Endogenous Indole-3-acetic Acid Levels Research Articles

Endogenous auxin and jasmonic acid levels are differentially modulated by abiotic stresses in rice

Abiotic stresses such as drought, salinity, and adverse temperatures are major limiting factors for plant growth and reproduction. Plant responses to these stresses are coordinated by arrays of regulatory networks including the induction of endogenous abscisic acid (ABA), a well documented phytohormone for stress responses. However, whether or how these abiotic stresses affect the endogenous biosynthesis or metabolism of other phytohormones remains largely unknown. Here, we report the changes of endogenous indole-3-acetic acid (IAA) and jasmonic acid (JA) levels and expression of genes related to the biosynthesis or signaling of these hormones in rice under various abiotic stress conditions. The IAA content was decreased after drought stress, but it was significantly increased under cold and heat stresses. And the auxin-regulated gravitropism of root tip was inhibited by cold stress. Many genes involved in the IAA biosynthesis and signaling were changed in transcript level under these stresses, and the changes were essentially in agreement with the change of endogenous IAA level. Interestingly, the endogenous JA content was increased markedly under drought and cold stresses, but it was reduced by heat stress. Accordingly, many genes involved in JA biosynthesis and signaling were induced by drought and cold treatment but these genes were significantly suppressed by heat stress. We concluded that endogenous levels of IAA and JA were differentially regulated by abiotic stresses in rice, implying diverse roles of these hormones in stress responses.

Enhancement of the differentiation of protocorm-like bodies of Dendrobium officinale to shoots by ultrasound treatment

An efficient micropropagation protocol has been developed for Dendrobium officinale, through protocorm-like bodies (PLBs). A correlation between enhanced differentiation of PLBs of D. officinale by ultrasound and changes in the levels of endogenous hormones and the antioxidant enzyme activities was described. Ultrasound treatments improved the conversion of PLBs of D. officinale to shoots. The highest conversion frequency of PLBs to shoots was obtained following the ultrasound treatment at 300W for 5min. Compared to the control, the enhanced conversion of PLBs to shoots following the ultrasound treatment was accompanied by an increase in the ratio of total cytokinins (CTKs) to indole-3-acetic acid (IAA), which was due to a decrease in the endogenous level of IAA and an increase in the endogenous level of total CTKs. Analysis of enzyme activities indicated that the increased endogenous level of total CTKs driven by ultrasound was associated with the inhibition of CTK decomposition by CTK oxidase (EC 1.4.3.6), while the decreased endogenous level of IAA was associated with the promotion of IAA decomposition by IAA oxidase (EC 1.10.3.3). In addition, ultrasound treatment increased the activities of superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6) and peroxidase (EC 1.11.1.7) in the conversion process of PLBs to shoots.

Endogenous auxin level is a critical determinant for in vitro adventitious shoot regeneration in potato (Solanum tuberosum L.)

Endogenous levels of indole-3-acetic acid (IAA) were determined in the internodal explants of Indian tetraploid potato cultivars (cvs) viz., Kufri Sutlej (K.Sutlej) and Kufri Giriraj (K.Giriraj). Seven fold higher level of endogenous IAA was recorded for cv K.Sutlej over cv K.Giriraj. As a result, perhaps there was a callusing response from the cut end of the internodal explants of both the cvs in the MS basal medium. The extent of callusing was relatively higher in cv K.Sutlej when compared to that in K.Giriraj. The callusing response was inhibitory to shoot morphogenesis. Inclusion of an established anti-auxin, 2,3,5-tri-iodobenzoic acid (TIBA) in the regeneration medium facilitated a high frequency adventitious shoot regeneration response with lower cytokinin levels. Murashige and Skoog (MS) medium containing TIBA at 2.5 mg l−1 and 0.25 mg l−1 zeatin evoked a 100% regeneration response (4.5 shoot buds per explant) in cv K.Sutlej within 20–25 days. However, in cv K.Giriraj, which had lower levels of endogenous IAA, 80% regeneration response (1.4 shoot buds per explant) was recorded in an extended period of 40–45 days on a medium containing 0.5 mg l−1 TIBA and 0.1 mg l−1 zeatin. Although, TIBA and zeatin induced shoot bud formation, it failed to support sustained growth of the regenerated shoots in cv K.Giriraj. Hence, 0.01 mg l−1 α-naphthaleneacetic acid (NAA) with a relatively higher concentration of zeatin (1.0 mg l−1) were used for sustained shoot regeneration (3.3 shoot buds per explant) within 25 days. From our results, it is evident that there was a difference in the requirement of exogenous zeatin levels required to induce a regeneration response in two cultivars of potato and this is attributed to the variable levels of endogenous IAA.

Role of Brassinosteroids, Ethylene, Abscisic Acid, and Indole-3-Acetic Acid in Mango Fruit Ripening

Rapid ripening of mango fruit limits its distribution to distant markets. To better understand and perhaps manipulate this process, we investigated the role of plant hormones in modulating climacteric ripening of ‘Kensington Pride’ mango fruits. Changes in endogenous levels of brassinosteroids (BRs), abscisic acid (ABA), indole-3-acetic acid (IAA), and ethylene and the respiration rate, pulp firmness, and skin color were determined at 2-day intervals during an 8-day ripening period at ambient temperature (21 ± 1°C). We also investigated the effects of exogenously applied epibrassinolide (Epi-BL), (+)-cis, trans-abscisic acid (ABA), and an inhibitor of ABA biosynthesis, nordihydroguaiaretic acid (NDGA), on fruit-ripening parameters such as respiration, ethylene production, fruit softening, and color. Climacteric ethylene production and the respiration peak occurred on the fourth day of ripening. Castasterone and brassinolide were present in only trace amounts in fruit pulp throughout the ripening period. However, the exogenous application of Epi-BL (45 and 60 ng g−1 FW) advanced the onset of the climacteric peaks of ethylene production and respiration rate by 2 and 1 day, respectively, and accelerated fruit color development and softening during the fruit-ripening period. The endogenous level of ABA rose during the climacteric rise stage on the second day of ripening and peaked on the fourth day of ripening. Exogenous ABA promoted fruit color development and softening during ripening compared with the control and the trend was reversed in NDGA-treated fruit. The endogenous IAA level in the fruit pulp was higher during the preclimacteric minimum stage and declined during the climacteric and postclimacteric stages. We speculate that higher levels of endogenous IAA in fruit pulp during the preclimacteric stage and the accumulation of ABA prior to the climacteric stage might switch on ethylene production that triggers fruit ripening. Whilst exogenous Epi-BL promoted fruit ripening, endogenous measurements suggest that changes in BRs levels are unlikely to modulate mango fruit ripening.

Erratum to: Levels of endogenous abscisic acid and indole-3-acetic acid influence shoot organogenesis in callus cultures of rice subjected to osmotic stress

Osmotic stress and endogenous hormone levels may have a role in shoot organogenesis, but a systematic study has not yet to investigate the links. We evaluated the changes of the endogenous indole-3-acetic acid (IAA) and abscisic acid (ABA) levels in rice (Oryza sativa L. cv. Tainan 5) callus during shoot organogenesis induced by exogenous plant growth regulator treatments or under osmotic stress. Non-regenerable callus showed low levels of endogenous ABA and IAA, with no fluctuation in level during the period evaluated. The addition of 100 μM ABA or 2 mM anthranilic acid (IAA precursor) into Murashige and Skoog basal induction medium containing 10 μM 2,4-D enhanced the regeneration frequency slightly, to 5 and 35%, respectively, and their total cellular ABA or IAA levels were increased significantly, correspondingly to the treatments. However, the regeneration frequency was greatly increased to 80% after treatment with 0.6 M sorbitol or 100 μM ABA and 2 mM anthranilic acid combined. Both treatments produced high levels of total cellular ABA and IAA at the callus stage, which was quickly decreased on the first day after transfer to regeneration medium. Thus, osmotic stress-induced simultaneous accumulation of endogenous ABA and IAA is involved in shoot regeneration in rice callus.

Chemical screening for promotion of adventitious root formation in Eucalyptus globulus

Eucalyptus globulusis one of the most profitable trees for pulp and paper industries due to its fast growth and short harvesting cycle. This species is easily pulped with high yield, and its fiber qualities are among the best for paper production. However, it is difficult to vegetatively propagate E. globulus.So far, we have developed “photoautotropic culture method” that promotes rooting percentage by feeding approximately three times higher level CO2 level (1000 µmol mol–1) and suitable culture condition [1]. However, several lines showed poor rooting percentage even in the higher CO2 conditions. Therefore, it was necessary to develop novel method for promotion of adventitious root (AR) formation in E. globulus. First, we measured endogenous levels of 20 kinds of hormones such as abscisic acid, auxins, cytokinines and gibberellins, at basal part of stem of easy-rooting line and poor-rooting line by UPLC-ESI-qMS/MS. As a result, the Indole-3-acetic acid (IAA) level of easy-rooting line was two times higher than that of the poor-rooting line, suggesting that endogenous IAA level may regulate ability of AR formation. Next, we focused on the cytochrome P450s that are involved in a vast array of reactions of many different metabolic pathways. Several triazole-containing chemical compounds have previously been shown act as efficient inhibitors of cytochrome P450 monooxygenases. A chemical library of triazole derivatives to find chemicals which have the effect of promoting AR formation was screened. Consequently, five compounds effectively promoted AR formation. Finally, we investigated how these chemicals affected the growth of Arabidopsis thaliana. Arabidopsis seedlings were grown on agar medium containing 1 μM selected chemicals. One of the selected chemicals, MA65 increased the number of roots in wild-type Arabidopsis seedlings and this phenotype was similar to a mutant superroot2 (sur2) [2]. The SUR2 gene encodes the cytochrome P450 CYP83B1, a modulator of auxin homeostasis. The amounts of endogenous IAA in 14-d-old Arabidopsis grown in the presence of 1 mM MA65 were analyzed. The IAA content was increased two-fold in the presence of MA65 as compared with untreated Arabidopsis. In addition, 1.0 cm explants of Arabidopsis stems were incubated for 7 d on MS medium containing 1 mMMA65. Stimulation of an AR formation was observed as compared to untreated samples. Taken together, MA65 may increase endogenous IAA level in a plant cell and promotes an AR formation.

The role of auxins in somatic embryogenesis of Abies alba

Abstract The somatic embryogenesis of conifers is a process susceptible to exogenous phytohormonal treatments. We report the effects of the synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4-D) and the auxin inhibitor p-chlorophenoxyisobutyric acid (PCIB) on the endogenous level of the auxin indole-3-acetic acid (IAA) and on the anatomical composition of early somatic embryos of Abies alba (European silver fir). The embryogenic suspensor mass (ESM) of Abies alba proliferated on a medium supplemented by 2,4-D as well as on an auxin-free medium. The endogenous level of IAA was significantly higher in the ESM cultivated on a medium supplemented by 2,4-D. The decrease in the endogenous level of IAA in the first week of maturation is one of the most important stimuli responsible for the subsequent development of embryos. However, suppression of IAA synthesis by an auxin inhibitor did not stimulate the development of embryos. The maturation of somatic embryos from the globular to the cotyledonary stage occurs when the concentration of endogenous auxin in the ESM (including the embryos) increases. Early somatic embryos proliferating on a medium supplemented by auxin had an increased probability of maturing successfully. Exogenous auxin treatment during maturation did not compensate for the auxin deficiency during proliferation.

Effect of putrescine on the conversion of protocorm-like bodies of Dendrobium officinale to shoots

The addition of 1.5 mM putrescine to the culture medium facilitated the conversion of protocorm-like bodies (PLBs) of Dendrobium officinale to shoots. The administration of 1.0 mM of the polyamine inhibitor, α-DL-difluoromethylarginine, decreased the conversion of PLBs to shoots. Compared to the control, the PLBs treated with 1.5 mM putrescine had higher levels of total intracellular free polyamines. The enhanced conversion of PLBs to shoots following the addition of putrescine was accompanied by an increase in the ratio of total cytokinins (CTKs) to indole-3-acetic acid (IAA). Analysis of enzyme activities indicated that the increased endogenous level of total CTKs driven by putrescine was associated with the inhibition of CTK decomposition by CTK oxidase, while the decreased endogenous level of IAA was associated with the promotion of IAA decomposition by IAA oxidase. In addition, putrescine increased the ratio of reduced glutathione to oxidized glutathione in the conversion process of PLBs to shoots.

Conversion of protocorm-like bodies of Dendrobium huoshanense to shoots: The role of polyamines in relation to the ratio of total cytokinins and indole-3-acetic acidindole-3-acetic acid

In the present paper, a correlation between enhanced conversion of protocorm-like bodies (PLBs) of Dendrobium huoshanense to shoots by free polyamines (PAs) and changes in the levels of endogenous hormones is described. The endogenous levels of free spermidine (Spd) and putrescine (Put) increased during the conversion of PLBs to shoots. The exogenous addition of PAs, mainly Spd or Put at 2.0mM, not only elevated the endogenous levels of PAs but also promoted the frequency of conversion of PLBs to shoots. As compared with control, the enhanced conversion of PLBs to shoots by exogenous PAs was accompanied by an increase in ratio of total cytokinins (CTKs) to indole-3-acetic acid (IAA), which was due to decrease in the endogenous level of IAA and increase in the endogenous level of total CTKs, including the levels of isopentenyladenine+isopentenyladenine 9-riboside and zeatin+zeatin riboside. Analysis of enzyme activities showed that the increased endogenous level of total CTKs by PAs was related to the inhibition of CTK decomposition by CTK oxidase, while the decreased endogenous level of IAA was related to the promotion of IAA decomposition by IAA oxidase. Addition of PA biosynthetic inhibitors, involving alpha-difluoromethylornithine for Put and methylglyoxal(bis)-guanylhydrazone for Spd and Spm, decreased the conversion of PLBs to shoots, the ratio of total CTKs to IAA, and the levels of endogenous Put and Spd. This inhibition could be partly reversed by the application of exogenous Put or Spd.

Changes in endogenous hormone levels and redox status during enhanced adventitious rooting by rare earth element neodymium of Dendrobium densiflorum shoot cuttings

The effects of neodymium nitrate (Nd 3+) on the adventitious rooting of Dendrobium densiflorum shoot cuttings were studied. The addition of Nd 3+ (5 μmol/L) to culture medium significantly increased rooting frequency. Histological investigation showed that Nd 3+ did not change the process of root initiation. Nd 3+ did not influence total endogenous cytokinin levels, but significantly increased the level of endogenous indole-3-acetic acid (IAA) in the base of shoot cuttings. Compared to the control, the ratio of IAA/cytokinins was very high in the Nd 3+ treatment. These results suggested that the enhanced rooting frequency may be related to the increase in endogenous IAA level in Nd 3+ treatment. Analysis of enzyme activities showed that the enhanced accumulation of the endogenous IAA by Nd 3+ should not be attributed to inhibition of IAA decomposition by IAA oxidase or promotion of cytokinin decomposition by cytokinin oxidase. Besides, Nd 3+ increased the ratio of reduced glutathione (GSH) and oxidized glutathione (GSSG) in the process of adventitious rooting while the ratio of ascorbate (ASC) to dehydroascorbate (DHA) was not affected.

Quantitation of endogenous levels of IAA, IAAsp and IBA in micro-propagated shoots of hybrid chestnut pre-treated with IBA

Endogenous levels of indole-3-acetic acid (IAA), indole-3-acetylaspartic acid (IAAsp) and indole-3-butyric acid (IBA) were measured during the first 8 d of in vitro rooting of rootstock from the chestnut ‘M3’ hybrid by high performance liquid chromatography (HPLC). Rooting was induced either by dipping the basal ends of the shoots into a 4.92-mM IBA solution for 1 min or by sub-culturing the shoots on solid rooting medium supplemented with 14.8-μM IBA for 5 d. For root development, the induced shoots were transferred to auxin-free solid medium. Auxins were measured in the apical and basal parts of the shoots by means of HPLC. Endogenous levels of IAA and IAAsp were found to be greater in IBA-treated shoots than in control shoots. In extracts of the basal parts of the shoots, the concentration of free IAA showed a significant peak 2 d after either root inductive method and a subsequent gradual decrease for the remainder of the time course. The concentration of IAAsp peaked at day 6 in extracts of the basal parts of shoots induced with 14.8-μM IBA for 5 d, whereas shoots induced by dipping showed an initial increase until day 2 and then remained stable. In extracts from basal shoot portions induced by dipping, IBA concentration showed a transient peak at day 1 and a plateau between day 2 and 4, in contrast to the profile of shoots induced on auxin-containing medium, which showed a significant reduction between 4 and 6 d after transferred to auxin-free medium. All quantified auxins remained at a relatively low level, virtually constant, in extracts from apical shoot portions, as well as in extracts from control non-rooting shoots. In conclusion, the natural auxin IAA is the signal responsible for root induction, although it is driven by exogenous IBA independently of the adding conditions.

Exogenously applied melatonin stimulates root growth and raises endogenous indoleacetic acid in roots of etiolated seedlings of Brassica juncea

Exogenous melatonin was applied to etiolated seedlings of wild leaf mustard (Brassica juncea) and the effect on root growth and endogenous indole-3-acetic acid (IAA) levels determined. The results show that 0.1microM melatonin has a stimulatory effect on root growth, while 100microM is inhibitory. Furthermore, the stimulatory effect was only detectable in young seedlings (2-d old). Older seedlings (4-d old) appear to be less susceptible to both the stimulatory and the inhibitory effect of melatonin. Exogenous application of 0.1microM melatonin also raised the endogenous levels of free IAA in roots, while higher concentrations had no significant effect. The specific mechanism that causes exogenous melatonin to increase the amount of free IAA in roots, paired with a stimulation of root growth, remains to be uncovered.

The Role of Aspterric Acid in Auxin-Regulated Reproductive Growth of Arabidopsis thaliana

Application of 100 microM aspterric acid (AA), a pollen growth inhibitor, with different concentrations of indole-3-acetic acid (IAA) results in the recovery of normal pollen development of Arabidopsis thaliana. Treatment with 100 microM AA plus 5 mM IAA significantly induced the normal seed production. Treatment with 100 microM N-1-naphthylphthalamic acid (NPA), a polar auxin transport inhibitor, did not reduce the pollen growth but inhibited seed production. 100 microM NPA plus 5 mM IAA did not induce any seed production. The endogenous level of IAA in stems and leaves of A. thaliana treated with 100 microM AA was similar to that of the untreated control. In contrast to AA treatment, the IAA level by the treatment with 100 microM NPA was about twice as much as that of the untreated control. These results suggest that AA affects the Arabidopsis reproductive growth without inhibiting IAA biosynthesis and transport.

Maturation of somatic embryos in two embryogenic cultures of Picea morrisonicola Hayata as affected by alternation of endogenous IAA content

Embryogenic culture lines T4 and T2 were initiated from two mature zygotic embryos of Picea morrisonicola Hay. Mature somatic embryos (SEs) were produced in culture line T2 but not in line T4 after 8-week abscisic acid (ABA) treatment. High performance liquid chromatography (HPLC) analysis of the endogenous indole-3-acetic acid (IAA) content has shown 7.5 times higher IAA production in T4 line than in T2 line during the proliferation phase. However, after ABA incubation the line T4 produced much less IAA than line T2. The application of an anti-auxin, 2,3,5-triiodobenzoic acid (TIBA) or 2-(4-chlorophenoxy)-2-methylpropionic acid (PCIB) induced culture line T4 to produce mature SEs. Both 1 μM TIBA and 5 μM PCIB increased the production of stage 2 SEs in T4 culture line when cultures were treated during the proliferation stage for 8 weeks. Occasionally cotyledonary (stage 3) SEs were even produced from treated T4 culture line. Both chemicals have also been demonstrated to significantly decrease the amount of IAA in the treated T4 and T2 embryogenic lines. However the decrease of the IAA level was not beneficial for SE production in the T2 embryogenic line. These results indicated the importance of endogenous IAA level in manipulating the process of SE maturation in spruce embryogenic cultures.

Possible correlation between high temperature-induced floret sterility and endogenous levels of IAA, GAs and ABA in rice (Oryza sativa L.)

In order to explore the possible physiological mechanism of high temperature induced sterility in rice, we examined the floret sterility and endogenous plant growth regulator contents in pollens of two hybrid rice cultivars Shanyou63 and Teyou559 that are tolerant and susceptible to high temperature, respectively. Indexes of floret sterility, pollen activity, and variation of endogenous indole-3-acetic acid (IAA), gibberellic acids (GAs), abscisic acid (ABA), free proline and soluble proteins in anthers were measured. We found that during the course of high temperature treatment, both cultivars exhibited a marked decrease in pollen activity, pollen germination and floret fertility; however, the high temperature tolerant Shanyou63 showed a much slower rate of decrease than the high temperature susceptible Teyou559. In addition, anthers of both cultivars displayed a decrease in the contents of IAA, GAs, free proline and soluble proteins but an increase in the ABA content. Yet compared to Teyou559, Shanyou63 retained significantly higher levels of free praline and GAs and a lower level of ABA, along with higher pollen vigour and pollen germination rate even after prolonged high temperature treatment. Our study suggests a possible correlation between pollen viability/floret sterility and high temperature-caused changes in IAA, GAs, ABA, free proline and soluble protein contents. The severity in these changes may reflect the variation of rice cultivars in their heat stress sensitivities for floret development.

Moniliophthora perniciosaproduces hormones and alters endogenous auxin and salicylic acid in infected cocoa leaves

Moniliophthora perniciosa is the causative agent of witches' broom disease in Theobroma cacao. Exogenously provided abscisic acid (ABA), indole-3-acetic acid (IAA), jasmonic acid (JA), and salicylic acid (SA) promoted mycelial growth, suggesting the ability of the pathogen to metabolize plant hormones. ABA, IAA, JA, and SA were found endogenously in the mycelium and in the fruiting body of the pathogen. The pathogen contained high amounts of SA in the mycelium (0.5+/-0.04 microg g(-1) DW) and IAA (2+/-0.6 microg g(-1) DW) in the basidiocarps. Growth of the mycelium in the presence of host leaves for 10 days did not affect ABA or JA content of the leaves but IAA and SA increased 2.5- and 11-fold, respectively. The amounts of IAA and SA in infected leaves increased beyond the levels of the uninfected leaves and suggest a synergistic response to host-pathogen interaction. The ability of M. perniciosa to produce and sustain growth in the presence of elevated endogenous IAA and SA levels during colonization indicates that these phytohormones contribute to its pathogenicity.

Light quality regulation of endogenous levels of auxin, abscisic acid and ethylene production in petioles and leaves of wild type and ACC deaminase transgenic Brassica napus seedlings

Brassica napus L. seedlings responded to low red to far-red (R/FR) ratio by elongating petioles and decreasing leaf expansion. These typical shade avoidance traits were correlated with significantly decreased endogenous indole-3-acetic acid (IAA) levels and significantly increased endogenous abscisic acid (ABA) levels and ethylene production. The transgenic (T) B. napus line bearing the bacterial ACC deaminase gene, did not respond to low R/FR ratio with altered petiole and leaf growth and less ethylene (especially by petioles) was produced. As with WT seedlings, T seedlings had significantly lower IAA levels in both petioles and leaves under low R/FR ratio. However, ABA levels of low R/FR ratio-grown T seedlings either increased (petioles) or were unaltered (leaves). Our results further suggest that low R/FR ratio regulates endogenous IAA levels independently of ethylene, but there may be an interaction between ABA and ethylene in leaf development.

Elongation rates and endogenous indoleacetic acid levels in roots of pea mutants differing in internode length

Growth of the primary root of 12 genotypes of peas (Pisum sativum) differing in their stem height was recorded for 14 days. The growth rate of roots of wild‐type tall, gibberellin (GA)‐deficient le dwarf or slender mutants (with la crys) was similar (3 cm day−1); that of severely GA‐deficient nana (na‐1) plants was 50% of wild‐type but elongation ceased after 8 days; moderately severe dwarf GA‐deficient lines ls‐1 and lh‐1 had a 15% reduction in elongation rate but displayed no time‐dependent slowing of the growth rate and brassinosteroid‐insensitive and ‐deficient dwarfs lka and lkb showed slightly decreased root elongation. GA (levels reported in Yaxley et al. 2001) is not substantially limiting to root growth until it is severely deficient. The terminal 3 cm of roots of tall plants contained about 25 or 35 ng g−1 fresh weight indole‐3‐acetic acid (IAA), depending on the genetic background, and le‐1 dwarfs were similar. Nana (na‐1) had less than 50% the level of IAA of tall, all the moderately severe dwarfs had reductions of about 30% and the slender plants had about 40% more IAA than the corresponding wild‐type. With the exception of slender plants, IAA level in the root tips correlated with root elongation. Root growth seems to be promoted by IAA within the range of the internal concentrations detected. Nana plants had a reduced amount of IAA and a lower root‐growth rate. Whereas external application of IAA always inhibits root growth, even at very low concentrations, root growth is not similarly inhibited by internal IAA as slender plants had the highest IAA level and growth rate similar to wild‐type, regardless of the shoot GA content.

The interaction of light quality and irradiance with gibberellins, cytokinins and auxin in regulating growth of Helianthus annuus hypocotyls

A reduced red to far-red (R/FR) light ratio and low photosynthetically active radiation (PAR) irradiance are both strong signals for inducing etiolation growth of plant stems. Under natural field conditions, plants can be exposed to either a reduced R/FR ratio or lower PAR, or to a combination of both. We used Helianthus annuus L., the sunflower, to study the effect of reduced R/FR ratio, low PAR or their combination on hypocotyl elongation. To accomplish this, we attempted to uncouple light quality from light irradiance as factors controlling hypocotyl elongation. We measured alterations in the levels of endogenous gibberellins (GAs), cytokinins (CKs) and the auxin indole-3-acetic acid (IAA), and the effect of exogenous hormones on hypocotyl growth. As expected, both reduced R/FR ratio and lower PAR can significantly promote sunflower hypocotyl elongation when given separately. However, providing the reduced R/FR ratio at a low PAR resulted in the greatest hypocotyl growth, and this was accompanied by significantly higher levels of endogenous IAA, GA1, GA8, GA20 and of a wide range of CKs. Providing a reduced R/FR ratio under normal PAR also significantly increased growth and again gave significantly higher levels of endogenous IAA, GAs and CKs. However, only under the de-etiolating influence of a normal R/FR ratio did lowering PAR significantly increase levels of GA1, GA8 and GA20. We thus conclude that light quality (e.g. the R/FR ratio) is the most important component of shade for controlling hypocotyl growth and elevated growth hormone content.

Stem fasciated, a Recessive Mutation in Sunflower (Helianthus annuus), Alters Plant Morphology and Auxin Level

Plant lateral organs such as leaves arise from a group of initial cells within the flanks of the shoot apical meristem (SAM). Alterations in the initiation of lateral organs are often associated with changes in the dimension and arrangement of the SAM as well as with abnormal hormonal homeostasis. A mutation named stem fasciated (stf) that affects various aspects of plant development, including SAM shape and auxin level, was characterized in sunflower (Helianthus annuus). F1, F2 and F3 generations were obtained through reciprocal crosses between stf and normal plants. For the genetic analysis, a chi2 test was used. Phenotypic observations were made in field-grown and potted plants. A histological analysis of SAM, hypocotyl, epicotyl, stem and root apical meristem was also conducted. To evaluate the level of endogenous indole-3-acetic acid (IAA), a capillary gas chromatography-mass spectrometry-selected ion monitoring analysis was performed. stf is controlled by a single nuclear recessive gene. stf plants are characterized by a dramatically increased number of leaves and vascular bundles in the stem, as well as by a shortened plastochron and an altered phyllotaxis pattern. By histological analysis, it was demonstrated that the stf phenotype is related to an enlarged vegetative SAM. Microscopy analysis of the mutant's apex also revealed an abnormal enlargement of nuclei in both central and peripheral zones and a disorganized distribution of cells in the L2 layer of the central zone. The stf mutant showed a high endogenous free IAA level, whereas auxin perception appeared normal. The observed phenotype and the high level of auxin detected in stf plants suggest that the STF gene is necessary for the proper initiation of primordia and for the establishment of a phyllotactic pattern through control of both SAM arrangement and hormonal homeostasis.