Discrimination between the red- and far-red-absorbing forms of phytochrome from Avena sativa L. by monoclonal antibodies

A set of rat monoclonal antibodies (ARC MAC 48 to 52 and 54 to 56), raised to phytochrome from dark-grown seedlings of Avena sativa L. was tested for the ability to discriminate between the red-absorbing (Pr) and far-red-absorbing (Pfr) forms of phytochrome by indirect enzyme-linked immunosorbent assay. MAC 50 bound more strongly to Pfr and MAC 49 and 52 showed preferential binding to Pr from extracts of dark-grown Avena seedlings; MAC 50 also bound more strongly to Pfr from brushite-purified phytochrome. The remainder of the monoclonal antibodies and a rabbit polyclonal antiphytochrome preparation did not discriminate between Pr and Pfr. The results provide evidence for conformational changes in defined regions of the phytochrome apoprotein upon photoconversion.Abbreviations ELISA enzyme-linked immunosorbent assay - FR far-red light - McAb monoclonal antibody(ies) - PBS phosphate-buffered saline - Pfr far-red-absorbing form of phytochrome - Pr red-absorbing form of phytochrome - R red light - PMSF phenylmethylsulphonylfluoride     

Phosphorus status of some saline and non-saline hydromorphic soils of the Niger Delta of Nigeria

Summary The phosphorus status of some mangrove and fresh-water hydromorphic soils of the Nigerian Niger Delta was evaluated by determining the relative abundance of various P forms and the P-sorption capacity indices. Total P was high in all soils ranging from 352 to 2055 mg/kg, with a mean of 1011 mg/kg. The saline mangrove-swamp soils had generally higher values than the fresh-water soils. Organic P formed about 34% of total P. The relative abundance of the inorganic P forms was in decreasing order, active P, occluded P and residual P. The relative distribution of active P followed the decreasing order, Fe–P, Al–P and Ca–P.The adsorption capacity was generally low in all soils. The amount of P sorbed from the addition of 150 mg/100g of soil ranged from zero to 13 mg/100g, giving an average of about 7% of added P sorbed.The abundance of active P and low content of occluded P were attributed to the poorly drained and unweathered nature of the soils. The low P adsorption suggests little capacity of the soils to fix P. The relatively high content of active P and the low P sorption capacity generally indicate high availability of P to plant in these soils.     

Folding pathway of a circular form of bovine pancreatic trypsin inhibitor

The pathway of unfolding and refolding of a circular form of bovine pancreatic trypsin inhibitor, in which the termini were linked together in a peptide bond, has been examined by trapping and identifying the disulphide-containing intermediates, as was done previously for the unmodified protein. The folding pathway of the circular protein was essentially the same as that of the unmodified inhibitor, although there were differences in the distribution of intermediates that accumulated and in the rates of some steps. The effects of the cross-link between the termini on the stabilities of the folding intermediates and the native state were determined by measuring the rates of the interconversions making up the folding transition, and comparing them with those measured for the unmodified protein. The major effect of the cross-link was to stabilize an intermediate containing two native disulphides, (30-51, 14-38), but lacking the disulphide nearest the termini, 5-55. The native conformation was not measurably stabilized by the cross-link, in spite of the expected reduction of entropy of the unfolded state, indicating that the native state of the circular protein had a slightly strained conformation. The stabilities of the major one-disulphide intermediates were not significantly affected by the cross-link, suggesting that the termini of bovine pancreatic trypsin inhibitor do not tend to interact during the early stage of folding.     

The functional role of molecular forms of acetylcholinesterase in neuromuscular transmission

The severity of poisoning following acetylcholinesterase (AChE) inhibition correlates weakly with total AChE activity. This may be partly due to the existence of functional and non-functional pools of AChE. AChE consists of several molecular forms. The aim of the present study was to investigate which of these forms will correlate best with neuromuscular transmission (NMT) remaining after partial inhibition of this enzyme. Following sublethal intoxication of rats with the irreversible AChE inhibitor soman, diaphragms were isolated after 0.5 or 3 h. It appeared that at 3 h after soman poisoning the percentage of G₁ increased, while those of G₄ and A₁₂ decreased. NMT was inhibited more strongly than in preparations obtained from the 0.5 h rats with the same level of AChE inhibition, but with a normal ratio of molecular forms. NMT correlated positively with G₄ as well as with A₁₂, but inversely with G₁. In vitro inhibition with the charged inhibitors DEMP and echothiophate resulted in higher levels of total AChE, relatively less G₁ and more G₄ and A₁₂ than after incubation with soman, but led to less NMT. Treatment of soman-intoxicated rats with the reactivating compound HI-6 resulted in preferential reactivation of A₁₂, persisting low levels of G₁ and concurrent recovery of NMT as compared with saline-treated soman controls with equal total AChE activity. Apparently, in rat diaphragm G₄ and A₁₂ are the functional AChE forms.     

Molecular forms of lipases and their localization in the fungus Rhizopus microsporus by immuno-electron microscopy

Several lipases differing in their molecular masses (24, 32, 43, 66 and 98 kDa and 28, 40, 45 and 69 kDa) were found in Rhizopus microsporus UzLT-4B and UzLT-5C, respectively. The lipases in each strain were immunologically related. Strain UzLT-5C grown on a medium with lipid substrate secreted lipases of 32, 66 and 98 kDa whereas strain UzLT-4B produced lipases of 45 and 69 kDa on the same medium. Immuno-electron microscopy indicated that the intracellular lipases were in peripheral zones of the hyphae, primarily in the periplasm and adjacent vesicles. Immobilization in Ca-alginate gel revealed unusual structures in the cell wall. These structures accumulated lipases and, apparently, exported the enzymes out of the cell.The authors are with the Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 142292, Pushchino, Moscow Region, Russia     

Biological species and morphological characteristics ofArmillaria mellea complex in Hokkaido:A. sinapina and two new species,A. jezoensis andA. singula

Three intersterility groups ofArmillaria mellea sensu lato were discovered by examining all pairwise combinations of monosporous isolates of basidiomes collected in Hokkaido. One of them, group IV, was identified asA. sinapina by mating it with tester strains. Two new species, groups III and V, were namedA. jezoensis andA. singula, respectively. Their morphological forms and the ecology of their basidiomes are described.     

有机物料对滨海盐渍土重金属根际效应的影响 Ⅰ.土壤内源Zn形态分布

研究了排水条件下施用腐熟有机物料、种稻改良滨海盐渍土内源Zn形态分布。结果表明,单淹水使土中各形态Zn一定程度上向生物有效性较低的Zn形态转化,有效态Zn降低。土壤盐分量不同,明显影响无定形氧化铁结合态、紧结有机态以及硅酸盐矿物态Zn变化。种稻不施有机物料,根际交换态和硅酸盐矿物态Zn亏缺;碳酸盐结合态、氧化锰结合态和无定形氧化铁结合态Zn富集。根际Zn形态转化强度大于非根际,其有效态Zn量接近临界值并高于非根际。有机物料利于根际内外土壤中硅酸盐矿物态Zn的转化,低盐土壤根际更强烈。随有机物料用量增加,促使根际硅酸盐矿物态、碳酸盐结合态及氧化锰结合态Zn向交换态、紧结有机态和无定形氧化铁结合态Zn转化,低盐土壤较明显。     

Determination of K forms in K-fertilized soils by electro-ultrafiltration

We confirmed the suitability of electro-ultrafiltration (EUF) for (a) determination of the distribution of potassium fertilizer among the various forms of potassium in soils with a predominance of micaceous minerals in their clay fraction, and (b) investigated the effects of the degree of openness of the dominant micaceous mineral and of incubation time on the kinetics of the EUF extraction of K from these soils.Samples of illitic, mixed-layer and vermiculitic soils from Galicia (N.W. Spain) were incubated at field capacity for 450 days with 0 (blank), 5,15 or 25 mg K (as KCl) per 100 g dry soil. After 1, 30, 150 and 450 days, subsamples were removed and repeatedly extracted using electro-ultrafiltration at low (20° C/200 V) and then high (80° C/400 V) temperature/voltage (6 and 10 five-minute extractions, respectively). Five different pools of K were identified: solution K (K_s), surface and internal K (collectively, K_p), slowly exchangeable K (K_e) and non-exchangeable K (K_i). The effects of increasing the incubation time depended on the dominant clay mineralogy: after 450 days, the K added to illitic soils was mostly solution K, whereas that added to vermiculitic soils was mostly internal K.For both low and high temperature/voltage EUF experiments, the extraction-time data were best fitted by the Elovich equation (extracted K=a+b ln t). The kinetic coefficient b depended on the incubation time and dominant clay mineral, and for given soil and incubation time increased linearly with the dose of added K.Abbreviations EUF Electroultrafiltration - K_s Solution potassium - K_p Easily exchangeable (surface + internal) potassium - K_e Slowly exchangeable potassium - K_i Non-exchangeable potassium     

Control of potato pale cyst-nematode, Globodera pallida, with a granular nematicide and partially resistant potatoes

The effects of oxamyl applied to the seedbed and growing partially resistant potatoes in controlling potato pale cyst-nematode, Globodera pallida, were assessed in eleven field experiments on sandy, peaty and silty loam soils in England from 1986 to 1988. Standardised procedures allowed valid comparisons to be made between data from the three experiments in 1987 and from the seven experiments in 1988. In soil moderately or heavily infested with G. pallida, oxamyl frequently increased tuber yields of susceptible cv. Désirée and of partially resistant potatoes and lessened crude nematode increase (Pf/Pi) significantly at some sites but not at others. Although 2.8 kg oxamyl ha“¹ often increased tuber yields significantly, it was significantly less effective than 5.6 kg in controlling G. pallida at one site and at another site G. pallida increased more than in untreated plots. Potatoes partially resistant to G. pallida were more effective that oxamyl in controlling nematode increase at some sites but not at others. Combined use of oxamyl and partially resistant potatoes was generally more effective in controlling G. pallida than either measure alone. As measured by a ‘control coefficient’ (weight of tubers over 40mm diameter (t ha“¹) 4-crude nematode increase (Pf/Pi)), the most effective integrated control of G. pallida was obtained by growing cvs Glenna, Morag or Santé in soil treated with 5.6 kg oxamyl ha^-1.     

Molecular forms in human serum of enzymes synthesizing DNA precursors and DNA

Both thymidine kinase (TK) and DNA polymerase (DNAp) are present in measurable amounts in human serum. Even though the use of TK as a clinical marker is rapidly increasing there has been no attempt to characterize the serum TK in a wider extent, i.e.; with respect to Mw or other biochemical parameters. Therefore sera with high TK or DNAp activities derived from patients with cytomegalo-virus (CMV) infection, B₁₂-deficiency and leukaemia were fractionated by gel exclusion chromatography. The TK activity eluted as two peaks, one major TK activity with an apparent molecular weight (Mw) or 730 kD and one minor TK activity corresponding to a Mw of 58 kD. The amount of TK activity at 58 kD varied between 7 and 23% of total activity, depending on the serum fractionated. The DNAp activity in sera from patients with malignant disease and B12 deficiency eluted as a single peak corresponding to a Mw of 240 kD. A DNAp with a different Mw (>1000 kD) was recovered from 1 of 3 investigated immunosuppressed patients with CMV infection. A similar pattern of enzyme forms was observed when sera were separated by glycerol gradient centrifugation.The effect of high salt and various reaction solution components on the enzymes were studied. The only condition found that affected the molecular forms of TK was the state of reduction. Incubation of sera with high concentrations of dithioerythritol (DTE) (400 mM) prior to separation transferred all serum TK to the 58 kD form, it also converted most of the serum DNAp from the 240 kD form to a smaller form (56 kD) without affecting the total recovery of enzymatic activity.The reaction product from both TK forms was exclusively monophosphate and none of the TK forms could efficiently utilize cytidine triphosphate as phosphate donor. The substrate kinetics of the small serum TK fraction was identical with those of an enzyme with similar size purified from proliferating HeLa cells, indicating that both serum TK activities are forms of TK 1, the proliferation associated cellular isozyme.