Acquisition and assimilation of gaseous ammonia as revealed by intracellular pH changes in leaves of higher plants

Atmospheric ammonia (NH₃) from various anthropogenic sources has become a serious problem for natural vegetation. Ammonia not only causes changes in plant nitrogen metabolism, but also affects the acid-base balance of plants. Using the pH-sensitive fluorescent dyes pyranine and esculin, cytosolic and vacuolar pH changes were measured in leaves of C₃ and C₄ plants exposed for brief periods to concentrations of NH₃ in air ranging from 1.33 to 8.29 mol NH₃ · mol^-1 gas (0.94–5.86 mg · m^-3). After a lag phase, uptake of NH₃ from air at a rate of 200 nmol NH₃ · m ^{- 2} leaf area · s^{- 1} into leaves of Zea mays L. increased pyranine fluorescence indicating cytosolic alkalinisation. The increase was much larger in the dark than in the light. In illuminated leaves of the C₃ plant Pelargonium zonale L. and the C₄ plants Z. mays and Amaranthus caudatus L., NH₃-dependent cytosolic alkalinisation was particularly pronounced when CO₂ was supplied at very low levels (16 or 20 mol CO₂ · mol^{- 1} gas, containing 210 mmol O₂ · mol^{- 1} gas). An increase in esculin fluorescence, which was smaller than that of pyranine, was indicative of trapping of some of the NH₃ in the vacuoles of leaves of Spinacia oleracea L. and Z. mays. Photosynthesis and transpiration remained unchanged during exposure of illuminated leaves to NH₃, yielding an influx of 200 nmol NH₃ · m^-2 leaf area · s^-1 for up to 30 min, the longest exposure time used. Both CO₂ and O₂ influenced the extent of cytosolic alkalinisation. At 500 mol CO₂ · mol^-1 gas the cytosolic alkalinisation was suppressed more than at 16 or 20 mol CO₂ · mol^-1 gas. The suppressing effect of CO₂ on the NH₃induced alkalinisation was larger in illuminated leaves of the C₄ plants Z. mays and A. caudatus than in leaves of the C₃ plant P. zonale. A reduction of the O₂ concentration from 210 to 10 mmol O₂ · mol ^-1 gas, which inhibits photorespiration, increased the NH₃induced cytosolic alkalinisation in C₃ plants. Suppression by CO₂ or O₂ of the alkaline pH shift caused by the dissolution and protonation of NH₃ in queous leaf compartments, and possibly by the production of organic compounds synthesised from atmospheric NH₃, indicates that NH₃ which enters leaves is rapidly assimilated if photosynthesis or photorespiration provide nitrogen acceptor molecules.This work was supported by the Biotechnology and Biological Sciences Research Council and the Deutsche Forschungsgemein-schaft within the framework of the research of Sonderforschun-gsbreich 251 of the University of Würzburg. We are grateful to Dr. B. Wollenweber (The Royal Veterinary and Agricultural University, Denmark) for discussions.     

Hox homeodomain proteins exhibit selective complex stabilities with Pbx and DNA.

Eight of the nine homeobox genes of the Hoxb locus encode proteins which contain a conserved hexapeptide motif upstream from the homeodomain. All eight proteins (Hoxb-1-Hoxb-8) bind to a target oligonucleotide in the presence of Pbx1a under conditions where minimal or no binding is detected for the Hox or Pbx1a proteins alone. The stabilities of the Hox-Pbx1a-DNA complexes vary >100-fold, with the proteins from the middle of the locus (Hoxb-5 and Hoxb-6) forming very stable complexes, while Hoxb-4, Hoxb-7 and Hoxb-8 form complexes of intermediate stability and proteins at the 3'-side of the locus (Hoxb-1-Hoxb-3) form complexes which are very unstable. Although Hox-b proteins containing longer linker sequences between the hexapeptide and homeodomains formed unstable complexes, shortening the linker did not confer complex stability. Homeodomain swapping experiments revealed that this motif does not independently determine complex stability. Naturally occurring variations within the hexapeptides of specific Hox proteins also do not explain complex stability differences. However, two core amino acids (tryptophan and methionine) which are absolutely conserved within the hexapeptide domains appear to be required for complex formation. Removal of N- and C-terminal flanking regions did not influence complex stability and the members of paralog group 4 (Hoxa-4, b-4, c-4 and d-4), which share highly conserved hexapeptides, linkers and homeodomains but different flanking regions, form complexes of similar stability. These data suggest that the structural features of Hox proteins which determine Hox-Pbx1a-DNA complex stability reside within the precise structural relationships between the homeodomain, hexapeptide and linker regions.     

生物复苏——大绝灭后生物演化历史的第一幕

生命史是一部生物界短期,快速剧变与长期,慢速稳定相互交替的历史。大绝灭（即集群绝灭）事件反映了全球环境的大突变,点断了地质历史中的生命记录及其发展历程,预示着生物界的演化出现了最有意义的飞跃,近年来尝试研究大绝灭后全球生物界的残存－复苏及其基本型式,并探索复苏的控制因素,标志着地质科学中一个重心的转移（即从大绝灭转向其后的生物残存与复苏的研究）。生物复苏揭示了大绝灭后生物演化历史的第一幕,其研究的     

Kinetics of retroviral production from the amphotropic ΨCRIP murine producer cell line

Rapidly expanding development and practice of gene therapy requires the availability of large quantities of high titer retroviral supernatants. One way to achieve high retroviral titers is through improved understanding of the kinetics of retroviral production and decay, and the subsequent development of improved cell culture methods. In the present study we investigated the effects of different operational modes on the retroviral production of the NIH 3T3 fibroblast derived amphotropic murine retroviral producing cell line pMFG/CRIP. Semi-continuous culture (exchange of 50% of medium volume daily) was found to promote cell growth and enhance retroviral production. The rapid medium exchange resulted in significantly larger amounts of high titer supernatants and an extended production phase as compared to the batch control cultures. The specific viral productivity of the pMFG/CRIP cells was in the range of 10 to 40 infectious viruses produced per thousand producer cells per day. The CV-1 African Green Monkey kidney cell line was used as the infection target. Lowering the serum level form 20% to 10% improved retroviral production slightly. However, at lower serum levels (1%, 5% and 10% (v/v)) growth of the producer cell line, and thus retroviral production, was directly proportional to the serum level. The half-life of the virus at 37°C was found to be 5.5 hours. Promoting the growth of producer cell lines can improve retroviral vectors titers and viral production. High cell density systems that allow for rapid cell growth and waste product removal are likely to be used to generate high-titer retroviral supernatants.     

鹅掌楸属植物的多糖壁前体和花粉管的生长

本文观察描述了中国鹅掌楸（Ｌｉｒｉｏｄｅｎｄｒｏｎｃｈｉｎｅｎｓｅ）和北美鹅掌楸（Ｌ．ｔｕｌｉｐｉｆｅｒａ）花粉在异已柱头萌发和花粉管生长期间多糖壁前体的发生、形态结构和生理功能．１、多糖壁前体在形态上有Ｐ－粒子（Ｐｏｌｙｓａｃｃｈａｒｉｄｅｐａｒｔｉｃｌｅｓ）,被膜小泡（ｃｏａｔｅｄｖｅｓｉｃｌｅ）和小泡（ｖｅｓｉｃｌｅ）三种。２、Ｐ－粒子于单核花粉期已经发生,至花粉管延伸期为发生高峰。多糖壁前体是在高尔基体,内质网和线粒体的相继、连续作用下,由淀粉质体、蛋白体和脂滴降解形成．３、Ｐ－粒子的形态随不同发育时期而变化,早期为成群的电子透明小泡,或为蛋白质束缚的挤压成多面体形,后期为内含颗粒或微纤丝的无被膜粒子或具刺被膜粒子。４、Ｐ－粒子移至管端．或融合或单个通过周质内质网（ＣＥＲ）,释放内容物参与管端壁的形成,被膜小池和小泡移至花粉管次顶端区向质膜外分泌,参与花粉管壁内层的形成,或移至管端,提供膜片。最后讨论了亲和性与超微结构特征的关系．     

肿瘤抑制基因APCmRNA在豚鼠脑中的分布

APC基因是1991年被发现的一类肿瘤抑制基因,它被定位于人第5号染色体5q21处。APC基因如发生缺失或突变,则易患直肠肿瘤,并伴有部分先天痴呆的病例。本工作在孟帆已获得的APC基因在豚鼠中的同源cDNA的基础上,完成了对它的亚克隆,并利用原位杂交和RNA酶保护分析的方法,对它在脑中的分布进行了研究。发现APCmRNA主要在海马、大脑和小脑中表达;嗅球中杂交信号稍弱,脑干中最弱。海马中阳性细胞主要是锥体细胞,小脑中则主要是内层颗粒细胞。在一个月大的豚鼠胚胎的脑中也观察到相似的表达型式。进一步的研究有助于我们更好地了解神经发育和先天痴呆发生的分子机制。     

一个恢复力受单基因控制的水稻CMS育性回复突变体

利用￣（６０）Ｃｏ－γ射线对具有印尼水田谷细胞质的籼稻细胞质雄性不育系Ⅱ－３２Ａ干种子进行诱变处理,获得了一育性回复突变体Ｔ２４。育性基因未纯合的突变体分离出可育株和完全不育株,比例为３∶１;其与Ⅱ－３２Ａ和珍汕９７Ａ测交,Ｆ１代分离出１∶１的可育株和不育株。育性稳定株系与Ⅱ－３２Ａ和珍汕９７Ａ杂交,Ｆ２分离成３∶１的可育株和不育株。表明其育性回复是由一对基因显性突变所致。这一突变体对不育系的育性恢复机制不同于明恢６３、２０９６４等恢复系,后者表现为两对显性恢复基因作用。未观察到Ｔ２４与亲本Ⅱ－３２Ａ除育性以外的其他性状的差异,因而两者构成育性恢复基因的近等基因系。本文还对不育系育性回复类型和Ｔ２４的理论意义与育种价值进行了讨论。     

钙调素对细胞周期的调节

RC3细胞是一种用真核表达载体1~(CaM)转染NIH 3T3细胞建成的可调钙凋素(Calmodulin,CaM)高表达细胞模型。通过分子杂交及蛋白免疫印迹方法证实在地塞米松(Dexamethasome,DXM)作用下,RC3细胞可高表达CaM。CaM的过表达使G_1期细胞减少,S期细胞增加;CaM拮抗剂三氟拉嗪(trifluoperazine,TFP)则使G_1期细胞增加,S期细胞减少。高表达CaM使细胞分裂指数提高,G_2期细胞减少,有丝分裂前期细胞增加,M中期细胞比例下降。而TFP处理则使分裂指数下降,G_2期细胞增加,M前期细胞减少,M中期细胞增加。实验结果表明CaM在G_1/S、G_2/M和M中期/M后期3个位点上对细胞周期进行调控;通过加速G_1至S期,G_2至M期和M中期至M后期的进程,使细胞倍增时间缩短,促进细胞增殖。本工作表明,RC3细胞作为CaM表达可调细胞模型,是研究细胞周期调控的有力工具。     

中药凌霄花的药理学考察

实验结果表明,凌霄花对离体猪冠状动脉条具有抑制收缩的作用;对大鼠血栓形成有抑制作用;能加快红细胞电泳,增加红细胞电泳率,使血液红细胞处于分散状态;对离体孕子宫能显著增强收缩活性,增加收缩频率及增强收缩强度。以上结果可初步说明,凌霄花行血去瘀的作用及其“孕妇慎用”的合理性。凌霄花的３种混淆品（白泡桐花,毛泡桐花,兰考泡桐花）的药理作用与凌霄花不同,故不能作凌霄花使用。     

精制蚕蛹油中碱液和过氧化值的控制

作者采用碱皂化的方法来达到精炼蚕蛹油的目的。实验表明,碱的浓度以１０％￣１３％比较合适,超碱量必须视酸值大小,通过小样试验来确定。精制油的过氧化值与皂化后中性油的洗涤温度、洗涤次数及搅拌速度有密切关系,必须严格控制。