海水胁迫对菠菜叶绿素代谢的影响

以耐海水品种‘荷兰3号’和海水敏感品种‘圆叶菠菜’为试验材料,采用营养液栽培,研究海水胁迫对菠菜叶绿素代谢的影响。结果表明,海水胁迫下,2个菠菜品种叶片的叶绿素a(Chl a)、叶绿素b(Chl b)和总叶绿素含量以及叶绿素合成前体———原叶绿素酸(Pchl)、镁原卟啉IX(Mg-proto IX)、原卟啉IX(Proto IX)和尿卟啉原III(UroIII)含量均明显降低,而胆色素原(PBG)和δ-氨基酮戊酸(ALA)积累,‘圆叶菠菜’的变化幅度大于‘荷兰3号’。海水胁迫下,‘荷兰3号’叶片的叶绿素酶(Chlase)活性无显著变化,胆色素原脱氨酶(PBGD)和尿卟啉原III合酶(UROS)活性在胁迫第3天显著下降,而‘圆叶菠菜’Chlase活性显著上升,PBGD和UROS活性显著下降。研究发现,在海水胁迫条件下,菠菜叶片的叶绿素合成代谢受阻,受阻位点位于PBG→UroIII的转化过程,其中‘圆叶菠菜’的受阻程度大于‘荷兰3号’;耐海水品种‘荷兰3号’叶片叶绿素含量降低主要由叶绿素合成代谢受阻引起,而海水敏感品种‘圆叶菠菜’叶绿素含量的降低则是由叶绿素合成受阻和叶绿素降解共同作用的结果。     

外源水杨酸对镉胁迫下甜瓜幼苗生长、光合作用和活性氧代谢的缓解效应

以甜瓜耐镉品种‘哈密绿’和镉敏感品种‘秀绿’为试验材料,在人工气候箱内采用基质栽培法,研究了外源水杨酸(SA)处理对镉(Cd)胁迫下甜瓜幼苗生长、光合作用和活性氧代谢的缓解效应。结果显示:与对照(CK)比较,Cd处理显著抑制了甜瓜幼苗的生长和光合作用,降低了叶绿素含量,抗氧化酶活性、脯氨酸(Pro)含量、丙二醛(MDA)含量和超氧阴离子(O2ˉ·)产生速率增加;在Cd胁迫下,SA处理可以有效促进甜瓜幼苗的生长,增加了叶绿素含量、净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)、气孔限制值(Ls)和水分利用效率(WUE),降低了胞间CO2浓度(Ci);提高了超氧化物歧化酶(SOD)、过氧化酶酶(POD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX),增强植株抗氧化能力,使Pro和可溶性蛋白含量升高,MDA含量和O2ˉ·产生速率下降,能有效抑制镉胁迫引起的膜脂过氧化伤害;镉敏感品种‘秀绿’变化幅度大于耐镉品种‘哈密绿’。研究结果说明,SA有利于甜瓜幼苗在Cd胁迫下活性氧代谢的提高和对光能的捕获与转换,促进了甜瓜幼苗的生长,降低Cd胁迫对甜瓜幼苗的抑制作用,且对镉敏感品种‘秀绿’效果大于耐镉品种‘哈密绿’。     

弱光胁迫对花生功能叶片RuBP羧化酶活性及叶绿体超微结构的影响

间作套种是我国主要的花生(Arachis hypogaea)种植方式之一。然而, 与单作相比, 在间作套种体系中, 花生截获的光能较少, 生长发育差, 产量低, 研究不同品种耐阴机理对选择适宜间作套种的花生品种具有重要意义。该研究用耐阴性不同的两个花生品种‘花育22号’ (强耐阴性)和‘白沙1016’ (弱耐阴性)为材料, 在大田条件下采用不同透光率遮阴网设置50%自然光强(中度弱光胁迫)和15%自然光强(严重弱光胁迫) 2个弱光处理, 从出苗期开始遮阴40天, 以自然光强为对照, 研究了弱光胁迫对花生功能叶片RuBP羧化酶活性和叶绿体超微结构的影响。结果表明: 光强为自然光照50%和15%的处理, ‘花育22号’ RuBP羧化酶活性与对照相比虽有降低, 但差异不显著, 而‘白沙1016’分别比对照低40.1%和59.4%, 显著低于对照。与对照相比, 50%自然光强下‘花育22号’叶绿体数不变, 叶绿体基粒数和基粒片层数显著增多, 叶绿体变长且发育完好, 15%自然光强下, 叶绿体数、基粒数和淀粉粒数显著减少, 叶绿体膜和基粒片层出现破损, 但叶绿体变长, 基粒片层数增加; ‘白沙1016’在50%自然光强下, 叶绿体数目和超微结构变化同‘花育22号’相似, 在15%自然光强下叶绿体变圆, 基粒数的降幅和基粒片层破损程度大于‘花育22号’且基粒片层数减少, 淀粉粒数增多。因此, 弱光胁迫特别是严重弱光胁迫条件下, 功能叶RuBP羧化酶活性降低幅度小、叶绿体超微结构受损程度低是‘花育22号’耐阴的光合生理基础。     

低氮胁迫对不同苦荞品种苗期生长和根系生理特征的影响

以2个耐低氮苦荞品种(‘迪庆苦荞’、‘广苦1号’)和2个不耐低氮苦荞品种(‘西荞1号’、‘黑丰1号’)为材料,采用盆栽试验研究了低氮胁迫对苦荞苗期生长和根系生理特性以及氮素利用的影响,以揭示苦荞耐低氮胁迫的生理响应机制。结果显示:(1)与正常供氮处理(15mmol/L)相比,低氮(0.5mmol/L)和极低氮(0.05mmol/L)胁迫处理下,苦荞幼苗的株高、茎粗、叶面积、地上部干重、根系平均直径、根系体积及根系表面积均呈下降趋势,主根长和根冠比呈升高趋势,但在低氮(0.5mmol/L)条件下各品种根系干重均有所增加,且耐低氮品种增加幅度较大(13.69%、19.26%)。(2)低氮胁迫使苦荞叶片的叶绿素含量、最大荧光产量(Fm)和最大光化学效率(Fv/Fm)以及根系活力、硝酸还原酶(NR)活性和可溶性蛋白含量显著降低,而叶片初始荧光产量(Fo)及根系超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性、丙二醛(MDA)、可溶性糖及游离脯氨酸含量呈升高趋势。(3)低氮胁迫使苦荞植株全氮含量及氮积累量显著降低,却使氮素吸收效率显著升高。研究表明,不同耐低氮苦荞品种对低氮胁迫的响应存在显著差异,在低氮胁迫下,耐低氮品种(‘迪庆苦荞’、‘广苦1号’)具有明显的生长优势,不仅农艺性状、叶片光合作用及根系活性受低氮胁迫影响小,而且具有较高的根系保护酶活性、渗透调节物质含量以及全氮量和氮积累量。     

低氧胁迫对不同耐性黄瓜品种根系抗氧化系统的影响

低氧胁迫下,两品种黄瓜幼苗根中活性氧、丙二醛(MDA)和抗氧化剂含量以及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和脱氢抗坏血酸还原酶(DHAR)活性均显著提高,耐低氧性较强的‘绿霸春4号’根中活性氧和丙二醛含量上升幅度小于耐低氧性较弱的‘中农8号’,而保护酶活性和还原型抗氧化剂含量提高幅度大于‘中农8号’。     

30个不结球白菜品种的耐弱光性鉴定

用人工遮荫技术对不同来源的30个不结球白菜品种进行弱光处理,以干物质减少百分率、叶片厚度减少百分率和叶绿素a/b减少百分率3个指标作为评价标准,用系统聚类分析方法,将30个供试不结球白菜品种按其耐弱光性分为3类,即耐弱光性强、耐弱光性中等和不耐弱光品种。其中耐弱光性强的品种6个,占全部样品的20%;耐弱光性中等的品种15个,占50%;不耐弱光的品种9个,占30%。方差分析结果表明,3类品种间的耐弱光性指标差异极显著。其中,品种暑绿、正大抗热青3号、热优2号、抗热605、矮王、华王属于耐弱光品种。     

盐胁迫下不同葡萄砧木的渗透调节及抗氧化能力

该研究以13种葡萄砧木品种为材料,采用盆栽试验用100 mmol/L NaCl的1/2 Hoaglands营养液浇灌处理,以葡萄砧木无盐胁迫为对照,待砧木长至20片完全展开叶后测定各品种叶片的相对电导率、渗透调节物质含量、MDA含量、活性氧含量、抗氧化酶活性等指标,探究不同葡萄砧木耐盐能力及其机制。结果显示：(1)至盐胁迫处理结束时,葡萄砧木 ‘188 08’、‘3309C’、‘140R’的处理组叶片近乎完全脱落,‘贝达’、‘5BB’、‘Dogridge’有少部分叶片脱落,而‘101 14’和‘110R’叶枯现象最少。(2)盐胁迫条件下,耐盐性较差的品种‘3309C’和‘140R’叶片相对电导率均大于90%,而耐盐性较强的品种‘101 14’和‘110R’的叶片相对电导率在60%左右;随着盐胁迫处理天数的增加,‘110R’的MDA含量呈明显的上升趋势。(3)盐胁迫条件下,耐盐性较强的品种‘101 14’和‘110R’积累了较多的可溶性蛋白、可溶性糖、脯氨酸等渗透调节物质,而耐盐性弱的品种‘3309C’则积累了较多的活性氧,引起明显的脂膜过氧化,严重影响砧木的正常代谢。(4)在盐胁迫条件下,耐盐品种‘101 14’的SOD、POD等抗氧化酶活性显著提高,有效清除体内过量的活性氧,从而维持细胞膜的稳定性。研究发现,葡萄砧木品种‘101 14’、‘110R’在盐胁迫下能积极积累较多渗透调节物质,增强自身抗氧化酶活性,表现出较强的渗透调节和抗氧化能力,有效抵御盐胁迫造成的伤害,可作为西北盐渍化地区葡萄栽培的砧木品种。     

干旱胁迫对不同抗旱性棉花品种抗氧化酶活性及基因表达的影响

为明确不同抗旱性棉花品种对干旱胁迫的适应性差异,该研究选用‘新陆早50号’(耐旱型)和‘新陆早27号’(干旱敏感型)为材料,分析干旱及复水处理下两材料的活性氧产生、电解质渗漏、抗氧化酶活性变化,并分析部分抗氧化酶基因、渗透调节基因和转录因子在干旱胁迫下的表达谱。结果显示:(1)棉花叶片的电导率和MDA含量随着干旱胁迫的加强逐渐增加,复水后恢复;在干旱胁迫处理6和8d,耐旱型品种‘新陆早50号’的抗氧化酶活性显著高于干旱敏感型品种‘新陆早27号’。(2)基因表达谱分析显示,棉花超氧化物歧化酶基因(GhSOD)仅在干旱胁迫下表达,且2个品种间差异不显著,GhBADH、GhNCED和GhP5Cs及转录因子GhPHD1、GhPHD5、GhPHD6和GhPHD10在‘新陆早50号’叶片中的表达量均高于‘新陆早27号’。研究表明,耐旱型棉花品种较干旱敏感型在干旱胁迫下其体内的活性氧含量、电导率和MDA含量较低,抗氧化酶活性较高,并且其抗旱相关基因表达量也更高。     

铝胁迫对豇豆幼苗根尖抗氧化系统的影响

选用豇豆铝敏感品种‘S3’和耐铝品种‘T6’为试验材料,研究不同耐铝性豇豆根系伸长和根尖活性氧代谢变化的差异,探讨铝胁迫下不同耐铝性豇豆在活性氧代谢上的差异及其与豇豆耐铝性的关系。结果表明:(1)随着铝处理浓度的增加,2个品种根系伸长均受到抑制,且‘S3’受到的抑制程度大于‘T6’。(2)随着处理铝浓度的升高,2个豇豆品种的根尖O2.-产生速率、H2O2含量、MDA含量及质膜透性都显著增加,且‘S3’的增加幅度大于‘T6’。(3)铝胁迫处理下,2个豇豆品种幼苗根尖超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性都有所上升,其中铝敏感品种‘S3’的SOD活性上升幅度比耐铝品种‘T6’高,而POD、CAT、APX的上升幅度比耐铝品种‘T6’低。研究认为,铝胁迫下铝敏感品种‘S3’内活性氧物质过量积累而导致氧化胁迫,使细胞的脂质过氧化程度加剧,最终严重影响根系的生长。     

低氧胁迫对苹果砧木幼苗生长与生理特性的影响

以苹果砧木‘平邑甜茶’(Malushupehensis Rehd.)和‘变叶海棠’(Malustoringoides Rehd.)为材料,采用营养液培养的方法,研究了低氧胁迫对苹果砧木幼苗生长和生理特性的影响。结果表明:低氧胁迫下苹果砧木幼苗新叶数减少,根系和最长新根长度下降,叶片中叶绿素a(Chla)、总叶绿素(Chl)和类胡萝卜素(Car)含量降低,并引起不同色素间比例的改变;根系活力先升后降并在胁迫中后期低于对照;根、叶中超氧阴离子(O2■)产生速率和过氧化氢(H2O2)含量持续增加,丙二醛(MDA)含量上升,膜透性增大,且根系变化幅度大于叶片;低氧耐性较强的‘平邑甜茶’除根系活力外其余指标变化幅度均小于耐性较弱的‘变叶海棠’。说明低氧胁迫影响了苹果砧木幼苗的根系活力和叶片光合色素水平,抑制了植株的生长,导致其体内活性氧自由基(ROS)过量积累,加重膜脂质过氧化作用,从而使植株受到伤害,且这种影响存在基因型和组织差异,根系和低氧耐性较弱的砧木受到的伤害较重。     

ent-Kaurene biosynthesis in a cell-free system from wheat (Triticum aestivum L.) seedlings and the localisation of ent-kaurene synthetase in plastids of three species

A cell-free system capable of converting [¹⁴C]geranylgeranyl diphosphate to ent-[¹⁴C]kaurene and to an unidentified acid-hydrolysable compound was obtained from the basal portions of 5-d-old shoots of wheat seedlings (Triticum aestivum L.). By means of marker enzyme activities, the synthesis of ent-kaurene and the unknown compound could be quantitatively assigned to a plastid fraction obtained by Percoll-gradient centrifugation of the homogenate. The enzyme activities were located within the plastids, probably in the stroma, because they withstood trypsin treatment of the intact plastids, and the plastids had to be broken to release the activity, which was then obtained in soluble form. Plastid membranes had no activity. Plastid stroma preparations obtained from pea (Pisum sativum L.) shoot tips and pumpkin (Cucurbita maxima L.) endosperm also yielded ent-kaurene synthetase activity, but did not form the unknown compound. The exact nature of the active plastids was not ascertained, but the use of methods for proplastid isolation was essential for full activity, and the active tissues are all known to contain high proportions of proplastids, developing chloroplasts or leucoplasts. We therefore believe that ent-kaurene synthesis may be limited to these categories. Mature chloroplasts from the wheat leaves did not contain ent-kaurene synthetase activity and did not yield the unknown component. Incorporation of [¹⁴C]geranylgeranyl diphosphate into ent-[¹⁴C]kaurene and the unknown component was assayed by high-performance liquid chromatography with on-line radiocounting. ent-[¹⁴C]Kaurene was identified by Kovats retention index and full mass spectra obtained by combined gas chromatography-mass spectrometry. The unknown component was first believed to be copalyl diphosphate, because it yielded a compound on acid hydrolysis, which migrated like copalol on high-performance liquid chromatography and gave a mass spectrum very similar to that of authentic copalol. However, differences in the mass spectrum and in retention time on capillary gas chromatography excluded identity with copalol. Furthermore, the unhydrolysed compound was not converted to ent-kaurene by a cell-free system from C. maxima endosperm as copalyl diphosphate would have been.Abbreviations ADH alcohol dehydrogenase - AMO 1618 2isopropyl-4-(trimethylammoniumchloride)-5-methylphenyl piperi-dine-1-carboxylate - BSA bovine serum albumin - DTT dithioth-reitol - GA_n gibberellin A_n - GAPDH NADP⁺-glyceraldehyde 3-phosphate dehydrogenase - GC-MS combined gas chromatography-mass spectrometry - GGPP all trans-isomer of geranyl-geranyl diphosphate - KS ent-kaurene synthetase - MDH malate dehydrogenase - MAA mevalonate activating activity - SOR shikimate oxidoreductase We thank Mrs. Gudrun Bodtke and Mrs. Dorothee Dasbach for able technical assistance, Prof. L.N. Mander (Australian National University, Canberra, Australia) for ent-[²H₂]kaurene and Dr. Yuji Kamiya (RIKEN, Saitama, Japan) for geranylgeraniol and copalol. The work was supported by the Deutsche Forschungsgemeinschaft.     

Identification and localisation of a nucleoporin-like protein component of the plant nuclear matrix

Salt-detergent extraction of purified plant nuclei yields a fraction enriched in putative structural proteins known as the nuclear matrix. Compared with mammalian nuclear matrices, which contain three major proteins, plant nuclear matrices are complex, containing at least 100 polypeptides. In order to characterise more fully the plant nuclear matrix we have used antibodies raised against both yeast (Saccharomyces cerevisiae) and mammalian (rat) nuclear pore proteins. We have shown that the nuclear matrix of carrot (Daucus carota L.) contains at least one nucleoporin-like protein of about 100 kDa which is immunologically related to both the yeast nuclear pore protein NSP1 and mammalian nucleoporins (p62). Antibody labelling of a variety of plant cells at the light-microscope and electron-microscope levels confirms that this antigen is located at the nuclear pores. This, to our knowledge, is the first identification of a nuclear pore protein in plants.Abbreviations IgG immunoglobulin G - kDa kilodaltons - DAPI 4,6-diamidino-2-phenylindole - FITC fluorescein isothioganate The authors would like to thank Dr. E. Hurt (European Molecular Biology Laboratory, Heidelberg, FRG) for antibodies against yeast nucleoporins, and Dr. L. Davis (Whitehead Institute for Biomedical Research, Cambridge, Mass., USA) for the monoclonal antibodies MAb 414 & 350. We thank Brian Wells for useful advice on electron microscopy. We also thank Peter Scott, Andrew Davis, and Nigel Hannant for photography, and Sue Bunnewell for development and printing of electronmicrographs.     

Autoradiographic studies on the role of plasmodesmata in the transport of gibberellin

Translocation of [¹⁴C]gibberellic acid into antheridial cells of Chara vulgaris L. was investigated in relation to the presence of symplasmic connections between the antheridium and the thallus. It was found that manubria, capitular cells, and antheridial filaments were about three-fold more strongly labelled in young antheridia connected to the thallus by plasmodesmata than in older antheridia in which spontaneous symplasmic isolation had occurred. Plasmolytically induced symplasmic isolation of young antheridia severely diminished the radioactivity of all the cells, down to the level characteristic for spontaneously isolated antheridia. It is concluded that plasmodesmata are the main channel of gibberellin transport into antheridia. The change in the character of symplasmic connections during the course of morphogenesis might, among other events, constitute a signal determining a shift of cell metabolism in a new direction, in response to a rapid change in gibberellin level.Abbreviations GA_(n) gibberellin (A_n) - GA₃ gibberellic acid - IAA indole-3-acetic acid This study was supported by the Polish Academy of Sciences research project CPBP 04.01.5.05.     

Effects of experimental infection with Eimeria bovis on the balance of sodium, potassium and water in calves

Balance trials were performed to investigate the effects of experimental Eimeria bovis coccidiosis on the metabolism of water, sodium and potassium in calves. Non-infected pair-fed controls and controls fed according to plan were included in the study to allow differentiation between the effects due to infection and due to changes in feed intake. Primary infection with 5 × 10⁴ (group A) or 1 × 10⁵ (group B) oocysts caused mild diarrhoea in three out of four group A calves and mild to severe haemorrhagic diarrhoea in all five group B calves. Losses of sodium and potassium via faeces tended to increase in the infected calves during patency and apparent digestibility (AD) of these minerals was comparably low. In the urine of the infected calves the Na/K-ratio decreased due to a reduced urinary excretion of sodium. The retention (RT) of sodium was particularly high in the calves that had received the higher oocyst dose. Potassium RT did not underlie significant changes during the course of coccidiosis. In the infected calves the plasma level of sodium was reduced transiently while the level of potassium remained fairly stable. Infections with the higher oocyst dose caused a distinct reduction of fluid excretion via urine which compensated for the increased faecal water losses during severe diarrhoea. Reinfection of the group A calves with 1 × 10⁵ oocysts did not cause any significant metabolic impairment. The results of this study indicate that although acute sublethal bovine coccidiosis alters electrolyte and water metabolism the overall balance of electrolytes and water is largely maintained by physiologic adaptation.     

Identification of keratins and analysis of their expression in carp and goldfish: comparison with the zebrafish and trout keratin catalog

With more than 50 genes in human, keratins make up a large gene family, but the evolutionary pressure leading to their diversity remains largely unclear. Nevertheless, this diversity offers a means to examine the evolutionary relationships among organisms that express keratins. Here, we report the analysis of keratins expressed in two cyprinid fishes, goldfish and carp, by two-dimensional polyacrylamide gel electrophoresis, complementary keratin blot binding assay, and immunoblotting. We further explore the expression of keratins by immunofluorescence microscopy. Comparison is made with the keratin expression and catalogs of zebrafish and rainbow trout. The keratins among these fishes exhibit a similar range of molecular weights and isoelectric points, with a similar overall pattern on two-dimensional gels. In addition, immunofluorescence microscopy studies of goldfish and carp tissues have revealed the expression of keratins in both epithelial and mesenchymally derived tissues, as reported previously for zebrafish and trout. We conclude that keratin expression is qualitatively similar among these fishes, with goldfish and carp patterns being more similar to each other than to zebrafish, and the cyprinid fishes being more similar to each other than to the salmonid trout. Because of the detected similarity of keratin expression among the cyprinid fishes, we propose that, for certain experiments, they are interchangeable. Although the zebrafish distinguishes itself as being a developmental and genetic/genomic model organism, we have found that the goldfish, in particular, is a more suitable model for both biochemical and histological studies of the cytoskeleton, especially since goldfish cytoskeletal preparations seem to be more resistant to degradation than those from carp or zebrafish. This work was supported by grants to J.M. from the Stiftung Rheinland Pfalz für Innovation (836-386261/138) and the Deutsche Forschungsgemeinschaft (Ma 843/5-1) and a grant to D.G. from the National Science Foundation (INT-0078261).     

Detection of lectins in nodulated peanut and soybean plants

The direct double-antibody enzymelinked immunosorbent assay system was used in the detection and measurement of seed lectins from peanut (Arachis hypogaea L.) and soybean (Glycine max L.) plants (PSL and SBL, respectively) that had been inoculated with their respective rhizobia. Concentrations of PSL dropped to undetectable levels in peanut roots at 9 d and stems and leaves at 27 d after planting; SBL could no longer be detected in soybean roots at 9 d and in stems and leaves at 12 d. A lectin antigenically similar to PSL was first detected in root nodules of peanuts at 21 d reaching a maximum of 8 g/g at 29 d then decreasing to 2.5 g/g at 60 d. There was no evidence of a corresponding lectin in soybean nodules.Sugar haemagglutination inhibition tests with neuraminidase-treated human blood cells established that PSL and the peanut nodule lectin were both galactose/lactose-specific. Further tests with rabbit blood cells demonstrated a second mannosespecific lectin in peanut nodule extracts that was not detected in root extracts of four-week-old inoculated plants or six-week-old uninoculated plants, although six-week-old root extracts from inoculated plants showed weak lectin activity. The root extracts from both nodulated and uninoculated plants contained another peanut lectin that agglutinated rabbit but not human blood cells. Haemagglutination by this lectin was, however, not inhibited by simple sugars but a glycoprotein, asialothyroglobulin, was effective in this respect.Abbreviations DAS double antibody sandwich - ELISA enzyme-linked immunosorbent assay - PBS phosphate-buffered saline - PSL peanut seed lectin - SBL soybean lectin     

Distribution of glucose/mannose-specific isolectins in pea (Pisum sativum L.) seedlings

We report on the distribution and initial characterization of glucose/mannose-specific isolectins of 4- and 7-d-old pea (Pisum sativum L.) seedlings grown with or without nitrate supply. Particular attention was payed to root lectin, which probably functions as a determinant of host-plant specificity during the infection of pea roots by Rhizobium leguminosarum bv. viciae. A pair of seedling cotyledons yielded 545±49 g of affinity-purified lectin, approx. 25% more lectin than did dry seeds. Shoots and roots of 4-d-old seedlings contained 100-fold less lectin than cotyledons, whereas only traces of lectin could be found in shoots and roots from 7-d-old seedlings. Polypeptides with a subunit structure similar to the precursor of the pea seed lectin could be demonstrated in cotyledons, shoots and roots. Chromatofocusing and isoelectric focusing showed that seed and non-seed isolectin differ in composition. An isolectin with an isoelectric point at pH 7.2 appeared to be a typical pea seed isolectin, whereas an isolectin focusing at pH 6.1 was the major non-seed lectin. The latter isolectin was also found in root cell-wall extracts, detached root hairs and root-surface washings. All non-seed isolectins were cross-reactive with rabbit antiserum raised against the seed isolectin with an isolectric point at pH 6.1. A protein similar to this acidic glucose/mannose-specific seed isolectin possibly represents the major lectin to be encountered by Rhizobium leguminosarum bv. viciae in the pea rhizosphere and at the root surface. Growth of pea seedlings in a nitrate-rich medium neither affected the distribution of isolectins nor their hemagglutination activity; however, the yield of affinity-purified root lectin was significantly reduced whereas shoot lectin yield slightly increased. Agglutination-inhibition tests demonstrated an overall similar sugar-binding specificity for pea seed and non-seed lectin. However root lectin from seedlings grown with or without nitrate supplement, and shoot lectin from nitrate-supplied seedlings showed a slightly different spectrum of sugar binding. The absorption spectra obtained by circular dichroism of seed and root lectin in the presence of a hapten also differed. These data indicate that nutritional conditions may affect the sugar-binding activity of non-seed isolectin, and that despite their similarities, seed and non-seed isolectins have different properties that may reflect tissue-specialization.Abbreviations IEF isoelectric focusing - MW molecular weight - pI isoelectric point - Psl1, Psl2 and Psl3 pea isolectins - SDSPAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis The authors wish to thank Professors L. Kanarek and M. van Poucke for helpful discussions.     

Spontaneous nodules induce feedback suppression of nodulation in alfalfa

A small subpopulation of alfalfa (Medicago saliva L.) plants grown without fixed nitrogen can develop root nodules in the absence of Rhizobium. Cytological studies showed that these nodules were organized structures with no inter- or intracellular bacteria but with the histological characteristics of a normal indeterminate nodule. Few if any viable bacteria were recovered from the nodules after surface sterilization, and when the nodular content was used to inoculate alfalfa roots no nodulation was observed. These spontaneous nodules were formed mainly on the primary roots in the region susceptible to Rhizobium infection between 4 and 6 d after seed imbibition. Spontaneous nodules appeared as early as 10 d after germination and emerged at a rate comparable to normal nodules. The formation of spontaneous nodules on the primary root suppressed nodulation in lateral roots after inoculation with R. meliloti RCR2011. Excision of spontaneous nodules at inoculation eliminated the suppressive response. Our results indicate that the presence of Rhizobium is not required for nodule organogenesis and the elicitation of feedback regulation of nodule formation in alfalfa.Abbreviation RT root tip This work was supported by an endowment to the Racheff Chair of Excellence of the University of Tennessee, and the Soybean Promotion Board, Haskinsville, Tenn., USA. We are indebted to Noel Gerahty for performing the acetylene-reduction assays, and Dr. E.T. Graham for allowing the use of microscope facilities.     

High-and low-intensity photosystems in Phycomyces phototropism: Effects of mutations in genes madA,madB, and madC

Sporangiophores of Phycomyces blakesleeanus Burgeff that have been grown in darkness and are then suddenly exposed to unilateral light show a two-step bending response rather than a smooth, monotonic response found in light-adapted specimens (Galland and Lipson, 1987, Proc. Natl. Acad. Sci. USA 84, 104–108). The stepwise bending is controlled by two photosystems optimized for the low-and high-intensity ranges. These two photosystems have now been studied in phototropism mutants with defects in genes madA, madB, and madC. All three mutations raise the threshold of the low-intensity (low-fluence) photosystem by about 10⁶-fold and that of the high-intensity (high-fluence) system by about 10³-fold. Estimates for the light-adaptation time constants of the low-and high-intensity photosystems show that the mutants are affected in adaptation. In the mutants, the light-adaptation kinetics are only slightly affected in the low-intensity photosystem but, for the high-intensity photosystem, the kinetics are considerably slower than in the wild type.Abbreviations WT wild type     

The distribution of 5-hydroxytryptamine in the gastrointestinal tract of reptiles,birds and a prototherian mammal

Summary The distribution of 5-hydroxytryptamine in the gut of several species of birds and reptiles, and of a prototherian mammal, the platypus, was studied using a monoclonal antibody. 5-Hydroxytryptamine-like immunoreactivity was found in enterochromaffin cells and, in birds, in thrombocytes. Immunoreactivity was not found in enteric neurons fixed immediately after dissection. A detailed study was made on one avian species, the budgerigar. Following incubation of intestine in physiological solution, immunore-activity was found in nerve fibres in the gut wall that was more marked after incubation with the monoamine oxidase inhibitor pargyline. These fibres took up exogenous 5-hydroxytryptamine. Similar fibres were found in the intestinal nerves and in perivascular plexuses on mesenteric arteries. Both the uptake of 5-hydroxytryptamine and the appearance of neuronal immunoreactivity after incubation were inhibited by the amine uptake inhibitors desmethylimipramine or fluoxetine. Fibres taking up 5-hydroxytryptamine were damaged by pretreatment with 6-hydroxydopamine. It was concluded that the fibres showing immunoreactivity after incubation were adrenergic fibres that had taken up 5-hydroxytryptamine released in vitro from enterochromaffin cells or thrombocytes. These, and more limited observations made on the other species, suggest that birds, reptiles and prototherian mammals lack enteric neurons that use 5-hydroxytryptamine as a transmitter substance.