Comparison of Growth Responses of Barnyard Grass (Echinochloa oryzoides) and Rice (Oryza sativa) to Submergence, Ethylene, Carbon Dioxide and Oxygen Shortage

Unlike germination of wheat (Triticum aestivum), millet (Eleucinecoracana), and sorghum (Sorghum caudatum), that of Echinochloaoryzoides (barnyard grass) and Oryza satwa (rice) was not inhibitedby poorly aerated solutions with 11 k Pa oxygen (equilibriumpartial pressure) or less In the dark, seedling shoots of riceincluded a coleoptile, and in Echinochloa, a mesocotyl alsoGrowth in fresh and dry weight of shoots was strongly depressedby poorly aerated solutions in both rice and Echinochloa butthe effects on extension differed in the two species in rice,coleoptile extension was promoted by solutions partly depletedof oxygen, and also by the absence of oxygen The stimulationin partly de-oxygenated solutions resulted from the combinedpromoting effects of small oxygen partial pressures, carbondioxide, ethylene and buoyant tension in contrast, these treatmentsneither promoted nor inhibited elongation by the Echinochloacoleoptile while severely inhibiting extension of the mesocotyl Overall, poorly aerated solutions lengthened the shoot of riceand shortened it in Echinochloa when compared with those submergedin well-aerated solutions These opposite effects were broughtabout by the same gaseous changes, i e oxygen shortage, elevatedethylene and carbon dioxide The effect on Echinochloa was almostentirely restricted to the mesocotyl, coleoptile extension beingremarkably insensitive to large increases in ethylene and carbondioxide, or to extreme oxygen shortage Seedlings of the twospecies thus have contrasting strategies for survival Stress, submergence, ethylene, oxygen shortage, carbon dioxide, adaptation, anaerobiosis, rice (Oryza sativa L), barnyard grass [Echinochloa oryzoides (Ard) Fritseh]     

A Critical Study of the Auxin Theory of Growth Regulation in the Mesocotyl of Avena sativa

A method of growing Avena seedlings is described, which allowsthem to be handled individually in darkness. Mesocotyls of seedlinge from which the tip of the coleoptileis repeatedly removed are as long as those of control plantsnot so decapitated. Mesocotyls of seedlings which are deseeded on the 3rd day ofgrowth, followed by decapitation of the cleoptile tip on the4th day, are, at 7 days old, as long as those of controls notso decapitated. When deseeded plants are decapitated, regeneration of auxinproduction occurs at the tip of the coleoptile stump. Where a reduction in the length of the mesocotyl results fromdecapitation, a wound reaction is probably concerned in additionto any auxin changes. Removal of the coleoptilar node causes a sharp decrease in thefinal length of the mesocotyl. Heating intact seedlings at 40° C. for 3 hours causes areduction in the length of the mesocotyl but not of the coleoptile.The effect of heating is not reversed by subsequent treatmentat low temperature, which instead appears to augment these effects. When seedlings are exposed to the action of KCN, iodoacetate,or anaerobic conditions, and illuminated while so exposed, perceptionof light takes place, resulting in a reduction in the lengthof the mesocotyl. Perception of light takes place in seedlings germinated at normaltemperatures, but maintained at low temprature during illuminationand also in seedlings grown for 6 weeks at 2° C. withoutany previous growth at normal temperature. Light perception takes place in embryos excised from dry grainand grown on a culture medium. No difference in free amino-acid content is apparent betweendark grown and illuminated seedlings. The effects of illumination survive a period of drying downand become apparent upon subsequent germination of the grainin darkness. The drying process itself causes an additionalreduction in mesocotyl length. It is concluded that auxin itself is not the primary reactantin the perception process, and that the growth of the mesocotylis probably controlled by the coleoptilar node and plumulargrowing point, rather than by auxin diffusing downward fromthe tip of the coleoptile.     

Sources of Free IAA in the Mesocotyl of Etiolated Maize Seedlings

Sources of free indole-3-acetic acid (IAA) for the mesocotyl of intact etiolized maize ((Zea mays L.) seedlings are evaluated. The coleoptile unit, which includes the primary leaves and the coleoptilar node, is the main source of free IAA for the mesocotyl. The seed and the roots are not immediate sources of IAA supply. Dependence of the apical growing region of the mesocotyl on the coleoptile unit as a source of free IAA is almost total. One-half or more of the supply of IAA comes from the coleoptile tip, the rest mainly from the primary leaves. Removal of the coleoptile tip results in inhibition of mesocotyl elongation. The hypothesis that growth of the mesocotyl is regulated by auxin supplied by the coleoptile is supported. Conjugated forms of IAA appear to play little part in regulating the levels of free IAA in the shoot.     

Comparative studies on auxin and fusicoccin actions on plant growth

Mode of action of FC was compared with that of auxin in differentexperimental systems and the following results were obtained.

1. FC, as well as auxin, primarily induced elongation of the epidermisof pea epicotyl segments, but it also promoted elongation ofthe inner tissue, as judged by its action in split stem tests,elongation of hollow-cylinder segments and elongation of unpeeledand peeled segments.
2. FC decreased the minimum stress relaxationtime (T₀) and increasedthe extensibility (mm/gr) of the epidermalcell wall of peaepicotyl segments, as did auxin.
3. FC failedto induce expansion growth of Jerusalem artichoketuber sliceswhen given alone or in combination with kinetinor gibberellicacid.
4. FC at concentrations lower than 10^–6 M, when givenwithauxin at concentrations lower than 0.03 mg/liter, promotedelongationof Avena coleoptile segments in an additive manner,to achievethe maximum elongation at higher concentrations.
5. An antiauxin, 2,4,6-trichlorophenoxyacetic acid, inhibitedtheelongation of Avena coleoptile segments due to auxin butnotthat due to FC.
6. Nojirimycin, an inhibitor of ß-glycosidases,inhibitedelongation of pea internode segments due not onlyto auxin butalso to FC.
7. At concentrations more than 10^–5MFC promoted root elongationof intact lettuce seedlings, whichwas inhibited by exogenousauxin.

From these results it is concluded that FC and auxin have acommon mechanism, which may involve hydrogen ion extrusion,leading to cell wall loosening and thus cell elongation. Thisgrowth is limited to the extent that the cells are capable ofelongating in response to hydrogen ions. Otherwise there isa definite difference in the mode of actions between FC andauxin, including the nature of cellular receptors for thesetwo compounds. (Received August 29, 1974; )     

The Analysis of Correlative Growth in the Etiolated Oat Seedling in Relation to Carbon Dioxide and Nutrient Supply

To overcome the reduced extension growth of the coleoptile whichoccurs when oats are grown in air enriched with 5 per cent.CO¹, plants have been provided with nutrients via the roots.2 per cent, sucrose, glucose or mannitol so applied furtherpromoted the mesocotyl and further depressed the coleoptile.Root growth was also depressed. To induce promotion of coleoptile growth by externally appliedsucrose, seedlings were heated in darkness at 40° C. for3 hours so restricting selectively the growth of the mesocotyl.Promotion of the coleoptile, however, was not observed. Application of mixed Na and K nitrates occasioned an immediategrowth promotion of doleoptile and leaves in both the presenceand absence of CO², and also a.much less pronounced promotionof the mesocotyl in CO²; there was no effect in air. This enhancedgrowth of the coleoptile and leaves was coupled with a correspondinglygreater dry weight and also with an increased outflow of reservesfrom the endosperm into the plumule. Thus, while externally applied sugars seemed not to reach thecoleoptile, those made available from the endosperm as a resultof improved nitrogen supply were rapidly translocated to it.Simultaneous provision to the roots of nitrate and sucrose didnot improve the absorption and translocation of sugar. An analysis of covariance has been computed using the mesocotyland coleoptile length data together with the outflow from theendosperm and the conclusions so derived are discussed in relationto the problem of growth integration in etiolated oat seedlings.     

Transport and Metabolism of Kinetin-8-14C in the Mesocotyl and Coleoptile of Zea mays L.

The movement and metabolism of kinetin-8-¹⁴C in the mesocotyland coleoptile of Zea mays have been investigated. The segments (10 mm) of coleoptile with and without an apexwere used. The presence of apex decreases the quantity of countswhich move basipetally in the segments. The quantity transported acropetally in the segments (apex incontact with receiver blocks) was about three times higher thanthat transported basipetally (apex in contact with donor blocks). Using 10 mm coleoptile segments excised I mm below the apex,there was no appreciable difference between acropetal and basipetalmovement. The transport of kinetin in the mesocotyl was studied with andwithout vascular tissue. The quantity transported in segmentswithout vascular tissue was always lower than in the segmentswith vascular tissue. The presence of the node between the coleoptile and mesocotyldoes not disturb the movement of kinetin in the basipetal position(node in contact with donor blocks). Thin layer chromatography of plant extracts after a 48-hourtransport period revealed several radioactive substances whichappeared to be adenine with R_f 0.5 and other components appearedat R_f 0.6.     

Red Light-inhibited Mesocotyl Elongation in Maize Seedlings: I. The Auxin Hypothesis

Red light-inhibited mesocotyl elongation, which occurs in intact Zea mays L. seedlings, was studied in excised segments which included the coleoptile (or parts therefrom) and apical centimeter of the mesocotyl. Experiments took into account, first, the ability of the segments to regenerate auxin supply sites, and, second, that auxin uptake can be greatly reduced if there is no cut surface, apical to the elongating cells, to act as a port of entry. In all cases, auxin completely reversed the inhibition of elongation by light. The results support the hypothesis that light regulates mesocotyl elongation by controlling auxin supply from the coleoptile. Sucrose concentration had no effect on auxin reversal of light-inhibited elongation, but relatively high concentrations of gibberellic acid (10 μm) could substitute for auxin in this system.     

Estimation of Free, Conjugated, and Diffusible Indole-3-acetic Acid in Etiolated Maize Shoots by the Indolo-alpha-pyrone Fluorescence Method

Procedures for estimating free indoleacetic acid (IAA extracted from tissue homogenates by aqueous acetone), conjugated IAA (extracted by aqueous acetone and hydrolyzed by 1 n KOH), and diffusible IAA (diffused from the excised tissue into water), in shoots of etiolated 3-day-old maize (Zea mays L. cv. GH 390) seedlings are described, the indolo-alpha-pyrone fluorescence method being used to assay IAA. The reliability of the procedure is shown by comparative IAA determinations of the extracts using the gas chromatography-mass spectrometry method in which the methyl ester, heptafluorobutyryl derivative of IAA is assayed using the selected-ion-monitoring technique with deuterated IAA as an internal standard. A 3-millimeter-long coleoptile tip, a coleoptile with its included leaves and nodal region (whole coleoptile), and a mesocotyl each contains 0.2, 1.7, and 1.5 nanograms of free IAA, respectively. The whole coleoptile and the mesocotyl contain slightly less conjugated IAA than their content of free IAA. IAA diffuses from the coleoptile tip at the rate of 1.0 nanograms per tip per hour; from the base of the whole coleoptile and a set of leaves excised from a coleoptile, IAA diffuses at the rate of 0.62 and 0.17 nanogram per plant part per hour, respectively. The data obtained support the classical assumption that the coleoptile tip produces IAA. It is also suggested that some IAA is decomposed during its downward transport in the coleoptile.     

The photoperceptive sites and the function of tissue light-piping in photomorphogenesis of etiolated oat seedlings*

Abstract. Responses to red light irradiation of discrete areas along the intact, etiolated oat seedling indicate that illumination of the region around the coleoptilar node results in maximal coleoptile growth stimulation and mesocotyl growth suppression. Quantitation of the fibre optic properties of these etiolated tissues shows that the amount of axially transmitted light is log linear as a function of distance for both the mesocotyl and coleoptile (plus primary leaf). Using the fibre optic properties of the tissues to predict the response of the etiolated seedling to defined illumination fields allows one to localize two sites of photoperception: although the mesocotyl response pattern can be explained by the action of a single site found near the top of the mesocotyl itself, the coleoptile response depends on irradiation of both the mesocotyl site and an additional site located just above the node. The very low- and the low-fluence responses of etiolated oats independently predict similar regions of the seedling as sites of photo-perception. The fibre optic properties of the seedling could allow the seedling to increase the effective light signal received by the photosensitive area significantly.     

The contribution of internode and mesocotyl tissues to root-to-shoot signalling of abscisic acid.

The xylem of first internode of runner bean and of previously etiolated maize mesocotyl segments was perfused with media containing abscisic acid (ABA) or abscisic acid glucose ester (ABA-GE) in concentrations as they occur under stress conditions. ABA-GE passed through the internode and mesocotyl segments unchanged. Within 10 min the concentration of ABA-GE(xyl) rose to a level similar to that in the external perfusion medium. By contrast, 30-40 min passed before the concentration of free ABA in the xylem sap [ABA(xyl)] reached the level in the external medium. When ABA-free media were used, ABA was released from the xylem parenchyma to the xylem vessels resulting in an [ABA(xyl)] of 13-23 nM (runner bean internode) or 1-6 nM (maize mesocotyl). The total perimeter and, hence surface area, of the xylem elements was measured microscopically and from these measurements it was estimated that, in both bean internodes and maize hpyocotyls, the flux of ABA to the xylem was 1 pmol m(-2) s(-1). The ABA efflux from the stem and mesocotyl parenchyma into the xylem could be increased when the tissues were treated with tetcyclacis, an inhibitor of ABA degradation, but also by changing the pH from its normal value of about pH 5.8 to pH 7.0 and by adding 100 mM NaCl to the perfusion medium. If 100 nM ABA was added to the perfusion medium the above treatments had only small effects on the release of ABA from the tissues into the xylem.     

Involvement of Abscisic Acid in Mesocotyl Growth in Etiolated Seedlings of a Foxtail Millet Dwarf Mutant

Etiolated seedlings of foxtail millet (Setaria italica Beauv.) dwarf mutant CH84113 were treated with various concentrations of abscisic acid (ABA), mefluidide, mannitol, or polyethylene glycol (PEG) 6000. It was found that these chemicals, at suitable concentrations, could increase mesocotyl length significantly, whereas these chemicals at higher concentrations had an inhibitory effect. Endogenous levels of ABA in mesocotyl were measured by enzyme-linked immunosorbent assay. It was found that endogenous ABA increased progressively in a chemical (ABA, mefluidide, mannitol, or PEG 6000) concentration-dependent manner, indicating that the effects of these chemicals on mesocotyl growth may be mediated by increased endogenous ABA levels. On the other hand, S-3307, an inhibitor of the oxidative reactions in gibberellin (GA) biosynthesis, inhibited the elongation of mesocotyl significantly. When ABA and GA₃ were applied simultaneously, the effect on mesocotyl growth was additive. These results imply that ABA and GA may control different processes in the regulation of mesocotyl growth. Received October 27, 1997; accepted May 11, 1998     

Effects of abscisic acid and its related compounds on rice seedling growth

Rice seedlings with the mesocotyl and coleoptile (the undeveloped leaves enclosed in the coleoptile) are here referred to as MC type seedlings and are considered to be suitable for deep sowing. We investigated the effects of abscisic acid (ABA) and several of its related compounds on the occurrence of MC type seedlings and on rice mesocotyl growth. Rice (Oryza sativa L. cv. JC 91) seedlings were grown on 0.8% agar medium in the presence or absence of various kinds of ABAs under aseptic conditions at 30 °C in the dark for 14 days. The activity of the R isomer of ABA (R-ABA) was slightly less than that of the naturally occurring S form (S-ABA) concerning the occurrence of MC type rice seedlings and the growth of the rice mesocotyl. In addition, the racemate of R-and S-ABA (RS-ABA) is less effective than R-ABA and S-ABA alone.Trans-ABA had no activity in relation to both percent occurrence of MC type seedlings and mesocotyl growth. The results of the present study suggest that the occurrence of MC type rice seedlings and the growth of rice mesocotyls were closely related to structure-activity relationships with analogs of ABA.     

Phytochrome-controlled extension growth of Avena sativa L. seedlings

The effects of continuous red and far-red light and of brief light pulses on the growth kinetics of the mesocotyl, coleoptile, and primary leaf of intact oat (Avena sativa L.) seedlings were investigated. Mesocotyl lengthening is strongly inhibited, even by very small amounts of P_fr, the far-red light absorbing form of phytochrome (e.g., by [P_fr]0.1% of total phytochrome, established by a 756-nm light pulse). Coleoptile growth is at first promoted by P_fr, but apparently inhibited later. This inhibition is correlated in time with the rupturing of the coleoptile tip by the primary leaf, the growth of which is also promoted by phytochrome. The growth responses of all three seedling organs are fully reversible by far-red light. The apparent lack of photoreversibility observed by some previous investigators of the mesocotyl inhibition can be explained by an extremely high sensitivity to P_fr. Experiments with different seedling parts failed to demonstrate any further obvious interorgan relationship in the light-mediated growth responses of the mesocotyl and coleoptile. The organspecific growth kinetics, don't appear to be influenced by P_fr destruction. Following an irradiation, the growth responses are quantitatively determined by the level of P_fr established at the onset of darkness rather than by the actual P_fr level present during the growth period.Abbreviation P_fr far-red light absorbing form of phytochrome     

Translocation of Radiolabeled Indole-3-Acetic Acid and Indole-3-Acetyl-myo-Inositol from Kernel to Shoot of Zea mays L.

Either 5-[³H]indole-3-acetic acid (IAA) or 5-[³H]indole-3-acetyl-myo-inositol was applied to the endosperm of kernels of dark-grown Zea mays seedlings. The distribution of total radioactivity, radiolabeled indole-3-acetic acid, and radiolabeled ester conjugated indole-3-acetic acid, in the shoots was then determined. Differences were found in the distribution and chemical form of the radiolabeled indole-3-acetic acid in the shoot depending upon whether 5-[³H]indole-3-acetic acid or 5-[³H]indole-3-acetyl-myo-inositol was applied to the endosperm. We demonstrated that indole-3-acetyl-myo-inositol applied to the endosperm provides both free and ester conjugated indole-3-acetic acid to the mesocotyl and coleoptile. Free indole-3-acetic acid applied to the endosperm supplies some of the indole-3-acetic acid in the mesocotyl but essentially no indole-3-acetic acid to the coleoptile or primary leaves. It is concluded that free IAA from the endosperm is not a source of IAA for the coleoptile. Neither radioactive indole-3-acetyl-myo-inositol nor IAA accumulates in the tip of the coleoptile or the mesocotyl node and thus these studies do not explain how the coleoptile tip controls the amount of IAA in the shoot.     

Mode of dinitroaniline herbicide action I. Analysis of the colchicine-like effects of dinitroaniline herbicides

Colchicine and a variety of dinitroaniline herbicides (DNHs)produce a similar pattern of inhibition of elongation, inductionof swelling in the elongation zone, depolarization of cell enlargement,and induction of multiple nuclei in corn seedling roots. However,a 1000-fold higher concentration of colchicine is needed toproduce effects quantitatively similar to those of oryzalin.Both colchicine- and DNH-inhibition of elongation start at about3 hr. Since these compounds cause swelling and inhibition ofelongation in -seedling roots, segments from the root elongationzone and intact roots in the presence of cell division inhibitors(all growing without cell division), it appears that the inhibitionof root elongation is caused in part by their effect on cellelongation independent of their effect on cell division. Sincethe growth (increase in fresh weight) of -seedling roots andexcised root segments is not inhibited by these compounds, theireffect on the polarity of cell enlargement must be fairly specific.Unlike colchicine, oryzalin applied to the roots did not causeany significant, visible effect on shoot (mesocotyl and coleoptile)growth. These organs are not resistant to oryzalin, for theIAA-induced elongation of coleoptile segments is inhibited whenthey are floated in oryzalin solutions. As expected, when coleoptilesegments are incubated in ¹⁴C-oryzalin, it is taken up rapidlyand not degraded. The failure of root-applied oryzalin to affectseedling shoot growth is due to lack of transport to the shoots. (Received June 14, 1977; )     

The Relationship between the Growth of the Primary Leaf and of the Coleoptile in Seedlings of Avena and Triticum

Partial inhibition of extension growth of the primary leaf occurswhen whole Triticum seedlings are immersed in aerated solutionsof IAA but is replaced by growth promotion when sucrose is addedto the external solution. In seedlings in which the coleoptilehas been excised, IAA increases the growth of the leaf bothwith and without additional sucrose. Inhibition of the leaf by moderate concentrations of IAA nolonger occurs when the seedling is detached from the endosperm.Sucrose added to the external solution raised the percentageelongation of the coleoptile almost to the level of that attainedin intact seedlings without additional carbohydrate. It alsoenabled the leaf to show a positive growth response with IAA. The results indicate that in intact seedlings treated with IAAthe growth of the primary leaf is markedly diminished owingto diversion of carbohydrate to the coleoptile if the growthof the latter is promoted as a result of the treatment. Whenthe competition of the coleoptile for carbohydrate is diminishedor eliminated, acceleration of the growth of the primary leafby IAA becomes apparent. In addition to the endogenous rhythm, with a period close to24 hours, induced in the growth-rate of the coleoptile whenseedlings of Avena are transferred from red light to darkness,a similar rhythm, with a slightly longer period, is inducedin the growth-rate of the primary leaf. This rhythm persistsin elongating leaves so long as they remain within the coleoptile.It can be recorded for at least 100 hours in deseeded seedlings. When intact seedlings of Avena are immersed for one hour inrelatively high concentrations of IAA and then transferred todistilled water for 18 hours, the elongation of the coleoptileis greater and the inhibition of the leaf is less than whenthey are transferred to humid air. Sections of the leaf of Triticum showed a slight increase inelongation in concentrations of IAA up to 5 mg./l., but no evidencewas obtained that sections of leaf and coleoptile exert any.influenceon each other's elongation when floated together on solutionsof IAA.     

Correlation Between Responses of Elongation of Wheat Coleoptile Segments to Abscisic Acid and Exogenous ATP

The time course of elongation and oxygen consumption of wheat coleoptile segments in 38μM ABA solution and 5 × 10-5 M 2,4-DNP solution were investigated. At the same time, effects of exogenous ATP (1 mM) on elongation and ABA response in wheat coleoptile segments were observed. Within the first hour of treatment, ABA and 2,4-DNP rapidly inhibited elongation in the coleoptile segments. Exogenous ATP stimulates elongation in coleoptile segments. When segments pretreated with ATP and then transfered into ABA solution, elongation in coleopfile segments were not inhibited. On the basis of the similarities of action pattern of ABA and 2,4-DNP and marked decrease of ABA inhibiting effect in presence of exogenous ATP, it is testified that uncoupling of respiration and oxidative phosphorylation is one of ABA important effects, so it is suggested that ABA inhibiting elongation of wheat coleoptile segments by decreasing tissue energy (ATP) levels necessary for synthetic reactions, including nucleic acid and protein, and associated with elongation.     

Growth, anatomy and morphology of the mesocotyl and the growth of appendages of the wild oat (Avena fatua L.) seedling

Morphological and histological studies were made on the mesocotyl and the emergence of seedlings of a nondormant strain (CS40) of wild oats (Avena fatua L.). The elongation of the mesocotyl was primarily responsible for the emergence of seedlings from deeper levels of soil. The mesocotyl of the seedling is here interpreted as the hypocotyl. The functionally suctorial scutellum together with coleoptile constitutes the first cotyledon and the first true-leaf is regarded as the second cotyledon. The development of tillers from scutellar and first-leaf buds depends on the depth at which level the seeds (caryopses) germinated and the seedlings emerged above the soil surface. The first-leaf axillary buds, regradless of depths, develop into dominant tillers. The scutellar buds, especially at greater depths, remain inhibited. At shallower levels, however, they develop into tillers. The scutellar buds, at deeper levels, behave as reserve ramets which feature adds to the success of the species as a weed in the agricultural prairies.