The effect of light on IAA metabolism in different parts of maize seedlings in correlation with their growth

The inhibitory effect of light on the growth of plantscorrelates with a decrease of free IAA content in their tissues andmight be mediated through changes of IAA metabolism. In different partsof Zea mays L. seedlings (roots, mesocotyls and coleoptiles)that respond to light with a different growth rate, the effect of lighton the formation of IAA metabolites was examined in feeding experimentswith ¹⁴C-IAA. In all tissues, IAA was taken up andmetabolised mainly into six compounds, four of them were tentativelyidentified as IAA-1-O-glucose (IAGlc), IAA-myo-inositol, indoleacetamide and IAA-aspartate (IAAsp). IAA was metabolised most slowly inthe roots. In coleoptiles and mesocotyls, IAGlc was the most abundantmetabolite, except for mesocotyls in the light. In roots, a relativelylarge amount of IAA was also metabolised into IAAsp. Light stimulatedthe rate of IAA metabolism in all tissues, but its effect on theconversion of IAA was exceptionally high in mesocotyls. In mesocotyltissue the conversion into IAAsp was greatly stimulated by light.Conversely, the content of IAGlc in mesocotyls was decreased by light.Since light inhibited mesocotyl growth significantly and specifically,it is possible that the high capacity of mesocotyls to synthesise IAAspin the light may have caused a depletion of free IAA, which then led toan inhibition of growth. In mesocotyls from the light-grown plants IAAconjugated into IAGlc was probably used for IAAspbiosynthesis.     

Effects of red light on the fluence–response relationship for pulse-induced phototropism of maize coleoptiles

Dark-adapted coleoptiles of maize (Zea mays L.) were treated with red light (3min at 10.5 μmol m^?2S^?1) and were Stimulated, after a dark interval, with a pulse of unilateral blue light to induce phototropism. Phototropic fluence-response curves were obtained in this way for different dark intervals. It was confirmed that the bell-shaped fluence-response curve for the first pulse-induced positive phototropism (FPIPP) shifts to higher fluences following the red-light treatment, the maximal shift being achieved at a dark interval of 2h. We found, however, that the two arms of the Fluence-response curve do not shift synchronously. The shift of the descending arm to higher fluences began at 15 min. The ascending arm showed a slight shift to lower fluences before a greater shift to higher flucnces. the change of the shift direction occurring at 30–40min. Accordingly, the fluence-response curve obtained for a 30 min dark interval was comparatively wide. Although dark-adapted coleoptiles showed only fPIPP, another bell-shaped fluence-response curve, representing the second pulse-induced positive phototropism (sPIPP), appeared gradually after the red-light treatment. These changes of the phototropic fluence– respnse curve following exposure to red light are likely to have adaptive values because they favour phototropism under brighter light.     

IAA对小麦胚芽鞘质膜蛋白磷酸化的影响

磷酸化／ 脱磷酸化机制是众多信号转导过程中的重要环节,很多信号物质被细胞受体识别后引发蛋白激酶和蛋白磷酸酶活性变化,通过磷酸化／ 脱磷酸化进一步调节多种酶活性而产生各种生理效应。在对生长素ＩＡＡ 的信号转导的研究中,发现ＩＡＡ 处理的小麦胚芽鞘质膜蛋白中蛋白激酶的活性和蛋白磷酸化程度都发生改变,并找到两种受到调节的蛋白激酶。钙离子通道抑制剂ＬａＣｌ３ 阻断了ＩＡＡ 的这种作用,表明Ｃａ２＋参与了ＩＡＡ的信号转导过程。     

Red light causes a reduction in IAA levels at the apical tip by inhibiting de novo biosynthesis from tryptophan in maize coleoptiles

When maize coleoptiles were unilaterally exposed to red light (7.9 μmol m⁻²s⁻¹ for 5 min), 3 h after treatment IAA levels in coleoptiles decreased in all regions, from top to basal, with levels about 60% of dark controls. Localized irradiation in the 5 mm top zone was sufficient to cause the same extent of IAA reduction in the tips to that in the tips of whole irradiated shoots. When coleoptiles were treated with N-1-naphthylphthalamic acid (NPA), an accumulation of IAA in the tip and a decrease of diffusible IAA from tips were simultaneously detected. IAA accumulation in red-light treated coleoptiles by NPA was much lower than that of dark controls. NPA treatment did not affect the content of conjugated IAA in either dark or light treated coleoptile tips. When ¹³C₁₁ ¹⁵N₂-tryptophan (Trp) was applied to the top of coleoptiles, substantial amounts of stable isotope were incorporated into free IAA in dark and red-light treated coleoptile tips. The ratio of incorporation was slightly lower in red-light treated coleoptile tips than that in dark controls. The label could not be detected in conjugated IAA. The rate of basipetal transport of IAA was about 10 mm h⁻¹ and the velocity was not affected by red light. These results strongly suggest that red light does not affect the rates of conversion of free IAA to the conjugate form or of the basipetal transport, but just reduces the IAA level in the tips, probably inhibited by IAA biosynthesis from Trp in this region.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

The response of dodder {Cuscuta spp.) seedlings to phytohormones under various light regimes

Phytohormones were screened for their effects on induction of coiling and prehaustoria in de-etiolated excised dodder seedlings under different light treatments. Zeatin stimulated coiling and the formation of prehaustoria under all light tested, even in darkness. A synergistic effect was observed with zeatin and far-red light (700–800 nm) but not with red light (600–700 nm) on the formation of prehaustoria. Application of indole-3acetic acid inhibited zeatin-induced coiling and prehaustoria development under blue (400–500 nm) and a mixture of red plus far-red light, but not in blue plus far-red light. Ethylene had no effect on coiling and prehaustoria development. Observations suggest that zeatin-induced coiling and prehaustoria development may be mediated by phytochrome.     

Blue light control of hypocotyl elongation in etiolated seedlings of Lactuca sativa (L.) cv. Grand Rapids related to exogenous growth regulators and endogenous IAA, GA3 and abscisic acid

Results of studies on the interaction of blue light (B) and exogenous applied plant hormones (IAA, GA3 and ABA) as well as inhibitors of their synthesis on the control of hypocotyl elongation in etiolated seedlings of Lactuca sativa (L.) cv. Grand Rapids are presented, and compared with endogenous GA3, IAA and ABA levels measured by capillary gas chromatography-mass spectrometry-selected ion monitoring (GC-MS-SIM). Hypocotyl elongation of etiolated seedlings was linearly inhibited by increasing the B fluence rate between 0 and 18.3 mol m–2 s–1. Both GA3 and IAA added to the incubation medium at different concentrations were able to eliminate partially the inhibition of growth caused by 7.2 mol m–2 s–1 B. When dark (D)-grown seedlings were treated with Ca-prohexadione, a specific inhibitor of gibberellin 3-hydroxylation, they showed a growth inhibition similar to under B. Also, a suppression of growth as in B was obtained when D-grown seedlings were treated with ABA 380 M. By lowering ABA levels with fluridone (an inhibitor of ABA synthesis) a partial reversion of hypocotyl growth inhibition was obtained in B-grown seedlings. While none of the growth promoters used were able to reverse completely the growth inhibition caused by B, a proper combination of GA3, IAA and (eventually) fluridone, abolished the B effects. Correspondingly, lower levels of GA3 and IAA and a higher concentration of ABA were measured by GC-MS-SIM in B-grown hypocotyls than in D-grown ones. These results support the hypothesis that hormones are implicated in mediation of B light-dependent inhibition of hypocotyl elongation, which seems to be the result of a balance among endogenous levels of growth promoting and growth inhibiting hormones.     

The effect of light on lipoxygenase activity in dwarf pea seedlings

Continuous illumination of 10-day-old etiolated dwarf pea seedlings caused an increase in lipoxygenase activity. At the same time the activity in both stem and leaf tissue decreased. The lipoxygenase isoenzymes of the whole seedling and separated leaf and stem tissue were affected differently by light. It is concluded that lipoxygenase is not involved directly in photosynthesis or chloroplast development.     

Action of red light on indole-3-acetic-acid status and growth in coleoptiles of etiolated maize seedlings

Brief irradiation of intact etiolated seedlings of maize (Zea mays L.) with red light (R; 30 W cm^-2, 10 min) reduces the amounts of diffusible and free (solvent-extractable) indole-3-acetic acid (IAA) obtainable from excised coleoptile tips. The effect is transient, the lowest level (30% of the dark control) occurring at about 3 h after irradiation. The free-IAA content of the whole coleoptile and the diffusible-IAA yield from the base of the same organ are similarly reduced, whereas the conjugated-IAA content of the coleoptile is not affected. These results support the view that R inhibits the production of IAA at the coleoptile tip. It is further shown that R inhibits biosynthesis of [³H]IAA from [³H]tryptophan supplied to the coleoptile tip. The shapes of the fluence-response curves obtained for the reduction of the diffusible-IAA yield by R and far-red light (FR) indicate the participation of two photoreactive systems. One has thresholds at 10^-3 W s cm² of R, five orders of magnitude less than the minimum required for the appearance of spectrophotometrically measurable far-red-absorbing form of phytochrome (Pfr) in vivo, and 10^-1 W s cm^-2 of FR; its response is linear to the logarithm of fluence exceeding five orders of magnitude. The other system is seen above 10² W s cm^-2 as an increase in the slope of the fluenceresponse curve; its response is FR reversible and related to the Pfr level of total photoreversible phytochrome. Both systems inhibit biosynthesis of IAA from tryptophan. Elongation of the coleoptile is stimulated by R; the stimulation is most apparent in the apical region, and is saturated with a fluence at which bo detectable pfr is formed. Farred light can also saturate this response. Since the endogenous IAA concentration in the coleoptile appears not to be in the inhibitory range, it is concluded that the stimulation of coleoptile elongation is not the result of changes in free-IAA levels.Abbreviations FR far-red light - IAA indole-3-acetic acid - Pfr phytochrome in the far-red-absorbing form - Pr phytochrome in the red-absorbing form - R red light     

Negative regulation of asparagine synthetase in the leaves of maize seedlings by light, benzyladenine and glucose

Asparagine synthetase (EC 6.3.5.4) activity was increased 4- and 8-fold when maize ( Zea mays L.) seedlings were kept in darkness for 24 h and 7 days, respectively; this increase was abolished by cycloheximide. Irradiation of the dark adapted seedlings with a pulse of red light resulted in a 4-fold decrease of the enzyme activity within 48 h, which was raised again following a far-red light pulse. Co-action of light and benzyladenine, reported for the light-inducible enzymes, was proved to hold also for the light-repressible asparagine synthetase. The induction of asparagine synthetase activity in the dark is abolished by glucose, suggesting the possible involvement of the enzyme in the contrae of metabolic fluxes of –carbon and nitrogen through assimilatory pathways.     

NaCl胁迫对甘薯苗期生长、IAA代谢的影响及其与耐盐性的关系

随NaCl胁迫浓度的提高 ,甘薯苗期株高和叶面积生长受抑制 ;叶片IAA水平下降 ,而IAA氧化酶和POD活性提高 .不耐盐品种生长受抑制程度及IAA水平下降幅度均大于中耐盐品种和耐盐品种 ,叶片IAA氧化酶和POD活性上升的幅度大于中耐盐及耐盐品种 .同时浓度NaCl胁迫还引起了叶片绿原酸含量的增加 ,中耐盐品种栗子香和不耐盐品种胜利百号在 170mmol·L-1的NaCl浓度胁迫下 ,绿原酸含量上升达到最高值 ,耐盐品种徐薯 18绿原酸含量达到最高值的浓度是 2 5 5mmol·L-1,而后随NaCl胁迫浓度的提高 ,绿原酸含量均有下降 ,但仍高于对照 .     

Transport of [3H]IAA label in gravistimulated primary roots of maize

The curvature of roots in response to gravity is attributed to the development of a differential concentration gradient of IAA in the top and bottom of the elongation region of roots. The development of the IAA gradient has been attributed to the redistribution of IAA from the stele to cortical tissues in the elongation region. The gravistimulated redistribution of IAA was investigated by applying [³H]IAA to the cut surface of 5 mm apical primary root segments. The movement of label from the stele-associated [³H]IAA into the root, tip, root cap, and cortical tissues on the top and bottom of the elongation region was determined in vertically growing roots and gravistimulated roots. Label from the stele moved into the region of cell differentiation (root tip) prior to accumulating in the elongation region. Little label was observed in the root cap. Gravistimulation did not increase the amount of label moving from the stele; but gravistimulation did increase the amount of label accumulating in cortical tissues on the lower side of the elongation region, and decreased the amount of label accumulating in cortical tissues on the upper side of the elongation region. Removal of the cap prior to or immediately following gravity stimulation rendered the roots partially insensitive to gravity and also prevented gravity-induced asymmetric redistribution of label. However, removal of the root cap following 30 min of gravistimulation did not alter root curvature or the establishment of an IAA asymmetry across the region of root elongation. These results suggest that a signal originating in the root cap directs auxin redistribution in tissues behind the root cap, leading to the development of an asymmetry of IAA concentration in the elongation region that in turn causes the differential growth rate in the elongation region of a graviresponding root.     

Immunohistochemical observation of indole-3-acetic acid at the IAA synthetic maize coleoptile tips

To investigate the distribution of IAA (indole-3-acetic acid) and the IAA synthetic cells in maize coleoptiles, we established immunohistochemistry of IAA using an anti-IAA-C-monoclonal antibody. We first confirmed the specificity of the antibody by comparing the amounts of endogenous free and conjugated IAA to the IAA signal obtained from the IAA antibody. Depletion of endogenous IAA showed a corresponding decrease in immuno-signal intensity and negligible cross-reactivity against IAA-related compounds, including tryptophan, indole-3-acetamide, and conjugated-IAA was observed. Immunolocalization showed that the IAA signal was intense in the approximately 1 mm region and the outer epidermis at the approximately 0.5 mm region from the top of coleoptiles treated with 1-N-naphthylphthalamic acid. By contrast, the IAA immuno-signal in the outer epidermis almost disappeared after 5-methyl-tryptophan treatment. Immunogold labeling of IAA with an anti-IAA-N-polyclonal antibody in the outer-epidermal cells showed cytoplasmic localization of free-IAA, but none in cell walls or vacuoles. These findings indicated that IAA is synthesized in the 0–2.0 mm region of maize coleoptile tips from Trp, in which the outer-epidermal cells of the 0.5 mm tip are the most active IAA synthetic cells.     

Effect of IAA content Modulators on peroxidase activity and on endogenous IAA during cold pretreatment of maize anthers prior to androgenesis

A cold pretreatment is usually applied to induce maize androgenesis. Peroxidase activity, including indole-3-acetic acid (IAA) oxidase activity, and endogenous IAA concentrations were followed during a cold pretreatment (14 days, 7°C) in anthers of two maize genotypes, Seneca 60 and DH5×DH7, respectively with a low or high androgenetic response. The most prominent result was the absence of a detectable IAA oxidase activity in DH5×DH7. Adding effectors of IAA-oxidase activity or IAA transport did not affect significantly the crude peroxidase activity of DH5×DH7 anthers while inducing a clear inhibition of androgenesis at higher concentrations. No strict correlation was found between IAA level and physiological response, the low responding variety having as much IAA as DH5×DH7. However, for DH5×DH7, every treatment that lowered the IAA level after 14 days of cold resulted in a decrease in androgenetic response.     

弱光胁迫对玉米产量及光合特性的影响

通过在玉米授粉前0～14d、授粉后1～14d、授粉后15～28d3个时期分别进行遮光处理,研究了弱光胁迫对两个玉米品种费玉3号和泰玉2号产量及光合特性的影响.结果表明:各时期弱光胁迫均使玉米产量降低,其中授粉前0～14d弱光处理的产量降幅最大,费玉3号对弱光胁迫的反应较泰玉2号敏感.弱光胁迫后籽粒灌浆高峰出现时间延迟、灌浆速率慢、积累量小;弱光胁迫开始的时间越早,籽粒达到最大灌浆速率的时间(Tmax)越晚.弱光胁迫期内,玉米Chl(a b)、Chla/b、光合速率(Pn)、光系统Ⅱ最大光化学效率(Fv/Fm)和光系统Ⅱ实际光化学效率(ФPSⅡ)显著下降,Chlb相对含量提高,胞间二氧化碳浓度(Ci)和非光化学淬灭系数(NPQ)则显著上升;胁迫结束后,玉米Chl(a b)、Chla/b、Pn、Fv/Fm、ФPSⅡ、Ci和NPQ逐渐恢复接近自然光照条件(CK)水平,而Chlb相对含量下降.表明非气孔因素是弱光胁迫下玉米光合速率降低的原因之一.     

Effect of salinity stress on growth,peroxidase and IAA oxidase activities in vigna seedlings

The present work was carried out with the aim of studying the effect of salinity stress on growth and IAA oxidizing system (i.e. peroxidase and IAA oxidase) in vigna (Vigna unguiculata L.) seedlings. The seedlings were treated with two concentrations of sodium chloride (NaCl) 0.1 M and 0.25 M. Length, fresh and dry weight were the parameters considered for growth. Salinity effect was distinct in fresh weight and dry weight of different organs. Peroxidase and IAA oxidase activities were measured at different time intervals for both cytoplasmic and wall bound fractions. Peroxidase activity was maximum at higher stress conditions bringing about the hypocotyl growth restriction. Thus there was a clear inverse correlation between elongation and peroxidase activity. IAA oxidase activity also showed a similar trend for both cytoplasmic and wall bound fractions. The role of IAA oxidizing system in defense mechanism in response to salinity stress is discussed.     

Rootless with undetectable meristem 1 encodes a monocot-specific AUX/IAA protein that controls embryonic seminal and post-embryonic lateral root initiation in maize

The maize (Zea mays L.) rum1‐R (rootless with undetectable meristems 1‐Reference) mutant does not initiate embryonic seminal roots and post‐embryonic lateral roots at the primary root. Map‐based cloning revealed that Rum1 encodes a 269 amino acid (aa) monocot‐specific Aux/IAA protein. The rum1‐R protein lacks 26 amino acids including the GWPPV degron sequence in domain II and part of the bipartite NLS (nuclear localization sequence). Significantly reduced lateral root density (approximately 35%) in heterozygous plants suggests that the rum1‐R is a semi‐dominant mutant. Overexpression of rum1‐R under the control of the maize MSY (Methionine SYnthase) promoter supports this notion by displaying a reduced number of lateral roots (31–37%). Functional characterization suggests that Rum1 is auxin‐inducible and encodes a protein that localizes to the nucleus. Moreover, RUM1 is unstable with a half life time of approximately 22 min while the mutant rum1‐R protein is very stable. In vitro and in vivo experiments demonstrated an interaction of RUM1 with ZmARF25 and ZmARF34 (Z. mays AUXIN RESPONSE FACTOR 25 and 34). In summary, the presented data suggest that Rum1 encodes a canonical Aux/IAA protein that is required for the initiation of embryonic seminal and post‐embryonic lateral root initiation in primary roots of maize.     

Soluble sugars of maize seedlings

The soluble sugars were determined in different parts of maize seedlings (seeds, roots and shoots), 0, 2, 4 and 6 days after sowing.     

Amino acid metabolism in the pedicel-placenta-chalazal region of the developing maize kernel

The pedicel-placenta-chalazal (PPCh) region is a heterogeneous maternal tissue located at the base of the endosperm. Its active role in the transport and metabolism of nitrogen compounds supplied to the endosperm has been shown by incorporation of labelled compounds, as well as by analysis of appropriate enzyme activities and the free amino acid pool. Within 30 min incubation, 60–70% of the label given in [¹⁴C]aspartate or [¹⁴C]glutamate was recovered in the organic acid fraction of the PPCh. Although the dry matter and protein content of the PPCh region was practically unchanged between days 15 and 30 after pollination, glutamate transaminase, alanine transaminase, NADP-malic enzyme and NAD-malate dehydrogenase were increased substantially. The PPCh region contains glutamine at a significantly higher concentration and alanine at a lower concentration than the endosperm. In contrast to glutamate synthase, the activity of glutamine synthetase per endosperm and per mg protein is significantly higher in the PPCh region. This implies involvement of these tissues in the transport of nitrogen compounds, mainly in the form of glutamine, as well as possible compartmentation of the glutamine-glutamate cycle in the developing maize kernel.     

Effects of phenolic substances on metabolism of exogenous indole-3-acetic acid in maize stems

Four-day-old stem segments of Zea mays L. cv. Seneca 60 were treated sequentially with phenolic substances and indole-3-acetic [2-¹⁴C] acid ([2-¹⁴C]IAA). Formation of bound IAA was rapid, but a pretreatment with p-coumaric acid, ferulic acid or 4-methylumbelliferone decreased the level of bound IAA. The decrease is not likely related to the effect of the phenolics on enzymic oxidation of IAA, since the level of free IAA was not limiting and the activity of ferulic acid in enzymic oxidation of IAA is different from that of p-coumaric acid and 4-methylum-belliferone. Apparently these compounds inhibited the formation of bound IAA and consequently caused an accumulation of free IAA. In contrast, caffeic acid, protocatechuic acid and 2,3-dihydro-2, 2-dimethyl-7-benzofuranol had little effect. After the uptake of IAA there was a slow but steady incorporation of the radioactivity into the 80% ethanol-insoluble, 1 M NaOH-soluble fraction. Phenolic substances also affected this process. The compounds which are cofactors of IAA-oxidase increased the incorporation while those which are inhibitors of IAA-oxidase decreased the incorporation. Results suggest that the phenolics also affected the enzymic oxidation of IAA in vivo in the same way as in vitro.