Transport of shoot- and cotyledon-applied indole-3-acetic acid to Vicia faba root

To clarify the participation of indole-3-acetic acid (IAA) originatingfrom the shoot in root growth regulation and the mechanism ofIAA translocation from shoot to root, the movement of ¹⁴C-IAAwhich was applied to the epicotyl or the cotyledon of Viciafaba seedlings was investigated. The radioactivity of IAA appliedto the cotyledon moved faster to the root tip than that appliedto the epicotyl. On the basis of the effect of 2,3,5-triiodobenzoic acid on IAAmovement, a comparison with ¹⁴C-glucose movement and autoradiographicexamination, the nature of IAA movement was concluded to bepolar transport from the epicotyl to the basal part of the roots,while IAA movement from the epicotyl to the cotyledon, fromthe basal part of roots to the apical part, and from the cotyledonto the epicotyl and to the root took place in the phloem. Theradioactivity from ¹⁴C-IAA applied to the cotyledon accumulatedin lateral root primordia and vascular bundles. These factssuggest that IAA produced in cotyledons may participate in theregulation of Vicia root development. (Received December 21, 1979; )     

Polar transport and content of indole-3-acetic acid in wounded sweet potato root tissues

In roots of sweet potato (Ipomoea batatas Lam. cv. Kokei 14),the metabolic response to wounding was remarkable in the proximalside and developed in the acropetal direction. We assumed thatthe polarity resulted from the increase in polar movement ofindoleacetic acid (IAA) (1977, Plant Physiol. 60: 563–566).Transport of IAA and change of the IAA level in the woundedtissue of sweet potato roots were investigated. Transport ofthe label from ¹⁴C-IAA was obviously polarized in the acropetaldirection. ¹⁴C-IAA administered to the wounded tissue was mainlymetabolized into two conjugates of IAA. The amount of IAA inthe wounded tissue, determined by the spectrofluorometric method,increased about 3-fold after 18 hr of incubation prior to thedevelopment of activities of some enzymes. The increase in IAAcontent was not affected with aseptic incubation, therefore,the possibility of IAA production by microorganisms on the woundedtissue was excluded. The results obtained strongly support ourhypothesis that IAA plays an important role in the metabolicresponse to wounding. (Received May 2, 1979; )     

The biosynthesis and translocation of 14C-IAA in Ricinus communis

The biosynthesis of ¹⁴C-IAA from ¹⁴C-tryptophan applied to abraded leaves of Ricinus communis and its subsequent export through the phloem were studied. Phloem sap was collected at intervals from incisions made in the stem below the IAA fed leaf. Any upward movement of label through the phloem or downward movement of phloem mobile compounds from leaves above the treated one were restricted by bark-ringing the plants.TLC and HPLC analyses of the collected sap indicate that some conversion of ¹⁴C-tryptophan to ¹⁴C-IAA had occurred. Subsequent GC-MS analysis of the HPLC purified samples of phloem sap revealed high levels of endogenous IAA transported from the fed leaf. The high ratio of unlabelled/labelled IAA in the phloem sap makes unequivocal confirmation by GC-MS of the predicted biosynthesis of ¹⁴C-IAA impossible. It is postulated that IAA is synthesised from tryptophan in mature leaves and exported to developing sink tissues with the flow of photoassimilates in the phloem.     

Auxin-promoted Transport of Metabolites in Stems of Phaseolus vulgaris L.: SOME CHARACTERISTICS OF THE EXPERIMENTAL TRANSPORT SYSTEMS

Indol-3yl-acetic acid (IAA) applied to sterns of Phaseolus vulgarisseedlings, decapitated above primary leaves, enhanced the mobilizationof ¹⁴C-metabolites to the treated stumps and this effect wasapparent within 3–6 h of applying the hormone. More than90 per cent of the total ¹⁴C-activity transported to the stumpswas detected in the alcohol-soluble extracts. In all treatments,less than 5 per cent of the ¹⁴C-photosynthate exported fromthe primary leaves was translocated upwards. Accumulation of¹⁴C-activity was also increased when the IAA was applied laterallyto intact internodes. This effect was obtained when ¹⁴C wassupplied either above or below the point of hormone application.By selective heat girdling, it was shown that the auxin affected¹⁴C transport when either the root ‘sink’ was removedor transpiratory flow of water through the treated internodewas maintained. Decapitated stems treated with plain lanolinfor 3 d were found to retain their responsiveness to auxin interms of enhanced metabolite transport. Heat-girdling experimentsand estimates of ¹⁴C transport velocity suggested that mostof the ¹⁴C movement was restricted to the phloem of treatedstumps. Similar effects of IAA on a transport in excised stemsegments of Phaseolus vulgaris were observed.     

In vitro binding of IAA to plasma membrane-rich fractions containing Mg++-activated ATPase from mung bean hypocotyls

Cell homogenates of dark-grown mung bean hypocotyls were fractionatedinto six fractions (L-0, L-l to L-5) by stepwise sucrose density-gradientcentrifugation. The majority (ca. 84%) of Mg⁺⁺-activated ATPase activity ofthe 10,000 x g pellet was localized in the L-0 (1.03 d 1.14)and L-l (1.14 d 1.16) fractions. Over 40% of the vesicularmembrane in the L-0 fraction and 60% of the L-l fraction couldbe stained with phosphotungstic acid (PTA)-chromic acid, a selectivestaining for the plant plasma membrane. In vitro binding of ¹⁴C-IAA to the fraction components was thegreatest in the L-l fraction among the six. The binding of ¹⁴C-IAAto the L-l fraction in vitro was markedly interfered with bythe presence of a high concentration of cold IAA (2 x 10^–4M).However, it was not affected by the IAA analogues IPA, IBA andIAN. This indicates that IAA highly specifically binds to theL-l fraction. In vitro specific binding of ¹⁴C-IAA to L-l andL-0 was decreased with an increasing acidity from pH 8.0 to5.0. In vitro binding of ¹⁴C-IAA to L-l and L-5 was furtherenhanced when these fractions were isolated from sections pretreatedwith 10^–5M cold IAA for 60 min ¹Present address: Institute for Plant Virus Research, 959 Aobacho,Chiba 280, Japan. (Received August 14, 1975; )     

Hormone-directed Transport of Assimilates in Decapitated Internodes of Phaseolus vulgaris L.

Application of ethylene, indole-3yl-acetic acid (IAA), 6-(benzylamino)-9-(2tetrahydropyranyl)-9-Hpurine (SD8339), or mixtures of IAA, gibberellic acid (GA),and cytokinins, increased the accumulation of ¹⁴C-activity indecapitated internodes of Phaseolus vulgaris seedlings. Differencesbetween treated and untreated tissues with respect to importof labelled assimilate were detected 3 h after application ofa mixture of IAA, GA, and SD8₃₃₉. In longer-term experimentseffects of the growth-regulator mixture on translocation oflabel were greater than those of IAA alone. Inhibitory effectsof abscisic acid on import of assimilate were counteracted bySD8₃₃₉. The ability of internode tissues to import ¹⁴C-photosynthatedeclines with time from decapitation, and a decrease in incorporationof ¹⁴C-leucine into protein was detected after 24 h. There wasan increase in protein and RNA synthesis in internodal tissuesfollowing a 2.5-h pre-treatment of decapitated internodes withIAA, GA, and SD8₃₃₉. Concentrations of 2, 3, 5-triiodobenzoicacid which inhibit ¹⁴C-IAA translocation stimulate protein synthesisin decapitated internodes, and augment the IAA-effect on importof ¹⁴C-photosynthate. ‘Hormone-directed’ assimilatetransport is discussed in relation to confounding effects ofgrowth responses and differential senescence of treated anduntreated tissues. It is suggested that accumulation of labelledassimilate in treated tissues results from effects of growthregulators on synthetic activities at the point of application.     

Auxin Transport in Secondary Tissues1

Auxin transport was investigated in excised stem segments ofNicotiana tabacum L. by the agar block technique using [1-¹⁴C]indol-3yl-acetic acid (IAA). The ability of the stems to transportauxin basipetally increased as secondary development proceeded;by contrast the ability of the pith to transport auxin declinedwith age. By separation of the stem tissues it was shown thatthe great majority of auxin transport took place in cells associatedwith the internal phloem and in cells close to the cambium;in both cases similar velocities of transport were found (c.5.0 mm h^–1 at 22°C). The effects of osmotic gradientson auxin transport through the internal phloem were investigated.IAA was found by chromatography to account for practically allthe radioactivity in receiver blocks and other extracts of stemsegments. The significance of these results is discussed.     

IAA Transport and Georeaction in the Flower Stalk of a Poppy, Papaver Rhoeas L.

The straightening process in the georeaction of the flower stalk(peduncle) of a poppy, Papaver Rhoeas L., was studied in relationto IAA transport in the peduncle and the receptacle. After thepeduncle had been formed, it first nodded. Cutting the flowerbud off at the nodding stage (stage 3–4 if the developmentis divided into 12 stages) caused rapid straightening of thenodding peduncle but the curvature was maintained if IAA wasapplied to the cut end of the decapitated peduncle. Applicationof 2,3,5-triiodobenzoic acid to the upper part of the pedunclejust below the flower bud at various growth stages caused immediatestraightening of the nodding peduncle. The IAA content in thepeduncle was lower at the straightening stages than at the noddingstage. Transport of ¹⁴C-IAA through the receptacle was lessat the straightening stages than at the nodding stage, whilethat through the peduncle was the same at these stages. Theseresults led to the conclusion that the straightening processof the poppy peduncle, which is considered to be a negativegeotropic response, is controlled by a decrease in the basipitaltransport of IAA from the flower bud to the peduncle throughthe receptacle. (Received April 12, 1982; Accepted September 13, 1982)     

Photoassimilate translocation in the petiole of Cyclamen and Primula is independent of lateral retrieval

Phloem translocation of photoassimilates between source andsink is considered to be linked with active retrieval of sugarsleaked to the vascular apoplast. This hypothesis was evaluatedby studying photo-assimilate movement in petioles of intactplants of Cyclamen persicum and Primula obconica in the presenceof inhibitors affecting sucrose retrieval (PCMBS, CCCP). Inhibitorsolutions were applied by rinsing locally isolated petiole bundlesor by injection into the petioe parenchyma. PCMBS and CCCP reduced[¹⁴C]sucrose retrieval from the petiole apoplast by the vascularcells and altered the distribution pattern of ¹⁴C-photoassimilateswithin the petiole tissues. However, these treatments did notaffect translocation through the petiole phloem. Evidence isprovided that the reagents were present in the vascular apoplastsurrounding the translocating phloem. It was concluded thatassimilate movement in the petiole of Cyclamen and Primula wasindependent of apoplastic retrieval. Key words: Cyclamen, Primula, phloem, transport, path, sucrose, retrieval     

TRANSPORT OF IAA AND ACC IN FLORAL ORGANS OF IPOMOEA NIL (CONVOLVULACEAE)

The involvement of the stamens as transporters of plant growth regulators in flowers was examined by measuring the movement of ¹⁴C-indole-3-acetic acid (IAA) and ^l4C-l-aminocyclopropane-1-carboxylic acid (ACC) through floral organs of Ipomoea nil. During the transport of ¹⁴C-IAA through isolated filament segments, the polar accumulation of ¹⁴C-IAA in receiver blocks increased with time during filament development, which correlated with polar efflux rates at older stages of filament development. An inhibitor of polar IAA transport, 2,3,5-triiodobenzoic acid, disrupted the polarity of auxin transport by reducing the movement of ¹⁴C- IAA from filaments into receiver blocks. Transport of both ¹⁴C-IAA and ^l4C-ACC through filaments into other floral organs also was monitored in isolated flower buds in the laboratory and intact buds in the greenhouse. In isolated and intact buds 21 hr before anthesis, substantially higher levels of isotope were recovered in corolla tissue when ¹⁴C-ACC was transported through the filaments than when ¹⁴C-IAA was transported from the filaments. In isolated buds, substantial levels of both isotopes accumulated in the pistil (69 hr and 45 hr before anthesis), but minimal amounts were observed in receptacle and calyx tissues (69 hr to 21 hr before anthesis). In intact buds, high levels of both isotopes were recovered in receptacle, calyx, and pistil tissues (69 hr to 21 hr before anthesis). The results from this study support the hypothesis that Ipomoea stamens are transporters for ACC and IAA to regulate ethylene production in the corolla and other floral tissues.     

Reversal of the Polarity of IAA Transport in the Leaf Sheath Base of Echinochloa colonum

The transport of [2-¹⁴C]IAA and its derived products has beeninvestigated in segments of the leaf sheath base of Echinochloacolonum. Segments orientated vertically in the normal uprightposition transport ¹⁴C basipetally at rates approximately 10times higher that acropetally. In horizontal segments the basipetalrates are lower, being only 1–4 times higher than acropetalrates. Segments orientated vertically in the inverted positiontransport ¹⁴C basipetally at approximately 1/10 of the acropetalrate. ¹⁴C in the receiver discs from vertically orientated segmentswas found to be present predominantly as [2-¹⁴C]IAA and itsderived metabolite. The levels of each of these compounds from upright and invertedvertical segments followed the same pattern as that of theirtotal ¹⁴C content.     

Hormone-directed Transport of Metabolites and its possible Role in Plant Senescence

The movement of metabolites and nutrients towards developingseeds, and their accumulation there, appears to play an importantrole in the regulation of senescence of the shoot in annualplants. The possibility that the directed-transport of nutrientstowards developing fruits is regulated by growth hormones hasbeen studied in French bean (Phaseolus vulgaris). Applicationof 3-indolyl acetic acid (IAA), in lanolin, to peduncles fromwhich the fruits had been removed, resulted in significantlygreater accumulation of ³²P (applied to the lower part of thestem) in the region of hormone application than in pedunclestreated with lanolin only. When gibberellic acid (GA) or kinetinwere applied alone to defruited peduncles they had no significanteffect on the accumulation of ³²P at the point of applicationof hormone, but when either was applied with IAA they greatlyenhanced the effect of the latter on movement of ¹⁴C-labelledphotosynthates from the leaves to the peduncles. It is suggestedthat hormone-directed transport may play an important role indirecting the movement of nutrients towards developing seeds,which are rich sources of endogenous hormones.     

Auxin-promoted Transport of Metabolites in Stems of Phaseolus vulgaris L: EFFECTS AT THE SITE OF HORMONE APPLICATION

The hypothesis that indol-3yl-acetic acid (IAA) stimulates solutemobilization in stems by increasing metabolite utilization (i.e.sink strength) and/or sieve tube unloading mechanisms was investigatedusing seedlings of Phaseolus vulgaris L. Sink strength was assessedby obtaining estimates of rates of growth, ¹⁴C incorporationinto protein, sucrose metabolism, and sucrose uptake. The dataobtained suggest that, whilst enhancing assimilate mobilization,IAA had no short term effect on the sink strength of the treatedstumps. In several experiments involving long distance transportof ¹⁴C-assimilates, the magnitude of auxin-promoted transportwas found to be insensitive to potential changes in sink strength.Furthermore, ¹⁴C transport profiles demonstrated that the siteof hormone action was not confined to its point of application.     

The Transport of Sugar, Water, and Ions into Developing Potato Tubers

Diurnal variations in the pattern of movement of sugars, water,and ions into developing tubers of the potato (Solanum tuberosumL.) were investigated. It was demonstrated using a recordingbalance that large increases in the fresh weight of tubers occurduring a dark period of reduced transpiration. Movement of assimilated¹⁴C did not reflect similar large changes and much of the weightchange observed is considered to be fluctuations in tuber watercontent. This water was shown to be moving predominantly throughthe xylem of the stolon by introducing labelled ions, ³²P and⁸⁹Sr into the plants. ³²P, which moves in both xylem and phloem,was transported to the tuber at a constant rate whereas ⁸⁹Sr,which behaves like calcium and is relatively immobile in thephloem, only moved into the tuber during the dark period. As well as the over-all long-term diurnal fluctuations severalsmaller rapid changes were recorded in the rate of water movement.Switching from darkness to light caused a transient increasefollowed by a rapid decrease in tuber weight. Switching fromlight to darkness caused a rapid increase in tuber weight. Insome experiments small oscillations in tuber weight were recorded.The possibility of these oscillations being directly relatedto cyclic changes in transpiration is considered. The resultsare discussed in relation to solute movement within plants.     

A Microautoradiographic Study of Auxin Transport in the Stem of Intact Pea Seedlings (Pisum sativum L.)

Soluble-compound microautoradiography was used to determinethe distribution of radioactivity in transverse sections ofintact dwarf pea stems (Pisum sativum L.) following the applicationof [³H]IAA to the apical bud. Near the transport front labelwas confined to the cambial zone of the axial bundles, includingthe differentiating secondary vascular elements. Fully differentiatedphloem and xylem elements remained unlabelled and no radioactivitywas detected in the leaf or stipule traces. Similar resultswere obtained in experiments with Vicia faba L. plants. Nearerthe labelled apical bud of the pea there was a more generaldistribution of label and evidence was found of free-space transportof radioactive material in the pith. When [³H]IAA was applied to mature foliage leaves the greatestconcentration of label was found in the differentiated phloemelements of the appropriate leaf trace and in the phloem ofthe adjacent axial bundles. Both basipetal and acropetal transportwas detected in this case. These results are consistent with the conclusions drawn fromearlier transport experiments which indicated that in the intactplant the long-distance basipetal transport of auxin from theapical bud takes place in a system which is separated from thephloem transport system and suggests that the vascular cambiumand its immediate derivatives may function as the normal pathwayfor the longdistance movement of auxin in the plant. The physiologicalsignificance of such a transport system for auxin is discussed.     

Distribution of Radioactivity from Exogenously Supplied [1-14C]Indol-3-yl-acetic Acid and [3, 4-3H (N)] Gibberellin A, in Geotropically-stimulated Picea abies (L.) Karst. Roots

The distribution of exogenously-supplied radioactive labelledindol-3-yl-acetic acid (IAA) and gibberellin A₁ (GA₁) in geotropicallystimulated roots of Norway spruce (Picea abies (L.) Karst.)has been demonstrated. Seedlings were positioned with theirroot tips in 2.1 x 10^–6 M [¹⁴C]IAA or 1.3 x 10^–8m ³H-GA₁ for 4 and 20 h, respectively. After geotropic stimulationfor 90 min in the horizontal position the root tips were cutlongitudinally in 50 µm thick sections, using a freeze-microtome.The radioactivity in the ¹⁴C-IAA treated roots occurred in higherconcentration in the lower than in the upper halves (ratio 1.25:1). A similar trend was observed in the [³H]GA₁-treated rootswhere the ratio lower: upper halves was 2.04: 1. The ratio ofradioactivity in right and left halves of vertical roots wasapproximately the same in roots supplied with [¹⁴C]IAA and [³H]GA₁(1.09: 1). The supplied radioactive compounds were analysed chromatographicallyafter extraction in methanol of 6 mm apical root segments. Onlya small fraction (7–8 per cent) of the supplied [¹⁴C]IAAwas revealed unchanged in the segments. The major part of thechromatographed, labelled compound has not been identified,but on basis of its R_F value it is suggested that it may beindol-3-acetyl-aspartic acid (IAAasp). The chromatographic analysis of the [³H]GA,-treated segmentsshowed that only small fractions of this gibberellin has beenconverted to other compounds. These results have been discussed and correlated with knowledgeof plant growth regulators and their participation in root geotropism. Picea abies, spruce, geotropism, gibberellin A₁, indol-3-yl-acetic acid, growth regulators, redistribution in roots     

Auxin-Promoted Transport of Metabolites in Stems of Phaseolus vulgaris L.: Auxin Dose-Response Curves and Effects of Inhibitors of Polar Auxin Transport

Transport of ¹⁴C-photosynthate in decapitated stems of Phaseolusvulgaris explants was dependent on the concentration of indole-3-aceticacid (IAA) applied to the cut surfaces of the stem stumps. Thephysiological age of the stem influenced the nature of the transportresponse to IAA with stems that had ceased elongation exhibitinga more pronounced response with a distinct optimum. Increasednutrient status of the explants had little influence on theshape of the IAA dose-response curve but increased, by two ordersof magnitude, the IAA concentration that elicited the optimalresponse. Applications of the inhibitor of polar auxin transport,1-(2-carboxyphenyl)-3-phenylpropane-1, 3-dione (CPD), affectedIAA-promoted transport of ¹⁴C-photosynthates. At sub-optimalIAA concentrations, CPD inhibited transport, whereas at supra-optimalIAA concentrations, ¹⁴C-photosynthate transport was marginallystimulated by CPD. Treatment with CPD resulted in a significantreduction in stem levels of [¹⁴C]IAA below the site of inhibitorapplication, while above this point, levels of [¹⁴C]1AA remainedunaltered. The divergent responses of auxin-promoted transportto CPD treatment are most consistent with a remote action ofIAA on photosynthate transport in the decapitated stems. Key words: Auxin, photosynthate, transport     

Transport of 14C-lableled indoleacetic acid in Vicia root segments

IAA transport in Vicia root segments was investigated for comparisonwith that in intact roots. Lanolin paste (1-mm-wide ring) oragar blocks (3?3?1.5mm), both containing IAA-2-¹⁴C were appliedto the surface or a cut end of the root segments, respectively;transported ¹⁴C was collected in receiver agar blocks placedon the cut end of the segments. When lanolin paste was appliedto 5-mm segments, basipetal transport of IAA predominated overacropetal transport. When agar blocks were applied to 1- and2-mm segments, the same was true; in longer segments (3 and5 mm long), however, basipetal movement occurred predominantlyat first but was surpassed by acropetal movement after 2–3hr. Among the segments tested (regions 2–4, 4–6and 8–10 mm from the tip), the most apical one showedthe distinctest predominancy of basipetal movement. The velocitiesof the acropetal and basipetal movement of the ¹⁴C were estimatedat 3–3.8 and 8–12 mm/hr, respectively. Autoradiographicstudy and the experiment in which wire was inserted longitudinallythrough the central part of the segments showed that basipetalmovement occurred mainly through the outer part of the rootsand acropetal movement mainly through the central cylinder.The present results were compatible with those obtained previouslywith intact roots. Some properties of polar movement, such asits specificity, inhibition by TIBA, and dependency on terneprature are described. (Received March 22, 1978; )     

The Polarity of Movement of Endogenously Produced IAA in Relation to a Gravity Perception Mechanism

The polarity of radioactive IAA transport in segments of theleaf sheath base of Avena fatua L. is reversed upon their inversionthrough 180° transport towards the apex being greater thantowards the base. Changes in rates of transport leading to thisreversal can be detected within 10–20 min, correlatingwith the timing of statolith movements from the base to theapex of statocyte cells and conforming to a proposed model forthe gravity control of auxin transport. Transport of H³-trytophan-derived H³-IAA was found to undergosimilar changes in polarity upon re-orientation as did exogenouslyapplied H³ or C¹⁴ IAA. It is concluded that the proposed modelalso relates to the movement of endogenously produced IAA.