Light- and Auxin-Induced Leaflet Opening in Detached Pinnae of Mimosa pudica

Leaflet pairs from detached pinnae of Mimosa pudica opened afterthe pinnae had been irradiated with light (2 W·m^–2)of 726 or 403 nm, whereas they remained almost closed with lightof 585 or 656 nm. Light-induced leaflet opening was observedonly in the daytime, from 6:00 to 16:00. Application of IAAat more than 30 µg/ml to the cut end of the pinna rachisesmade the leaflets open even in darkness with almost constantlag times of about 100 min which were independent of the concentration.NAA and 2,4-D also made the leaflet open at lower concentrationsthan IAA. Auxin-induced leaflet opening showed diurnal variation.Application of IAA for 2 to 6 min, depending on the concentration,was enough to open the leaflets. Autoradiography showed thatIAA was transferred from the cut end of a rachis throughouta pinna within 4 min. ¹Present address: Biological Institute, Faculty of Science,Kobe University, Kobe 657, Japan. (Received September 24, 1982; Accepted March 4, 1983)     

Maintenance of the diurnal petiolar movement by exogenous indole-3-acetic acid under day-night cycles in Mimosa pudica

Effects of plant hormones on the diurnal movement of the petiole of Mimosa pudica L. were examined under the conditions of day-night cycles. Surgical removal of the leaf at the middle of the petiole led to gradual loss of the movement. Aqueous solutions of indole-3-acetic acid (IAA) applied as a pulse each day 10 μl at (10⁻⁶−10⁻⁵ M ) or continuously (10⁻⁸−10⁻⁶ M ) to the cut end of the petiole maintained a diurnal movement that has the same phase of oscillation as that shown in the intact leaf. Pulse treatments given at different times of the day had no effect on the phase of the oscillation. Higher concentrations of the acid gave larger amplitudes. Other hormones such as gibberellic acid, kinetin and [R,S]-abscisic acid or 1-aminocyclopropane-1-carboxylic acid did not show diurnal movement-maintaining activities of IAA. Gibberellic acid disturbed the phase of the diurnal movements and kept the petiole elevated at very high positions. A possible action mechanism of IAA is discussed.     

Effects of Jasmonic Acid on Motor Cell Physiology in Mimosa pudica L. and Cassia fasciculata Michx

When applied to pulvini of Mimosa pudica, jasmonic acid (JA)affected neither proton fluxes nor the membrane potential ofthe motor cells. When added to leaflets of Cassia fasciculata,JA increased the rate of dark-induced pulvinar movements ina concentration-dependent manner. This effect was observed withinas little as 15 min after a 1-h treatment that preceded theinducing signal. Treatments in buffered media at acidic pH resultedin the greatest physiological responses. Light-induced pulvinarmovements were considerably reduced under the same conditions.With continuous illumination, JA induced a closing movementof the leaflets in a concentrationdependent manner. These resultsare discussed in relation to the ionic changes in the pulvinarmotor cells and in relation to results obtained previously upontreatment of Cassia plants with ABA. Although ABA and JA havesimilar physiological effects on the dark-induced closure, theydiffer in the type of response elicited by brief treatment andwith respect to light-induced opening. (Received September 27, 1993; Accepted January 15, 1994)     

Growth-regulator Interactions in the Growth of the Shoot System of Avena sativa Seedlings: I. THE GROWTH OF THE FIRST INTERNODE

1. Segments, 3.5 mm. long, cut from the first internode of Avenasativa seedlings grown in complete darkness respond to bothauxins and gibberellic acid by accelerated extension. 2. The optimum concentration of indole-3-acetic acid (IAA) is10 p.p.m. and of gibberellic acid (GA) is 0.1 p.p.m. 3. The degree of stimulation relative to the growth of controlsegments is affected by the inclusion in the segement of thenode between the internode and coleoptile. Thus the gibberellineffect is greatly increased while the IAA effect is decreased.The optimal concentrations are not affected by inclusion ofthe node. 4. These results can best be explained in terms of the supplyby the node tissue of an endogenous auxin which is necessaryfor the expression of GA action. 5. Numerous factorial experiments demonstrated that there isno detectable interaction between applied IAA and GA in thepromotion of first-internode extension. This implies that thepostulated endogenous auxin which synergized GAA action in (4)is either an active form of IAA produced only in the node tissueor is a completely different auxin. 6. No synergism of growth-promotive action can be detected betweenGA and the two synthetic auxins I-naphthylacetic acid and 2,4-dichlorophenoxyaceticacid. 7. p-chlorophenoxy-iso-butyric acid (PCIB) anc 2,4,6-trichlorophenoxyaceticacid (2,4,6-T) act as weak auxins and thus antagonize competitivelythe promotive action of GA. 8. The anti-auxin -(I-naphythyl-methyl-sulphide)propionic acid(NMSP) antagonizes competitively the promotive action of bothIAA and GA. 9. The facts under (5)–(8) suggest that auxins and GAare acting at the same growth-promotion centres and may competefor them. 10. Growth inhibitions are induced by high concentrations ofPCIB, 2,4,6-T and NMSP. The inhibitions produced by PCIB and2,4,6-T are both synergized by supra-optimal concentrationsof IAA while that of NMSP is synergized by supra-optimal concentrationsof both IAA and GA. This similarity of the effects of IAA andGA suggests that their inhibition actions also are of a closelysimilar nature.     

EFFECTS OF GROWTH SUBSTANCES ON FROND AND FLOWER PRODUCTION IN LEMNA GIBBA G3

Frond and flower production in a long-day duckweed, Lemna gibbaG3, growing under different photoperiodic conditions in presenceand absence of varied concentrations of applied IAA, KIN andGA was investigated. The predominant actions of IAA, KIN and GA were revealed tobe respectively the depression of flowering, the promotion offrond multiplication and the enhancement of flowering. The rateof increase in FL % as an index of the concentration of floralstimulus in the tissues was curtailed greatly by IAA, to a lesserextent by KIN and little by GA. GA at 10^–5 M apparentlyreduced the critical day length (12 hr) and the induction period(48 hr) by 2 and 4 hr, respectively. No vegetative growth waspromoted by KIN in the light. On the basis of these and relevant findings the sites of actionof the growth substances examined in the flowering process ofthe duckweed were suggested. (Received May 30, 1965; )     

Site of photo-reception to opening response in Mimosa leaflets

A pair of leaflets excised from Mimosa pinna and floated onwater for a day, responded to light and mechanical stimuli.Only when a small spot of light was shone on the abaxial surfaceof pulvinule, did the opening response of the leaflets occur.No translocatable effect from suchirradiation was observed. ¹ Present address: Biological Institute, Faculty of Science,Kobe University, Kobe 657, Japan. (Received May 17, 1973; )     

Phototropic leaf movements and photosynthetic performance in an amphibious fern, Marsilea quadrifolia

Diurnal phototropism has not been reported in ferns. In this study we found that the four leaflets of the amphibious fern Marsilea quadrifolia are capable of adjusting their leaflet angle and leaflet azimuth in response to changes in the position of the sun’s direct beam, exhibiting more diaphototropic movements (orienting the plane of the lamina perpendicular to incident light) in the morning and late afternoon, and more paraphototropic movements (orienting the plane of the lamina parallel to incident light) at noon. In addition, by cutting off the leaflet lamina and covering portions of leaflets with black tape, the junction between the leaflet and petiole was found to be responsible for light reception. Among the light spectrum investigated, blue light was the most effective at inducing diaphototropism. The role of diurnal phototropism in enhancing carbon return and ameliorating photoinhibition was also evaluated. It was concluded that diurnal phototropic leaf movement represents one of the plastic responses enabling this amphibious fern to grow under terrestrial conditions.     

Comparative Effects of lndole-3-acetic acid, Abscisic acid, Gibberellic acid and 6-Benzylaminopurine on the Dark- and Light-induced Pulvinar Movements in Cassia fasciculata Michx

Effects of plant hormones were examined on the dark- and light-inducedmovements of Cassia fasciculata. Indole-3-acetic acid (IAA),gibberellic acid (GA₃) and 6-benzylaminopurine (6-BAP) inhibitedthe scotonastic movement whereas abscisic acid (ABA) enhancedit. After brief treatments (5 to 30 min), the ABA effect wasinhibitory rather than promotional. Hormonal treatment in theacidic range gave the best physiological response for ABA, butthe greatest efficiency of IAA, GA₃ and 6-BAP was obtained withpH values close to neutrality. Three to 5 h were needed beforeexpression of the physiological effect triggered by GA₃ and6-BAP, while 5 min treatments were sufficient for IAA and ABA.Light-induced movements were largely enhanced by IAA and slightlyby GA₃ but inhibited by 6-BAP and ABA. The results are discussedin relation to the ionic changes in the pulvinar motor cells,regulating leaflet movements. Key words: Abscisic acid, auxins, cytokinins, gibberellic acid, pulvinar movements     

Resemblance of growth substances to metal chelators with respect to their actions on duckweed growth

Action patterns of IAA, KIN and GA on the growth of Lemna gibbaG3 are similar to those reported for agents chelating both cupricand ferric ions. Relatively high doses of growth substances,e.g.10^–6 M IAA or KIN and ^–4 M GA, inhibit developmentof photoperiodically induced flower buds and antagonisticallypromote frond multiplication; whereas, at relatively low doses,e.g. 10^–9 M IAA or KIN and ^–5 M GA, they acceleratethe process occurring in the latter half of the induction periodto enhance flower induction. Complex forming abilities of IAA,KIN and GA with cupric and ferric ions are demonstrated spectrophotometrically.Moreover, the ferrous ion-dependent oscillatory change in reproductiveand vegetative photophilies of duckweed is eliminated by KINbut not by IAA and GA. Of the three growth substances tried,KIN alone shows an affinity for ferrous ions. (Received April 5, 1971; )     

Responses of Stomata to IAA and Fusicoccin at the Opposite Phases of an Entrained Rhythm

The stomata of Commelma communis showed reduced opening responsesto light and low CO₂ concentrations during the night phase oftheir entrained circadian rhythm. Increased supplies of potassiumions, and treatments with indol-3-ylacetic acid and fusicoccin,failed to promote opening during the night phase to a levelequivalent to that in the day phase. The inability of fusiccocinto overcome the suppression of opening during the night phasecontrasts with its ability to counteract the closure inducedby agents such as CO₂, darkness and abscisic acid. It is concludedthat there are at least two basic mechanisms by which the turgorof guard cells can be regulated, one which is susceptible tooverriding control by fusicoccin and another which is unaffectedby fusicoccin. Several previous studies had shown a positive correlation betweenmalate in the epidermis (mainly located in guard cells) andstomatal opening. In the present experiments the aperture/malatecorrelation was broken in epidermis treated with fusicoccinduring the night phase of the rhythm. The amount of malate presentexceeded that associated with the same stomatal aperture inthe day phase. Possible explanations are (1) that fusicoccinstimulates similar proton fluxes out of the guard cells duringboth phases of the rhythm, but an unknown factor imposes a restrictionon stomatal opening during the night phase; (2) that there arelower proton fluxes in the night phase (limited, for example,by a reduced supply of ATP) but chloride availability or transportis reduced to an even greater extent so that a larger productionof malate in the guard cells is required. Key words: Stomata, IAA, Fusicoccin, Rhythms     

Role of growth regulators in the bean hypocotyl hook opening response

Summary The opening of the hypocotyl hook in bean seedlings is due to a rapid elongation of cells on the inner side of the hook elbow. Red light promotes hook opening by inducing this cell elongation.Opening is inhibited by low concentrations of indoleacetic acid (IAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), and higher concentrations of these auxins cause a closure of the hook. In darkness, opening is induced slightly by p-chlorophenoxyisobutyric acid (PCIB), whereas in red light this auxin antagonist promotes opening only when IAA is added simultaneously to inhibit opening.The amount of diffusible auxin released by the hook tissue is not affected by red illumination that is sufficient to induce maximal hook opening.Gibberellic acid (GA) promotes the hook opening. The magnitude of its effect is, however, rather small, especially in darkness. (2-Chloroethyl)-trimethylammonium chloride (CCC) and 2-isopropyl-4-(trimethylammonium-chloride)-5-methylphenyl piperidine-1-carboxylate (Amo-1618) inhibit hook opening in red light, and this inhibition is completely overcome by addition of GA.Cytokinins and abscisic acid at rather high concentrations inhibit hook opening in light but produce no significant effect in darkness.Hook opening is promoted by Ca⁺⁺ and K⁺, and notably by Co⁺⁺ and Ni⁺⁺.It is concluded that 1. endogenous gibberellin assists in hook opening, but light does not act by changing the gibberellin level; 2. light does not act by decreasing the endogenous auxin level; and 3. cytokinins or abscisic acid do not seem to have a special role in the response.     

An Ecophysiological Comparison of Measurements of the Diurnal Rhythm of the Leaf Elongation and Changes of the Leaf Thickness of Salt-resistant Dicotyledonae and Monocotyledonae

Rozema, J., Arp, W., van Diggelen, J., Kok, E. and Letschert,J. 1987. An ecophysiological comparison of measurements of thediurnal rhythm of the leaf elongation and changes of the leafthickness of salt-resistant Dicotyledonae and Monocotyledonae.—J.exp. Bot. 38: 442–453. The continuous measurement of leaf elongation and leaf thicknesswith the use of a rotation potentiometer set up revealed a rapidand sensitive reaction of halophytic plants to conditions affectingthe plant's water relations. At increased salinity (450 molm^–3 NaCl) the rate of leaf elongation decreased both inAster tripolium and in Sparlina anghca. Increased shrinkageduring the day and a long period for recovery swelling at nightin leaves of Aster iripolium at increased salinity illustratesthat water shortage is part of the cause of salinity-inducedgrowth reduction. All dicotyledonous species analysed (Aster tripolium, A triplexhastata, A. littoralis, Suaeda maritima and Beta vulgaris) showeda day/night ratio of the leaf elongation rate lower than 1,while this ratio was higher than or equal to 1 in Monocotyledons(Spartina anglica, Juncus gerardii, J. maritimus, Festuca rubrassp. litoralis, Elymus pycnanthus). With the exception of Triglochinmaritima none of the monocotyledonous halophytes tested (Sparlinaanglica, Juncus gerardii, J. maritimus, Festuca rubra ssp. litoralis,Elymus pycnanthus) exhibited a diurnal rhythm of leaf thicknesschanges, such as was observed for all dicotyledonous speciesstudied (Aster tripolium, Atriplex hastata, A. littoralis, Salicorniabrachyslachya, Suaeda maritima, Glaux maritima, Odontites vernassp. serotina). The diurnal pattern of the leaf elongation rateand the leaf thickness changes can be explained by variationof photosynthetic rate and transpiration water losses by stomatalclosure in the dark and opening in the light such as shown forthe dicotyledon species Glaux maritima. This difference betweendicot and monocot species in diurnal variation of the leaf elongationrate and leaf thickness may partly be explained in terms ofthe different position of the growth zone and possibly by adifference in elasticity of the tissue of halophytic monocotyledonsand dicotyledons. The consequences of these differences arediscussed. Key words: Leaf elongation rate, leaf thickness, water relations, salt resistance, Dicotyledonae, Monocotyledonae     

Simulation of acetylcholine action by {beta}-indole acetic acid in inducing diurnal change of floral response to chilling under continuous light in Lemna gibba G3

IAA induced a diurnal change in floral response to chillingof Lemna gibba G3 under continuous light, as has been reportedfor ACh. This IAA effect was antagonized by an antiauxin, T-CA.NAA and 2,4-D had no influence on the temperature sensitivityof the duckweed. ACh-like reagents were examined with respectto the sensitivity to their antagonists. Atropine, an ACh-specificantagonist, inhibited the action of IAA. The IAA treatment,however, decreased the ACh content of the plant by ca. 30%.Thus, IAA and ACh are surmised to attack the same or similaraction site(s) independently. (Received July 19, 1978; )     

与变化光环境关联的刺槐光合气体交换和复叶运动

在夏季的晴天阳生叶和阴生叶方位角和方向角的日变化均很小 ,但是叶片的中脉角存在显著的日变化差异 ,这种叶片垂直方向上的显著运动与躲避中午的强光胁迫有关。而在阴天 ,阳生叶和阴生叶方位角、方向角和中脉角的日变化均很小。这说明变化的光环境是引起刺槐复叶运动的主要原因 ,而节律性运动不是刺槐复叶运动的诱导因素。机械固定叶片的控制实验显示 :机械固定叶片的光合速率、气孔导度和蒸腾速率均显著低于对照叶片 ,而机械固定叶片温度明显高于对照叶片 ,这可能是由于机械固定叶片和对照叶片两者在光辐射接受量上的差异造成的。光诱导下的刺槐复叶运动是植株维持最佳生理状态的形态调节表现。     

Control of Hypocotyl Hook Angle in Phaseolus mungo L. Role of Growth Substances

Effects of growth hormones on the hook angle and light responseof Phaseolus mungo L. hypocotyl hooks are described and theresults are discussed with reference to the functions of otherparts of the seedling in controlling the growth and shape ofthe hook. Apically applied IAA (indolyl acetic acid) prevented hook openingin decapitated seedlings in the dark and in all the red-irradiatedseedlings. [¹⁴C]IAA experiments showed that only a small quantityof IAA (2–6 ?g per hook) was required to produce theseeffects, and that transport of IAA through the hook was negligibleand unaffected by red irradiation. ABA (abscisic acid) had little effect on the hooks or theirlight response. Applied ethylene and IAA-induced ethylene slightly closed thehooks, but only slightly reduced light-induced opening. IAAreduced the effect of ethylene in the dark, but after irradiationthe hooks appeared more sensitive to the ethylene in the presenceof IAA, resulting in light-induced hook closure. Basally applied kinetin (6-furfurylaminopurine) prevented decapitatedhooks from opening in the dark, especially when GA₃ (gibberellicacid) was also present. Some combinations of kinetin and GA₃(with high kinetin concentrations) also prevented light-inducedopening, but combinations with lower kinetin concentrationsallowed almost as much opening as was found in intact hooks. It is proposed that the terminal parts act by regulating thesupply of cytokinins and gibberellins from the basal parts,and that IAA does not mediate this funotion in this species. The results are compared with those reported for other species.     

Diurnal Patterns of Transport and Accumulation of Minerals in Fruiting Plants of Lupinus angustifolius L.

Fluctuations in mineral elements id xylem (tracheal) sap, fruitphloem sap, leaflets and dmloping fruits were studied in a fieldpopulation of Lupinus angustifolius L. by three-hourly samplingover a 39 h period. Elements usually reached maximum contentsor concentrations at or near noon, minimum levels during thenight. Amplitudes of diurnal fluctuations in minerals lay withinthe range ±4–33 per cent of the mean content ofleaflets, and ±17–157 per cent of the mean concentrationsin xylem and phloem sap. Most minerals elements fluctuatcd inphase with daily changes in sugar level of phloem sap and drymatter and carbohydrate fluctuations of leaflets, suggestinga coupling of translocation of photosynthate and minerals fromthe leaflets. Rates of import of minerals by shoots wereestimatedfrom shoot transpiration and mineral concentrations in trachealsap. Average day time rates of import of most elements were12–25 times those at night. Translocation of minerals,nitrogen and carbon to fruits also exhibited diurnal periodicity,average rates of import king three to seven times higher inthe day than at night. A model of transport based on the carbonand water economy of the fruit suggested that P, K, Fe, Zn,Mn and Cu were imported predominantly by phloem. Estimates ofvascular import accounted for 87–104 per cent of the fruit'sactual increment of these elements. Na and Ca were gauged tobe imported mainly by xylem, Mg almost equally by xylem andphloem. However, large discrepancies existed for these threeelements between estimated vascular import and actual intakeby the fruit. Lupinus angustifolius L., mineral transport, accumulation, fruits, xylem sap, phloem sap, transpiration     

A Comparative Study of the Effects of Some Growth Regulators on the Biochemistry of Oryza sativa L.

A comparative study was made on the effects of indol-3yl-aceticacid (IAA), maleic hydrazide (MH), naphthalene acetic acid (NAA),and gibberellic acid (GA) at a concentration of 1o^-1 mg I^-1,on the growth and metabolism of Oryza sativa L., cv. Bhasamanik.All the growth substances excepting NAA caused promotion ofroot elongation and increased the number of roots formed buthad very little effect on shoot growth. NAA was found to havea retarding effect on the general growth of the plant, but alsoincreased the number of roots formed. At the concentration used IAA caused enhanced protein synthesisin parallel with increased root growth. However, with MH nosuch direct relationship could be observed between root growthand protein synthesis at the early stage, but stimulation ofroot growth was directly related to increased protein synthesisat the later stage. Despite the adverse effect of NAA on thegrowth of the plant, protein synthesis was not affected. Infact a general increase in the protein content of the plantwas observed. In plants treated with GA increased sugar contentand enhanced protein synthesis were noted. The levels of free amino acids were found to be affected markedlyby the application of the growth regulators. With NAA treatmentan accumulation of asparagine in the shoot and with GA an increasein the level of alanine in the root were noted. Aspartic aciddecreased in the roots, but increased in the shoots of treatedplants. Glutamic acid was lower in plants treated with IAA,MH, and NAA, while treatment with GA increased it in the shootat the tillering stage. In the shoots of IAA-, MH-, and NAA-treatedplants an increase in the quantity of serine was noted whereasunder the action of GA both serine and glycine decreased.     

The Effect of Growth Hormones on Cell Division and Expansion in Liquid Suspension Cultures of Acer pseudoplatanus

The effects on cell division and cell size of indole-3-aceticacid (IAA), gibberellic acid (GA), and kinetin were studiedin liquid suspension cultures of cambial cells derived fromAcer pseudoplatanus. It was shown that all three hormones promotecell division and that the effects of both GA and kinetin areadditive to those of IAA, but the effects of GA and kinetintogether are not additive. Treatment with IAA resulted in anincrease of mean cell size (indicating that cell expansion ispromoted), but after GA or kinetin treatment the mean cell sizewas smaller, indicating that little cell expansion had takenplace after each division. The results are discussed in relationto previous work on the effects of hormones in the intact cambiumand to current theories on the interactions of growth hormones.     

Changes in enzyme activity during elongation and tuberization of stolons of Solanum tuberosum L. cultured in vitro

Isolated stolons of Solanum tuberosam L. were cultured in vitroin the presence of kinetin, which induced tuber initiation orgibberellic acid which inhibited initiation. Progressive changes in enzyme activity, at the locus of tuberinitiation, were monitored at specified intervals. In the presenceof kinetin soluble invertase activity was decreased with timewhereas gibberellic acid (GA) evoked substantial increases inactivity. Acid phosphatase activity was enhanced by GA but changedonly slightly under tuber inducing conditions. Enzymic hydrolysisof glucose-6-phosphate and fructose-6-phosphate decreased duringthe course of tuber induction but increased in the presenceof GA. In contrast hydrolysis of 3'AMP was stimulated undertuber inducing conditions. GA evoked substantial increases in peroxidase activity duringthe initial stages of incubation while under tuber inducingconditions increased activity was only observed after 5 days.Substantially higher levels of IAA oxidase activity were associatedwith tuber initiation. RNase activity decreased with time undertuber inducing conditions but showed an initial stimulationin the presence of GA. These results are discussed with referenceto the role of these enzymes in carbohydrate metabolism andthe regulation of hormone levels. (Received February 29, 1972; )     

Effects of Light Spectral Quality on Morphogenesis and Source–Sink Relations in Radish Plants

The accumulation of dry matter and the content of major phytohormones in the aboveground and underground plant parts, as well as light curves and the diurnal course of photosynthesis in the leaves were studied in radish (Raphanus sativusL.) plants of different ages that were grown under red (RL) or blue (BL) light. As seen from the rapid increase in plant biomass, the development of storage organs (hypocotyl or tap root) started on the 14th day after the emergence of seedling of the BL plants and on the 21st day for the RL plants. Conversely, RL stimulated biomass accumulation in the aboveground parts (petioles and stems) already in the early stages of plant development. Light spectral quality only slightly affected the activity and the diurnal course of photosynthesis. The GA content was ten times higher in the aboveground parts of the RL plants than those of the BL plants. The hypocotyl of the BL plants contained much higher amounts of cytokinins and IAA than that of the RL plants. The specific responses of the source–sink relations to the light quality were related to the distribution of various phytohormones between the aboveground and underground parts of the plants: RL increased the content of gibberellins (GA) in the aboveground parts of plants, thus increasing their sink activity, whereas BL stimulated the synthesis of cytokinins and IAA in the hypocotyl and enhanced its development. Light quality-specific morphogenetic responses were reversed when plants were treated with exogenous GA or paclobutrazol, an inhibitor of GA synthesis. The treatment of the BL plants with exogenous GA stimulated petiole and hypocotyl elongation and induced stem formation. The treatment of the BL plants with paclobutrazol led to shortened petioles, the flattening of the storage organ, and the disappearance of the stem.