叶施NO对NaCl胁迫下红砂幼苗叶片和根系中氮代谢酶及营养物质的影响

以当年生红砂(Reaumuria soongorica)幼苗为材料,采用盆栽实验,考察叶面喷施不同浓度(0、0.01、0.10、0.25、0.50、1.00 mmol·L^-1)NO供体硝普钠 (SNP) 对NaCl(300 mmol·L^-1)胁迫下红砂根、叶中可溶性蛋白、游离氨基酸和硝态氮含量,以及谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、硝酸还原酶(NR)活性的影响,并采用主成分分析和隶属函数法筛选NO对NaCl胁迫缓解效应的氮代谢指标和最佳NO浓度,以探讨外源NO对NaCl 胁迫下红砂缓解效应的氮代谢响应机制。结果表明：(1)在300 mmol·L^-1 NaCl胁迫处理下,红砂幼苗根、叶中可溶性蛋白、硝态氮含量以及GS、GOGAT、NR活性均比对照显著下降。(2)外源NO能显著提高盐胁迫下红砂叶、根中GS、GOGAT、NR活性和硝态氮含量,增加根中可溶性蛋白和游离氨基酸含量。(3)NR和GOGAT活性可用于评价NO对NaCl胁迫下红砂幼苗的缓解作用,外源NO(SNP)对红砂幼苗在NaCl胁迫下的缓解效果强弱表现为0.25 mmol·L^-1> 0.50 mmol·L^-1> 0.10 mmol·L^-1> 1.00 mmol·L^-1> 0.01 mmol·L^-1。研究发现,300 mmol·L^-1 NaCl胁迫显著抑制了红砂幼苗氮代谢,外源NO(SNP)有助于提高盐胁迫下红砂NR活性,加快硝态氮转化为铵态氮,促进红砂叶片和根中GS/GOGAT对转化物的同化,从而增强红砂幼苗的耐盐性,并以0.25 mmol·L^-1SNP处理时缓解作用最佳;NR和GOGAT活性可作为NO缓解盐胁迫的评价指标。     

外源Spd对盐碱胁迫下番茄幼苗氮代谢及主要矿质元素含量的影响

采用水培方法,研究了盐碱与Spd处理对两品种番茄(中杂9号和金棚朝冠)幼苗氮代谢及主要矿质元素含量的影响.结果表明:盐碱胁迫下,番茄幼苗干生物量显著减少,植株生长受到抑制;叶片和根系硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合成酶(GOGAT)活性及硝态氮(NO3--N)、全N、全K、Ca2+、Mg2+含量显著降低,铵态氮(NH4+-N)、Na+含量显著增加;两品种叶片及中杂9号根系谷氨酸脱氢酶(GDH)活性显著升高,金棚朝冠根系GDH活性变化不显著;叶片全P含量显著降低,根系全P含量显著升高(金棚朝冠)或无显著变化(中杂9号).Spd处理通过增强NR、GS、GOGAT活性提高了植株对NH4+的同化利用率,有效缓解了盐碱胁迫导致的氮代谢紊乱,进而促进不同器官对P、K、Ca、Mg、Na的吸收、释放或转运,在一定程度上维持了各元素之间的相对平衡,从而增强植株对逆境的适应能力.此外,盐碱对中杂9号的抑制作用及外源Spd对其氮代谢紊乱和营养失衡的缓解作用高于金棚朝冠.     

外源褪黑素对高温胁迫条件下黄瓜幼苗氮代谢的影响

以‘津优4号’（热敏感型）和‘美国保尔’（耐热型）黄瓜幼苗为试材,研究了叶面喷施褪黑素对高温胁迫条件下黄瓜幼苗氮代谢的影响。结果显示,高温胁迫下,（1）两种黄瓜幼苗硝态氮含量先升高后降低,‘津优4号’总氮和氨态氮含量先下降后持续升高,而‘美国保尔’总氮和氨态氮含量持续上升;（2）两种黄瓜幼苗硝酸还原酶（NR）活性均先上升后下降,而谷氨酰胺合成酶（Gs）、谷氨酸合成酶（GOGAT）和谷氨酸脱氢酶（GDH）均持续下降,‘美国保尔’的4种酶活性下降幅度显著低于‘津优4号’。研究结果表明,叶面喷施褪黑素可有效缓解高温胁迫对NR、GS、GOGAT和GDH的抑制作用,显著增加硝态氮含量,降低氨态氮含量,减轻氨态氮积累对黄瓜幼苗造成的毒害作用,增强高温胁迫条件下黄瓜幼苗氮素的代谢能力,减轻高温胁迫对黄瓜幼苗造成的伤害,提高黄瓜幼苗抵御高温胁迫的能力。     

亚低温条件下外源褪黑素对甜瓜幼苗氮代谢及渗透调节物质的影响

以甜瓜品种‘羊角酥瓜’幼苗为试材,利用人工气候室进行亚低温处理(昼/夜18℃/12℃)6 d,研究外源褪黑素(MT)对亚低温条件下甜瓜幼苗叶片氮代谢酶[硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)和谷氨酸脱氢酶(GDH)]活性、叶片总氮、铵态氮、硝态氮和渗透调节物质含量的影响.结果表明:与对照相比,亚低温处理降低了甜瓜总氮、硝态氮及硝酸还原酶活性,增加了铵态氮含量,抑制了甜瓜幼苗的生长.外源MT可显著提高亚低温下甜瓜幼苗氮代谢酶的活性,尤其可显著提高叶片GS和GOGAT活性,从而有效降低铵态氮含量;外源MT还可以提高叶片脯氨酸、可溶性蛋白质和可溶性糖的含量,进而降低了亚低温对细胞膜的伤害,表现为甜瓜叶片相对电导率和丙二醛(MDA)含量较低.总之,在亚低温条件下,外源MT在一定程度上可通过降低甜瓜叶片铵态氮含量和促进渗透调节物质的积累,降低膜质过氧化水平,从而增加其对亚低温的适应性.     

亚低温条件下外源褪黑素对甜瓜幼苗氮代谢及渗透调节物质的影响

以甜瓜品种‘羊角酥瓜’幼苗为试材,利用人工气候室进行亚低温处理(昼/夜18 ℃/ 12 ℃)6 d,研究外源褪黑素（MT）对亚低温条件下甜瓜幼苗叶片氮代谢酶\[硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)和谷氨酸脱氢酶(GDH)\]活性、叶片总氮、铵态氮、硝态氮和渗透调节物质含量的影响.结果表明：与对照相比,亚低温处理降低了甜瓜总氮、硝态氮及硝酸还原酶活性,增加了铵态氮含量,抑制了甜瓜幼苗的生长.外源MT可显著提高亚低温下甜瓜幼苗氮代谢酶的活性,尤其可显著提高叶片GS和GOGAT活性,从而有效降低铵态氮含量;外源MT还可以提高叶片脯氨酸、可溶性蛋白质和可溶性糖的含量,进而降低了亚低温对细胞膜的伤害,表现为甜瓜叶片相对电导率和丙二醛(MDA)含量较低.总之,在亚低温条件下,外源MT在一定程度上可通过降低甜瓜叶片铵态氮含量和促进渗透调节物质的积累,降低膜质过氧化水平,从而增加其对亚低温的适应性.     

外源Spd对盐碱胁迫下番茄幼苗氮代谢及主要矿质元素含量的影响

采用水培方法,研究了盐碱与Spd处理对两品种番茄（中杂9号和金棚朝冠）幼苗氮代谢及主要矿质元素含量的影响.结果表明： 盐碱胁迫下,番茄幼苗干生物量显著减少,植株生长受到抑制;叶片和根系硝酸还原酶（NR）、谷氨酰胺合成酶（GS）、谷氨酸合成酶（GOGAT）活性及硝态氮（NO₃^--N）、全N、全K、Ca²⁺、Mg²⁺含量显著降低,铵态氮（NH₄⁺-N）、Na⁺含量显著增加;两品种叶片及中杂9号根系谷氨酸脱氢酶（GDH）活性显著升高,金棚朝冠根系GDH活性变化不显著;叶片全P含量显著降低,根系全P含量显著升高（金棚朝冠）或无显著变化（中杂9号）.Spd处理通过增强NR、GS、GOGAT活性提高了植株对NH₄⁺的同化利用率,有效缓解了盐碱胁迫导致的氮代谢紊乱,进而促进不同器官对P、K、Ca、Mg、Na的吸收、释放或转运,在一定程度上维持了各元素之间的相对平衡,从而增强植株对逆境的适应能力.此外,盐碱对中杂9号的抑制作用及外源Spd对其氮代谢紊乱和营养失衡的缓解作用高于金棚朝冠.     

分蘖期冷水胁迫下施氮量对寒地粳稻氮代谢和籽粒蛋白质含量的影响

以寒地粳稻东农428、龙稻7和松粳10为试验材料,研究分蘖期冷水胁迫下不同施氮量处理对功能叶片氮代谢关键酶活性、籽粒蛋白质含量以及产量的影响,并探讨功能叶片氮代谢关键酶活性与籽粒蛋白质含量以及产量的关系。研究结果表明,分蘖期冷水胁迫使NR,GS活性及籽粒蛋白质含量升高,GOGAT和GDH活性下降。分蘖期冷水胁迫下,增加施氮量使各品种功能叶酶活性及籽粒蛋白质含量均缓慢升高,其中NR、GS和GOGAT活性在N150处理下最高,GDH活性和籽粒蛋白质含量在N175处理下最高,可知,高施氮量不利于寒地粳稻抗冷。不同品种在冷水胁迫下酶活性表现不一,其中耐冷型品种东农428在各处理下氮代谢关键酶保持较高生理活性,龙稻7次之,松粳10最弱。分蘖期冷水胁迫下,纯氮施用量为100 kg/hm2,其产量表现最佳,3个品种表现一致。     

氮素形态对黄檗幼苗生长及氮代谢相关酶类的影响

通过改变水培溶液中NH4+-N和NO3--N的比例, 研究了不同氮素形态对黄檗(Phellodendron amurense)幼苗生长及氮代谢相关酶类的影响。结果表明, 硝态氮比例较高的营养供给比铵态氮比例较高的营养供给有利于黄檗幼苗的生长, 叶片叶绿素含量和可溶性蛋白含量也高。在NH4+-N/NO3--N为25/75 时黄檗幼苗具有最大生物量。在铵态氮比例大的营养供给下, 黄檗幼苗的谷氨酰胺合成酶(GS)活性增强,而在硝态氮比例大的营养供给下幼苗的硝酸还原酶(NR)活性则较高, 叶片中的硝态氮较低。营养液的氮素形态及其组成通过影响GS与NR的活性而调控黄檗幼苗的氮素代谢。     

纯化腐植酸对氮胁迫下黄瓜幼苗生长和氮代谢的影响

为研究纯化腐植酸(PHA)在不同水平氮胁迫下对黄瓜植株生长和氮代谢的影响,探明PHA对逆境胁迫的缓解作用机制,采用水培方式,选用新泰密刺为供试品种,以正常氮水平(11 mmol·L^-1 NO₃^-)为对照,进行低氮(1.0 mmol·L^-1 NO₃^-)和高氮(101 mmol·L^-1 NO₃^-)胁迫处理,研究纯化腐植酸对氮胁迫下黄瓜幼苗生长和氮代谢的影响.结果表明: 低氮和高氮胁迫均抑制了黄瓜幼苗生长,株高、茎粗、叶面积、干物质积累量均低于正常氮水平处理.施用纯化腐植酸促进了正常氮水平及低氮胁迫下黄瓜干物质积累,在高氮胁迫下差异不显著.PHA影响了黄瓜幼苗NO₃^-的吸收,呈低氮下促进、高氮下抑制吸收的趋势;PHA显著降低了低氮、高氮胁迫下根系和叶片中的铵态氮含量;与正常氮水平相比,低氮、高氮胁迫下根系和叶片的硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、谷氨酸脱氢酶(GDH)活性及根系中亚硝酸还原酶(NiR)活性均显著降低,PHA不同程度地提高了NR、NiR、GS、GOGAT、GDH活性,还提高了根系和叶片中游离氨基酸、可溶性蛋白的含量.综上,添加PHA缓解了氮胁迫对黄瓜幼苗生长的抑制作用.     

不同氮素形态及其配比对盐胁迫下紫苏生理特性的影响

以60 mmol·L~(-1)NaCl处理紫苏幼苗为对象,研究不同氮素形态配比对盐胁迫下紫苏生长、叶绿素含量、可溶性蛋白质、抗氧化酶和氮素代谢相关酶活性的影响,为进一步研究盐胁迫环境下紫苏生产中氮肥的合理调控提供理论依据。结果表明:非盐胁迫下,当NH_4~+-N/NO_3~--N=25∶75时,紫苏地上部分的鲜重和干重、总叶绿素含量、硝酸还原酶(NR)和谷氨酰胺合成酶(GS)活性、可溶性蛋白含量和抗氧化酶活性都显著高于其他氮素处理;在盐胁迫下,紫苏生长受到抑制,叶绿素含量降低,可溶性蛋白和MDA含量增加,抗氧化酶活性升高,氮代谢相关酶NR和GS活性也升高; NaCl胁迫下,当NH_4~+-N/NO_3~--N=25∶75时,紫苏叶片的叶绿素含量、SOD活性、CAT活性和可溶性蛋白含量最高,MDA含量最低;当NH_4~+-N/NO_3~--N=50∶50时,POD活性最高;叶片中NR和GS活性都随着硝态氮比例的增加不断升高,并在NH_4~+-N/NO_3~--N=0∶100达到最大值,酰胺态氮处理组的NR和GS活性均低于全硝处理,但高于全铵处理组;不同氮素形态及配比对盐胁迫下紫苏叶绿素含量、氮素代谢和抗逆调节指标、产量影响显著;研究发现,在盐胁迫下,铵硝混合配施比单独施用全硝、全铵和酰胺态氮效果好,并且NH_4~+-N/NO_3~--N=25∶75处理更有利于维持抗氧化酶、氮代谢酶活性,缓解盐胁迫对紫苏幼苗生长的抑制,促进鲜重和干重的增加,从而维持较高的耐盐性。     

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.     

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.     

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.     

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     

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.     

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     

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).     

The measurement of ethylene binding and metabolism in plant tissue

Methods are described for the rigorous measurement of C₂H₄ metabolism and C₂H₄ binding in plant tissue. Comparisons are drawn between the results obtained using other methods and those which emerge from our studies, indicating that significant misapprehensions may have arisen in relation both to the distribution of metabolism and binding.