Endogenous Indole-3-acetic Acid Levels Research Articles

Identification and Fine Mapping of a Mutant pv‐nts Gene Responsible for Dwarf and Thin Stem of Snap Bean (Phaseolus vulgaris L.)

ABSTRACTPlant height is one of the most important traits in the breeding of snap bean, which determines the planting density and the resistance to failure, which in turn affects its yield and quality. Therefore, studying the molecular mechanism of plant height in snap bean is an important measure to realize the improvement of snap bean. We obtained a dwarf, thin‐stemmed mutant nts (normal to slim) from a dwarf variety of snap bean A18 via 60CO‐γ‐ray treatment. We conducted phenotypic observations, endogenous hormone content measurements and genetic analyses of the dwarfing thin‐stemmed mutant nts and explored gene localization and candidate gene screening for the mutant trait in snap bean. Compared with wild‐type A18, nts mutant exhibited various abnormal phenotypes such as dwarfing, reduced branching, thinner top stem, reduced plant width, reduced number of leaf area and thousand grain weight and increased content of hormones such as indole‐3‐acetic acid (IAA) in the top stem. Genetic analysis indicated that the mutant trait was controlled by a single recessive nuclear gene, and the mutant gene was named pv‐nts. pv‐nts was localized to a 361.1‐kb segment on chromosome 8 as revealed by bovine serum albumin (BSA) hybrid pool sequencing and simple sequence repeat (SSR) molecular markers. Phvul.008G106300 (encoding LAX2 protein) was identified as a candidate gene corresponding to pv‐nts via gene function annotation. Phvul.008G106300 was significantly upregulated in nts mutant compared with wild‐type A18, which was consistent with the results of endogenous IAA levels. Sequence analysis showed that the Phvul.008G106300 gene in nts had a base mutation that altered the protein structure, thus causing the dwarfed, thin‐stemmed phenotype of cauliflower. Taken together, these results contribute to a better understanding of the genetic basis of dwarfing traits in snap bean.

Integrated metabolomics and transcriptomics analysis highlight key pathways involved in the somatic embryogenesis of Darjeeling tea.

Darjeeling tea is a globally renowned beverage, which faces numerous obstacles in sexual reproduction, such as self-incompatibility, poor seed germination, and viability, as well as issues with vegetative propagation. Somatic embryogenesis (SE) is a valuable method for rapid clonal propagation of Darjeeling tea. However, the metabolic regulatory mechanisms underlying SE in Darjeeling tea remain largely unknown. To address this, we conducted an integrated metabolomics and transcriptomics analysis of embryogenic callus (EC), globular embryo (GE), and heart-shaped embryo (HE). The integrated analyses showed that various genes and metabolites involved in the phenylpropanoid pathway, auxin biosynthesis pathway, gibberellin, brassinosteroid and amino acids biosynthesis pathways were differentially enriched in EC, GE, and HE. Our results revealed that despite highly up-regulated auxin biosynthesis genes YUC1, TAR1 and AAO1 in EC, endogenous indole-3-acetic acid (IAA) was significantly lower in EC than GE and HE. However, bioactive Gibberellin A4 displayed higher accumulation in EC. We also found higher BABY BOOM (BBM) and Leafy cotyledon1 (LEC1) gene expression in GE along with high accumulation of castasterone, a brassinosteroid. Total flavonoids and phenolics levels were elevated in GE and HE compared to EC, especially the phenolic compound chlorogenic acid was highly accumulated in GE. Integrated metabolome and transcriptome analysis revealed enriched metabolic pathways, including auxin biosynthesis and signal transduction, brassinosteroid, gibberellin, phenylpropanoid biosynthesis, amino acids metabolism, and transcription factors (TFs) during SE in Darjeeling tea. Notably, EC displayed lower endogenous IAA levels, conducive to maintaining differentiation, while higher IAA concentration in GE and HE was crucial for preserving embryo identity. Additionally, a negative correlation between bioactive gibberellin A4 (GA4) and IAA was observed, impacting callus growth in EC. The high accumulation of chlorogenic acid, a phenolic compound, might contribute to the low success rate in GE and HE formation in Darjeeling tea. TFs such as BBM1, LEC1, FUS3, LEA, WOX3, and WOX11 appeared to regulate gene expression, influencing SE in Darjeeling tea.

A New Function of MbIAA19 Identified to Modulate Malus Plants Dwarfing Growth.

The primary determinants of apple (Malus) tree architecture include plant height and internode length, which are the significant criteria for evaluating apple dwarf rootstocks. Plant height and internode length are predominantly governed by phytohormones. In this study, we aimed to assess the mechanisms underlying dwarfism in a mutant of Malus baccata. M. baccata dwarf mutant (Dwf) was previously obtained through natural mutation. It has considerably reduced plant height and internode length. A comparative transcriptome analysis of wild-type (WT) and Dwf mutant was performed to identify and annotate the differentially expressed genes responsible for the Dwf phenotype using RNA-seq and GO and KEGG pathway enrichment analyses. Multiple DEGs involved in hormone signaling pathways, particularly auxin signaling pathways, were identified. Moreover, the levels of endogenous indole-3-acetic acid (IAA) were lower in Dwf mutant than in WT. The Aux/IAA transcription factor gene MbIAA19 was downregulated in Dwf mutant due to a single nucleotide sequence change in its promoter. Genetic transformation assay demonstrated strong association between MbIAA19 and the dwarf phenotype. RNAi-IAA19 lines clearly exhibited reduced plant height, internode length, and endogenous IAA levels. Our study revealed that MbIAA19 plays a role in the regulation of dwarfism and endogenous IAA levels in M. baccata.

Biocompatible Fluorescent Biosensor Reveals the Level and Distribution of Indole-3-Acetic Acid Signals in Plants.

To fully understand the function of the phytohormone indole-3-acetic acid (IAA) in regulating plant growth and development, we need to monitor their levels and distribution with high spatial and temporal resolution. In this work, an anthracene-based fluorescent biosensor for IAA was prepared using bovine serum albumin (BSA) as a bio-template. The single linear oxygen (1O2) specifically produced by IAA catalyzed with horseradish peroxidase (HRP) turns on the fluorescence of the probe, enabling specific trace sensing of IAA in the presence of multiple structural analogues. The presence of the bio-template BSA extends the biocompatibility of the probe, enabling visual monitoring of the level and distribution signal of endogenous IAA of plants in the field of bioimaging. In addition, the strategy has shown potential for application in portable paper-based sensors and in vivo fluorescent flower culture. This work provides a technical and theoretical basis for exploring the growth regulatory mechanisms of IAA in plants at the molecular level.

Whole Genome Analysis of SLs Pathway Genes and Functional Characterization of DlSMXL6 in Longan Early Somatic Embryo Development.

Strigolactones (SLs), a new class of plant hormones, are implicated in the regulation of various biological processes. However, the related family members and functions are not identified in longan (Dimocarpus longan Lour.). In this study, 23 genes in the CCD, D27, and SMXL family were identified in the longan genome. The phylogenetic relationships, gene structure, conserved motifs, promoter elements, and transcription factor-binding site predictions were comprehensively analysed. The expression profiles indicated that these genes may play important roles in longan organ development and abiotic stress responses, especially during early somatic embryogenesis (SE). Furthermore, GR24 (synthetic SL analogue) and Tis108 (SL biosynthesis inhibitor) could affect longan early SE by regulating the levels of endogenous IAA (indole-3-acetic acid), JA (jasmonic acid), GA (gibberellin), and ABA (abscisic acid). Overexpression of SMXL6 resulted in inhibition of longan SE by regulating the synthesis of SLs, carotenoids, and IAA levels. This study establishes a foundation for further investigation of SL genes and provides novel insights into their biological functions.

The thiol-reductase activity of YUCCA6 enhances nickel heavy metal stress tolerance in Arabidopsis.

Anthropogenic activities cause the leaching of heavy metals into groundwater and their accumulation in soil. Excess levels of heavy metals cause toxicity in plants, inducing the production of reactive oxygen species (ROS) and possible death caused by the resulting oxidative stress. Heavy metal stresses repress auxin biosynthesis and transport, inhibiting plant growth. Here, we investigated whether nickel (Ni) heavy metal toxicity is reduced by exogenous auxin application and whether Ni stress tolerance in Arabidopsis thaliana is mediated by the bifunctional enzyme YUCCA6 (YUC6), which functions as an auxin biosynthetic enzyme and a thiol-reductase (TR). We found that an application of up to 1 µM exogenous indole-3-acetic acid (IAA) reduces Ni stress toxicity. yuc6-1D, a dominant mutant of YUC6 with high auxin levels, was more tolerant of Ni stress than wild-type (WT) plants, despite absorbing significantly more Ni. Treatments of WT plants with YUCASIN, a specific inhibitor of YUC-mediated auxin biosynthesis, increased Ni toxicity; however yuc6-1D was not affected by YUCASIN and remained tolerant of Ni stress. This suggests that rather than the elevated IAA levels in yuc6-1D, the TR activity of YUC6 might be critical for Ni stress tolerance. The loss of TR activity in YUC6 caused by the point-mutation of Cys85 abolished the YUC6-mediated Ni stress tolerance. We also found that the Ni stress–induced ROS accumulation was inhibited in yuc6-1D plants, which consequently also showed reduced oxidative damage. An enzymatic assay and transcriptional analysis revealed that the peroxidase activity and transcription of PEROXIREDOXIN Q were enhanced by Ni stress to a greater level in yuc6-1D than in the WT. These findings imply that despite the need to maintain endogenous IAA levels for basal Ni stress tolerance, the TR activity of YUC6, not the elevated IAA levels, plays the predominant role inNi stress tolerance by lowering Ni-induced oxidative stress.

Auxin plays a role in the adaptation of rice to anaerobic germination and seedling establishment.

Auxin is well known to stimulate coleoptile elongation and rapid seedling growth in the air. However, its role in regulating rice germination and seedling establishment under submergence is largely unknown. Previous studies revealed that excessive levels of indole-3-acetic acid(IAA) frequently cause the inhibition of plant growth and development. In this study, the high-level accumulation of endogenous IAA is observed under dark submergence, stimulating rice coleoptile elongation but limiting the root and primary leaf growth during anaerobic germination (AG). We found that oxygen and light can reduce IAA levels, promote the seedling establishment and enhance rice AG tolerance. miRNA microarray profiling and RNA gel blot analysis results show that the expression of miR167 is negatively regulated by submergence; it subsequently modulates the accumulation of free IAA through the miR167-ARF-GH3 pathway. The OsGH3-8 encodes an IAA-amido synthetase that functions to prevent free IAA accumulation. Reduced miR167 levels or overexpressing OsGH3-8 increase auxin metabolism, reduce endogenous levels of free IAA and enhance rice AG tolerance. Our studies reveal that poor seed germination and seedling growth inhibition resulting from excessive IAA accumulation would cause intolerance to submergence in rice, suggesting that a certain threshold level of auxin is essential for rice AG tolerance.

Growth-Promoting Endophytic Fungus (Stemphylium lycopersici) Ameliorates Salt Stress Tolerance in Maize by Balancing Ionic and Metabolic Status.

Climate change is a major cause of the world's food security problems, and soil salinity is a severe hazard for a variety of crops. The exploitation of endophytic fungi that are known to have a positive association with plant roots is preferred for improving plant growth, yield, and overall performance under salt stress. The current study thus rationalized to address how salt stress affected the growth, biochemical properties, antioxidant capacity, endogenous indole-3-acetic acid (IAA), and the ionic status of maize associated with endophytic fungus (Stemphylium lycopersici). According to the findings, salt stress reduced chlorophyll a and b, total chlorophyll, total protein, sugars, lipids, and endogenous IAA levels. Enhanced values of chlorophyll a/b ratio, carotenoids, secondary metabolites (phenol, flavonoids, and tannins), antioxidant enzyme activity (catalase, ascorbate peroxidase), proline, and lipid peroxidation were noticed in maize plants under salt stress. Increased ionic content of Na+, Cl−, Na+/K+, and Na+/Ca2+ ratio, as well as decreased Ca2+, K+, Mg2+, N, and P contents, were also found in salt-stressed maize plants. In comparison to the non-saline medium, endophytic association promoted the antioxidant enzyme activities (798.7 U/g protein; catalase activity, 106 U/g protein; ascorbate peroxidase activity), IAA content (3.47 mg/g FW), and phenolics and flavonoids (88 and 1.68 μg/g FW, respectively), and decreased MDA content (0.016 nmol/g FW), Na+ ion content (18 mg/g dry weight), Cl− ion (16.6 mg/g dry weight), and Na+/K+ (0.78) and Na+/Ca2+ (1.79) ratios, in maize plants under salt stress, whereas Ca2+, K+, Mg2+, N, and P contents were increased in maize plants associated with S. lycopersici under salt stress. Current research exposed the role of S. lycopersici as an effective natural salt stress reducer and maize growth promoter; hence, it can be used as a biofertilizer to ameliorate salt stress tolerance in crops along with better growth performance in saline regions.

Uniconazole Augments Abscisic Acid in Promoting Somatic Embryogenesis in Cotton (Gossypium hirsutum L.).

During somatic embryogenesis (SE), somatic cells initiate embryogenic development under appropriate conditions. Uniconazole, a plant growth regulator, was found to inhibit the proliferation of callus but promoted the conversion of callus into an embryogenic callus (EC) in cotton. The supplementation of uniconazole in the culture medium significantly suppressed the endogenous auxin [indole acetic acid (IAA)] level in callus tissues in both the callus initiation and proliferation stage but enhanced the abscisic acid (ABA) level only in the callus proliferation stage. Exogenous ABA and uniconazole showed cooperative effects on promoting the differentiation rate of callus into EC. These findings were verified by RNA-seq analysis, which elucidated that the genes involved in the IAA biosynthesis, metabolism, and signaling, and ABA metabolism pathways were regulated by uniconazole during the callus development and SE. Overall, the results suggest that uniconazole could modulate callus proliferation and callus differentiation rate by regulating the endogenous levels of IAA and ABA.

Determination of age-dependent endogenous indole-3 acetic acid (IAA) level in different organs of tomato plants

Indole-3-acetic acid (IAA), a phytohormone, is a crucial modulator of plant growth and development. As increased levels of endogenous IAA is responsible for inhibitory effects on plants, knowing its concentration would be helpful for controlling exogenous IAA input. An optimum endogenous IAA level is required for proper growth and development of plant with good health status. Hence, in this study, we have detected and measured IAA present in tomato root, shoot, leaves and fruits at two different ages (25 days and 50 days) by Reversed-Phase High Performance Liquid Chromatography (RP-HPLC), a suitable chromatographic method for separating molecules based on hydrophobicity. We observed that the endogenous level of IAA was significantly higher in roots, shoots, leaves and fruits at 50-days old plant compared to 25-days old mature plant (p-value <0.01). These results suggested that the IAA level may increase with the increase of the age of tomato plants. The tomato plants showed good vigor with IAA range 0.3-3.0 µg/g.F.W (25 days) and 3.4-7.5 µg/g.F.W (50 days) of age, respectively, implying that IAA might be used as a possible plant health indicator. Further study is needed to manipulate the IAA concentration for adequate plant growth and development.

Transcriptomic analyses provide insight into adventitious root formation of Euryodendron excelsum H. T. Chang during ex vitro rooting

Euryodendron excelsum H. T. Chang, a critically endangered species endemic to China, is a source of valuable material for the furniture and construction industries. However, this species has some challenges associated with rooting during in vitro propagation that have yet to be resolved. In this study, we optimized rooting and conducted a transcriptomic analysis to appreciate its molecular mechanism, thereby promoting the practical application of in vitro propagation of E. excelsum, and providing technical support for the ecological protection of this rare and endangered species. Results showed that ex vitro rooting performed the highest rooting percentage with 98.33% at 25 days. During ex vitro rooting, there was a wide fluctuation of endogenous levels of indole-3-acetic acid (IAA) and hydrogen peroxide (H2O2) at the stage of root primordia formation. Transcriptome analysis revealed multiple differentially expressed genes (DEGs) involved in adventitious root (AR) development. DEGs involved in plant hormone signal transduction, such as genes encoding auxin-induced protein, auxin-responsive protein, and auxin transporter-like protein, and in response to H2O2, oxidative stress, abiotic and biotic stimuli were significantly up- or down-regulated by ex vitro treatment with 1 mM indole-3-butyric acid (IBA). Our results indicate that ex vitro rooting is an effective method to induce AR from E. excelsum plantlets during micropropagation. DEGs involved in the plant hormone signal transduction pathway played a crucial role in AR formation. H2O2, produced by environmental stimulation, might be related to AR induction as a result of the synergistic action with IBA, ultimately regulating the level of endogenous IAA. Under ex vitro rooting, a synergistic action between H2O2 produced by environmental stimulation and IBA played crucial role in the regulation of AR formation from E. excelsum plantlets during micropropagation.

Dominant Fungal Epiphytes Promote Growth of the Invasive Plant Ipomoea cairica Through Hormone Interactions

Plant growth-promoting microbial symbionts have been found in many plants, but their association with invasive plant Ipomoea cairica remains largely unknown. The present study aimed to screen such microbes from the fungal epiphyte community of I. cairica. A total of 858 culturable isolates grouped into 20 morphotypes associated with 10 genera were recovered from healthy leaves of I. cairica. According to morphological and molecular identification, morphotypes Ep2 with 51.74% and Ep19 with 39.86% relative abundance in the epiphyte community were determined as dominant and identified as Fusarium fujikuroi and Fusarium oxysporum. In vitro analysis revealed that both epiphytes were active in producing indole-3-acetic acid (IAA) but not gibberellin-3 (GA3). Compared to F. oxysporum, F. fujikuroi accumulated significantly higher IAA concentration in its culture. The belowground biomass of I. cairica cuttings was significantly increased by F. oxysporum culture extract. Besides belowground biomass, F. fujikuroi culture extract significantly increased the aboveground biomass, and the number and total length of secondary shoots. In vivo inoculation showed that the conidial suspension of F. oxysporum significantly increased endogenous IAA level, the aboveground and belowground biomass of I. cairica cuttings, while the conidial suspension of F. fujikuroi slightly elevated endogenous IAA level only resulting in a significantly improved aboveground biomass. Our results revealed both of the dominant epiphytes promoted the growth of I. cairica through hormone interactions.

Estimation of endogenous indole-3-acetic acid in leaf explants and a protocol for direct regeneration in Lilium cv. Pavia

A method for regeneration via direct organogenesis in Lilium cv. Pavia using leaf primary explants has been established. Outer leaves from in vitro plantlets of 10 to 12 cm were used as the primary explant for direct organogenesis. Shoots were directly regenerated from leaf explants within 25 d of culture on Murashige and Skoog (MS) augmented with 4.5 μM 2, 4-dichlorophenoxyacetic acid (2, 4-D), 5.4 μM α-naphthaleneacetic acid (NAA), and 2.2 μM 6-benzylaminopurine (BAP). Different regions of the same leaf explant exhibited varied responses. The acropetal 3 to 4 cm of the leaf (L1) produced the maximum number of shoots (75%) indicating its high regeneration ability whereas the response of middle portion (L2) and basipetal portion of the leaf (L3) exhibited a significantly decreasing trend of shoot induction with 37.5 and 12.5%, respectively. HPLC analysis of endogenous indole-3-acetic acid (IAA) concentration established significantly higher concentration in the acropetal segments (L1). The in vitro regenerated plants were rooted in half strength MS media and were successfully hardened and field transferred. Our study established that different regions of the same leaf produce significantly different responses for shoot induction and this is attributed to the variable levels of endogenous IAA. This quick method of direct organogenesis can be applied for commercial multiplication of this important hybrid.

Tomato auxin biosynthesis/signaling is reprogrammed by the geminivirus to enhance its pathogenicity.

Tomato leaf curl New Delhi virus-derived AC4 protein interacts with host proteins involved in auxin biosynthesis and reprograms auxin biosynthesis/signaling to help in viral replication and manifestation of the disease-associated symptoms. Perturbations of phytohormone-mediated gene regulatory network cause growth and developmental defects. Furthermore, plant viral infections cause characteristic disease symptoms similar to hormone-deficient mutants. Tomato leaf curl New Delhi Virus (ToLCNDV)-encoded AC4 is a small protein that attenuates the host transcriptional gene silencing, and aggravated disease severity in tomato is correlated with transcript abundance of AC4. Hence, investigating the role of AC4 in pathogenesis divulged that ToLCNDV-AC4 interacted with host TAR1 (tryptophan amino transferase 1)-like protein, CYP450 monooxygenase-the key enzyme of indole acetic acid (IAA) biosynthesis pathway-and with a protein encoded by senescence-associated gene involved in jasmonic acid pathway. Also, ToLCNDV infection resulted in the upregulation of host miRNAs, viz., miR164, miR167, miR393 and miR319 involved in auxin signaling and leaf morphogenesis concomitant with the decline in endogenous IAA levels. Ectopic overexpression of ToLCNDV-derived AC4 in tomato recapitulated the transcriptomic and disruption of auxin biosynthesis/signaling features of the infected leaves. Furthermore, exogenous foliar application of IAA caused remission of the characteristic disease-related symptoms in tomato. The roles of ToLCNDV-AC4 in reprogramming auxin biosynthesis, signaling and cross-talk with JA pathway to help viral replication and manifest the disease-associated symptoms during ToLCNDV infection are discussed.

Formation of the Secondary Abscission Zone Induced by the Interaction of Methyl Jasmonate and Auxin in Bryophyllum calycinum: Relevance to Auxin Status and Histology.

The interaction of methyl jasmonate (JA-Me) and indole-3-acetic acid (IAA) to induce the formation of the secondary abscission zone in the middle of internode segments of Bryophyllum calycinum was investigated in relation to auxin status and histology. When IAA at 0.1% (w/w, in lanolin) was applied to the segments, the formation of the secondary abscission zone at a few mm above the treatment in the apical direction was observed. On the contrary, IAA at 0.5% (w/w, in lanolin) did not induce the formation of the secondary abscission zone. JA-Me at 0.5% (w/w, in lanolin) applied to the middle of internode segments kept in the normal (natural) or inverted positions also induced the formation of the secondary abscission zone below and above parts of the treatment. IAA at 0.5% applied to the cut surface of the upper part of the segments completely prevented the formation of the secondary abscission zone induced by JA-Me. Simultaneous application of IAA 0.5% with JA-Me 0.5% in the middle part of the internode segments induced the formation of the secondary abscission zone at 10 mm to 12 mm above the treatment. Histological analyses indicated that the formation of the secondary abscission zone was characterized by the presence of newly synthesized cell plates that resulted from periclinal cell division within one layer of mother cells in stems. The effects of IAA (0.1%) and JA-Me (0.5%) on the formation of the secondary abscission zone were histologically similar. Comprehensive analyses of plant hormones revealed that the balance of the endogenous levels of IAA in both sides adjacent to the abscission zone was significantly disturbed when the secondary abscission formation was induced by the application of IAA. These results strongly suggest that an auxin gradient is important in the formation of the secondary abscission zone in the internode segments of B. calycinum, and IAA gradient results from polar IAA transport from the application site. IAA is important in the regulation of formation of the secondary abscission zone induced by JA-Me. Further possible mechanisms of the formation of the secondary abscission zone in the internode segments of B. calycinum are also discussed in the interaction of JA-Me and IAA.

Colonization of endophyte Acremonium sp. D212 in Panax notoginseng and rice mediated by auxin and jasmonic acid.

Endophytic fungi can be beneficial to plant growth. However, the molecular mechanisms underlying colonization of Acremonium spp. remain unclear. In this study, a novel endophytic Acremonium strain was isolated from the buds of Panax notoginseng and named Acremonium sp. D212. The Acremonium sp. D212 could colonize the roots of P. notoginseng, enhance the resistance of P. notoginseng to root rot disease, and promote root growth and saponin biosynthesis in P. notoginseng. Acremonium sp. D212 could secrete indole-3-acetic acid (IAA) and jasmonic acid (JA), and inoculation with the fungus increased the endogenous levels of IAA and JA in P. notoginseng. Colonization of the Acremonium sp. D212 in the roots of the rice line Nipponbare was dependent on the concentration of methyl jasmonate (MeJA) (2-15 μmol/L) and 1-naphthalenacetic acid (NAA) (10-20 μmol/L). Moreover, the roots of the JA signaling-defective coi1-18 mutant were colonized by Acremonium sp. D212 to a lesser degree than those of the wild-type Nipponbare and miR393b-overexpressing lines, and the colonization was rescued by MeJA but not by NAA. It suggests that the cross-talk between JA signaling and the auxin biosynthetic pathway plays a crucial role in the colonization of Acremonium sp. D212 in host plants.

Genome-Wide Identification of the Auxin Response Factor (ARF) Gene Family and Their Expression Analysis during Flower Development of Osmanthus fragrans

Auxins have long been implicated in many aspects of plant growth and development. Auxin response factors (ARFs) are important proteins in auxin-mediated pathways and they play key roles in plant physiological and biochemical processes, including flower development. Endogenous indoleacetic acid (IAA) levels were measured and ARFs were studied in the flowers during the developmental stages in order to further elucidate the role of auxin in flower development of Osmanthus fragrans. A systematic analysis of OfARFs was conducted by carrying out a genome-wide search of ARFs. A total of 50 ARF genes (OfARFs) were detected and validated from the Osmanthus fragrans genome. Furthermore, a comprehensive overview of the OfARFs was undertaken, including phylogenetic relationship, gene structures, conserved domains, motifs, promoters, chromosome locations, gene duplications, and subcellular locations of the gene product. Finally, expression profiling, while using transcriptome sequencing from a previous study and quantitative real-time PCR (qRT-PCR), revealed that many OfARF genes have different expression levels in various tissues and flower developmental stages. By comparing the expression profiles among the flower developmental stages, and the relationship between ARFs and endogenous IAA levels, it can be supposed that OfARFs function in flower development of O. fragrans in an auxin-mediated pathway.

Auxin acts as a downstream signaling molecule involved in hydrogen sulfide-induced chilling tolerance in cucumber.

This report proves a cross talk between H2S and IAA in cold stress response, which has presented strong evidence that IAA acts as a downstream signal mediating the H2S-induced stress tolerance in cucumber seedlings. We evaluated changes in endogenous hydrogen sulfide (H2S) and indole-3-acetic acid (IAA) emission systems, and the interactive effect of exogenous H2S and IAA on chilling tolerance in cucumber seedlings. The results showed that chilling stress increased the activity and relative mRNA expression of L-/D-cysteine desulfhydrase (L-/D-CD), which in turn induced the accumulation of endogenous H2S. Similarly, the endogenous IAA system was triggered by chilling stress. We found that 1.0mM sodium hydrosulfide (NaHS, an H2S donor) significantly enhanced the activity of flavin monooxygenase (FMO) and relative expression of FMO-like proteins (YUCCA2), which in turn elevated endogenous IAA levels in cucumber seedlings. However, IAA had little effects on activities of L-/D-CD and endogenous H2S levels. H2S-induced IAA production accompanied by increase in chilling tolerance, as shown by the decrease in stress-induced electrolyte leakage (EL) and reactive oxygen species (ROS) accumulation, and increase in gene expressions and enzyme activities of photosynthesis. 1-naphthylphthalamic acid (NPA, an IAA polar transport inhibitor) declined H2S-induced chilling tolerance and defense genes' expression. However, scavenging of H2S had a little effect on IAA-induced chilling tolerance. These results suggest that IAA acting as a downstream signaling molecule is involved in the H2S-induced chilling tolerance in cucumber seedlings.

Global transcriptome analyses reveal that infection with chrysanthemum stunt viroid (CSVd) affects gene expression profile of chrysanthemum plants, but the genes involved in plant hormone metabolism and signaling may not be silencing target of CSVd-siRNAs

Abstracts Chrysanthemum stunt viroid (CSVd) is a pathogen that induces stunted plant growth as a typical disease symptom in infected chrysanthemum plants. Because of its simple RNA genomic structure, the mechanism of how CSVd provokes pathogenicity in host plants remains elusive. Since small RNAs generated from CSVd genomic RNA accumulated in infected plants ( Tsuro et al., 2013 ), it has been suggested that forced gene silencing against intrinsic genes by CSVd-small interfering (si)RNAs is involved in its pathogenicity. In this study, we revealed that CSVd infection induced global fluctuation of gene expression in plants. Hypothetical CSVd-siRNAs (22 nucleotides long) from (+) and (−) strands of the CSVd RNA genome were surveyed to examine if they could target the down-regulated gene sequences. Six genes were identified as target candidates, but the causal link between their functions and disease symptoms was remained unclear. In contrast, the expression levels of genes involved in the biosynthesis of gibberellic acid (GA) and cytokinin (CK), indole-3-acetic acid (IAA) transport and cell wall growth in infected plants were down-regulated, but mRNAs of those genes were not found as possible target sequences of CSVd-siRNAs. On the other hand, endogenous levels of GA, CK, IAA, and abscisic acid and the rate of IAA movement in the stem were not different between healthy and infected plants. Growth promotion of infected plants by GA treatment was poorer than that of healthy plants, suggesting that CSVd infection suppressed the GA-signaling pathway. This was further supported by the findings of a reduced response of GA-regulated genes to GA treatment and up-regulation of the DELLA-protein gene, a negative regulator of GA-signaling, in infected plants. Thus, the down-regulation of GA responsiveness and inhibition of cell wall expansion are major causes of the stunted growth of CSVd-infected chrysanthemum plants.

Insertion of a transposon-like sequence in the 5'-flanking region of the YUCCA gene causes the stony hard phenotype.

Melting-flesh peaches produce large amounts of ethylene, resulting in rapid fruit softening at the late-ripening stage. In contrast, stony hard peaches do not soften and produce little ethylene. The indole-3-acetic acid (IAA) level in stony hard peaches is low at the late-ripening stage, resulting in low ethylene production and inhibition of fruit softening. To elucidate the mechanism of low IAA concentration in stony hard peaches, endogenous levels of IAA and IAA intermediates or metabolites were analysed by ultra-performance liquid chromatography-tandem mass spectrometry. Although the IAA level was low, the indole-3-pyruvic acid (IPyA) level was high in stony hard peaches at the ripening stage. These results indicate that YUCCA activity is reduced in ripening stony hard peaches. The expression of one of the YUCCA isogenes in peach, PpYUC11, was suppressed in ripening stony hard peaches. Furthermore, an insertion of a transposon-like sequence was found upstream of the PpYUC11 gene in the 5'-flanking region. Analyses of the segregation ratio of the stony hard phenotype and genotype in F1 progenies indicated that the transposon-inserted allele of PpYUC11, hd-t, correlated with the stony hard phenotype. On the basis of the above findings, we propose that the IPyA pathway (YUCCA pathway) is the main auxin biosynthetic pathway in ripening peaches of 'Akatsuki' and 'Manami' cultivars. Because IAA is not supplied from storage forms, IAAde novo synthesis via the IPyA pathway (YUCCA pathway) in mesocarp tissues is responsible for auxin generation to support fruit softening, and its disruption can lead to the stony hard phenotype.