Studies in Stomatal Behaviour: IV. THE WATER-RELATIONS OF THE EPIDERMIS

1. It is shown that a dry external atmosphere exerts the followingeffects on stomatal movement:

1. A striking accelaration ofclosure in darkness.
2. A slight acceleration of opening in light.
3. If the water-supply to the leaf is impaired, an inabilitytomaintain full opening in the light.

Conversely, a saturatedexternal atmosphere induces sluggishness of movement and a tendencyto incomplete closure in darkness.

1. These results are consideredto support La Rue's contentionthat the epidermal water-supplyis drawn solely by lateral movementfrom the main veins, andnot from the underlying mesophyll.The stomatal phenomena themselvesdo not appear capable of anysimple explanation based on currentknowledge of guard-cellphysiology.
2. The biological significanceof these results is discussed, withparticular reference tothe problem of xeromorphic structures,for which a new interpretationis suggested.

     

The Metabolism of Glucose and Acetate in Aspergillus niger

1. The metabolism of a citric-acid-forming strain of A. nigerwhengrown on a glucose-acetate medium has been investigated.
2. Acetate greatly accelerated the rate of utilization of glucose.
3. Citric acid production from glucose was much increased bythepresence of acetate.
4. The formation of oxalate from glucose-acetatecultures was muchless than from acetate alone.
5. In some cultureslarge amounts of glucose and acetate were consumedbut no acidicproducts were formed.

     

Factors Controlling Flowering of the Chrysanthemum: I. THE EFFECTS OF PHOTOPERIOD AND TEMPORARY CHILLING

1. Experiments are described which indicate that a temporary exposureto low temperature (vernalization) hastens inflorescence budinitiation in the Chrysanthemum, as measured by the time tothe macroscopic appearance of the bud and also by the numberof leaves produced. This effect is found in both long-and short-dayconditions.
2. In the absence of vernalization plants kept inshort day assumea diageotropic growth habit and remain vegetativefor long periods,frequently for much more than one year. Unvernalizedlong-dayplants also remain vegetative but have a normal geotropicreaction.
3. While the day-length effect is less important forinflorescencebud initiation, the opening and further developmentof budsformed in long day depend normally on subsequent day-lengthtreatment.
4. A vernalization period of only three weeks appearsto be fullyadequate.
5. The low-temperature treatment may begiven discontinuously,and evidence to hand appears to indicatethat it is more effectiveif given during the dark phase thanduring the light phase.Hence de-vernalization by temperaturesof about 20–25°C. does not appear to take place.
6. There is evidence that little or none of the stimulus is carriedover from one year to another.
7. The results are discussed inrelation to the auxin metabolismof the plant and also withregard to the absence in the literatureof previous mentionof the cold requirement.

The author is indebted to Professor F. G. Gregory and to Mr.F. J. Richards for their stimulating interest and helpful suggestionsin the course of this work, and to Dr. M. Holdsworth for providinghim with the results of earlier unpublished work. Messrs. H. Woolman Ltd. kindly supplied some of the plant materialused.     

PHYSIOLOGICAL AND BIOCHEMICAL CHANGES OF CARROT ROOT CALLUS DURING SUCCESSIVE CULTURES

1. The longer the period of stock culture, the more remarkableis the growth inhibition by 8-azaguanine in callus.
2. Chloramphenicol,5-methyltryptophane and mitomycin C exert greaterinhibitionon growth in CCL than in CCS.
3. Bud formation is inhibited bysome concentrations of chloramphenicolwithout accompanyinginhibition of the growth.
4. Cell size and the contents of RNA,DNA, protein and lipid percell of CCL are greater than thoseof CCS, respectively. Thecontents per cell of RNA and lipidin "mitochondrial fraction"are higher in CCL than in CCS.
5. Incorporationof guanine-8-¹⁴C into RNA of CCS occurs rapidlyin the first12 hr and slows down thereafter, but that in CCL-RNAincreasessteadily for 16 hr. This difference in rate of theincorporationafter 12 hr between CCS and CCL is principallydue to the differencein rate of the incorporation into RNAof nuclear, mitochondrialand soluble fractions.

1. The rate of RNA breakdown in CCL wasnot so great as the rateof synthesis.
2. 8-azaguanine (10^–3and 10^–4M) inhibits incorporationof guanine-8.¹⁴C intoRNA of both CCS and CCL during 14 hr,but thereafter (up to25 hr) it inhibits the incorporation intoCCL-RNA alone leavingthat into CCS-RNA unaffected.

1. In CCL 510^–5M 8.azaguaninedoes not affect total radioactivityincorporated into bulk RNA,but inhibits incorporation intoRNA of "mitochondrial fraction".

(Received December 23, 1964; )     

EFFECT OF SUCROSE ON FLOWER INITIATION OF PHARBITIS NIL IN ASEPTIC CULTURE

1. Pharbitis plants were cultured on a nutrient medium containing5% sucrose or no sucrose. They were subjected to various lightconditions at temperatures of 10? or 20?.
2. All the plants culturedon the sucrose-medium at 10? initiatedflower buds independentlyof the light to which the plants wereexposed.
3. Plants culturedon the sucrose-free medium at 10? initiatedflower buds undershort photoperiods, but did not under continuousillumination.
4. Plants cultured on the sucrose-medium at 20? initiated flowerbuds in total darkness or under short photoperiods regardlessof the intensity and quality of the light, but those subjectedto continuous illumination did not.
5. Plants cultured on thesucrose-free medium at 20? initiatedflower buds under shortphotoperiods but did not initiate themunder continuous illumination.

(Received June 9, 1960; )     

An Examination of a Method of Auxin Assay using the Growth of Isolated Sections of Avena Coleoptiles in Test Solutions

1. Methods of auxin assay using the Avena coleoptile are discussed.
2. A review is given of experimental procedure and evaluationofresults in the straight-growth method using isolated sectionsof coleoptiles in test solutions.
3. Possible sources of variationin the straight-growth methodare investigated and discussed.
4. A revised experimental procedure for the straight-growth methodis described.

The author wishes to thank Professor E. Ashby for his adviceand encouragement during the course of the experiments. Thanks are also due to Mr. D. Payne, a technical assistant inthe Botany Department, for his assistance in some of the assays.     

CHEMICAL CONTROL OF MORPHOGENESIS IN MOSS PROTONEMA

1. The protonema of the moss, Funaria hygrometrica, grows continuouslyin calcium-free liquid media.
2. The growth was promoted by additionof oxalate, although themorphogenesis resulting in formationof gametophytic buds onthe protonema was suppressed by theaddition.
3. Calcium oxalate promoted the growth of protonema,while at ahigh concentration (10^–2 M) it caused the formationofclumped protonema (falsebuds).
4. Addition of plant growthhormones, such as IAA, NAA, 2,4-D andgibberellin retarded thegrowth of protonema, while 2,4-D ata low concentration stimulatedthe growth of protonema.
5. Kinetin greatly stimulated the formationof gametophytic budsin the protonema, but these buds were foundto be morphologicallyand physiologically abnormal.

(Received January 29, 1965; )     

FLOWER-PROMOTING EFFECT OF GIBBERELLIN IN WINTER WHEAT AND BARLEY

1. Vernalized and unvernalized seeds of barley were sown in greenhouseunder short day or long day condition. At the second leaf stage,a half of the plants in each plot was sprayed with gibberellin.Plants grown from vernalized seeds headed at about the sametime under long day condition, whether they were treated withgibberellin or not. Under short day condition, however, gibberellinpromoted heading. With plants grown from the unvernalized seeds,gibberellin promoted heading under long day condition whereasit delayed heading under short day condition. Thus the substitutionof gibberellin for long day effect and for vernalization effectwas shown.
2. Immature embryos growing on developing ear of wheatand barleywere vernalized with low temperature combined withor withoutgibberellin application. Plants grown from the seedstreatedat their early ripening stage with low temperature combinedwith gibberellin gave earlier heading, associated with the decreasedleaf number, than the plants from seeds treated with low temperaturealone. This effect of gibberellin lasted for a half-year ofseed storage.
3. Gibberellin was applied to germinating seedsunder vernalizationtreatment. Gibberellin promoted floweringin varieties whichpossess high requirement for chilling vernalization,when thechilling was insufficient to satisfy the high requirement.However,the effect of gibberellin was obscure with varietiespossessingrelatively low requirement for vernalization.

(Received September 15, 1964; )     

Physiological Studies in the Mucorales: PART I THE PHOTOTROPISM OF SPORANGIOPHORES OF PHYCOMYCES BLAKESLEEANUS

Previous views on the physical basis of phototropism in Phycomycessporangiophores are briefly discussed.

1. It was confirmed thatunilaterally illuminated sporangiophoresimmersed in liquidparaffin show strong negative phototropism.
2. Elongation growthceased and no phototropic response took placeunder anaerobicconditions.
3. By focusing a fine beam of light on to one edgeof the growingzone of a sporangiophore, leaving the other sidein darkness,it was established that greater elongation tookplace in theilluminated zone, the sporangiophore tending tobend out ofthe beam. Rapid reversal of the curvature followedwhen theillumination was transferred to the opposite edge ofthe sporangiophore.

Wassink and Boumann's suggestion that phototropism can be initiatedby a one-quantum-per-cell process is criticized in the lightof this result and other work by Castle.     

Gametogenesis and Embryogeny of Tamarix ericoides Rottl

1. The haploid chromosome number in Tamarix ericoides Rottl., reportedfor the first time, is twelve.
2. The archesporial cell is single,rarely double.
3. There is no tetrad formation, division of themegaspore mother-cell;instead, a four nucleate embryo-sac results.
4. The nuclei are originally arranged in a cruciform type, butlater form a I+3 arrangement.
5. As a result of the third division,the embryo-sac is again four-nucleate,with two haploid nucleiat the micropylar end and two triploidnuclei at the chalazalend.
6. The cells of the embryo-sac are formed at the end ofthe fourthdivision, the embryo-sac thus representing the sixteennucleatecondition.
7. The development of the embryo-sac of Tamarixericoides conformsto the Fritillaria-type.
8. The endospermis free nuclear, and the embryo is characteristicin developinga massive suspensor and a large pad of cellulartissue fromthe proximal cell.
9. Double embryo-sacs and the occurrence ofpolyembryony are reportedfor the first time in the family.

     

The Isolation of l-Quinic Acid from the Apple Fruit

1. A method is described by means of which several grammes ofl-quinicacid were isolated from young fruits of the WorcesterPearmainapple stored for a number of days in nitrogen.
2. Theessential stages in the method are the removal of colouringmatter from extracts of the frozen ground pulp tissue of thefruit with charcoal, the removal of amino acids by absorptionon cation exchange resin, and the separation of the organicacid from sugars by adsorption of the former on anion exchangeresin. Finally, the quinic acid was separated from malic acidby fractional displacement from the anion exchange resin.
3. Thecharacterization of the isolated quinic acid, and of malicacidalso isolated from the fruits, is described.
4. Citric acid isshown to be formed by oxidation of quinic acidwith hot hydrogenperoxide.
5. The possible function of quinic acid in the applefruit andin plants generally is discussed.

     

Studies with Radioactive Tracers in Plant Nutrition: II. THE ESTIMATION OF RADIOACTIVE TRACERS

Part I Tracer methods can yield valid results only if the behaviourof the tracer within the plant is indistinguishable from thatof the normal element. The extent to which radioactive isotopessatisfy this requirement must be considered from the followingviewpoints:

1. their chemical and physical bahaviour before disintegration;
2. the effects and substances formed by their disintegration;
3. the effects of the radiation they emit.

Whereas, chemically and physically, isotopes with the exceptionof H¹ and H³ satisfy this requirement, they may cause significantradiation damage even when supplied in low concentration ifthey are accumulated metabolically. The investigation of possibleradiation effects is therefore an essential preliminary to allphysiological investigations. The value of tracer methods is considered. Part II Equipment and laboratory procedures for the electronic assayof P³² are described. For the assay of samples containing P³²a wet ashing method and the subsequent use of a liquid countingtube is preferred. Improvements of equipment to suit the requirementsof plant nutritional studies are considered. Sources of experimentalerror are examined. On account of the decay of radioactive isotopes,the time factor may be of special importance in tracer work.Its effect on both experimental design and on laboratory proceduresis considered. It is stressed that the scope and replicationof large experiments, particularly if they are of long duration,may be subject to serious limitations. The importance of simplifyingthe equipment used in tracer assay and increasing its reliabilityis emphasized.     

The Peltier Effect and its Use in the Measurement of Suction Pressure

1. A method is described by means of which the suction pressureof living material may be measured by observations on its equilibriumvapour pressure.
2. It employs a very fine thermocouple whosetip is cooled by meansof the Peltier effect, below the dew-point.It collects a minuteamount of dew, and can thus be used asan exceedingly fine ‘wet-bulb’thermometer. As suchit is convenient to use, quick in action,and causes very littledisturbance to the system under observation.
3. The thermocoupleis calibrated by means of salt solutions ofknown strength.The scale is fairly linear.
4. The method possesses the advantagesof giving the suction pressureby direct observation ratherthan by interpolation; of speedin the actual measurement; ofnot subjecting the tissue to immersion;and of permitting organslike leaves, otherwise difficult tohandle, to be easily dealtwith. It also opens possibilitiesoutside the range of the conventionalmethods.

     

CHANGES IN ORGAN FORMING CAPACITY OF CARROT ROOT CALLUSES DURING SUBCULTURES

1. The formation of buds and roots in seven strains of carrotrootcallus successively cultured for various periods on a mediumcontaining WHITE'S inorganic salts, sucrose, 2,4-D and yeastextract was investigated. 2,4-D completely suppressed organformation during stock subculturing. It was confirmed that theorgan forming capacity of the callus in a 2, 4-D-free test mediumdiminishes and finally completely disappears with prolongedperiods of previous subculturing of the callus.
2. IAA promotedthe root formation. Yeast extract, casein hydrolysateand aminoacids mixture promoted the bud formation of callusesat earlieststeps of subculturing (Phase I).
3. At next steps of subculturing(Phase II), IAA-dependent rootforming capacity of calluseswas lost although the bud formingcapacity induced by yeastextract, casein hydrolysate and aminoacids mixture was retained.
4. At further advanced steps of subculturing (Phase III), yeastextract induced only root formation, while casein hydrolysatestill could induce buds. IAA and amino acids mixture did notaffect the organ formation.
5. No organ formation was observedin calluses subcultured over38 months under any conditionsattempted (Phase IV).
6. Single cells or small cell clumps obtainedfrom the callus subculturedfor 2 months formed only roots onthe medium containing IAAand formed buds and roots on the mediumcontaining yeast extract.
7. These differences in organ formingcapacity and in responsestowards various factors are interpretedto reflect the changesin physiological states of the callusduring successive cultureson the stock culture medium.

(Received December 24, 1964; )     

The Effect of Prolonged Chilling on Water Movement and Radial Growth in Trees

1. The woody shoots of young saplings of Fraxinus.excelsior andAcer Pseudo-platanus in pots were subjected to continuous coolingto about 2° C. during the growth season, with the resultthat radial growth was almost completely inhibited throughoutthe woody stem.
2. The chilling did not adversely affect extensiongrowth exceptthat it was later in commencement and proceededmore slowly.
3. If the temperature around the stem is loweredfrom 2° C.to 0° C., water conduction is cut down tosuch an extentas to cause wilting of the leafy shoots; turgidityis recoveredwhen the temperature is again raised to 2°C.
4. This wilting effect is discussed particularly in relationtothe part played by living cells in the upward movement ofwaterin the wood.

     

Microbiological Synthesis of Fat: PRELIMINARY SURVEY OF THE FAT-PRODUCING MOULDS

1. A survey of potential fat-producing moulds has shown that atleast 40 strains from 10 species are of interest.
2. These havebeen grown on five different media and A. nidulans,P. spinu-losum,P. javanicum, P. piscarium, P. flavo-cinereum,P. oxalicum,A. flavus, A. flavipes have shown the most promise.
3. The fatcontent on felt weight was maximal at 39?7 per cent,with A.flavipes and 34?6 per cent, with F. lini, and on usedsugarwas maximal at 9?3 per cent, with A. flavipes and 6?7per cent,with A. nidulans.
4. The potential value of micro-organisms asfat producers is discussedwith reference to the moulds.

     

STUDIES ON PROTEIN SYNTHESIS IN TOMATO COTYLEDONS AND LEAVES. II. INTERMEDIATE STAGES OF PROTEIN SYNTHESIS

1. The activities of RNase, ATPase and proteolytic enzymes inextractsof tomato leaves have been investigated with regardto detrimentaleffects on protein synthesis. The use of coppersulphate asan inhibitor of RNase is described.
2. The effectof microbial contamination on amino acid uptake hasbeen studiedin relationship with cell-free systems, and thehigh activitynoted underlines the necessity for aseptic procedures.
3. Methodsfor the preparation of cell-free systems capable ofamino acidactivation and incorporation into protein in an asepticenvironmentare discussed.

¹Present address: Dept. of Horticultural Science, Universityof Wisconsin, Madison, Wisconsin, U.S.A.     

The Morphology and Anatomy of Pellia epiphylla considered in Relation to the Mechanism of Absorption and Conduction of Water

1. In Pellia epiphylla water travels in the form of capillary filmsbetween the rhizoids over the under surface, being partly absorbedthere. The remainder passes over the surface and is retainedin the region of the antheridia by the incurved and wavy marginsof the thallus.
2. The antheridia, oospheres, segmenting oospores,and sporogoniaare largely supplied with water from the exterior.This wateris retained in the narrow crevices between antheridialcavityand antheridium, between the flaps of the involucre,and betweenthese and the long necks of the archegonia. Travellingdownthe necks of the archegonia, it reaches the oospheres,and islater available for the developing sporophyte in theform offilms retained between the foot of the segmenting embryoandthe calyptra.
3. In the sporophyte, this water, absorbedby the foot largelyfrom external sources, travels up the seta,into the elaterophore,out to the elaten, and ultimately tothe spores.
4. The sporophyte, therefore, is much less dependenton the gametophytefor supplies of water and raw materials thanmight be expected.

The writer wishes to take this opportunity of thanking ProfessorF. A. Mockeridge for suggesting this work and for her valuableadvice and criticism during its progress.     

INFLUENCE OF LIGHT ON ACETATE UTILIZATION IN GREEN EUGLENA

1. The kinetics of adaptation to exogenous acetate, measured asthe ability to stimulate respiration, has been studied in Euglenapreviously grown with autotrophic nutrition.
2. The continuedpresence of light significantly inhibits the fulldevelopmentof respiratory adaptation to acetate.
3. Carbon fixed in photosynthesisis routed almost exclusivelyinto protein when acetate is present.Acetate incorporated withconcomitant photosynthesis largelyenters lipid and polysaccharide,with only a small fractionincorporated into protein.

(Received December 25, 1964; )     

The Extension Growth-Time Relationship for Avena Coleoptile Sections

1. The growth rates of coleoptile segments supplied with indole-3-aceticacid is not constant with time, but, when the IAA concentrationis high, decreases very rapidly.
2. With sufficiently high concentrationof IAA, the initial rapidgrowth may be eventually followedby shrinkage of the tissue.
3. The relation between initial rateof growth and auxin concentrationis not significantly differentfrom hyperbolic.
4. The significance of these facts in relationto kinetics of auxinaction is discussed.