sugarcane ubiquitin-conjugating enzyme,ScUBc E2）并探索其在激素信号通路和甘蔗与黑穗病互作过程中的作用,选择黑穗病菌胁迫下甘蔗抑制消减杂交文库（Suppression subtractive hybridization,SSH）中注释为泛素结合酶的差异表达EST序列为探针,结合电子克隆技术和RT-PCR技术,以甘蔗cDNA为模板进行泛素结合酶基因克隆.对克隆获得的序列进行生物信息学分析,并利用qRT-PCR技术分析该基因在甘蔗根、蔗髓、叶、芽中的组织特异性表达以及在黑穗病菌、茉莉酸甲酯（Methyl jasmonate,Me JA）、脱落酸（Abscisic acid,ABA）和水杨酸（Salicylic acid,SA）胁迫下的表达情况.最终克隆得到一条长度为699 bp的甘蔗泛素结合酶基因（ScUBc E2;Gen Bank accession number：KJ577594.1）,该基因包含长度为447 bp的完整开放读码框,编码148个氨基酸.生物信息学分析结果显示,ScUBc E2编码的蛋白分子量（Mr）为16.507×10^3;无信号肽,为碱性不稳定的亲水蛋白;包含4个α螺旋、4个β折叠和一些无规则卷曲;第15位氨基酸为泛素化位点,74-89位氨基酸为活性位点;在进化过程中与高粱泛素结合酶基因的亲缘关系最近.qRT-PCR分析结果表明,ScUBc E2基因组成型表达,但在芽中的表达量最高;其表达在甘蔗感黑穗病基因型ROC22中受到黑穗病菌胁迫的抑制,在黑穗病抗病基因型YC05-179中则先被抑制,后被诱导;ScUBc E2基因受MeJA及SA诱导表达,对ABA胁迫的响应不明显.本研究表明,甘蔗ScUBc E2基因在抗病基因型和感病基因型甘蔗中存在不同表达模式,可能参与甘蔗与黑穗病菌的互作过程,有望为抗病育种分子标记提供潜在基因资源;同时,该基因受MeJA和SA外源激素胁迫后的表达模式,可为泛素-蛋白酶体途径及激素调控的信号转导在甘蔗与黑穗病菌互作过程中的作用提供一定的理论依据.     

微囊藻毒素对真菌和哺乳动物细胞毒害效应的研究

主要探讨微囊藻毒素对真菌和哺乳动物细胞的毒害效果,以及其作为水体生理毒害评价模式细胞的可能。通过毒性试验和单细胞凝胶电泳实验,利用栗酒裂殖酵母细胞（S.pombe）,人肝癌细胞（HepG2）和人脑星形胶质母细胞癌细胞（U87）3种生物模型评估MC对细胞DNA的损伤以及对细胞内氧化自由基水平的影响。并利用栗酒裂殖酵母的基因缺失体,判断MC所诱导的酵母细胞氧化应激路径。研究表明MC通过氧化路径诱导DNA损伤;缺失wis1△和pap1△基因的S.pombe细胞无法诱导ROS生产,wis1△和pap1△缺失型S.pombe细胞在40 mg·L-1-1MC暴露24 h下细胞增殖率仅下降7.3%、7.7%;氧化损伤是MC损伤S.pombe的主要方式;HepG2细胞和U87细胞较S.pombe细胞适于研究、评估MC的毒性。     

雨生红球藻3个株系的生长及对光盐反应初探

对雨生红球藻3个株系（No.3、No.712和No.797)的生长、对诱导虾青素形成的光盐因子的反应进行了比较，结果表明：No.3、No.712和No.797的生长速率分别为0．128d^-1,0.137d^-1和0.144d^-1；外加盐的量较大（4gL^-1)时导致雨生红球藻细胞数的减少，而外加盐浓度较低（2gL^-1)时，细胞数的反应因株系和光照的不同而有所差异；盐和强光照均导致静细胞比例显著增加；盐的加入仅在弱光照下诱导雨生红球藻3个株系的虾青素累积，在强光照下却导致件株系虾青素含量的下降，强光照下不外加盐时No.3、NO.712和No.797藻液的虾青素含量无显著差异，分别为2.15、2.77和2．45mg L^-1。图1表2参12     

青蒿试管苗开花及用花器官为外植体诱导丛生芽生产青蒿素

将培养于MS培养基上5wk的青蒿试管苗置于光周期13，光强6000lx，温度为26℃（昼）和22℃（夜）的条件下，成功地诱导其开花，利用不同发育阶段的花蕾和花器官为外植体诱导丛生芽，发现不同浓度（ρ）的6－BA对丛生芽的诱导率和青蒿素的生物合成有重要影响。结果表明：以花蕾为外植体诱导丛生芽的最佳培养基是：MS附加4.0mg L^-1的6－BA和0.05mg L^-1的NAA；但当ρ（6－BA）在0－0.5mg L^-1之间时，有利于青蒿素的合成ρ（6－BA）=0.5-4mg L^-1时抑制青蒿素的生命合成，丛生芽中促进青蒿素合成的最佳ρ（6－BA）为0.5mg L^-1，用HPLC法测定发现用此法得到的丛生芽青蒿素的含量比直接利用叶片诱导的丛生芽青蒿素含量高1倍。     

雪松聚球藻对重金属镉的抗性和解毒作用

培养液中不同浓度（c）的重金属镉（Cd）对雪松聚藻（Synechococcuscedrorum）有相当的致毒作用，当c（Cd）＞2μmol／L时，藻体的生长和光合作用均明显受到抑制，c＞10μmol／L时，藻细胞出现解体，光合放氧量几近0,然而，当按梯度逐级提高培养基中c（Cd）时，雪松聚球藻则相应产生对有毒金属Cd的抗性，在即使c（Cd）达20μmol／L的培养基中亦生长良好．为探讨该藻对重金属致毒的抗性产生机理及其解毒作用，对藻体蛋白和胞内Cd进行了分离检测，结果表明：Cd在进入藻细胞后，与胞内多肽结合形成一种大分子量的色素蛋白复合体，和一种低分子量的金属结合蛋白，特别是后一种蛋白在藻细胞中的含量明显受外界Cd的诱导，并起解毒作用．     

苜蓿培养细胞抗羟脯氨酸变异体的筛选和特性分析

以返地卫星搭载的苜蓿（ＭｅｄｉｃａｇｏｓａｔｉｖａＬ．）种子为实验材料，取其无菌苗下胚轴切段诱导愈伤组织，该愈伤组织经正筛选法，获得抗脯氨酸类似物Ｌ羟脯氨酸（Ｈｙｐ）的变异细胞系（Ｈｙｐｒ）．Ｈｙｐｒ脱离选择压力３ｍｏ后，对Ｈｙｐ的抗性仍比对照强，ＦＷ细胞内游离脯氨酸ｗ＝０．７２ｍｇ／ｇ，是ＣＫ的３．４３倍，同时还具有对ＮａＣｌ和ＰＥＧ的交叉抗性．和ＣＫ相比，Ｈｙｐｒ愈伤组织可溶性蛋白质ＳＤＳＰＡＧＥ图谱出现两条新多肽（Ｍｒ≈５６×１０３、３２×１０３）；其过氧化物酶同工酶谱中酶活性明显高于对照，并出现１条新带，酯酶同工酶酶谱中亦出现３条新带．这些特性分析表明，该变异细胞系对Ｈｙｐ的抗性是稳定的．     

Successional changes in plant resistance and tolerance to herbivory

Despite considerable research on plant defenses, we know very little about how temporal changes in the environment may influence resistance and tolerance levels, or the costs and benefits of these defense strategies for long-lived plant species. We hypothesized that, in successional habitats, predictable environmental changes should favor strong plasticity in defense phenotypes and that the costs, benefits, and levels of tolerance and resistance will change with environmental context. Using a widely distributed, old-field perennial, late goldenrod (Solidago altissima), we conducted a field experiment to test these predictions. We planted goldenrod genets exhibiting varying levels of resistance and tolerance into three early-successional and three late-successional fields (approximately three and 15 years in age, respectively) and experimentally measured resistance and tolerance levels and their associated costs and selection coefficients. We found a significant effect of successional stage but no effect of genotype or stage-genotype interaction on defense levels. Genets planted in early-successional fields appeared to be more resistant and less tolerant to herbivory than those same genets planted in late-successional fields. There were significant trade-offs between resistance and tolerance in early-successional fields but not in late-successional fields. Each late-successional field exhibited a significant cost or selection gradient for resistance, but there was no general pattern of resistance costs or selection gradients specific to a successional stage class. In contrast, there was evidence of stage-specific costs of tolerance; late-successional fields exhibited significant costs of tolerance whereas early-successional fields did not. There was no evidence of direct selection for or against tolerance in either stage. Our results suggest that defense phenotypes might change in qualitative ways during succession. High resistance in early stages may be attributed to associational effects of the early-successional community, reducing the probability of damage, and despite a cost of tolerance in late stages, tolerance may be beneficial in mitigating the effects of both herbivory and environmental stresses (i.e., low light availability) that limit fitness in these fields. This study provides experimental evidence that succession can strongly influence defense phenotypes and promote temporal variability in relative resistance and tolerance levels.     

Mutualism as reciprocal exploitation: African plant-ants defend foliar but not reproductive structures

The foundation of many plant-ant mutualisms is ant protection of plants from herbivores in exchange for food and/or shelter. While the role of symbiotic ants in protecting plants from stem- and leaf-feeding herbivores has been intensively studied, the relationship between ant defense and measures of plant fitness has seldom been quantified. We studied ant aggression, damage by herbivores and seed predators, and fruit production among Acacia drepanolobium trees occupied by four different acacia-ant species in an East African savanna. Levels of ant aggression in response to experimental disturbance differed strongly among the four species. All four ant species recruited more strongly to new leaf growth on host plants following disturbance, while recruitment to developing fruits was on average an order of magnitude lower. Host plants occupied by more aggressive ant species suffered significantly less vegetative damage from leaf-feeding insects, stem-boring beetles, and vertebrate browsers than host plants occupied by less aggressive ant species. However, there were no differences among fruiting host plants occupied by different ant species in levels of seed predation by bruchid seed predators. Fruit production on host trees was significantly correlated with tree stem diameter but not with the identity of resident ants. Our results demonstrate that defense of host plants may differ substantially among ant species and between vegetative and reproductive structures and that fruit production is not necessarily correlated with high levels of aggression by resident ants.     

三种菊科入侵植物的生长与化学防御的关系研究

“生活史理论”认为,植物可利用的资源总量是有限的,在植物的不同功能之间存在着此消彼长的权衡关系。入侵植物的生长和化学防御一般优于本地植物,那么其生长与化学防御之间是否存在权衡及其权衡关系怎样,目前尚不清楚。以广东省3种菊科入侵植物[三裂叶蟛蜞菊（Wedelia trilobata （L.） Hitchc.）、飞机草（Eupatorium odoratum）和薇甘菊（Mikania micrantha）]为研究对象,并分别以近缘或伴生的本地植物[蟛蜞菊（Wedelia chinenses）、华泽兰（Eupatorium chinense）和鸡矢藤（Paederia scandens）]为对照,研究入侵植物的生长特性（相对生长率和比叶面积）与化学防御物质（缩合单宁和总酚）含量,并基于这2种光合碳分配的主要形式,探讨入侵植物生长与化学防御之间的权衡关系。结果表明：3种入侵植物的相对生长率均高于本地对照种;薇甘菊的比叶面积大于对照种,而其他2种无明显优势。薇甘菊和三裂叶蟛蜞菊的缩合单宁显著高于对照种,飞机草的总酚含量高于对照种。我们的结果显示,入侵植物的生长和化学防御均优于本地植物,但它们的碳同化能力相近;因此,入侵植物特殊的内在资源分配与利用机制可能是其成功入侵的关键。     

微量元素对红豆杉菌根菌生长的影响

微量元素对植物的生长发育有着十分重要的意义。对太行山野生红豆杉(Taxus chinensis var.mairei)菌根菌菌丝在分别加有硫酸锌、钼酸铵以及硫酸亚铁的PDA培养基中的生长发育状况进行了试验研究。所得数据同PDA对照组进行比较分析后,结果表明:Mo元素对红豆杉菌根菌菌丝的生长发育有很好的促进作用;Fe元素明显地促进红豆杉菌根菌菌丝的生长发育;Zn元素在一定范围内对红豆杉菌根菌菌丝的生长发育有一定的抑制作用。实验结果揭示出红豆杉菌根菌生长发育的一些营养因素。     

Induced responses to herbivory in the Neotropical ant-plant association between Azteca ants and Cecropia trees: response of ants to potential inducing cues

Plant defense against herbivores often involves constitutive and inducible mechanisms of resistance. Obligate ant-plants, which provide food and housing for ants, are thought to primarily rely on ants for defense against herbivores. This form of plant defense has largely been viewed as static. We have been investigating the dynamic nature of Azteca ants as an inducible defense of Cecropia trees. Ants rapidly recruit to and patrol sites of foliar damage. We propose that Azteca ants can be viewed as an inducible defense for Cecropia trees because of their sensitivity to cues associated with herbivory, their rapid and aggressive recruiting ability, and their reclaimable and redeployable nature as a plant defense. In this study, we examine ant behavior following plant damage, and the potential cues that indude ant recruitment. We found that ants present on leaves when the plant is damaged leave the damaged leaf and recruit other ants to it, presumably by laying recruitment trails. Volatile leaf cues associated with herbivory were important in eliciting an induced response in two experiments. However, we found that cues associated with a congeneric plant elicited a much stronger ant response than conspecific cues. Although the type of leaf damage (gaping wounds versus leaf edge wounds) did not affect the level of ant recruitment, the extent of damage did. Leaves with one hole punched showed a 50% increase in ants, while leaves with five holes punched in them elicited a 100% increase in ant numbers. In sum, it appears that multiple plant-related cues associated with herbivory are involved in induction of ant recruitment in the Cecropia-Azteca system. We discuss the generality of ant responses to herbivory in obligate ant-plant systems, and in facultative ant-plant associations, which may be more common. Received: 23 March 1998 / Accepted after revision: 5 July 1998     

中国红豆杉内生放线菌En-1的分离、鉴定及其次生代谢物的研究

采用选择性分离方法对药用植物红豆杉Taxus chinensis嫩枝和叶进行内生菌分离,得到一株放线菌En-1,经鉴定为链霉菌Streptomyces sp.;并对该菌进行液体培养条件优化、次生代谢物初筛及其抗菌活性的研究。通过添加宿主植物浸出物至En-1液体培养中以考察宿主对后者生长的影响,结果表明红豆杉叶浸出物能促进En-1的体外生长;而且En-1次生代谢物对5株供试菌中的黑曲霉具有抑制活性。通过En-1菌培养基优化并联合波谱法检测其发酵产物,显示8#配方（淀粉25 g.L-1,KNO3 1 g.L-1,K2HPO4.3H2O 0.5 g.L-1,NaCl 0.5 g.L-1,FeSO4.7H2O 0.01 g.L-1,MgSO4.7H2O 0.5 g.L-1）培养液中,En-1所产的次生代谢物最具结构多样性;同时8#配方也有利于该菌产生抗菌活性产物。本研究提示红豆杉内生放线菌可作为寻求药物先导化合物的资源菌。     

What enables coexistence in plant communities? Weak versus strong species traits and the role of local processes

Explaining the coexistence of species that basically depend on the same resources has been a brainteaser for generations of ecologists. Different mechanisms have been proposed to facilitate coexistence in plant communities, where space is an important resource. Using a stochastic cellular automaton simulation model we analyze - separately and in combination - the influence of different species traits and processes which alter local competition on the coexistence of plant species over a fixed time horizon. We show that different species traits operate on different time scales in competition. We therefore suggest the concept of weak versus strong traits according to short- or long-term exclusion of species differing in these traits. As a consequence, highly non-linear trade-offs between weak and strong traits can result in communities. Furthermore, we found that trade-offs based on physiological species traits such as plant lifetime, dispersal range and plant growth, did not support broad and long-term coexistence—further processes such as density-dependent mortality and light-dependent colonization were necessary. This suggests that coexistence in plant communities requires (stabilizing) local processes to support the (equalizing) trade-offs in species traits.