Transport Inhibitor 2,3,5-triiodobenzoic Acid Research Articles

Corrigendum to: Auxin is required for pollination-induced ovary growth in Dendrobium orchids.

In Dendrobium and other orchids the ovule becomes mature long after pollination, whereas the ovary starts growing within two days of pollination. The signalling pathway that induces rapid ovary growth after pollination has remained elusive. We placed the auxin antagonist ±-(p-chlorophenoxy) isobutyric acid (PCIB) or the auxin transport inhibitor 2,3,5-triiodobenzoic acid (TIBA) on the stigma, before pollination. Both treatments nullified pollination-induced ovary growth. The ovaries also did not grow after similar stigma treatment with 1-methylcyclopropene (1-MCP), AgNO3 (both inhibitors of ethylene action), aminooxyacetic acid (AOA) or CoCl2 (which both inhibit ethylene synthesis), before pollination. Pollination could be replaced by placement of the auxin naphthylacetic acid (NAA) on the stigma. All mentioned inhibitors nullified the effect of NAA, indicating that if auxin is the initiator of ovary growth, it acts through ethylene. The results suggest that the pollination effect on ovary growth requires auxin (at least auxin transport and maybe also auxin signalling), and both ethylene synthesis and ethylene action.

Comparative effects of auxin transport inhibitors on rhizogenesis and mycorrhizal establishment of spruce seedlings inoculated with Laccaria bicolor.

We compared the effects of two auxin transport inhibitors (2,3,5-triiodobenzoic acid (TIBA) and 1-N-naphthylphthalamic acid (NPA)) on rhizogenesis and mycorrhizal establishment of Picea abies L. (Karst.) seedlings inoculated with Laccaria bicolor S238N (Maire) Orton. Inoculation of seedlings with L. bicolor under in vitro conditions strongly increased host root and shoot growth. Although TIBA had no effect on taproot growth, NPA decreased taproot growth and deformed the root apex into a globular shape in both non-inoculated seedlings and seedlings inoculated with L. bicolor. Inoculation with L. bicolor strongly increased lateral rhizogenesis of the seedlings, and application of 100 microM indole-3-acetic acid (IAA) partially reproduced this effect. Although TIBA completely inhibited the stimulatory effect of L. bicolor on lateral root formation, NPA inhibited it only partially. Both TIBA and NPA counteracted the effect of exogenous IAA on lateral rhizogenesis. Inoculation with L. bicolor significantly increased shoot growth and seedling dry biomass, whereas application of exogenous IAA had no effect on either parameter. There was no effect of NPA on shoot growth and biomass production. The presence of TIBA completely prevented the development of ectomycorrhizal structures (mantle and Hartig net). In the presence of NPA, the number of seedlings colonized by the fungus was reduced and the degree of development of ectomycorrhizal structures was variable, but not completely prevented. In medium lacking tryptophan, neither TIBA nor NPA inhibited the release of IAA produced by L. bicolor in pure culture. When 100 microM tryptophan was added to the medium, TIBA significantly increased the amount of IAA released by the fungus, whereas NPA had no significant effect. We conclude that fungal IAA plays an important role in plant rhizogenesis and in the establishment of ectomycorrhizal symbiosis.

Ectomycorrhizal fungi and exogenous auxins influence root and mycorrhiza formation of Scots pine hypocotyl cuttings in vitro.

We studied the ability of the ectomycorrhizal (ECM) fungi, Pisolithus tinctorius (Pers.) Coker and Couch and Paxillus involutus (Batsch) Fr. (Strain H), to produce indole-3-acetic acid (IAA) and to affect the formation and growth of roots on Scots pine (Pinus sylvestris L.) hypocotyl cuttings in vitro. Effects of indole-3-butyric acid (IBA) and the auxin transport inhibitor, 2,3,5-triiodobenzoic acid (TIBA), on rooting and the cutting-fungus interaction were also studied. Both fungi produced IAA in the absence of exogenous tryptophan, but the mycelium and culture filtrate of Pisolithus tinctorius contained higher concentrations of free and conjugated IAA than the mycelium and culture filtrate of Paxillus involutus. Inoculation with either fungus or short-term application of culture filtrate of either fungus to the base of hypocotyl cuttings enhanced root formation. Inoculation with either fungus was even more effective in enhancing root formation than treatment of the hypocotyl bases with IBA. Fungal IAA production was not directly correlated with root formation, because rooting was enhanced more by Paxillus involutus than by Pisolithus tinctorius. This suggests that, in addition to IAA, other fungal components play an important role in root formation. Treatment with 5 microM TIBA increased the rooting percentage of non-inoculated cuttings, as well as of cuttings inoculated with Pisolithus tinctorius, perhaps as a result of accumulation of IAA at the cutting base. However, the marked reduction in growth of Pisolithus tinctorius in the presence of TIBA suggests that the effects of TIBA on rooting are complicated and not solely related to IAA metabolism. The high IAA-producer, Pisolithus tinctorius, formed mycorrhizas, and the IBA treatment increased mycorrhizal frequency in this species, whereas TIBA decreased it. Paxillus involutus did not form mycorrhizas, indicating that a low concentration of IAA together with other fungal components were sufficient to stimulate formation and growth of the roots, but not the formation of ECM symbiosis.

Adventitious rooting: examining the role of auxin in an easy- and a difficult-to-root plant

Therooting responses of cuttings of difficult-to-root lilac (Syringavulgaris) and easy-to-root forsythia(Forsythia × intermedia)were compared. The rooting ability of lilac cuttings declined over the growingseason (May–June). There was also a decline in the initial concentrationof free IAA at the base of the cuttings, but there was not a tight relationshipbetween basal IAA concentration and rooting ability. Polar auxin transportability was measured in lilac and forsythia during the period of maximum growthby [3H]IAA application to stem internodal tissue. Transport abilitydeclined in lilac over this time period, particularly in terms of transportintensity and percentage of [3H]IAA transported. In contrast thechanges in polar auxin transport ability in forsythia were less marked. Thisdifference between species was maintained in winter hardwood cuttings, withforsythia tissue showing greater polar auxin transport ability than lilac. Theimportance of polar auxin transport for adventitious rooting was demonstratedinboth lilac and forsythia softwood cuttings by use of the polar transportinhibitor 2,3,5-triiodobenzoic acid (TIBA). Overall the results indicate thatdifferences in polar auxin transport ability between lilac and forsythiacontribute to differences in rooting ability.

Proteoid Root Development of Phosphorus Deficient Lupin is Mimicked by Auxin and Phosphonate

White lupin (Lupinus albus L.) develops proteoid (cluster) roots in response to phosphorus deficiency. Proteoid roots are composed of tight clusters of rootlets that initiate from the pericycle opposite protoxylem poles and emerge from every protoxylem pole within the proteoid root axis. Auxins are required for lateral root development, but little is known of their role in proteoid root formation. Proteoid root numbers were dramatically increased in P-sufficient (+P) plants by application of the synthetic auxin, naphthalene acetic acid (NAA), to leaves, and were reduced in P-deficient (−P) plants by the presence of auxin transport inhibitors [2,3,5-triiodobenzoic acid (TIBA) and naphthylphthalamic acid (NPA)]. While ethylene concentrations in the root zone were 1.5-fold higher in −P plants, there was no effect on proteoid root numbers of the ethylene inhibitors aminoethoxyvinvylglycine (AVG) and silver thiosulphate. Phosphonate, which interferes with plant perception of internal P concentration, dramatically increased the number of proteoid root segments in +P plants. Activities of phosphoenolpyruvate carboxylase (PEPC), malate dehydrogenase (MDH) and exuded acid phosphatase in proteoid root segments were not different from +P controls when NAA was applied to +P lupin plants, but increased to levels comparable to −P plants in the phosphonate treatment. Addition of TIBA or NPA to −P plants reduced PEPC and MDH activity of −P proteoid roots to levels found in +P or −P normal root tissues, but did not affect acid phosphatase in root exudates. These results suggest that auxin transport from the shoot plays a role in the formation of proteoid roots during P deficiency. Auxin-stimulated proteoid root formation is necessary, but not sufficient, to signal the up-regulation of PEPC and MDH in proteoid root segments. In contrast, phosphonate applied to P-sufficient white lupin elicits the full suite of coordinated responses to P deficiency

Effects of Auxin Transport Inhibitors on Gibberellins in Pea

The effects of the auxin transport inhibitors 2,3,5-triiodobenzoic acid (TIBA), 9-hydroxyfluorene-9-carboxylic acid (HFCA), and 1-N-naphthylphthalamic acid (NPA) on gibberellins (GAs) in the garden pea (Pisum sativum L.) were studied. Application of these compounds to elongating internodes of intact wild type plants reduced markedly the endogenous level of the bioactive gibberellin A1 (GA1) below the application site. Indole-3-acetic acid (IAA) levels were also reduced, as was internode elongation. The auxin transport inhibitors did not affect the level of endogenous GA1 above the application site markedly, nor that of GA1 precursors above or below it. When plants were treated with [13C,3H]GA20, TIBA reduced dramatically the level of [13C,3H]GA1 recovered below the TIBA application site. The internodes treated with auxin transport inhibitors appeared to be still in the phase where endogenous GA1 affects elongation, as indicated by the strong response to applied GA1 by internodes of a GA1-deficient line at the same stage of expansion. On the basis of the present results it is suggested that caution be exercised when attributing the developmental effects of auxin transport inhibitors to changes in IAA level alone.

Effects of growing environments, planting date, and stem treatments on sandersonia tuber weight and secondary tuber development

Sandersonia (Sandersonia aurantiaca Hook) tuber growth and secondary tuber development were assessed in two growing environments (a greenhouse and outdoors), at three planting dates (1 October, 1 December, and 1 February) and with six stem pruning treatments. Tubers of c. 1 g were used. The pruning treatments consisted of cutting the stem just above the second flower when 50% of the stems had the second flower open, or 2, 4, 6, and 8 weeks later. In another experiment the effect of stem tipping, stem pruning, flower removal, and an auxin transport inhibitor (2,3,5‐triiodobenzoic acid (TIBA)) applied 3 weeks after the second flower opened were assessed. Tuber growth and secondary tuber development were affected by the date of planting, with both tuber weight and incidence of secondary tubers declining significantly in the later planting dates. Environmental factors including temperature, radiation, and day length may have influenced the rate and type of tuber growth. Stem pruning, removal of the growing tip plus flowers, or applying TIBA reduced secondary tuber numbers and had little effect on tuber size. This study suggests that it is possible to reduce the presence of undesirable secondary tubers by manipulating the growing environment or by developing appropriate cultural practices.

The role of the shoot apex in controlling rhizogenesis in vitro

This paper reports that rhizogenesis in woody plant species in vitro was mediated through the basipetal transport of auxin from the shoot apex. This can directly induce roots in easy-to-root species such as Betula pendula, but was dependent upon an interaction with exogenous auxin in more difficult-to-root species such as Daphne cneorum, and to a lesser extent in Quercus robur. Shoot apex removal reduced rhizogenesis in Quercus, and inhibited it in Daphne, even in the presence of exogenous auxin, whereas rooting in Betula was unaffected. That basipetally transported auxin modulates rhizogenesis was demonstrated by the inhibition of root induction in Betula shoots by the auxin transport inhibitor 2,3,5-triiodobenzoic acid (TIBA), and by the substitution of indole-3-acetic acid (IAA) for a bud in Betula internodal sections.

Control of Internode Length in Pisum sativum (Further Evidence for the Involvement of Indole-3-Acetic Acid).

The effects of altered endogenous indole-3-acetic (IAA) levels on elongation in garden pea (Pisum sativum L.) plants were investigated. The auxin transport inhibitors 2,3,5-triiodobenzoic acid (TIBA) and 9-hydroxyfluorene-9-carboxylic acid (HFCA) were applied to elongating internodes of wild-type and mutant lkb plants. The lkb mutant was included because elongating lkb internodes contained 2- to 3-fold less free IAA than those of the wild type. In the wild type, TIBA reduced both the IAA level and internode elongation below the site of application. Both TIBA and HFCA strongly promoted the elongation of lkb internodes and also raised IAA levels above the application site. The synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4-D) also markedly increased internode elongation in lkb plants and virtually restored petioles and tendrils to their wild-type length. In contrast, treatment of wild-type plants with TIBA, HFCA, or 2,4-D caused little or no increase in elongation above the application site. The ethylene synthesis inhibitor aminoethoxyvinylglycine also increased stem elongation in lkb plants, and combined application of HFCA and aminoethoxy-vinylglycine restored lkb internodes to the wild-type length. It is concluded that the level of IAA in wild-type internodes is necessary for normal elongation, and that the reduced stature of lkb plants is at least partially attributable to a reduction in free IAA level in this mutant.

The quiescent center in roots of maize: initiation, maintenance and role in organization of the root apical meristem

Using roots of maize, we tested the hypothesis that the origin and maintenance of the quiescent center (QC) are a consequence of polar auxin supply. Exposing roots to the polar auxin transport inhibitor 2,3,5-triiodobenzoic acid (TIBA), or to low temperature (4 °C, with subsequent return to 24 °C), enhances mitotic frequency within the QC. In both treatments, the QC most typically is activated at its distal face, and the protoderm/dermatogen undergoes several periclinal divisions. As a result, the root body penetrates and ruptures the root cap junction and the characteristic “closed” apical organization changes to “open”. A QC persists during these changes in apical organization, but it is diminished in size. The data from the TIBA-treated roots suggest a role for auxin in the origin and maintenance of the QC, and further, that alterations in QC dimensions are a consequence of polar auxin supply. We hypothesize that the root cap, and specifically the root cap initials, are important in regulating polar auxin movements towards the root apex, and hence are important in determining the status of the QC.

The Control of Lateral Root Development in Cultured Pea Seedlings. II. Root Fasciation Induced by Auxin Inhibitors

AbstractLateral root development in cultured seedlings of Pisum sativum (cv. Alaska) was modified by the application of auxin transport inhibitors or antagonists. When applied either to replace the root tip or beneath the cotyledonary node, two auxin transport inhibitors, 2,3,5‐triiodobenzoic acid (TIBA) and 3,3a‐dihydro‐2‐(p‐methoxyphenyl)‐8H‐pyrazolo[5,1‐α]isoindol‐8‐one (DPX‐1840), increased cell division activity opposite the protoxylem poles. This resulted in the formation of masses of cells, which we are calling root primordial masses (RPMs), 2 to 3 days after treatment. RPMs differed from lateral root primordia in that they lacked apical organization. Some roots however developed both RPMs and lateral roots indicating that both structures were similar in terms of the timing and location of cell division in the pericycle and endodermis leading to their initiation. Removal of the auxin transport inhibitors allowed many of the RPMs to organize later into lateral root primordia and to emerge in clusters. When the auxin, indoleacetic acid (IAA) was added to the growth medium along with DPX‐1840, 3 ranks of RPMs now in the form of fasciated lateral roots emerged from the primary root. The auxin antagonist, p‐chlorophenoxy‐isobutyric acid (PCIB), also induced RPM formation. In contrast to DPX‐1840 treatment, the addition of IAA during PCIB treatment caused normal lateral root development.