The Absorption of Ions by Excised Root Systems: II. OBSERVATIONS ON ROOTS OF BARLEY GROWN IN SOLUTIONS DEFICIENT IN PHOSPHORUS, NITROGEN, OR POTASSIUM

1. Barley plants were grown in complete culture solution and indeficiencies of phosphorus, nitrogen, or potassium for a periodof about 6 weeks. Excised roots of these plants were treatedwith a complete, aerated culture solution at 25? C. for varyingperiods of time, and the changes in respiration rate, phosphorus,nitrogen, potassium, sugars, and starch contents measured.
2. Therewere changes in fresh weight and dry weight of the excisedrootsduring treatment. The dry weight decreased with time butthewater-content changes were variable. There was a gain orlossof water by the roots according to the treatment.
3. In all casesthe deficient roots increased in content of theelement in whichthey were originally deficient. The roots ofthe plants suppliedwith full nutrient usually decreased incontent of phosphorus,nitrogen, and potassium, but exceptionsoccurred and the reasonsare discussed.
4. In most of the experiments described simultaneousloss of oneion and gain of another occurred.
5. Nitrogen-deficientroots accumulated nitrate when exposed toa complete nutrientsolution, and some of this was assimilatedwith formation ofprotein. Under similar conditions nitrogen-richroots decreasedin nitrogen content and proteolysis took place.
6. There wasa rapid fall in sucrose and reducing sugar contentof the excise'roots. The starch content was initially verysmall and showedlittle change with time.
7. The respiration rate declined withtime in all treatments exceptwhere a nitrogen deficiency existed.Here the respiration rateincreased to a maximum value at about8 hours and then fell.This increase in rate is attributed toprotein synthesis. Noevidence of a ‘salt respiration’was observed evenwhen active uptake of phosphorus or potassiumwas occurring.
8. In most instances the carbon dioxide evolvedin respirationgreatly exceeded the carbon dioxide equivalentof the sugarconsumed in the same period. Exceptions were foundwith thenitrogen-deficient roots where less carbon dioxidewas evolvedthan the equivalent of sugar consumed. It is probablethat apart, at least, of the sugar unaccounted for was usedin proteinsynthesis.
9. Where the carbon dioxide of respirationwas in excess of theequivalent of sugar consumed, protein oramino-acid is the mostprobable substrate. Respiration rateis found to be relatedboth to nitrogen and sugar content.

     

THE INFLUENCE OF HIGH CONCENTRATIONS OF AMMONIUM AND SODIUM MOLYBDATES ON FLAX, SOYBEAN AND PEAS GROWN IN NUTRIENT SOLUTIONS CONTAINING DEFICIENT OR EXCESS IRON

Ammonium molybdate supplying 20 or 40 p.p.m. Mo prevented chlorosis caused by low iron supply in young flax plants, but sodium molybdate was effective only at the higher concentration. Temporary darkening of the green colour of the shoots was also produced by 40 p.p.m. Mo in iron-deficient soybean and pea plants, but was soon followed by more severe chlorosis. Symptoms of molybdenum toxicity always developed when 40 p.p.m. Mo were given, whether or not the intensity of chlorosis was reduced. With an increase in iron supply, a reduction in molybdenum toxicity symptoms was confirmed in soybean and peas. In flax the higher level of iron eventually proved excessive unless it was combined with 40 p.p.m. Mo. High molybdenum thus seemed able to counteract both iron deficiency and toxicity in this plant.
High iron reduced the molybdenum content (p.p.m./d.m.) of both shoot and root in soybean, peas and also in flax provided the iron was not excessive. High molybdenum usually reduced the iron content of the shoot, but markedly increased it in the root. Molybdenum-induced chlorosis could thus be partly attributed to inhibition in iron translocation, but the beneficial effect of high molybdenum or high iron on colour was not obviously correlated with the analytical data.     

Effectiveness of Lotus Root Nodules: III. EFFECT OF COMBINED NITROGEN ON NODULE EFFECTIVENESS AND FLAVOLAN SYNTHESIS IN PLANT ROOTS

When grown in a nutrient solution containing combined nitrogen(NH₄NO₃), Lotus pedunculatus and L. tenuis seedlings inoculatedwith a fast-growing strain of Rhizoblum (NZP2037) did neitherdevelop root nodules nor develop flavolans in their roots. Incontrast, the roots of nodulated seedlings growing in a nitrogen-freenutrient solution contained flavolans. Flavolan synthesis coincidedwith root nodule development on these plants. When added as a single dose, high concentrations of NH₄NO₃ (5and 10 mg N per plant) stimulated the growth of L. pedunculatusplants but suppressed nodulation and nitrogen fixation. In contrastthe continued supply of a low concentration of NH₄NO₃ (1?0 mgN d^–1 per plant) stimulated nitrogen fixation by up to500%. This large increase in nitrogen fixation was associatedwith a large increase in nodule fresh weight per plant, a doublingof nodule nitrogenase activity, and a lowering of the flavolancontent of the plant roots. The close relationship between nitrogendeficiency, nodule development, and flavolan synthesis in L.pedunculatus meant that it was not possible (by nitrogen pretreatmentof plants) to alter the ineffective nodule response of a Rhizobiumstrain (NZP2213) sensitive to the flavolan present in the rootsof this plant.     

STUDIES ON SUBMICROSCOPIC ASPECTS OF MINERAL DEFICIENCIES. II. NITROGEN,POTASSIUM, SULFUR,PHOSPHORUS, AND MAGNESIUM DEFICIENCIES IN THE SHOOT APEX OF BARLEY

Marinos, Nicos G. (Waite Agric. Res. Inst., Adelaide, S.A., Australia.) Studies on submicroscopic aspects of mineral deficiencies. II. Nitrogen, potassium, sulfur, phosphorus, and magnesium deficiencies in the shoot apex of barley. Amer. Jour. Bot. 50(10): 998–1005. Illus. 1903.—The elongation of the shoot apex of barley was retarded in the absence of N, P, or Mg, while the lack of S or K did not affect its growth as compared with the controls. Ultrastructural changes in cells of the terminal 100μ of the apical dome either appeared concurrently with, or preceded, measurable effects on apical elongation. These changes included: (1) formation of many dense intramitochondrial granules and a statistically significant reduction of the length of mitochondrial tubules; (2) accumulation in the cytoplasm of dense manganophilic inclusions which appeared to be formed by the Golgi bodies (dictyosomes); (3) starch accumulation, only in P-dcficient plants, in a region of the apex that is normally free of starch. The similarities in the symptoms induced by N, P, and Mg deficiencies reflect the metabolic interrelationships between these elements. On the basis of the time of appearance of these symptoms and their distribution in the longitudinal axis of the apical dome, it is concluded that altered mitochondrial function is an early response of cells to the lack of certain essential elements.     

The Structure and Development of Trifolium subterraneum L. Root Nodules: I. IN PLANTS GROWN AT OPTIMAL ROOT TEMPERATURES

The structure of effective nodules on Trifolium subterraneumL. grown at a root temperature favourable for nitrogen fixationwas examined by light microscopy. Rhizobium were released intohost cells from vesicles on infection threads which were closelyassociated with the host cell nucleus. Enlargement into bacteroidforms was rapid and synchronous in similarly-aged host cells,but groups of rhizobia close to the nucleus sometimes did notdevelop into bacteroids. Rhizobium changed in shape from smallrods to more elongate and then to irregularly swollen formsbefore finally becoming coccoid. On degeneration clusters ofbacteroids became un evenly stained before coalescing at thecentre of the host cell and finally disintegrating. Small vegetativerods multiplied amongst the degenerating bacteroids and in thegreatly enlarged intercellular spaces; host cell walls becamefolded and distorted and sometimes broke before collapsing ontoeach other.     

The Structure and Development of Trifolium subterraneum L. Root Nodules: II. IN PLANTS GROWN AT SUB-OPTIMAL ROOT TEMPERATURES

Cold root temperature affected infection thread proliferation,cell invasion, and release of Rhizobium and the subsequent developmentof this infection in Trifotium subterraneum. These events werealso modified by both host cultivar and bacterial strain. At7 °C bacteroid development was only substantial with strainTA1, with either sparsely or abundantly nodulating lines ofthe host. At 11 °C strain SU297 also readily formed effective,bacteroid-filled nodules with both lines. Strain 0403 formeda few bacteroids with the abundant line only at 7 °C andreadily formed bacteroids with the sparse line only at 19 °C.At 15 °C 0403 nodules were effective on abundant lines,but mostly ineffective on sparse lines. The development of Rhizobium rods into bacteroicis and theirsubsequent degeneration wa slower at low temperatures with bothstrains. Low root temperatures favoured the deposition of starchthroughout the nodule. At higher temperatures, when bacteroidswere more active in nitrogen fixation, starch was mostly confinedto a narrow band of the youngest bacteroid filled cells andto the zone of bacteroid degeneration.     

GROWTH AND NUTRITION OF TIMOTHY (PHLEUM PRATENSE L.): IV. THE EFFECT OF NITROGEN, PHOSPHORUS AND POTASSIUM SUPPLY ON GROWTH DURING THE FIRST YEAR

In a sand-culture experiment lasting 21 weeks plants of timothy grown from seed were maintained at all combinations of three levels of nitrogen, phosphorus and potassium. The number and weight of tillers, leaves and ears, and the weight of roots, were determined at the end of the experiment, and for some of the treatments at intervals of 4 weeks. All three nutrients had significant effects, especially N which at its lowest level of supply tended to mask the effect of the other two elements. Potassium influenced tiller numbers least, especially those of primary tillers, but in the presence of high concentrations of N and P it had a large effect on leaf area and dry weight. Relative growth and net assimilation rates responded to varying nutrient supply only in the early stages of growth, so that in general variations in dry weight were associated with nutrient effects on leaf area.     

THE ACCUMULATION OF ELECTROLYTES : III. BEHAVIOR OF SODIUM,POTASSIUM, AND AMMONIUM IN VALONIA

When 0.001 M NH₄Cl is added to sea water containing Valonia macrophysa there seems to be a rapid penetration of undissociated NH₃ (or NH₄OH) which raises the pH value of the sap so that the thermodynamic potential of KOH becomes greater inside than outside and in consequence K leaves the cell: NaOH continues to go in because its thermodynamic potential is greater outside than inside. NH₄Cl accumulates, reaching a much higher concentration inside than outside. This might be explained on the ground that NH₃, after entering, combines with a weak organic acid produced in the cell whose anion is exchanged for the Cl^- of the sea water, or (more probably) the organic acid is exchanged for HCl.     

Devlopmental Physiology of Sugar-Beet: II. EFFECTS OF TEMPERATURE AND NITROGEN SUPPLY ON THE GROWTH, SOLUBLE CARBOHYDRATE CONTENT AND NITROGEN CONTENT OF LEAVES AND ROOTS

Plants were grown at temperatures of 15 and 25 ?C with two ratesof nitrogen supply. The changes in dry weight, leaf area, cellnumber, mean cell volume, soluble carbohydrate, and total nitrogenconcentration of the cotyledons, the first and second pair oftrue leaves, and the storage root were measured. Changes incell number and cell volume of the first pair of true leavesand storage root of plants were also measured at 11, 18, 25,and 32 ?C. Leaf growth before unfolding was chiefly by increase in cellnumber and after unfolding by increase in mean cell volume,while the growth of the storage root was almost entirely byincrease in cell number. The rates of cell division and cellexpansion were fastest at 25 ?C, but the initially high ratesof cell division in the terminal bud and in individual leavesdecreased rapidly and greater rates were maintained at the sub-optimaltemperatures, i.e. 15 and 18 ?C. After an initial period ofslow growth, the first-formed leaves grew faster and becamelarger at 15 than at 25 ?C. Leaves were produced, unfolded,grew faster, and became larger with increase in the externalconcentration of nitrogen, because cells divided and expandedfaster, so that nitrogen increased the number and size of cells. Sugar concentration was greater at 15 than at 25 ?C in leavesbut not in the storage root. Sugar concentration in the petiolesof the first and second pair of true leaves increased to 1.2and 2.0 per cent fresh weight respectively. Decreased nitrogensupply temporarily increased the sugar concentration of cotyledonpetioles and the seedling hypocotyl, but later decreased itin the leaves and storage root. Nitrogen concentration was greaterin the leaves and storage root at 15 than at 25 ?C with thelarger nitrogen supply. Nitrogen concentrations were similarin young leaves of all treatments but as the size of leavesincreased nitrogen concentrations decreased most rapidly at25 ?C with the smaller nitrogen supply. It is suggested that when increased leaf production and storage-rootgrowth occurs at temperatures below the growth optimum (25 ?C),they may be due to an effect of increased carbohydrate supplyon cell division and sugar storage.     

STUDIES ON THE ANOMALOUS VISCOSITY AND FLOW-BIREFRINGENCE OF PROTEIN SOLUTIONS : III. CHANGES IN THESE PROPERTIES OF MYOSIN SOLUTIONS IN RELATION TO ADENOSINETRIPHOSPHATE AND MUSCULAR CONTRACTION

1. An investigation of the physicochemical properties of myosin has been carried out. Prepared under standard conditions, the ratio of flow-birefringence to protein concentration is uniform. The effect of electrolytes, pH, and urea on the flow-birefringence and viscosity (relative and anomalous) of myosin has been examined. 2. Decrease or abolition of flow-birefringence does not necessarily imply far reaching denaturation, since such effects can be reversed by a variety of means. 3. When a myosin solution is treated with adenosinetriphosphate, its flow-birefringence is decreased (average 48 per cent), its anomalous viscosity is retained, and its relative viscosity is decreased (average 14 per cent). The full effect of adenosinetriphosphate is obtained at 0.004 M; a molarity very much less than that of other substances which decrease the flow-birefringence of myosin. 4. The changes in the physicochemical properties of myosin brought about by adenosinetriphosphate are spontaneously reversible, and are connected with the enzymatic action of the protein as adenosinetriphosphatase. 5. Effects similar to those of adenosinetriphosphate on the physicochemical properties of purified myosin have been obtained so far only with inosinetriphosphate. 6. Inorganic phosphate is split off by myosin from inosinetriphosphate as well as from adenosinetriphosphate. Inorganic triphosphate is split by 1 to 2 per cent solution of three times precipitated myosin. 7. Adenosinediphosphate and inorganic triphosphate act as competitive inhibitors with adenosinetriphosphate, blocking the fall of flow-birefringence. 8. The implications of the results, and the conception of active enzymic groups attached to proteins participating in cell structure, whether contractile or non-contractile, are discussed in relation to present views on muscle physiology and other biological problems.     

Nutrient Supply and the Growth of the Seminal Root System in Barley: III. COMPENSATORY INCREASES IN GROWTH OF LATERAL ROOTS, AND IN RATES OF PHOSPHATE UPTAKE, IN RESPONSE TO A LOCALIZED SUPPLY OF PHOSPHATE

Responses to a localized supply of phosphate were studied inbarley grown in continuous flow solution culture. Root systemswere either uniformly supplied with 50 µM phosphate (controls)or the same solution was supplied to only a 4 cm or 2 cm lengthof a seminal root (localized supply), the remainder of the rootsystem receiving a nutrient solution lacking phosphate. Little development of laterals occurred on those parts of theroot system receiving no phosphate from the external solution,while an increase in the number and extension of laterals tookplace in the 4 cm zone enriched with phosphate. Compared withsimilar zones on controls, the total length of laterals wasincreased 15-fold in 21 d plants. In addition, rates of ³²P-phosphateuptake and translocation to shoots per unit root weight werehigher than in controls by a factor of 2?5–5?0. Furtherincreases in the growth of lateral roots, and rates of phosphateuptake, were induced when the segment initially supplied withphosphate was restricted to only 2 cm. These localized modifications to root growth and uptake of phosphatelargely compensated for the deficient supply of phosphate tothe remainder of the root system. After an early period of retardedgrowth and phosphate stress, the relative growth rate of plantsand the concentration of phosphate in shoots were restored tolevels similar to that of the controls. The manner in which the supply of phosphate may control rootdevelopment, and the nature of the co-ordination between rootgrowth, phosphate uptake, and shoot growth, are discussed.     

Effects of Mechanical Impedance on Root Growth in Barley (Hordeum vulgare L.): III. OBSERVATIONS ON THE MECHANISM OF RESPONSE

The effects of mechanical stress on whole root systems was investigatedusing beds of solid glass spheres (ballotini) continuously suppliedwith aerated nutrient solution. As noted in earlier experiments,increased mechanical impedance slowed root extension and alteredcell size and number; it also caused distortion of the rootapex, stimulated growth of lateral shoot meristems, and inducedthe formation of nodal roots. The development of lateral branchroots was enhanced and where root axes curved around ballotinilateral roots formed preferentially on the outer (convex) sidewhereas root hairs developed on the inner (concave) side. After roots were relieved from mechanical stress at least 3d elapsed before the rate of extension growth equalled thatof unimpeded plants. When intact Zea mays root apices first made contact with ballotinitheir elongation was slowed by 70% for about 10 min; where rootcaps were removed before the roots made contact, no such effectswere seen. We discuss the general nature of the mechanism of response tomechanical stress.     

Studies on the Growth in Culture of Plant Cells: VII. EFFECTS OF KINETIN ON THE CARBOHYDRATE AND NITROGEN METABOLISM OF ACER PSEUDOPLATANUS, L CELLS GROWN IN SUSPENSION CULTURE

Aspects of the carbohydrate and nitrogen metabolism of Acerpseudcplatanus, L cell suspension cultures grown on a syntheticmedium containing 2 per cent glucose and 1.0 mg/l 2,4-dichlorophenoxyaceticacid and kinetin either at 0.25 mg/l (low kinetin) or at 2.5mg/l (high kinetin) are described. High kinetin inhibits growthas measured by increase in cell number, packed-cell volume,and cell dry weight. Although not inhibitory to glucose utdization,high kinetin inhibits the O₂ uptake of the cells. Such cellscontain only a trace amount of fructose and their rate of O₂uptake can be raised to that of the low kinetin cells by a periodof fructose feeding. The O₂ uptake of both kinds of cell issensitive to malonate but the stimulation of O₂ uptake inducedby bis(hexafiuoroacetonyl)-acetone (‘1799’) at 0.2mM is much less with the high-kinetin than the low-kinetin cells.The enzymes phosphoglucoseiseomerase and glucose-6-phosphatedehydrogenase are much less active in the high-kinetin cells.Mitochondria isolated from both kinds of cells show good respiratorycontrol although slightly lower values for Q_O2(N), ADP/O ratioand control ratio are recorded with mitochondria from the highkinetin cells. Kinetin at 2.5 mg/l slightly reduces the ADP/Oratio of isolated mitochondria but at 4.0 mg/l their responseto ADP is completely suppressed. Extracellular hemicelluloseformed in presence of high kinetin has a reduced content ofgalactose and xylose and an increased content of glucose. Theseobservations indicate that the inhibition of respiration byhigh kinetin is mainly due to suppression of glucose conversionto other sugars rather than to inhibition of glycolysis or terminalrespiration. High kinetin decreases the rate of protein but not of amino-acidsynthesis. Suppression of the synthesis of particular proteinsmay be an important factor responsible for the reduced cellyield of the cultures in presence of high kinetin. The significanceof these observations to our understanding of the critical metaboliceffects of cytokinina is discussed. Acer pseudoplatanus cells release amino acids into their culturemedium early in the period of batch culture and largely reabsorbthem as incubation proceeds.     

The Effect of Micro-organisms on the Absorption of Inorganic Nutrients by Intact Plants: II. UPTAKE AND UTILIZATION OF PHOSPHATE BY BARLELY PLANTS GROWN UNDER STERILE AND NON-STERILE CONDITIONS

The uptake of phosphate by barley plants grown under sterileand non-sterile conditions has been compared from solutionsranging in concentration between 0.001 to 10.0 ppm P. If specialprecautions are not taken to exclude microorganisms their activityon or in plant roots greatly modifies the absorption and utilizationof phosphate in plants, especially when the external concentrationis low. More phosphate is incorporated into nucleic acids, phospholipids,and phosphoproteins in the roots; at 0.001 ppm P these compoundsaccount for over 60 per cent of the phosphate absorbed comparedwith about 15 per cent in plants grown under sterile conditionsand even at 1.0 ppm P there is a twofold difference. The immobilizationof phosphate in the roots reduces its transfer to the shoots;at 0.001 ppm P, 2 as compared with 20 per cent of the phosphateabsorbed is transferred to the shoots. This effect decreaseswith increasing concentration but small differences are stillapparent up to about 0.5 ppm P. A further effect of micro-organisms is apparent when plantsare transferred from dilute solutions to phosphate-free solutions.Under non-sterile conditons there is an almost tenfold increasein the loss to these solutions of previously absorbed phosphate. It is evident that conclusions on the mechanism of absorptionof phosphate, based on experiments in which the actions of micro-organismshave been ignored, require reinvestigation.