Role of Cation and Anion Uptake in Salt-stimulated Elongation of Lettuce Hypocotyl Sections

The role of cation and anion uptake in salt-stimulated growth of light-grown, GA₃-treated lettuce (Lactuca sativa L.) hypocotyl sections was investigated. Potassium chloride (10 mm) causes a 2-fold increase in the growth rate of GA₃-treated hypocotyl sections without affecting the growth rate of sections incubated in the absence of GA₃. Salt uptake is the same in both treatments, and furthermore the uptake of cation and anion is stoichiometric during the first 24 hours under all incubation conditions. The importance of the anion for cation uptake is demonstrated in experiments with benzenesulfonate⁻ and iminodiacetate²⁻. When K⁺ and Na⁺ are supplied only as the benzenesulfonate and iminodiacetate salts, growth and cation uptake are markedly reduced compared to KCl and NaCl. Calculation of the osmotic potential of salt-treated sections based on measurement of K⁺ and Cl⁻ uptake suggests that the observed increase in tissue osmolality is a result of salt uptake. Similarly, uptake of ions can account for the shift in water potential when sections are incubated in 10 mm KCl. We conclude that the change in growth rate of light-grown, GA₃-treated sections caused by the addition of KCl or NaCl to the incubation medium results solely from decreased water potential of the tissue due to ion uptake.     

Similarity between Cytokinin and Blue Light Inhibition of Cucumber Hypocotyl Elongation

The cytokinin benzyladenine inhibited endogenous hypocotyl elongation in intact etiolated seedlings of cucumber (Cucumis sativus L.). In hypocotyl segments, the inhibitory effect of benzyladenine on growth was clearly detectable in the presence of indoleacetic acid. Fusicoccin-induced elongation was unaffected by the presence of cytokinin. The effect of cytokinin on elongation of the segments was determined by measuring changes in fresh weight, a linear function of extension growth. The effect of benzyladenine on hypocotyl growth was at least as large in segments prepared from red-light-grown seedlings as in those from seedlings grown in total darkness. A comparison was made between the inhibitory effects of cytokinin and blue light. The use of the calcium chelator ethyleneglycol-bis(β-aminoethyl ether)-N, N′-tetraacetic acid indicated that calcium ions are required for manifestation of benzyladenine-induced inhibition.     

Brassinosteroid Stimulation of Hypocotyl Elongation and Wall Relaxation in Pakchoi (Brassica chinensis cv Lei-Choi)

Hypocotyl elongation of pakchoi (Brassica chinensis cv Lei-Choi) was stimulated by applying 300 ng of brassinosteroid (2[alpha],3[alpha],22[beta],23[beta]-tetrahydroxy-24[beta]-methyl-B-homo-7- oxa-5[alpha]-cho-le stan-6-one, BR) in 1 [mu]L of 50% ethanol to the apex of hypocotyls. BR had its greatest effect on elongation of the apical 3-mm region below the cotyledonary node (75% stimulation) between 6 and 18 h after treatment. Stress/strain (Instron) analysis of this 3-mm region revealed that plastic and elastic components of extension were not significantly different between BR-treated and control seedlings. In pressure-block experiments, the initial rate of relaxation was 2-fold faster in BR-treated plants as compared with controls, whereas after 125 min the total amount of relaxation and the relaxation rate were the same for the two treatments. Osmotic pressure of cell sap expressed from this 3-mm region showed a large decrease (28%) in BR-treated seedlings compared to the controls. We conclude that BR stimulates growth in pakchoi by accelerating the biochemical processes that cause wall relaxation, without inducing a large change in wall mechanical properties.     

Further Comparisons between Carbowax 6000 and Mannitol as Suppressants of Cucumber Hypocotyl Elongation

Stirring largely eliminated the greater effectiveness of Carbowax 6000 compared to mannitol in reducing cucumber (Cucumis sativus L.) hypocotyl section elongation. Stirring increased elongation in water but reduced elongation in growth regulator solution. Shrinkage of cells without plasmolysis in hypertonic Carbowax solutions indicates that Carbowax 6000 did not penetrate cell walls. Sections prevented from elongating during pretreatment exposure to saturated air grew as much as those not pretreated. Sections pretreated with isotonic Carbowax responded similarly, but sections pretreated with isotonic mannitol grew less.     

IAA和GA3在调控豌豆黄化苗茎切段伸长生长中的相互作用

IAA和GA3均能促进豌豆黄化苗茎切段的伸长。IAA效应可以为GA的合成抑制剂S-3307抑制,GA3效应同样也为IAA的运输抑制剂TIBA所抑制,并且分别再施用GA3和IAA后,抑制效应又能有所解除。观察顶端切半茎切段的结果表明,IAA主要促进茎切段表皮细胞的伸长,而GA3可能主要促进内部组织细胞的伸长。观察切段横纵切片的结果则显示,IAA促进皮层细胞的伸长和增大,而GA3只促进皮层细胞的伸长。这些结果说明两者是通过不同的作用部位和方式共同调节豌豆茎切段伸长生长的。     

表油菜素内酯对黄瓜幼苗下胚轴过氧化物酶和IAA氧化酶活性的影响

黄瓜幼苗用0.1～1 ppm表油菜素内酯(epiBR)处理1～3d后,下胚轴中过氧化物酶活性明显低于对照;随着处理浓度的增加和处理时间的延长,与对照之间的差别愈趋增大。当浓度高于1ppm时,过氧化物酶的活性不再继续降低。表油菜素内醋对过氧化物酶活性的这种抑制作用需经约1O h的滞后期。IAA氧化酶的活性变化与过氧化物酶相似,epiBR处理时间愈长酶活性增加愈趋缓慢。 经IAA处理的下胚轴,过氧化物酶和IAA氧化酶的活性变化与对照无明显差异。这提示油菜素内酯与IAA促进生长可能是通过不同的作用方式。     

Touch-Sensitive NADH Oxidase Activity of Pea and Cucumber Tendrils and Soybean Hypocotyl Sections

The cell surface reduced nicotinamide adenine dinucleotide (NADH) oxidase activity of soybean stems and of pea and cucumber tendrils responded to touch with a several-fold increase in activity. The increase in NADH oxidase persisted for 20 min or longer, and further touch stimulation during this period did not alter activity. With soybean sections, the specific activities in response to touch approximated those achieved maximally by auxin. Where the NADH oxidase was fully stimulated by 2,4-d, the NADH oxidase failed to respond further to touch. The findings indicate that the NADH oxidase of the plant cell surface is involved in the growth response to touch and in tendril coiling.     

Effect of IAA on Growth and Soluble Cell Wall Polysaccharides Centrifuged from Pine Hypocotyl Sections

Auxin-induced elongation and cell wall polysaccharide metabolism were studied in excised hypocotyl sections of ponderosa pine (Pinus ponderosa) seedlings. Sections excised from hypocotyls of ponderosa pine elongate in response to the addition of auxin. The neutral sugar composition of the extracellular solution removed from hypocotyl sections by centrifugation was examined. In cell wall solution from freshly excised sections, glucose, galactose, xylose, and arabinose make up more than 90% of the neutral sugars, while rhamnose, fucose, and mannose are relatively minor components. The neutral sugar composition of the polysaccharides of the pine cell wall solution is both qualitatively and quantitatively similar to that of pea. Following auxin treatment of pine hypocotyls, the neutral sugar composition of the cell wall changes; glucose, xylose, rhamnose, and fucose increase by nearly 2-fold relative to controls in buffer without auxin. These changes in neutral sugars in response to auxin treatment are similar to those found in pea, with the exception that in pea, rhamnose levels decline in response to auxin treatment.     

1-萘氨甲酰苯甲酸、外源IAA和GA3对拟南芥根的伸长和向重力性反应的影响

拟南芥幼苗用1-萘氨甲酰苯甲酸(NPA)、IAA和GA3处理后测定根的伸长和向重力性弯曲的结果表明,低浓度(0.001μmol·L-1)IAA和(0.01~1μmol·L-1)GA3促进根的伸长和向重力性弯曲,高浓度(0.01~10μmol·L-1)IAA和(10~100μmol·L-1)GA3的则相反。NPA在总体上是抑制根的伸长和向重力性弯曲,但低浓度(0.4μmol·L-1)的NPA有促进根伸长的趋势。低浓度的IAA和GA3均拮抗NPA对根伸长的影响,且低浓度的GA3对根伸长的促进作用并不依赖IAA。     

Adventitious Root Formation in Bean Hypocotyl Cuttings in Relation to IAA Translocation and Hypocotyl Anatomy

Adventitious roots develop in bean hypocotyl cuttings in fourrows parallel to and between the four pairs of vascular bundles,in contrast to their irregular development in petiole and epicotylcuttings where the distribution pattern of xylem bundles isalso irregular. Auxin, applied acropetally or basipetally, increases the numberof roots but does not alter the pattern of their emergence.Similarly, [2-¹⁴C]IAA which is translocated mainly via the vascularbundles (but also laterally towards the cortex and pith) accumulatesin the root-forming areas irrespective of how it was applied. Consequently, it is deduced that the adventitious root developmentin bean hypocotyls is induced by IAA accumulation, but thatIAA does not affect or modify the predetermination of rootingsites.     

拟南芥复制因子C亚基3在黄化苗下胚轴伸长生长中的作用

拟南芥遮光培养2．5d时,rfc3-1突变体黄化幼苗的下胚轴平均长度约比野生型植株黄化幼苗的下胚轴长27．5％。观察表明,相对于野生型复制因子C亚基3（replication factor C3,AtRFC3）基因突变体的下胚轴表皮细胞,特别是上部靠近子叶部分的表皮细胞,单细胞长度变长。将野生型RFC3基因转染到rfc3-1后,突变体恢复野生型表型,进一步说明RFC3在黄化苗的下胚轴伸长生长中有作用。     

Comparison of Auxin-induced and Acid-induced Elongation in Soybean Hypocotyl

Acid-induced growth was compared to auxin-induced growth. After a transient pH 4-induced increase in the elongation rate was completed, auxin could still induce an enhanced rate of elongation in soybean (Glycine max) hypocotyl segments. This auxin response occurred both when the medium was changed to pH 6 before auxin addition, and when the auxin was added directly to the pH 4 medium. This postacid response to auxin was persistent, and quite unlike a postacid response to acid, which was again shortlived. One mm N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (pH 7) inhibited the first response to auxin (the first response to auxin being similar to the acid response), but not the second response. This did not appear to be simply a hydrogen ion neutralizing effect, however, since a 50-fold increase in buffer concentration at pH 6 did not inhibit the first response. Decrease in the pH of the external medium, previously shown to occur with excised soybean hypocotyl segments, was not affected by auxin. Furthermore, this pH drop, during which the cells appear to be adjusting their external pH to about 5.4, did not result in an increased rate of elongation. Addition of auxin after the equilibrium pH had been attained did not alter the pH, but it did increase the rate of elongation, eliciting a normal auxin response. It was concluded that hydrogen ions do not mediate in long term auxin-induced elongation in soybean hypocotyl.     

Adaptive Growth Responses to Osmotic Stress of Hypocotyl Sections of Vigna unguiculata: Roles of the Xylem Proton Pump and IAA

The growth responses to osmotic stress of hypocotyl sectionsof Vigna unguiculata were studied by the xylem perfusion method.Hypocotyl sections shrank upon exposure to osmotic stress. Sectionsshowed no adaptive responses to osmotic stress when they werein an IAA-depleted condition as a result of perfusion with solutionsthat lacked IAA for 3–4 h. The correlation between thegrowth rate and the membrane potential of the xylem/symplastboundary (V^px) was very limited in the absence of IAA. By contrast,hypocotyl sections showed distinct adaptive responses to osmoticstress after perfusion with solutions that contained 10 µMIAA. In the presence of IAA, V^px increased in the negative directionand growth resumed in spite of the osmotic stress. The growthrate was closely correlated with the xylem membrane potential.Hyper-polarization of the membranes of the xylem/symplast boundaryalways preceded the recovery of growth under osmotic stress.It appears that IAA is essential for the adaptive recovery ofgrowth under osmotic stress and, moreover, that the xylem protonpump plays an indispensable role in modulating the growth ofhypocotyl sections. This result confirms prediction of an earliersimulation study using the apoplast canal model [Katou and Furumoto(1986) Protoplasma 133: 174, Katou and Enomoto (1991) PlantCell Physiol. 32: 343]. (Received June 27, 1996; Accepted October 28, 1996)     

Light-Regulated Hypocotyl Elongation Involves Proteasome-Dependent Degradation of the Microtubule Regulatory Protein WDL3 in Arabidopsis

Light significantly inhibits hypocotyl cell elongation, and dark-grown seedlings exhibit elongated, etiolated hypocotyls. Microtubule regulatory proteins function as positive or negative regulators that mediate hypocotyl cell elongation by altering microtubule organization. However, it remains unclear how plants coordinate these regulators to promote hypocotyl growth in darkness and inhibit growth in the light. Here, we demonstrate that WAVE-DAMPENED 2–LIKE3 (WDL3), a microtubule regulatory protein of the WVD2/WDL family from Arabidopsis thaliana, functions in hypocotyl cell elongation and is regulated by a ubiquitin-26S proteasome–dependent pathway in response to light. WDL3 RNA interference Arabidopsis seedlings grown in the light had much longer hypocotyls than controls. Moreover, WDL3 overexpression resulted in overall shortening of hypocotyl cells and stabilization of cortical microtubules in the light. Cortical microtubule reorganization occurred slowly in cells from WDL3 RNA interference transgenic lines but was accelerated in cells from WDL3-overexpressing seedlings subjected to light treatment. More importantly, WDL3 protein was abundant in the light but was degraded through the 26S proteasome pathway in the dark. Overexpression of WDL3 inhibited etiolated hypocotyl growth in regulatory particle non-ATPase subunit-1a mutant (rpn1a-4) plants but not in wild-type seedlings. Therefore, a ubiquitin-26S proteasome–dependent mechanism regulates the levels of WDL3 in response to light to modulate hypocotyl cell elongation.     

Effects of Ca2+ and Mg2+ on Elongation and H+ Secretion of Vigna radiata Hypocotyl Sections

The effects exerted by divalent cations Ca²⁺ and Mg²⁺ on spontaneous,IAA- and H⁺-induced elongation were investigated with excisedmung bean hypocotyl segments. Low concentrations of calcium(<1 mM) were sufficient to inhibit spontaneous and IAA-inducedelongation of abraded segments without any lag time whereaseffects of magnesium were not so evident. The initial growthrate could be restored, at least partly, with EDTA or acidicsolutions. Both calcium and magnesium enhanced H⁺ excretionbut Mg²⁺ was a more potent activator. Divalent cations alsoinduced simultaneously an inhibition of growth and a stimulationof proton release. Such contrasting effects are discussed withrespect to the functions of the cuticle. (Received May 1, 1984; Accepted September 10, 1984)     

Evaluation of a Bioassay for Cytokinins Using Soybean Hypocotyl Sections

The dose-response curves of several cytokinins were investigatedin a soybean hypocotyl bioassay. Zeatin riboside, zeatin-O-ß-D-glucoside,dihydrozeatin, and dihydrozeatin riboside produced linear responsesparallel to that for zeatin. The hypocotyl section assay wassuperior to the conventional soybean callus assay because theresponse (log₁₀ transformed data) was linear, exhibited lowvariability, and was more reproducible and more sensitive. Theassay was quicker to perform and required less cytokinin.