Hormones andCuscuta development: interaction of cytokinin and indole-3-acetic acid in the growth and curvature of subapical stem segments

Cuscuta stem (vines) exhibits two modes of growth—longitudinal elongation forming free-hanging vines, or coiling growth to twine around the host. The elongation zone of free-hanging vine extended up to 160 mm from the stem apex and in vivo growth rate (during 8 h of growth) was maximal in the 20-to-40-mm region. While gibberellic acid (GA₃) or fusicoccin (FC) could maintain (GA₃) or enhance (FC) the growth rate of apical (10 or 25 mm) segments, indole-3-acetic acid (IAA) (10 M) induced growth only in subapical (5–160 mm) segments. In vitro growth rate induced by IAA (10 M) was similar to the in vivo growth rate up to 40 mm. Thereafter, up to 100 mm, IAA induced growth rate exceeded in vivo growth. p ]Subapical segments (13 mm) from 5- to 40-mm regions responded to a cytokinin (BA, Z, or iP) or to low IAA (0.1 M) with curved growth, whereas the segments grew straight in the presence of high IAA (10 M). Curvature (measured as the angle subtended at the center of the circle of which the segment formed an arc) induced by BA and low (0.1 M) IAA was greater than either added separately. Besides, segments induced to curve in BA + low-IAA solution could be made to straighten out by transferring to a solution containing high IAA (10 M) with or without BA. Thus in vivo patterns of straight and coiling growth could be mimicked reversibly in vitro by adjusting the relative concentrations of cytokinin and auxin; low auxin and cytokinin induced coiling growth, whereas high auxin and cytokinin induced straight growth. p ]Beyond 40 mm, BA had no growth-promoting or curvative-inducing effect.Cuscuta vine segments thus showed sequential sensitivity to applied hormones, the apical region (0–25 mm) to GA₃, the subapical (5–40 mm) region to BA and IAA and the region beyond (40–160 mm) to IAA alone.     

The effects of IAA and tetcyclacis on tuberization in potato (Solanum tuberosum L.) shoot cultures in vitro

Potato (Solanum tuberosum cv. Désirée) shoots grown in vitro in continuous darkness or in long days (LDs), were used to investigate indole-3-acetic acid (IAA) effects on stolon initiation and tuber formation, combining IAA with increased or decreased gibberellin levels. An increased gibberellin (GA) level was achieved by the applying 1 μM GA₃, while decreased gibberellin level was presumably realized by the adding 3 μM tetcyclacis (Tc). About 15% of potato shoots developed stolons both in LDs and in darkness. Stolon initiation was stimulated by GA₃ in darkness and by Tc in LDs. Tuber formation was strongly inhibited in LDs and by GA₃ both in light and darkness, but stimulated in darkness at low GA level. Exceptionally, tuber formation occurred in LDs at the highest Tc concentrations, in about 25% of explants. Indole-3-acetic acid alone stimulated stolon formation in LDs, both in the presence or absence of GA₃. IAA alone also stimulated tuber formation in dark-grown shoots, but could not overcome the inhibitory effect of LDs. Indications that, depending on their concentration ratio, IAA may interact with GA₃ in different tuberization phases, have been discussed. Radomir Konjević—Deceased in July 2006.     

Effects of prohexadione on cambial and longitudinal growth and the levels of endogenous gibberellins A1, A3, A4, and A9 and indole-3-acetic acid in Pinus sylvestris shoots

Prohexadione, a gibberellin (GA) biosynthesis inhibitor, was applied in ethanol around the circumference at the midpoint of the previous year terminal shoot of dormant Pinus sylvestris seedlings. After cultivating the seedlings under environmental conditions favorable for growth for 10 weeks, longitudinal and cambial growth were measured, and the endogenous levels of GA₁, GA₃, GA₄, GA₉, and indole-3-acetic acid (IAA) were determined by combined gas chromatography-mass spectrometry, using deuterated GAs and [¹³C₆]IAA as internal standards. Prohexadione application inhibited elongation and xylem and phloem production in the current year terminal shoot and xylem production in the previous year terminal shoots. Concomitantly, in both ages of shoots the cambial region contents of GA₁; GA₃, and GA₄ were decreased, whereas the level of GA₉ was increased. However, the IAA content was not altered in the terminal bud on the current year terminal shoot or in the cambial region of the current year or previous year terminal shoots. The results provide additional evidence that: (1) GAs are involved in the regulation of cambial growth, as well as longitudinal growth, in Pinus sylvestris shoots; (2) they act directly, rather than indirectly, by altering the IAA level; and (3) the GA₉ GA₄ GA₁ pathway is a major route of GA biosynthesis in conifer species.Abbreviations GA gibberellin - IAA indole-3-acetic acid - HPLC high performance liquid chromatography - GC gas chromatography - SIM selected ion monitoring - MS mass spectrometry     

(Aminooxy)acetic acid inhibits petunia growth and gibberellin- and cytokinin-stimulated growth in bioassays

(Aminooxy)acetic acid (AOA) was applied to greenhouse-grown petunias and was used in bioassays for three plant growth hormones so that its growth regulator properties could be studied. In greenhouse studies foliar sprays of 4.8–12 mm AOA inhibited vegetative growth of petunia seedlings (Petunia xhybrida Vilm. White Flash). When gibberellin A ₃ (GA₃) was applied to shoot tips previously treated with AOA, plant growth was stimulated, but there was no AOA x GA₃ interaction. Some changes in petunia leaf morphology induced by AOA were reversed by GA₃. AOA inhibited elongation of corn coleoptile segments (Zea mays L. B73 x Mol7) whether or not 10 m indole-3-acetic acid (IAA) was present, but there was no AOA x IAA interaction. AOA reduced lettuce hypocotyl (Lactuca sativa L. Grand Rapids) elongation induced by GA₃ and radish cotyledon (Raphanus sativus L. Champion) expansion induced by benzyladenine (BA). We propose that AOA interferes with postsynthetic metabolism of plant hormones during cell elongation induced by GA₃ and cell expansion induced by BA.Abbreviations AOA (aminooxy)acetic acid - GA₃ gibberellin A₃ - IAA indole-3-acetic acid - BA benzyladenine     

Changes in jasmonate and gibberellin levels during development of potato plants (Solanum tuberosum)

Among the multiple environmental signals and hormonal factors regulatingpotato plant morphogenesis and controlling tuber induction, jasmonates (JAs)andgibberellins (GAs) are important components of the signalling pathways in theseprocesses. In the present study, with Solanum tuberosum L.cv. Spunta, we followed the endogenous changes of JAs and GAs during thedevelopmental stages of soil-grown potato plants. Foliage at initial growthshowed the highest jasmonic acid (JA) concentration, while in roots the highestcontent was observed in the stage of tuber set. In stolons at the developmentalstage of tuber set an important increase of JA was found; however, in tubersthere was no change in this compound during tuber set and subsequent growth.Methyl jasmonate (Me-JA) in foliage did not show the same pattern as JA; Me-JAdecreased during the developmental stages in which it was monitored, meanwhileJA increased during those stages. The highest total amount of JAs expressed asJA+Me-JA was found at tuber set. A very important peak ofJA in roots was coincident with that observed in stolons at tuber set. Also, aprogressive increase of this compound in roots was shown during the transitionof stolons to tubers. Of the two GAs monitored, gibberellic acid(GA₃) was the most abundant in all the organs. While GA₁and GA₃ were also found in stolons at the time of tuber set, noothermeasurements of GAs were obtained for stolons at previous stages of plantdevelopment. Our results indicate that high levels of JA and GAs are found indifferent tissues, especially during stolon growth and tuber set.     

Hormone-induced protection of sunflower photosynthetic apparatus against copper toxicity

The effects of excess Cu as affected by the application of exogenous hormones (gibberellic acid - GA₃ and indole-3-acetic acid - IAA) with respect to sunflower (Helianthus annuus L.) growth, physiology, and metabolism were studied. Application of 100 M IAA lessened the toxic effects of 80 M Cu in roots indicating greater root length and root hair formation, while addition of 100 M GA₃ ameliorated the toxic effect mainly to the shoot. The content of photosynthetic pigments significantly declined under Cu stress, whereas application of hormones led to a substantial preservation of chlorophylls and carotenoids. Under Cu stress, the rate constant of energy trapping by photosystem 2 (PS2) reaction centres (RCs) was reduced as a result of physical dissociation of the light-harvesting complex (LHC) from PS2 core, while application of IAA and especially GA₃ resulted in stability of the LHC of PS2 RCs. The drop in net photosynthetic (PN) and transpiration (E) rates with preserved or slightly reduced variable to maximum fluorescence ratio (F_v/F_m) in the presence of 80 M Cu could be explained by a possible inhibition of the enzymatic processes in the Calvin cycle. Application of 100 M IAA and 100 M GA₃ lessened Cu effects mainly on P _N. Water use efficiency was also improved under hormone exposure.     

Effect of relative hormone concentration on auxin-gibberellin interaction in correlative inhibition of axillary buds

Summary Indole-3-acetic acid (IAA) applied to the fully elongated second internode of decapitated Phaseolus multiflorus plants always inhibited axillary bud elongation at concentrations down to 100 g/g lanolin, whereas gibberellic acid (GA₃) enhanced bud elongation at concentrations down to 1000 g/g lanolin. Lower concentrations than these of either IAA or GA₃ were without significant effect. All possible combinations of IAA and GA₃ within the concentration range 10¹ to 10⁵ g/g lanolin were antagonistic; IAA tending to inhibit, and GA₃ promote, axillary bud elongation growth. Treatment of an elongating internode with the hormones resulted in an increase in inhibition of bud growth by IAA in the presence of GA₃.     

Effect of plant growth regulators on in vitro fiber development from unfertilized and fertilized Egyptian cotton ovules

Unfertilized and fertilized ovules of Gossypium barbadense Giza 45 (extra long staple variety) were used to study the effect of plant growth substances (auxins, gibberellins and cytokinins) on in vitro fiber initiation and development. Kinetin, alone did not increase total fiber unit (TFU) of unfertilized ovules, while an increase in TFU value occurred when a constant level of IAA and GA₃ were used either separately or in combination in the liquid medium. GA₃ used alone, produced a higher TFU value than that produced by IAA, whilst, IAA with a constant level of GA₃ (5 M) produced the highest value of TFU. GA₃ with a constant level of IAA (5 M) produced a lower TFU value. Kinetin reduced the stimulatory effect of IAA and GA₃ on TFU value when used in combination with either substance. In fertilized ovules, the highest level of TFU was reached when IAA, with a constant level of GA₃, was added to the medium, whilst its lowest level was obtained when IAA was used alone. Estimation of in vitro fiber production, as well as the effect of growth substances used in different concentrations on in vitro fiber initiation and development from unfertilized and fertilized ovules of Egyptian cotton varieties Gossypium barbadense Giza 45 are discussed.     

Gibberellic acid induces cytoplasmic acidification in maize coleoptiles

Indole-3-acetic acid (IAA), fusicoccin and weak acids all lower the cytoplasmic pH (pH_i) and induce elongation growth of maize (Zea mays L.) coleoptiles. Gibberellic acid (GA₃) also induces elongation growth and we have used confocal laser scanning microscopy to study the effects of GA₃ on pH_i employing the pH-indicator dyes, 2,7-bis(2-carboxyethyl)-5-(and-6) carboxyfluorescein and carboxy-semi-naphthorhodafluor-1. We confirm that GA₃ induces growth significantly in light-grown but only slightly or not at all in dark-grown coleoptiles. The growth induced by IAA treatment was similar in light- and dark-grown coleoptiles. The pH_i decreased by up to 0.6 units during the first 7 min of GA₃ or IAA treatment of both light- and dark-grown coleoptiles. Gibberellic acid inhibited IAA-induced growth of dark-grown coleoptiles. Hence, in dark-grown coleoptiles GA₃ may activate either directly or indirectly reactions that interfere with the signalling pathway leading to elongation growth. The possible role of pH_i in growth is discussed.Abbreviations ABA abscisic acid - AM acetoxymethyl ester - BCECF 2,7-bis(2-carboxyethyl)-5-(and-6) carboxyfluorescein - [Ca²⁺]_i cytoplasmic free calcium - GA_(n) gibberellin A_(n) - GA₃ gibberellic acid - IAA indole-3-acetic acid - PGR plant growth regulator - pH_i cytoplasmic pH - Pipes piperazine-N,N-bis[2-ethanesulfonic acid] - Snarf-1 carboxy-semi-naphthorhodafluor-1 We thank Dr R. King (CSIRO, Canberra) for providing the GA₁ and T. Phillips for processing the photographic material. H.R. Irving was supported by an Australian Research Council Research Fellowship and the work was supported by an Australian Research Council grant.     

STRUCTURAL INVESTIGATIONS OF ASEXUAL REPRODUCTION IN NEPHROLEPIS EXALTATA AND PLATYCERIUM BIFURCATUM

Nephrolepis exaltata cv. Bostoniensis, the Boston fern, exhibits extreme stem dimorphism. The plant has orthotropic, dictyostelic shoots which bear pinnatifid leaves and plagiotropic, protostelic stolons which are aphyllous. Vegetative reproduction occurs by budding from primary and secondary stolons. Secondary stolons arise exogenously from derivatives of the apical cell of the primary stolon, whereas root primordia develop endogenously. Shoots develop in vivo when a creeping stolon makes contact with the substrate via extensive root proliferation. When stolon segments are excised and grown in vitro, secondary stolon primordia expand and initiate leaf primordia, forming new leafy shoots. In Platycerium bifurcatum, the staghorn fern, asexual propagation occurs on ageotropic roots ramifying among the basal nest fronds. Root bud initiation is marked by root tip hypertrophy following cortical parenchyma expansion. Root apical cell derivatives produce the bud apex; the root apical cell remains separate from the developing root bud. Superficially, vegetative reproduction in Nephrolepis and Platycerium appears to involve unusual organs. However, both ferns exhibit leafy bud development from distinct sites of origin, not from undetermined primordia or from direct transformation of root to shoot. Thus, distinctness of organ types is maintained in these two ferns and no evidence for interconvertibility of organ types has been found.     

In vitro culture of vanhoutte's spirea explants from ‘secondary cultures’ and dormant stems forced in solutions containing plant growth regulators

Explants from new growth of forced dormant stems and secondary cultures of Vanhoutte's spirea were cultured on Linsmaier and Skoog (L.S.) medium containing benzyladenine (BA), indoleacetic acid (IAA), thidiazuron (TDZ), and zeatin. The dormant stems were forced by immersing their basal portions in forcing solutions containing 626 µM 8-hydroxyquinoline citrate (8-HQC) and 2% sucrose. BA and gibberellic acid (GA₃) were also added into the forcing solutions to determine if explants obtained from the new growth will benefit from this treatment when culturedin vitro.L.S. medium supplemented with 5 µM BA alone, 5 µM BA plus 1 or 5 µM IAA, and 0.5 or 0.75 µM TDZ alone produced the best shoot proliferation for both sources of explants. BA and GA₃ appeared to be taken up from the forcing solution by the new softwood growth. BA in the forcing solution stimulatedin vitro shoot proliferation in different degrees depending on the period of treatment, while GA₃ caused lessin vitro shoot proliferation. It is proposed that forcing solutions containing plant growth regulators (P.G.R.) are a useful approach for manipulating responses of plant tissues culturedin vitro.     

In vitro establishment and multiplication of Nymphaea hybrid ‘James Brydon’

Rhizome tips were the most suitable explants for in vitro plant regeneration and multiplication of Nymphaea hybrid James Brydon on Murashige and Skoog medium containing different concentrations and combinations of indole-3-acetic acid, 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid, 6-benzyladenine (BA), kinetin, 2-isopentenyladenine (2iP) and gibberellic acid (GA₃). A combination of 2iP, BA, and NAA strongly favored induction of shoot buds and shoot proliferation. Pretreatment of shoot cultures at 8°C for 30 days or with 14.4 or 28.9 M GA₃ for 15 days did not improve shoot multiplication. A 16-h photoperiod with photosynthetic photon flux of 30 mol m^-2 s^-1 was found to be the optimum light condition for shoot growth and multiplication. Multiple shoots produced well developed root systems within 4 weeks after transfer to a plant growth regulator-free medium containing activated charcoal.Abbreviations BA 6-benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - 2iP 2-isopentenyladenine - MS Murashige and Skoog medium - NAA 1-naphthaleneacetic acid     

Gibberellin-induced ethylene production and its effect on cowpea epicotyl elongation

The effect of gibberellin A₁ (GA₁) on production of ethylene by cowpea (Vigna sinensis cv Blackeye pea no. 5) epicotyl explants and its relationship to epicotyl elongation was investigated. The explants were placed upright in water and incubated in sealed culture tubes or in large jars. GA, and IAA in ethanol solution were injected into the subapical tissues of the decapitated epicotyls. Cowpea epicotyl explants elongated after GA but not after IAA treatment, and they were very sensitive to exogenous ethylene. As little as 0.14 1/1 ethylene reduced significantly GA₁-induced epicotyl elongation.Treatment with GA₁ induced the production of ethylene which began 10 h after GA application, showed a peak at about 22 h and then declined. The yield of ethylene was proportional to the amount of GA, injected. The inhibition of epicotyl elongation in closed tubes was avoided by absorbing ethylene released with Hg(Cl0₄)₂ , or by adding AVG to the incubation solution to inhibit ethylene production. Treatment with IAA elicited a rapid production of ethylene which ceased about 10 h after application. The effects of IAA and GA₁ on ethylene production were additive.Abbreviations AVG aminoethoxyvinylglycine 2-amino-4-(2-aminoethoxy)-trans-3butenoic acid - ACC 1-aminocyclopropane-1-carboxylic acid - GA gibberellin - IAA indole-3-acetic acid     

The effect of plant growth regulator treatments on the levels of ethylene emanating from excised turgid and wilted wheat leaves

Abscisic acid (ABA) inhibits the production of ethylene induced by water stress in excised wheat leaves and counteracts the stimulatory effect of 6-benzyladenine (BA) on this process. The stimulatory effect of BA and the inhibitory effect of ABA were equally pronounced whether external or endogenous ethylene levels were determined. When leaves were sprayed or floated on solutions of BA, indole-3-acetic acid (IAA), gibberellic acid (GA₃), or ABA, the relative activities of these growth regulators on stress-induced ethylene at 10^-4 mol l^-1 were BA>IAA >GA₃>controls>ABA. In non-stressed leaves, however, where the levels of ethylene produced were 2–20 times smaller, the relative activities were IAA >BA>GA₃>controls>ABA. The effects of BA and ABA spray treatment on water stress induced ethylene were closely similar whether the solutions were applied 2 or 18 h prior to the initiation of water stress. The relationships between the levels of endogenous growth regulators in the plant and ethylene release induced by water stress are discussed.Abbreviations BA 6-benzyladenine - IAA indole-3-acetic acid - GA₃ gibberellic acid - ABA abscisic acid - GLC gas-liquid chromatography - _leaf leaf water potential     

Improvement of in vitro propagation of apricot cultivar ‘Bebecou’

Factors affecting successful establishment in vitro, rapid proliferation and rooting of apricot cultivar ‘Bebecou’ were studied. Ethanol and NaOCl were applied in several combinations for disinfection; chilling, plant growth regulators BA, IAA and GA₃, antibiotics, different culture vessels and systems of subculture were evaluated for the optimization of shoot proliferation and the auxins NAA and IBA were assessed for root induction. The highest number of new microshoots/explant (18.7) was obtained in a culture medium supplemented with 2.2 μM BA+0.57 μM IAA after 300 h of chilling. The effect of GA₃ (11.4 μM) on shoot proliferation was positive in combination with 4.4 or 8.9 μM BA. Shoot length and productivity were highest at 2.2 μM BA+11.4 μM GA₃+0.57 μM IAA and at 2.2 μM BA+0.57 μM IAA, respectively and decreased as cytokinin concentration increased. The antibiotic ‘Na-cefotaxime’ had a minimal impact on shoot growth when used at the lowest concentration (250 mg l⁻¹). Subculture every 2 weeks in a medium supplemented with 2.2 μM BA and 0.57 μM IAA was more efficient for shoot induction than alternation of 20 days culture in a propagation medium supplemented with 2.2 μM BA and 10 days culture in an elongation medium supplemented with 1.1 μM BA and 5.71 μM IAA. The highest number of roots/shoot (8.1) was recorded at 19.6 μM IBA.     

海岸单叶蔓荆沙埋胁迫下碳水化合物变化与其耐沙埋的关系

选择烟台海岸沙地抗沙埋强的单叶蔓荆(Vitex trifolia var.simplicifolia)为试材,在自然环境条件下根据单叶蔓荆匍匐茎长度进行了轻度(1/3茎长)、中度(2/3茎长)和重度半埋以及全埋处理。在沙埋20d后,测定了不同沙埋处理下匍匐茎各段上匍匐茎长度、枝条高度、不定根长度,以及可溶性糖、淀粉、纤维素含量,以探讨单叶蔓荆碳水化合物变化和转化在其耐沙埋中作用。结果显示,在轻度、中度半埋和全埋下单叶蔓荆匍匐茎长度均显著大于对照,被沙埋匍匐茎处有大量不定根生成;同时,可溶性糖和淀粉含量增高和纤维素含量下降,尤其是生长最快的匍匐茎顶部(如轻度半埋),茎中可溶性糖较低、淀粉增加最多,纤维素最低。但是被重度半埋和全埋的匍匐茎生长较少,茎中纤维素含量较多、淀粉含量较少。研究表明,沙埋是一种胁迫,它损伤叶片、扰乱碳水化合物代谢平衡。但它又是胁迫信号使植物产生适应性反应,它使未遭沙埋的匍匐茎顶端通过加速碳水化合物转化、分解纤维素、提高淀粉和可溶性糖含量,为顶端生长提供能量和营养,以加速匍匐茎快速生长摆脱沙埋。同时沙埋部位枝叶通过分解其纤维素,产生更多的可溶性糖和淀粉为匍匐茎不定根生长提供能量。因此,沙埋后匍匐茎内碳水化合物的转化是其快速生长和摆脱沙埋的能量来源而在其适应沙埋生长中起重要作用。单叶蔓荆对沙埋的适应性反应表现了其具有表型可塑性特性,该特性是其沙埋后维护匍匐茎顶部快速生长、不定根形成、碳水化合物转化以及具有较高抗沙埋能力的关键。     

High efficiency plant regeneration from cotyledons of watermelon (Citrullus vulgaris Schrad)

Cotyledons of various ages from seedlings of eight watermelon (Citrullus vulgaris) cultivars were cultured on MS medium supplemented with different combinations of phytohormones. High frequency shoot regeneration (60.0–92.0%) was induced from 5-day-old cotyledons of cultivars cultured on MS medium containing 5.0 mg/l 6-benzylaminopurine (BA) and 0.5 mg/l indole-3-acetic acid (IAA). Multiple shoot buds elongated on MS medium containing 0.2 mg/l kinetin (KT) and 5–10 shoots per expiant could be recovered depending on the cultivars. Elongated shoots rooted on MS medium with 0.1 mg/l -naphthalene acetic acid (NAA). Zeatin riboside (ZT) had a similar efficiency as BA in shoot induction, and it was significantly more functional than 2-isopentenyladenine (2iP) or kinetin (KT). Cotyledons from 5-day-old seedlings were the most responsive to shoot induction.Abbreviation BA 6-benzylaminopurine - GA₃ gibberellic acid - IAA indole-3-acetic acid - 2iP 2-isopentenyladenine - KT kinetin - MS Murashige and Skoog (1962) - NAA -naphthalene acetic acid - ZT zeatin riboside     

Internode length in Pisum sativum L. The kinetics of growth and [3H]gibberellin A20 metabolism in genotype na Le

The relationship between shoot growth and [³H]gibberellin A₂₀ (GA₂₀) metabolism was investigated in the GA-deficient genotype of peas, na Le. [17-¹³C, ³H₂]gibberellin A₂₀ was applied to the shoot apex and its metabolic fate examined by gas chromatographic-mass spectrometric analysis of extracts of the shoot and root tissues. As reported before, [¹³C, ³H₂]GA₁, [¹³C, ³H₂]GA₈ and [¹³C, ³H₂]GA₂₉ constituted the major metabolites of [¹³C, ³H₂]GA₂₀ present in the shoot. None of these GAs showed any dilution by endogenous ¹²C-material. [¹³C, ³H₂]GA₂₉-catabolite was also a prominent metabolite in the shoot tissue but showed pronounced isotope dilution probably due to carry-over of endogenous [¹²C]GA₂₉-catabolite from the mature seed. In marked contrast to the shoot tissue, the two major metabolites present in the roots were identified as [¹³C, ³H₂]GA₈-catabolite and [¹³C, ³H₂]GA₂₉-catabolite. Both of these compounds showed strong dilution by endogenous ¹²C-material. Only low levels of [¹³C, ³H₂]GA₁, [¹³C, ³H₂]GA₈, [¹³C, ³H₂]GA₂₀ and [¹³C, ³H₂]GA₂₉ accumulated in the roots. It is suggested that compartmentation of GA-catabolism may occur in the root tissue in an analogous manner to that shown in the testa of developing seeds. Changes in the levels of [1,3-³H₂]GA₂₀ metabolites over 10 d following application of the substrate to the shoot apex of genotype na Le confirmed the accumulation of [³H]GA-catabolites in the root tissues. No evidence was obtained for catabolic loss of [³H]GA₂₀ by complete oxidation or conversion to a methanol-inextractable form. The results indicate that the root system may play an important role in the regulation of biologically active GA levels in the developing shoot of Na genotypes of peas.Abbreviations GA_n gibberellin A_n - GC-MS gas chromatography-mass spectrometry - HPLC high-pressure liquid chromatography     

A Possible Role for Indoleacetic Acid, Low Temperature, and Phospholipid Metabolism in the Induction of GA(3) Responsiveness in GA(3) Insensitive (Rht3-Containing) Dwarf Wheat Aleurone

Preincubation of dwarf, Rht3-containing deembryonated seed for 4 hours in 342 nanomolar indoleacetic acid (IAA) induced maximum sensitivity to GA₃. In addition, the 4-hour IAA pretreatment caused a 2-fold increase in total phospholipids which coincided identically on a temporal basis with the induced GA₃ sensitivity. Changes in absolute levels of individual phospholipids and their acyl groups were recorded and compared with the changes observed in several Rht-containing aleurone tissues which were induced to develop GA₃ sensitivity by exposure to low temperature (5°C). Several distinct similarities between all tissues were recorded as they develop GA₃ sensitivity. One parameter, the percentage phospholipid composition, was quite similar in all tissues after they had become maximally sensitive to GA₃, suggesting that there is at least one membrane phospholipid composition which is particularly responsive to GA₃. The results indicate that (a) the basis of the GA₃ insensitivity of the Rht mutation resides in an aberrant phospholipid/fatty acid composition and/or metabolism; (b) exposure to low temperature (5°C) for 20 hours or longer, or 342 nanomolar IAA for 4 hours or longer reverses or corrects the genetic lesion, enabling the tissue to adopt a GA₃ responsive membrane composition. Finally, an hypothesis is discussed which indicates that IAA may play a controlling role in the mobilization of endospermal reserves, at least in Rht3-containing wheat aleurone.