外源NO提高小麦幼苗抗旱性的生理机制

以小麦(Triticum aestivum L.)品种'豫麦49'为材料,采用50μmol·L-1SNP(NO供体)处理自然干旱和PEG模拟干旱下的小麦幼苗,分析外源NO对水分胁迫下小麦幼苗相对含水量、光合速率、细胞膜透性以及茎叶中关键性离子含量的影响.结果显示:自然干旱10 d后,对照组幼苗几乎全部枯死,而50μmol·L-1SNP处理幼苗并未发生枯死,其幼苗在旱后复水2 d后能完全恢复正常生长;在25%PEG-6000模拟干旱条件下,50μmol·L-1SNP处理也能明显改善受胁迫小麦幼苗长势.50μmol·L-1SNP处理使模拟干旱胁迫下小麦叶片的相对含水量显著提高15.98%,净光合速率(Pn)、气孔导度(Gs)分别显著提高47.11%和42.86%,而胞间CO2浓度(GI)显著降低了8.19%;也使小麦组织浸出液电导率显著降低30%,茎叶的K 含量显著增加24.55%(P<0.01).研究发现,外源NO既可通过降低小麦幼苗叶片蒸腾来维持较高的叶片相对含水量,缓解因干旱缺水对植株的伤害;又可增加K 在茎叶中积累,减轻干旱胁迫对小麦幼苗细胞膜伤害,维持干旱胁迫下小麦幼苗较高的光合速率,以确保植株正常生长和有机物质积累.     

一氧化氮对杨树耐旱性的影响

探讨了外源NO对水分胁迫下杨树叶片质膜相对透性、叶片光合作用和氧化伤害保护酶的影响.结果表明,NO供体硝普钠(sodiumnitroprusside,SNP)能提高杨树叶片的含水率,在水分胁迫(PEG6000渗透液处理)下,能缓解叶片的水分丢失.NO对杨树叶片光合作用具有双重性,低浓度SNP(200、500μmol·L^-1)能促进叶片的光合,高浓度SNP(1000、2000μmol·L^-1)则明显抑制叶片的光合.较短时间水分处理胁迫(1h))的杨树叶片SOD和POD活性显著高于较长时间(3h)水分胁迫下叶片的酶活性.经SNP处理后,各处理组POD、SOD活性明显上升.同时,随SNP浓度的增加,POD和SOD活性表现出先上升后下降的趋势.外源NO可通过诱导POD和SOD活性的上升,延缓活性氧的积累,从而减轻水分胁迫对杨树的伤害,增强树木的耐旱能力.     

外源NO对酸雨胁迫下龙眼幼苗生理特性的影响

研究了外源NO供体硝普钠(SNP)对pH3.0酸雨胁迫下龙眼幼苗叶绿素含量、抗氧化酶活性和渗透调节物质含量的影响.结果表明:酸雨胁迫下,龙眼幼苗叶片中的超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性降低,叶绿素、蛋白质和可溶性糖含量下降,丙二醛含量升高,呈现出一定的毒害效应.外源NO对酸雨胁迫下龙眼幼苗的生理代谢的作用具有双重性,0.1 ～0.5.mmol·L-1SNP显著提高叶绿素、可溶性糖和蛋白质含量,增强SOD、POD和CAT活性,降低丙二醛含量;其中0.5 mmol·L-1 SNP处理效果最好,叶绿素、可溶性糖和蛋白质含量分别比单独酸雨胁迫处理增加了76.0％、107.0％和216.1％,SOD、POD和CAT活性分别提高150.0％、350.9％和97.1％,丙二醛含量降低了46.4％.低浓度外源NO能通过刺激抗氧化酶活性来清除活性氧,减轻氧化胁迫,缓解酸雨胁迫对龙眼幼苗的毒害作用;而高浓度外源NO对酸雨胁迫的缓解作用减弱.     

渗透胁迫下外源茉莉酸甲酯对油菜种子生理特性的影响

采用PEG模拟水分胁迫的方法,研究在5%PEG-6000胁迫下,不同浓度(0.025、0.05、0.1 mmol·L-1)茉莉酸甲酯(MeJA)对油菜种子萌发、幼苗生长、叶片氧化损伤和保护酶活性等生理生化特性的影响.结果显示:0.1mmol·L-1MeJA处理能显著缓解5%PEG-6000的胁迫伤害,使油菜种子发芽率、发芽势、发芽指数和活力指数分别显著提高62.3%、11.2%、55.2%、126.3%,主根长度降低45.0%,不定根数提高59.4%,但对株高无显著影响;同时叶片MDA含量降低20.6%,SOD、CAT、POD和APX活性分别提高100.0%、109.1%、452.6%和134.8%.研究表明,MeJA在一定程度上能够缓解渗透胁迫的抑制作用,提高SOD等保护酶的活性,缓解渗透胁迫造成的氧化损伤,有效促进渗透胁迫下油菜种子的萌发和幼苗生长.     

外源NO调控小麦幼苗生长与生理的浓度效应

在室内人工控制条件下,研究100-2000 μmol·L-1 5个浓度梯度的SNP(外源NO供体)处理后小麦幼苗的生长、水分生理和光合能力的动态变化,探讨外源NO优化调控小麦幼苗生长生理的最适浓度.结果表明,不同的SNP浓度对小麦幼苗地上部和根系的生长具有不同的促进或抑制作用,低浓度(100～200 μmol·L-1)SNP明显增加了根系的可溶性糖、游离氨基酸等渗透调节物质含量,但对根系的干物质积累影响不大.浓度超过200 μmol·L-1的SNP处理后,根系的干物质积累降于对照以下,渗透调节物质含量也开始降低.低浓度SNP处理(100 μmol·L-1)明显改善了小麦叶片的光合性能从而促进地上部干物质与渗透调节物质的积累,且增加了叶片的水势和渗透势.另外,叶片的气体交换参数(Pn、Gs)和叶绿素荧光参数(qP、NPQ)对SNP的反应也表现出明显的浓度效应.因此,外源NO对小麦幼苗生长生理的调控具有一定的剂量效应,100 μmol·L-1SNP对小麦幼苗生长生理的调控作用最明显.     

外源一氧化氮对镉胁迫下玉米幼苗根生长及氧化伤害的影响

以玉米幼苗为材料,通过在镉处理的同时补充外源一氧化氮(NO)供体硝普钠(SNP)及其类似物[K3Fe(CN)6]、以及NO消除剂,分析NO对植物耐镉性的影响,探讨NO在植物逆境胁迫响应中的作用及其机理。结果显示:添加20μmol·L-1 SNP能显著降低镉引发的玉米幼苗根生长抑制及根尖内源镉的积累,减少电解质的渗漏以及超氧化物自由基(O2.-)和过氧化氢(H2O2)的上升幅度,抑制超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)活性的增加,进一步提高镉胁迫下谷胱甘肽还原酶(GR)的活性。SNP的上述效应可被NO消除剂2-(4-羧基-2-苯基)-4,4,5,5-四甲基咪唑-1-氧-3-氧化物(cPTIO)所逆转,而SNP类似物K3Fe(CN)6的应用对上述反应几乎无影响,说明该反应具有NO特异性。研究表明,外源NO能够显著缓解镉胁迫对玉米幼苗生长造成的伤害,该缓解作用主要是通过降低植株体内内源镉积累和减轻镉诱发的氧化伤害来实现的。     

外源NO对UV-B胁迫下红豆杉抗氧化系统的影响

为探讨一氧化氮(nitric oxide,NO)对紫外线-B( UV-B)辐射胁迫下植物抗氧化系统的影响,以盆栽5年生南方红豆杉(Taxus chinensis var.mairei)幼苗为材料,硝普钠(sodium nitroprusside,SNP)为外源NO供体,设置CK(对照)、SNP(+0.1 mmol·L-1SNP)、UV-B(+4.22 kJ·m-2·d-1 UV-B)及UV-B+ SNP(+0.1 mmol·L-1 SNP+4.22 kJ·m-2·d-1UV-B)4个处理,研究外源NO对UV-B胁迫下南方红豆杉幼苗针叶过氧化氢(H2O2)含量、脂质过氧化程度及抗氧化物质含量的影响.结果表明:UV-B胁迫显著提高了南方红豆杉针叶H2O2及MDA含量(P＜0.05),施加外源NO降低UV-B胁迫下针叶H2O2及MDA含量,提高紫杉醇、类黄酮及类胡萝卜素等抗氧化物质含量(P＜0.05);各处理对抗氧化酶活性影响不同,SNP处理显著提高针叶中CAT和POD活性(P＜0.05),UV-B和SNP+UV-B处理均提高针叶中POD活性,降低CAT活性和APX活性(P＜0.05).本研究证实,外源NO可提高UV-B胁迫下植物抗氧化酶活性和抗氧化物质含量,降低其H2O2含量及脂质过氧化程度,从而在一定程度上缓解UV-B胁迫对植物的伤害.     

外源IAA对NaHCO3胁迫下黄瓜幼苗光合特性和抗氧化系统的影响

以黄瓜‘津研四号’幼苗为试材,采用Hoagland营养液栽培,研究了不同浓度(0、0.01、0.1、1和10μmol·L-1)IAA处理对50 mmol·L-1 NaHCO3胁迫下黄瓜幼苗光合特性及抗氧化系统的影响。结果表明,碱胁迫对黄瓜幼苗的生长有抑制作用,0.01~1μmol·L-1外源IAA处理可显著增加黄瓜幼苗的生物量;使叶中Na+积累降低,K+积累增加,且IAA的缓解效果具有浓度效应。叶绿素a、叶绿素b和类胡萝卜素含量提高,净光合速率(Pn)和气孔导度(Gs)增加,以1μmol·L-1 IAA处理的效果最好。添加1μmol·L-1外源IAA显著提高了碱胁迫下黄瓜叶中超氧化物歧化酶(SOD)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、脱氢抗坏血酸还原酶(DHAR)和谷胱甘肽还原酶(GR)的活性及还原型抗坏血酸(AsA)和谷胱甘肽(GSH)的含量,降低了碱胁迫诱导的活性氧积累和膜脂过氧化反应;而10μmol·L-1外源IAA处理则加剧碱胁迫对黄瓜幼苗的危害。     

外源NO对铜胁迫下番茄幼苗根系构型及其超微结构的影响

采用营养液水培的方法,以“改良毛粉802F1”番茄为材料,硝普钠（sodiumnitroprusside,sNP）为一氧化氮（N0）供体,研究外源N0对铜胁迫下番茄幼苗根系构型及其超微结构的影响。结果表明,50μmol·L-1的铜胁迫下,外施100μmol·L-1 SNP能够显著增加番茄幼苗植株的生物量、株高和茎粗,提高根系活力,改善根系构型中的根长度、根平均直径、根表面积和根体积,缓解番茄幼苗亚细胞结构（细胞核、线粒体、叶绿体、液泡、核膜）的改变,维持番茄幼苗组织结构的稳定,减缓铜胁迫对植株生长的抑制作用,添加NO清除剂牛血红蛋白后,能显著消除NO的缓解效果。     

外源NO 对Cd 胁迫下娃娃菜种子萌发及幼苗生长的保护效应

为探讨外源一氧化氮(NO)对镉(Cd)胁迫下娃娃菜(Brassica pekinensis)的保护效应, 采用0、50、100、200、500 和1000 μmol·L–1 的外源NO 供体硝普钠(SNP)对100 和200 mg·L–1 CdCl2 胁迫下的娃娃菜种子进行处理, 分析其对种子萌发、幼苗生长及生理指标的影响。结果表明: Cd 胁迫抑制娃娃菜种子萌发和幼苗生长, 幼苗叶片叶绿素(Chl)和类胡萝卜素(Car)含量显著降低, 丙二醛(MDA)含量和相对电导率(REC)显著上升, 幼苗叶片膜脂过氧化加速。不同浓度的SNP处理, 不仅能提高Cd 胁迫下娃娃菜种子的发芽率、发芽势、发芽指数和活力指数, 促进幼苗生长, 而且能抑制MDA含量上升, 降低叶片REC, 增加Chl 和脯氨酸(Pro)含量, 增强抗氧化酶活性。外源NO 可缓解Cd 胁迫对娃娃菜种子萌发及幼苗生长的抑制作用, 缓解Cd胁迫引起的膜脂过氧化, 提高抗Cd能力, 其中以200 μmol·L–1 SNP对Cd胁迫的缓解效果较好。     

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.     

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.     

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     

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.     

Observations on enzymatically isolated,living and fixed embryo sacs in several angiosperm species

A technique has been developed for isolating embryo sacs (ESs) by enzymatic maceration. Ovules were macerated in a mixture of pectinase, cellulase and, in some cases, snailase and pectolyase Y-23. The ovular tissues were removed and the ESs were isolated in toto. Embryo sacs were isolated from both fixed and fresh ovules of Antirrhinum majus L., Helianthus annuus L. and Nicotiana tabacum L. Fluorochromasia by fluorescein diacetate showed that the ESs isolated from fresh ovules were viable. The method has promise for various histochemical and cell-physiological studies and quite possibly also for in-vitro culture of ESs.Abbreviations ES embryo sac - FDA fluorescein diacetate - FPA formalin-propionic acid 50% alcohol (5:5:10, by vol.) - H33258 Hoechst 33258     

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