基于高通量测序的闽楠幼林根际土壤丛枝菌根真菌群落变化

【背景】闽楠是珍贵用材树种。阐明闽楠根际土壤微生物特征,对科学经营闽楠人工林有指导作用。【目的】明确闽楠人工林恢复过程中根际土壤丛枝菌根群落随林龄的变化特征。【方法】采用高通量Illumina MiSeq测序的方法,评估不同林龄闽楠幼林根际土壤丛枝菌根真菌(Arbuscular Mycorrhiza Fungi,AMF)群落的多样性和组成,揭示造林后闽楠根际土壤真菌群落对时间序列中土壤养分变化的响应特征。【结果】从3个林龄闽楠根际土壤中共得到342 555条有效序列,共得到253个AMF-OTU,分属于1门1纲5目9科14属。α多样性指数表明,丛枝菌根真菌群落的Chao1、Shannon指数随着林龄的增加而增加,但不同林龄间差异不显著。β多样性指数表明,在闽楠幼龄阶段,不同林龄间AMF群落组成差异明显。根据Bray-Curtis相异度分析阐明AMF群落组成的相似性与林龄相关。种水平物种丰度与土壤因子的相关性分析表明,影响AMF群落丰度的主要土壤因子是pH、全钾、硝态氮和铵态氮。铵态氮与优势AMF分子虚拟种Glomus-Franke-A1-VTX00076、Glomus-Franke-A1-VTX00269的丰度呈显著正相关。【结论】随着林龄的增加,闽楠幼龄根际土壤丛枝菌根真菌的种类组成发生了显著变化,影响AMF特性的主要土壤因子是pH、全钾、硝态氮和铵态氮。这些发现将有助于揭示闽楠幼林生态系统和土壤微生物多样性的相互关系和机制,为生物多样性-生态系统功能实验提供理论依据。     

内蒙古中部地区马铃薯根际和根系丛枝菌根真菌类群的多样性

【背景】丛枝菌根真菌(arbuscular mycorrhizal fungus,AMF)是菌根真菌中分布最广泛的一类真菌,能够与90%以上的植物形成菌根共生体,并通过调节宿主体内的代谢活动增强植物的抗逆性。【目的】揭示内蒙古马铃薯主产区土壤中AMF的结构与组成,解析马铃薯不同发育阶段以及连作对AMF类群的影响。【方法】以内蒙古中部地区大井村、徐麻夭村和红格尔图村采集的马铃薯根系及根际土为材料,通过PCR扩增建立18S rRNA基因文库,研究不同地点、不同马铃薯生育阶段以及连作对马铃薯根系内和根际土中的AMF类群组成和多样性的影响。【结果】大井村和红格尔图村马铃薯根际土中的AMF的类群多样性优于徐麻夭村,其中Glomus为大井村和红格尔图村马铃薯根际土样本中AMF的优势菌群,而徐麻夭村样本根际土中的优势类群为Diversispora。马铃薯根系中的AMF多样性的研究结果表明,3个供试地点根系内AMF类群的类型没有显著差异,但是AMF类群的比例却不同。大井村根系样本中AMF的优势菌为Diversispora,而Rhizophagus是红格尔图村和徐麻夭村根系样本中的优势AMF类群。同一供试地点马铃薯苗期和块茎膨大期根际土中的AMF类群的类型多于块茎形成期,其中Glomus是马铃薯不同发育阶段根际土中的优势类群;而Rhizophagus是3个不同发育阶段根系内的优势类群。2015和2016年马铃薯连作地块中根际土和根系内AMF类群的类型完全相同,但是不同的AMF类群的相对丰度随着马铃薯的连作呈现出显著的变化。其中,根际土样本中Glomus和Archaeospora类群的相对丰度随着连作有所上升,相应的Entrophospora和Diversispora相对丰度有所下降;根系样本中AMF的优势类群随着连作从Glomus转变为Rhizophagus。AMF多样性测定结果表明,连作对马铃薯根际土中AMF多样性的影响显著高于对根系内类群的影响。【结论】AMF类群的多样性会因土壤组成成分、马铃薯生育期和连作效应而呈现不同变化。     

典型农田土壤中丛枝菌根真菌-根际细菌互作及与氮磷利用的关系

在农业生态系统中,土壤微生物是土壤-作物系统养分循环的重要驱动力,其中丛枝菌根真菌(Arbuscularmycorrhizalfungi,AMF)能够促进作物对养分的吸收,适应逆境胁迫。【目的】进一步揭示AMF和根际细菌群落的跨界网络互作,挖掘与作物氮磷利用显著相关的关键微生物类群,揭示关键类群的生态网络特征。【方法】利用Illumina测序技术对3种典型农田旱地土壤(黑土、潮土和红壤)中AMF和根际细菌群落结构进行分析;构建互作网络并利用偏冗余分析、相关性分析探究了与氮磷利用相关的潜在关键类群。【结果】3种土壤中AMF与根际细菌均以正相互作用为主。不同土壤中AMF与根际细菌互作关系差异明显,在红壤中跨界互作最为密切,其中球囊霉属真菌(Glomus)与根际细菌中的放线菌(Actinobacteria)和变形菌(Proteobacteria)之间的交互作用最多。而在黑土中主要体现为根际细菌的界内互作。与氮磷利用率显著相关的关键微生物类群主要属于球囊霉属真菌、放线菌和α变形菌。【结论】典型旱地土壤中AMF与根际细菌的正相互关系对作物氮磷利用有潜在促进作用,关键类群在有机质和养分贫乏的红壤中可能起到更重要的作用。     

连作花魔芋软腐病株与健株根域丛枝菌根真菌群落多样性

【目的】解析不同连作年限花魔芋软腐病株、健株根域的丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)群落多样性。【方法】使用AMF 18S SSU rRNA基因特异引物AMV4.5NF/AMDGR对正茬及连作2年和3年的软腐病株、健株魔芋根系和根际土壤DNA扩增建库,通过高通量测序和生物信息学分析探究魔芋软腐病与其根域AMF群落多样性的关系。【结果】魔芋根系具有明显的AMF菌丝、泡囊和丛枝等结构。在相同连作年限条件下,健株根系AMF总侵染率、侵染强度和孢子密度均显著高于病株(P<0.05);在不同连作年限条件下,病株根系AMF总侵染率和侵染强度随连作年限延长而降低。从所有样品中共鉴定到9属53种AMF,其中有49个已知种和4个新种。球囊霉属(Glomus)和类球囊霉属(Claroideoglomus)是AMF群落的优势属,其AMF种分别占总AMF种数的41.5%和26.4%;丰度最高的Paraglomus sp.VTX00308是所有样品的共有种。连作、软腐病及二者的交互作用显著影响根系AMF群落的Shannon指数和Simpson指数及根际土壤AMF的Chao1指数(P<0.05)。通过丰度差异分析发现6个在连作软腐病发生后丰度差异显著的AMF种(P<0.05);NMDS分析表明,不同连作年限的魔芋软腐病株与健株之间的根域AMF菌种组成、相对丰度和群落结构存在差异。相关性分析表明,软腐病发病率和病情指数与魔芋根系和根际土壤AMF的Shannon指数、根系AMF的Chao1和Simpson指数以及AMF总侵染率、侵染强度和孢子密度极显著负相关(P<0.01)。【结论】比对健株,连作魔芋软腐病株根际土壤AMF孢子密度以及根系AMF侵染率、种数和多样性均降低,其群落结构显著改变。     

山西历山珍稀药用植物AM真菌资源与土壤因子的关系

以山西历山自然保护区暴马丁香、连香树、南方红豆杉和领春木4种珍稀药用植物为材料,研究其根际土壤中丛枝菌根真菌(AMF)的菌根结构类型和种属分布,同时分析土壤因子与其侵染率和孢子密度的关系。结果表明:(1)暴马丁香菌根类型为中间型(I-型),连香树和领春木为重楼型(P-型),南方红豆杉为疆南星型(A-型);4种植物根际共鉴定AMF 27种,分别隶属于球囊霉属(Glomus)、无梗囊霉属(Acaulospora)、盾巨孢囊霉属(Scutellos-pora)、多孢囊霉属(Diversispora)和原囊霉属(Archaeospora)5属,其中Glomus为优势属。(2)pH与暴马丁香和领春木的侵染率和孢子密度呈正相关,但与连香树和南方红豆杉呈负相关;速效磷与暴马丁香、南方红豆杉和领春木的侵染率和孢子密度呈正相关,但与连香树呈负相关;碱解氮、有机质与暴马丁香、连香树和领春木的侵染率和孢子密度呈正相关,但与南方红豆杉呈负相关。(3)4种药用植物的菌根侵染率主要受其根际土壤碱解氮的影响,而根际AMF孢子密度主要受根际土壤pH制约。可见,历山自然保护区内AMF资源丰富,多样性程度也较高;宿主植物不同,土壤因子对其侵染率和孢子密度的影响也不同。     

云南岩溶断陷盆地土地利用方式对丛枝菌根真菌群落结构的影响

【背景】丛枝菌根真菌(arbuscular mycorrhizal fungus,AMF)是一类能与绝大部分植物形成共生关系的真菌,在植被演替和生态恢复过程中发挥重要作用。【目的】了解云南岩溶断陷盆地不同土地利用方式对AMF群落结构的影响。【方法】采用Illumina HiSeq 2500高通量测序技术对云南岩溶断陷盆地林地、灌丛和草地等3种土地利用方式土壤样品的AMF群落结构进行分析。【结果】草地总磷(total phosphorus,TP)、速效钾(available potassium,AK)、交换性镁(exchangeable magnesium,E-Mg)、电导率(electrical conductivity,EC)和土壤温度(temperature,T)显著高于林地和灌丛,林地总氮(total nitrogen,TN)和有机碳(soil organic carbon,SOC)含量显著高于灌丛和草地。TN、SOC、AK和EC是AMF群落结构最主要的影响因子。草地样品的Chao1、ACE、Shannon和Observed species指数显著高于林地和灌丛样品,尽管Simpson指数差异不显著,但是也表现出了类似的变化趋势。9个土壤样品共产生953个AMF-OTU,鉴定出AMF 3个纲4个目9个科13个属。Glomeromycetes和Paraglomeromycetes为优势纲;Glomus和Paraglomus为优势属,其次为Claroideoglomus、Acaulospora和Diversispora。Glomus和Diversispora主要分布在草地和灌丛中,随着演替的进行其相对丰度有所下降,而Paraglomus的相对丰度逐渐上升。【结论】云南岩溶断陷盆地土地利用方式的AMF群落结构具有显著差异,土壤理化因子是重要的影响因素。     

土壤铅和锌对植物根际丛枝菌根真菌分布的影响

从秦岭凤县铅锌污染区4种植物根际共鉴定出球囊霉属(Glomus)丛枝菌根真菌(Arbuscular mycorrhizal ungi,AMF)12种,其中缩球囊霉(G.constrictum)是该区域的优势种;缩球囊霉、副冠球囊霉(G.coronatum)、苏格兰球囊霉(G.caledonium)和聚丛球囊霉(G.aggregatum)对铅锌污染具有较强的耐性,而地球囊霉(G.geospo-rum)、台湾球囊霉(G.formosanum)、地表球囊霉(G.versiforme)和两型球囊霉(G.dimorphicum)对铅锌污染的耐性较弱.相关分析表明,土壤Pb、Zn、速效P浓度和pH较低时,对AMF丰度有一定的促进作用,当Pb、Zn、速效P浓度和pH较高时,对AMF丰度为抑制作用,AMF丰度与高浓度Pb呈极显著负相关,与高浓度速效P呈显著负相关.通径分析表明,Pb是影响秦岭重金属污染区AMF丰度的主要因素,其直接和间接作用都较大,而pH、速效P和Zn主要通过Pb的间接作用来影响AMF丰度.     

云南金顶铅锌矿区丛枝菌根真菌多样性的研究

对云南金顶铅锌矿区丛枝菌根真菌(Arbuscular mycorrhizal fungi,AMF)资源进行了调查,从32种植物的83个根际土壤样本中分离鉴定出5属36种丛枝菌根真菌,其中无梗囊霉属Acaulospora 5种、内养囊霉属Entrophospora 2种、巨孢囊霉属Gigaspora 1种、球囊霉属Glomus 24种及盾巨孢囊霉属Scutellospora 4种。球囊霉属和无梗囊霉属为金顶铅锌矿区中丛枝菌根真菌的优势属,沃克球囊霉Glomus walkeri是我国的新记录种。近明球囊霉Glomus claroideum、明球囊霉G.clarum、缩球囊霉G.constrictium、地球囊霉G.geosporum、摩西球囊霉G.mossaea、膨胀球囊霉G.pansihalos和疣突球囊霉G.verruculosum是金顶铅锌矿区的优势种;金顶铅锌矿区土壤中丛枝菌根真菌的孢子密度为495-11175个/100g土,平均3368±291(SE,标准误差)个/100g土,每种植物根际土壤中丛枝菌根真菌的物种丰富度为2-20种,平均11.5种;金顶铅锌矿区植物根际土壤中较高的AMF孢子密度和物种丰富度说明AMF对重金属污染具有较强的抗(耐受)性。     

土壤因子对府谷清水川流域砒砂岩区刺槐和沙棘AMF的影响

应用相关性分析和通径分析研究了府谷清水川流域砒砂岩区土壤因子对刺槐(Robinia pseudoacacia)和沙棘(Hippophae rhamnoides)根际丛枝菌根真菌(AMF)的影响.结果表明:AMF可以与刺槐和沙棘形成良好的共生关系;从两种植物根际土中共分离鉴定出AMF 13种,其中摩西囊霉(Glomus mosseae)、地球囊霉(Glomus geospo-rum)和缩球囊霉(Glomus constrictum)为优势种;AMF平均侵染率为90.6%,平均孢子密度为559.1个?100 g-1干土;AMF的孢子密度、侵染率和种类之间没有显著的相关性;刺槐和沙棘根际AMF侵染率均与pH呈显著正相关,在刺槐根际,pH和有效氮通过直接作用影响侵染率,有机质和有效磷通过间接作用影响侵染率;在沙棘根际,pH和有效磷通过直接作用影响侵染率,有机质和有效氮通过间接作用影响侵染率;在沙棘根际,有机质和pH主要是通过直接作用影响AMF种类,有效氮主要通过有效磷的间接作用影响AMF种类.     

铜尾矿白羊草各生长阶段根际土壤丛枝菌根真菌群落特征

以北方铜业铜矿峪矿十八河尾矿坝主要恢复植被白羊草为研究对象，分析重金属污染环境下，白羊草不同生长阶段根际土壤中丛枝菌根真菌（Arbuscular mycorrhizal fungi，AMF）群落结构与多样性的变化特征，研究各生长阶段白羊草AMF群落结构与环境因子的相互关系。结果发现，白羊草不同生长阶段根际土壤的理化性质和酶活性均存在显著差异。白羊草各生长阶段根际土壤中具有相同的优势科球囊霉科，但AMF群落组成在各个生长阶段的结构存在明显不同。白羊草各生长阶段主要AMF物种对生态因子的响应表现为：幼苗期根际土壤中球囊霉科主要受土壤养分和碳氮比的影响，分蘖期根际土壤中多样孢囊霉科的主要影响因子为土壤pH，成熟期根际土壤中，双型囊霉科双型囊霉科与土壤过氧化氢酶显著相关，类球囊霉科与土壤脲酶显著正相关。总体看来，白羊草各生长阶段根际土壤AMF群落特征受到的生态因子影响具有明显差异。这有助于进一步认识污染环境下AMF的群落特征及其关键影响因子，为铜尾矿生态恢复过程中发掘和利用菌种资源提供科学依据，从而提高矿区生态修复效率。     

Role for dopamine in malonate-induced damage in vivo in striatum and in vitro in mesencephalic cultures

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.     

2018年中国植物科学若干领域重要研究进展

2018年中国植物科学继续呈现快速发展的态势, 我国科学家在国际植物科学主流学术刊物发表论文数量大幅增加, 取得了多项具有重要影响的成果。调控植物生长-代谢平衡实现可持续农业发展入选2018年度中国科学十大进展; 中国被子植物区系进化历史研究入选2018年度中国生命科学十大进展。以水稻为代表的农作物和果蔬等经济作物研究在国际上已呈现出明显的优势, 若干领域已从“追赶”状态跨越到“领跑”地位。该文对2018年中国科学家在植物科学若干领域取得的重要研究成果进行了概括性评述, 旨在全面追踪和报道当前中国植物科学领域的发展前沿和热点, 展示中国科学家所取得的杰出成就。     

2017年中国植物科学若干领域重要研究进展

2017年中国植物科学继续保持高速发展态势, 重大成果频出, 具体表现在中国植物学家在国际顶级学术期刊发表的文章数量平稳上升。中国植物科学领域的研究工作者成果精彩纷呈, 如新型广谱抗病机制的发现、水稻广谱抗病遗传基础及机制和疫霉菌诱发病害成灾机制研究等。2017年中国生命科学领域十大进展评选中, 有两项植物科学领域的研究成果入选。水稻生物学、进化与基因组学和激素生物学等领域学科发展突出。另外, 值得一提的是, 长期从事高等植物与代谢途径调控分子网络研究和水稻品种设计育种的李家洋院士的研究成果“水稻高产优质性状形成的分子机理及品种设计”荣获2017年国家自然科学一等奖。这一具有重大国际影响的开创性贡献标志着中国植物科学在该领域的国际科学前沿居于引领和卓越地位。该文对2017年中国本土科学家在植物科学若干领域取得的重要研究成果进行了系统梳理, 旨在全面追踪和报道当前中国植物科学领域发展的最新前沿动态, 与广大读者共同分享我国科学家所取得的辉煌成就。     

我国葫芦科植物离体培养研究进展

葫芦科植物包括多种瓜类蔬菜,对其进行离体培养研究具有重要的理论和实践意义。综述了国内在葫芦科植物器官培养、体细胞胚胎发生、花药培养、原生质体培养和体细胞杂交及离体遗传转化等方面取得的研究进展,并对葫芦科植物离体培养、遗传转化与育种的前景作了展望。     

SSTR5 ablation in islet results in alterations in glucose homeostasis in mice

Somatostatin (SST) peptide is a potent inhibitor of insulin secretion and its effect is mediated via somatostatin receptor 5 (SSTR5) in the endocrine pancreas. To investigate the consequences of gene ablation of SSTR5 in the mouse pancreas, we have generated a mouse model in which the SSTR5 gene was specifically knocked down in the pancreatic beta cells (betaSSTR5Kd) using the Cre-lox system. Immunohistochemistry analysis showed that SSTR5 gene expression was absent in beta cells at three months of age. At the time of gene ablation, betaSSTR5Kd mice demonstrated glucose intolerance with lack of insulin response and significantly reduced serum insulin levels. Insulin tolerance test demonstrated a significant increase of insulin clearance in vivo at the same age. In vitro studies demonstrated an absence of response to SST-28 stimulation in the betaSSTR5Kd mouse islet, which was associated with a significantly reduced SST expression level in betaSSTR5Kd mice pancreata. In addition, betaSSTR5Kd mice had significantly reduced serum glucose levels and increased serum insulin levels at 12 months of age. Glucose tolerance test at an older age also indicated a persistently higher insulin level in betaSSTR5Kd mice. Further studies of betaSSTR5Kd mice had revealed elevated serum C-peptide levels at both 3 and 12 months of age, suggesting that these mice are capable of producing and releasing insulin to the periphery. These results support the hypothesis that SSTR5 plays a pivotal role in the regulation of insulin secretion in the mouse pancreas.     

2018年中国植物科学若干领域重要研究进展

2018年中国植物科学继续呈现快速发展的态势, 我国科学家在国际植物科学主流学术刊物发表论文数量大幅增加, 取得了多项具有重要影响的成果。调控植物生长-代谢平衡实现可持续农业发展入选2018年度中国科学十大进展; 中国被子植物区系进化历史研究入选2018年度中国生命科学十大进展。以水稻为代表的农作物和果蔬等经济作物研究在国际上已呈现出明显的优势, 若干领域已从“追赶”状态跨越到“领跑”地位。该文对2018年中国科学家在植物科学若干领域取得的重要研究成果进行了概括性评述, 旨在全面追踪和报道当前中国植物科学领域的发展前沿和热点, 展示中国科学家所取得的杰出成就。     

我国葫芦科植物离体培养研究进展

葫芦科植物包括多种瓜类蔬菜,对其进行离体培养研究具有重要的理论和实践意义.综述了国内在葫芦科植物器官培养、体细胞胚胎发生、花药培养、原生质体培养和体细胞杂交及离体遗传转化等方面取得的研究进展,并对葫芦科植物离体培养、遗传转化与育种的前景作了展望.     

Changes in aquatic vegetation in Rhine floodplain streams in Alsace in relation to disturbance

Abstract. Changes are described in aquatic vegetation in oligotrophic, groundwater-fed Rhine floodplain streams in Alsace (eastern France), resulting from disturbance. Disturbance factors include changes in nutrients, either permanent ones - effluent from a waste water treatment plant or trout hatcheries - or periodic ones: flooding. Regular inputs of high levels of phosphate and ammonia modified the macrophyte vegetation in these streams. The floristic composition, which was characteristic of oligotrophic waters upstream of the eutrophicated sector, changed to that of a eutrophic situation as originally found downstream. Periodic disturbance by floods which normally occur once a year, irregularly eutrophicates the small streams, causing the development of a mixture of eutrophic and oligotrophic species. Six macrophyte communities are distinguished, indicating different trophic levels. The aquatic vegetation is adapted to the variations of phosphate and ammonia levels. Hence, aquatic macrophytes can be used as bio-indicators of fluctuations in water nutrient levels in relation to the type of disturbance.     

Apoptosis in experimental myocardial infarction in situ and in the perfused heart in vitro

We report the appearance of apoptotic cells in experimental myocardial infarction (rabbit heart) in in situ and in vitro preparations. Apoptosis was recognized by intravital staining with Hoechst 33342 (Ho342), by nick-end labeling (TUNEL) and by DNA laddering. A steady rise in the relative number of apoptotic cardiomyocytes (apoptotic index) was noted in in situ preparations. Apoptosis was first noted 6 h after the onset of ischemia with its highest value occurring after 72 h. Apoptotic nuclei were absent in remote areas of the left and right ventricles. Apoptotic nuclei within the infarcted area showed diminished intensity of Ho342 fluorescence. Three days after ischemia, a border zone adjacent to the infarcted area consisting of apoptotic macrophages was recognized. A novel finding was the appearance of apoptotic cardiomyocytes in the isolated perfused ischemic heart. Occurring as early as 50 min after the onset of ischemia, a high apoptotic index was present adjacent to the ligature placed around the coronary artery. This observation provides the opportunity to selectively examine factors leading to apoptosis in the ischemic heart under controlled experimental conditions.     

Morphine-potentiated platelet aggregation in in vitro and platelet plug formation in in vivo experiments

The detailed mechanisms underlying morphine-signaling pathways in platelets remain obscure. Therefore, we systematically examined the influence of morphine on washed human platelets. In this study, washed human platelet suspensions were used for in vitro studies. Furthermore, platelet thrombus formation induced by irradiation of mesenteric venules with filtered light in mice pretreated with fluorescein sodium was used for an in vivo thrombotic study. Morphine concentration dependently (0.6, 1, and 5 microM) potentiated platelet aggregation and the ATP release reaction stimulated by agonists (i.e., collagen and U46619) in washed human platelets. Yohimbine (0.1 microM), a specific alpha(2)-adrenoceptor antagonist, markedly abolished the potentiation of morphine in platelet aggregation stimulated by agonists. Morphine also potentiated phosphoinositide breakdown and intracellular Ca(2+) mobilization in human platelets stimulated by collagen (1 microg/ml). Moreover, morphine (0.6-5 microM) markedly inhibited prostaglandin E(1) (10 microM)-induced cyclic AMP formation in human platelets, while yohimbine (0.1 microM) significantly reversed the inhibition of cyclic AMP by morphine (0.6 and 1 microM) in this study. The thrombin-evoked increase in pH(i) was markedly potentiated in the presence of morphine (1 and 5 microM). Morphine (2 and 5 mg/g) significantly shortened the time require to induce platelet plug formation in mesenteric venules. We concluded that morphine may exert its potentiation in platelet aggregation by binding to alpha(2)-adrenoceptors in human platelets, with a resulting inhibition of adenylate cyclase, thereby reducing intracellular cyclic AMP formation followed by increased activation of phospholipase C and the Na(+)/H(+) exchanger. This leads to increased intracellular Ca(2+) mobilization, and finally potentiation of platelet aggregation and of the ATP release reaction.