Auxin Homeostasis Research Articles

Physcomitrium patens Mutants in Auxin Conjugating GH3 Proteins Show Salt Stress Tolerance but Auxin Homeostasis Is Not Involved in Regulation of Oxidative Stress Factors.

Salt stress is among the most challenging abiotic stress situations that a plant can experience. High salt levels do not only occur in areas with obvious salty water, but also during drought periods where salt accumulates in the soil. The moss Physcomitrium patens became a model for studying abiotic stress in non-vascular plants. Here, we show that high salt concentrations can be tolerated in vitro, and that auxin homeostasis is connected to the performance of P. patens under these stress conditions. The auxin levels can be regulated by conjugating IAA to amino acids by two members of the family of GH3 protein auxin amino acid-synthetases that are present in P. patens. Double GH3 gene knock-out mutants were more tolerant to high salt concentrations. Furthermore, free IAA levels were differentially altered during the time points investigated. Since, among the mutant lines, an increase in IAA on at least one NaCl concentration tested was observed, we treated wild type (WT) plants concomitantly with NaCl and IAA. This experiment showed that the salt tolerance to 100 mM NaCl together with 1 and 10 µM IAA was enhanced during the earlier time points. This is an additional indication that the high IAA levels in the double GH3-KO lines could be responsible for survival in high salt conditions. While the high salt concentrations induced several selected stress metabolites including phenols, flavonoids, and enzymes such as peroxidase and superoxide dismutase, the GH3-KO genotype did not generally participate in this upregulation. While we showed that the GH3 double KO mutants were more tolerant of high (250 mM) NaCl concentrations, the altered auxin homeostasis was not directly involved in the upregulation of stress metabolites.

HSP90 affects root growth in Arabidopsis by regulating the polar distribution of PIN1.

Auxin homeostasis and signaling affect a broad range of developmental processes in plants. The interplay between HSP90 and auxin signaling is channeled through the chaperoning capacity of the HSP90 on the TIR1 auxin receptor. The sophisticated buffering capacity of the HSP90 system through the interaction with diverse signaling protein components drastically shapes genetic circuitries regulating various developmental aspects. However, the elegant networking capacity of HSP90 in the global regulation of auxin response and homeostasis has not been appreciated. Arabidopsis hsp90 mutants were screened for gravity response. Phenotypic analysis of root meristems and cotyledon veins was performed. PIN1 localization in hsp90 mutants was determined. Our results showed that HSP90 affected the asymmetrical distribution of PIN1 in plasma membranes and influenced its expression in prompt cell niches. Depletion of HSP90 distorted polar distribution of auxin, as the acropetal auxin transport was highly affected, leading to impaired root gravitropism and lateral root formation. The essential role of the HSP90 in auxin homeostasis was profoundly evident from early development, as HSP90 depletion affected embryo development and the pattern formation of veins in cotyledons. Our data suggest that the HSP90-mediated distribution of PIN1 modulates auxin distribution and thereby auxin signaling to properly promote plant development.

New insights into the hormonal regulation of silicon-supplied sorghum plants challenged with Colletotrichum sublineolum

Importance of sorghum as staple food worldwide is unquestionable and silicon (Si) supply has decreased the foliar symptoms of anthracnose, caused by Colletotrichum sublineolum, through potentiation of defense responses and preservation of photosynthetic apparatus. This study investigated the role of Si in cytological changes as well as in the expression of genes involved in auxin metabolism (GH3.1) and in signaling pathways mediated by salicylic acid (SA) (BTHip), jasmonic acid (JA)/ethylene (ET) (LOX), and JA (Jar, AOS 1, and ACCox) by real-time polymerase chain reaction in the leaves of plants non-supplied (-Si) or supplied (+Si) with Si and non-inoculated or inoculated with C. sublineolum. Sheath cells from +Si plants were less colonized by fungal hyphae due to intense phenolics and lignin deposition and high hydrogen peroxide production. Foliar lesions for +Si plants sprayed with an ET signaling inhibitor, ET biosynthesis inhibitor, and JA biosynthesis inhibitor were bigger compared to +Si plants sprayed with water. For +Si plants sprayed with ET and JA signaling inhibitors, foliar lesions were smaller compared to -Si plants sprayed with these same compounds. Suppression of auxin biosynthesis, transporter, and signaling by specific inhibitors resulted in smaller lesions. Expressions of GH3.1, BRHip, LOX, Jar, AOS 1, and ACCox were greater for +Si plants compared to -Si plants during the infection process of C. sublineolum. This study highlighted the potential of Si to boost sorghum resistance to anthracnose through a multistep process involving hormone-signaling pathways, especially regarding auxin metabolism and homeostasis, hydrogen peroxide generation, and plant cell wall strengthening.

Dose-dependent responses of Arabidopsis thaliana to zinc are mediated by auxin homeostasis and transport

Zinc is an essential element for plant; however, in excess it could cause adverse effects on plant growth and productivity. Plants have evolved various regulatory mechanisms to optimize their growth and development at different zinc concentrations. In this work, we elucidated the role of auxin homeostasis and transport in the dose-dependent responses to zinc in Arabidopsis thaliana. The results showed that zinc had dose-dependent effects on both vegetative and reproductive growth of Arabidopsis. At the concentration of 50, 100 and 350 μM, zinc significantly affected the length of primary root and the size of root meristem. This root response to Zn were associated with differentially regulation of endogenous auxin, as demonstrated by the DR5:GUS reporter line and in vivo hormone measurements. Exogenous IAA could alleviate the Zn-induced short-root phenotype by affecting auxin accumulation in plants, while Arabidopsis mutants with deficient auxin synthesis or transport mechanisms exhibited hypersensitivity to Zn. In conclusion, the effect of zinc on root growth of Arabidopsis thaliana is regulated by differences in auxin synthesis and transport.

Sedentary Plant-Parasitic Nematodes Alter Auxin Homeostasis via Multiple Strategies.

Sedentary endoparasites such as cyst and root-knot nematodes infect many important food crops and are major agro-economical pests worldwide. These plant-parasitic nematodes exploit endogenous molecular and physiological pathways in the roots of their host to establish unique feeding structures. These structures function as highly active transfer cells and metabolic sinks and are essential for the parasites’ growth and reproduction. Plant hormones like indole-3-acetic acid (IAA) are a fundamental component in the formation of these feeding complexes. However, their underlying molecular and biochemical mechanisms are still elusive despite recent advances in the field. This review presents a comprehensive overview of known functions of various auxins in plant-parasitic nematode infection sites, based on a systematic analysis of current literature. We evaluate multiple aspects involved in auxin homeostasis in plants, including anabolism, catabolism, transport, and signalling. From these analyses, a picture emerges that plant-parasitic nematodes have evolved multiple strategies to manipulate auxin homeostasis to establish a successful parasitic relationship with their host. Additionally, there appears to be a potential role for auxins other than IAA in plant-parasitic nematode infections that might be of interest to be further elucidated.

Iso-anchorene is an endogenous metabolite that inhibits primary root growth in Arabidopsis.

Carotenoid-derived regulatory metabolites and hormones are generally known to arise through the oxidative cleavage of a single double bond in the carotenoid backbone, which yields mono-carbonyl products called apocarotenoids. However, the extended conjugated double bond system of these pigments predestines them also to repeated cleavage forming dialdehyde products, diapocarotenoids, which have been less investigated due to their instability and low abundance. Recently, we reported on the short diapocarotenoid anchorene as an endogenous Arabidopsis metabolite and specific signaling molecule that promotes anchor root formation. In this work, we investigated the biological activity of a synthetic isomer of anchorene, iso-anchorene, which can be derived from repeated carotenoid cleavage. We show that iso-anchorene is a growth inhibitor that specifically inhibits primary root growth by reducing cell division rates in the root apical meristem. Using auxin efflux transporter marker lines, we also show that the effect of iso-anchorene on primary root growth involves the modulation of auxin homeostasis. Moreover, by using liquid chromatography-mass spectrometry analysis, we demonstrate that iso-anchorene is a natural Arabidopsis metabolite. Chemical inhibition of carotenoid biosynthesis led to a significant decrease in the iso-anchorene level, indicating that it originates from this metabolic pathway. Taken together, our results reveal a novel carotenoid-derived regulatory metabolite with a specific biological function that affects root growth, manifesting the biological importance of diapocarotenoids.

Auxin regulated metabolic changes underlying sepal retention and development after pollination in spinach

BackgroundPollination accelerate sepal development that enhances plant fitness by protecting seeds in female spinach. This response requires pollination signals that result in the remodeling within the sepal cells for retention and development, but the regulatory mechanism for this response is still unclear. To investigate the early pollination-induced metabolic changes in sepal, we utilize the high-throughput RNA-seq approach.ResultsSpinach variety ‘Cornel 9’ was used for differentially expressed gene analysis followed by experiments of auxin analog and auxin inhibitor treatments. We first compared the candidate transcripts expressed differentially at different time points (12H, 48H, and 96H) after pollination and detected significant difference in Trp-dependent auxin biosynthesis and auxin modulation and transduction process. Furthermore, several auxin regulatory pathways i.e. cell division, cell wall expansion, and biogenesis were activated from pollination to early developmental symptoms in sepals following pollination. To further confirm the role auxin genes play in the sepal development, auxin analog (2, 4-D; IAA) and auxin transport inhibitor (NPA) with different concentrations gradient were sprayed to the spinach unpollinated and pollinated flowers, respectively. NPA treatment resulted in auxin transport weakening that led to inhibition of sepal development at concentration 0.1 and 1 mM after pollination. 2, 4-D and IAA treatment to unpollinated flowers resulted in sepal development at lower concentration but wilting at higher concentration.ConclusionWe hypothesized that sepal retention and development might have associated with auxin homeostasis that regulates the sepal size by modulating associated pathways. These findings advanced the understanding of this unusual phenomenon of sepal growth instead of abscission after pollination in spinach.

RGB1 Regulates Grain Development and Starch Accumulation Through Its Effect on OsYUC11-Mediated Auxin Biosynthesis in Rice Endosperm Cells.

RGB1, a subunit of heterotrimeric G protein, plays important roles in regulating grain size and weight of rice. However, the molecular mechanisms underlying controlling grain filling process by G protein are still largely unclear. In the present study, we show that RGB1 controls not only the grain size but also the grain filling process. Knock-down of RGB1 significantly delayed grain development and reduced starch accumulation and grain weight, which was closely related to the delayed and the lower expression of genes encoding sucrose metabolism and starch biosynthesis related enzymes during grain filling stage. Suppression of RGB1 expression also resulted in the lower auxin content in grains, which was correlated with the lower expression of OsNF-YB1 and OsYUC11 during grain filling stage. Further biochemical evidence showed that OsYUC11 expression was under control of OsNF-YB1 by its interaction with promoter of OsYUC11. Taken together, we propose that RGB1 controls rice grain development and grain filling process by changing auxin homeostasis in endosperm cells. OsNF-YB1, which acts as a key downstream effector of RGB1, interacts directly with the promoter of OsYUC11 and stimulates the OsYUC11 expression, thereby regulating auxin biosynthesis and starch accumulation and grain size.

AUXIN RESPONSE FACTOR 18-HISTONE DEACETYLASE 6 module regulates floral organ identity in rose (Rosa hybrida).

The phytohormone auxin plays a pivotal role in floral meristem initiation and gynoecium development, but whether and how auxin controls floral organ identity remain largely unknown. Here, we found that auxin levels influence organ specification, and changes in auxin levels influence homeotic transformation between petals and stamens in rose (Rosa hybrida). The PIN-FORMED-LIKES (PILS) gene RhPILS1 governs auxin levels in floral buds during floral organogenesis. RhAUXIN RESPONSE FACTOR 18 (RhARF18), whose expression decreases with increasing auxin content, encodes a transcriptional repressor of the C-class gene RhAGAMOUS (RhAG), and controls stamen-petal organ specification in an auxin-dependent manner. Moreover, RhARF18 physically interacts with the histone deacetylase (HDA) RhHDA6. Silencing of RhHDA6 increases H3K9/K14 acetylation levels at the site adjacent to the RhARF18-binding site in the RhAG promoter and reduces petal number, indicating that RhARF18 might recruit RhHDA6 to the RhAG promoter to reinforce the repression of RhAG transcription. We propose a model for how auxin homeostasis controls floral organ identity via regulating transcription of RhAG.

Brassinosteroid-BZR1/2-WAT1 module determines the high level of auxin signalling in vascular cambium during wood formation.

The tight regulation of local auxin homeostasis and signalling maxima in xylem precursor cells specifies the organising activity of the vascular cambium and consequently promotes xylem differentiation and wood formation. However, the molecular mechanisms underlying the local auxin signalling maxima in the vascular cambium are largely unknown. Here, we reveal that brassinosteroid (BR)-activated WALLS ARE THIN1 (WAT1) facilitates wood formation by enhancing local auxin signalling in the vascular cambium in Solanum lycopersicum. Growth defects and low auxin signalling readouts in the BR-deficient tomato cultivar, Micro-Tom, were associated with a novel recessive allele, Slwat1-copi, created by the insertion of a retrotransposon in the last exon of the SlWAT1 locus. Molecular and genetic studies by generating the gain-of-function and loss-of-function tomato mutants revealed that SlWAT1 is a critical regulator for fine tuning local auxin homeostasis and signalling outputs in vascular cambium to facilitate secondary growth. Finally, we discovered that BR-regulated SlBZR1/2 directly activated downstream auxin responses by SlWAT1 upregulation in xylem precursor cells to facilitate xylem differentiation and subsequent wood formation. Our data suggest that the BR-SlBZR1/2-WAT1 signalling network contributes to the high level of auxin signalling in the vascular cambium for secondary growth.

MicroRNAs modulate ethylene induced retrograde signal for rice endosperm starch biosynthesis by default expression of transcriptome

Control of stage specific spike in ethylene production at anthesis has been a vauable route to potentially enhance genetic ceiling for grain filling of rice spikelet. A number of genes controlling ethylene homeostasis and starch synthesis have been identified so long, but lack of credible information on master modulation of gene expression by miRNAs and their target genes associated with hormonal dynamics obfuscate mechanisms controlling genotype difference in quantum of grain filling. The confusion accounts for consequent shrinkage of options for yield manipulation. In a two by two factorial design, miRNA regulation of spikelet specific grain development in low against high sterile recombinant inbred lines of rice Oryza sativa L. namely CR 3856-62-11-3-1-1-1-1-1-1 (SR 157) and CR 3856-63-1-1-1-1-1-1-1 (SR 159) respectively, and inferior verses superior spikelets were compared during first 10 days after anthesis. Grain filling was poorer in SR159 than SR157 and inferior spikelets in the former were most vulnerable. Between the cultivars, overall expression of unique miRNAs with targets on ethylene pathway genes was higher in SR159 than SR157 and the situation was opposite for auxin pathway genes. Precision analysis in psTarget server database identified up-regulation of MIR2877 and MIR530-5p having Os11t0141000-02 and Os07t0239400-01 (PP2A regulatory subunit-like protein and ethylene-responsive small GTP-binding proteins) and MIR396h having Os01t0643300-02 (an auxin efflux carrier protein) and Os01t0643300-01 (a PIN1-like auxin transport protein), as targets with highest probability at anthesis and 5 days after anthesis respectively, in the inferior spikelet and the fold change values of DGE matched with pattern of gene expression (relative transcript level) in the qRT-PCR studies conducted for relevant miRNAs and protein factors for ethylene and auxin signalling. In conclusion, epigenetic regulation of both auxin and ethylene homeostasis control grain filling of rice spikelet was established, but evidences were more robust for the latter.

Allelopathic root inhibition and its mechanisms

Allelopathy represents a valuable biochemical strategy in plant-plant interactions among different plants, e.g. among crops and weeds. It is an important strategy in the colonization of many invasive alien plants. Allelopathic plants affect the growth of other plants in the vicinity through the release of secondary metabolites (allelochemicals) into the soil. In particular, many allelochemicals suppress the root growth of target plants, but little is known about the mechanism involved in root growth inhibition. In this review, we will highlight the mechanism of root suppression involving: (i) Alterations in auxin homeostasis affecting polar auxin transport and root gravitropism, (ii) Biochemical and physiological processes in inhibited roots associated with oxidative stress due to direct production and accumulation of reactive oxygen species or suppression of antioxidative response and (iii) The ultrastructural modifications in root tip exposed to allelochemicals that drastically suppress the cell division and eventually lead to shorter roots of target plants.

The Diverse Salt-Stress Response of Arabidopsis ctr1-1 and ein2-1Ethylene Signaling Mutants Is Linked to Altered Root Auxin Homeostasis.

We explored the interplay between ethylene signals and the auxin pool in roots exposed to high salinity using Arabidopsis thaliana wild-type plants (Col-0), and the ethylene-signaling mutants ctr1-1 (constitutive) and ein2-1 (insensitive). The negative effect of salt stress was less pronounced in ctr1-1 individuals, which was concomitant with augmented auxin signaling both in the ctr1-1 controls and after 100 mM NaCl treatment. The R2D2 auxin sensorallowed mapping this active auxin increase to the root epidermal cells in the late Cell Division (CDZ) and Transition Zone (TZ). In contrast, the ethylene-insensitive ein2-1 plants appeared depleted in active auxins. The involvement of ethylene/auxin crosstalk in the salt stress response was evaluated by introducing auxin reporters for local biosynthesis (pTAR2::GUS) and polar transport (pLAX3::GUS, pAUX1::AUX1-YFP, pPIN1::PIN1-GFP, pPIN2::PIN2-GFP, pPIN3::GUS) in the mutants. The constantly operating ethylene-signaling pathway in ctr1-1 was linked to increased auxin biosynthesis. This was accompanied by a steady expression of the auxin transporters evaluated by qRT-PCR and crosses with the auxin transport reporters. The results imply that the ability of ctr1-1 mutant to tolerate high salinity could be related to the altered ethylene/auxin regulatory loop manifested by a stabilized local auxin biosynthesis and transport.

Function of histone H2B monoubiquitination in transcriptional regulation of auxin biosynthesis in Arabidopsis

The auxin IAA is a vital plant hormone in controlling growth and development, but our knowledge about its complicated biosynthetic pathways and molecular regulation are still limited and fragmentary. cytokinin induced root waving 2 (ckrw2) was isolated as one of the auxin-deficient mutants in a large-scale forward genetic screen aiming to find more genes functioning in auxin homeostasis and/or its regulation. Here we show that CKRW2 is identical to Histone Monoubiquitination 1 (HUB1), a gene encoding an E3 ligase required for histone H2B monoubiquitination (H2Bub1) in Arabidopsis. In addition to pleiotropic defects in growth and development, loss of CKRW2/HUB1 function also led to typical auxin-deficient phenotypes in roots, which was associated with significantly lower expression levels of several functional auxin synthetic genes, namely TRP2/TSB1, WEI7/ASB1, YUC7 and AMI1. Corresponding defects in H2Bub1 were detected in the coding regions of these genes by chromatin immunoprecipitation (ChIP) analysis, indicating the involvement of H2Bub1 in regulating auxin biosynthesis. Importantly, application of exogenous cytokinin (CK) could stimulate CKRW2/HUB1 expression, providing an epigenetic avenue for CK to regulate the auxin homeostasis. Our results reveal a previously unknown mechanism for regulating auxin biosynthesis via HUB1/2-mediated H2Bub1 at the chromatin level.

Systematical characterization of YUCCA gene family in five cotton species, and potential functions of YUCCA22 gene in drought resistance of cotton

Auxin plays an essential role in the normal growth and development of plants. The tryptophan aminotransferase of Arabidopsis (TAA)/YUCCA (YUC) enzymes functions as flavin monooxygenases (FMOs), which are the key enzymes in auxin synthesis. However, comprehensive characterization of the YUC gene family in cotton has not yet been completed. In order to reveal the functions of YUC genes in cotton, we systematically identified and characterized the YUC gene family in five species of cotton including analysis of the member, characterization, intragenomic duplication, and collinear relationships. Transcriptomic analysis indicates that GhYUCs participate in abiotic stress processes in cotton. The GhYUC22 gene that is responsive to polyethylene glycol (PEG) and abscisic acid (ABA) was functionally identified via virus-induced gene silencing (VIGS). Subsequent natural drought simulation experiments and physiological index data demonstrated that partial silencing of GhYUC22 gene endowed cotton with drought resistance by modulating indole-3-acetic acid (IAA) and ABA signaling pathways. In conclusion, our findings support the role of that GhYUC22 gene in modulating both endogenous auxin and ABA homeostasis and regulating the drought resistance in cotton.

Molecular Bases for the Regulation of Adventitious Root Generation in Plants.

The formation of adventitious roots (ARs) is an ecologically and economically important developmental process in plants. The evolution of AR systems is an important way for plants to cope with various environmental stresses. This review focuses on identified genes that have known to regulate the induction and initiation of ARs and offers an analysis of this process at the molecular level. The critical genes involved in adventitious rooting are the auxin signaling-responsive genes, including the AUXIN RESPONSE FACTOR (ARF) and the LATERAL ORGAN BOUNDARIES-DOMAIN (LOB) gene families, and genes associated with auxin transport and homeostasis, the quiescent center (QC) maintenance, and the root apical meristem (RAM) initiation. Several genes involved in cell wall modulation are also known to be involved in the regulation of adventitious rooting. Furthermore, the molecular processes that play roles in the ethylene, cytokinin, and jasmonic acid signaling pathways and their crosstalk modulate the generation of ARs. The crosstalk and interaction among many molecular processes generates complex networks that regulate AR generation.

S-nitrosoglutathione Reductase-Mediated Nitric Oxide Affects Axillary Buds Outgrowth of Solanum lycopersicum L. by Regulating Auxin and Cytokinin Signaling

Auxin and cytokinin are two kinds of important phytohormones that mediate outgrowth of axillary buds in plants. How nitric oxide and its regulator of S-nitrosoglutathione reductase (GSNOR) take part in auxin and cytokinin signaling for controlling axillary buds outgrowth remains elusive. We investigated the roles of GSNOR during tomato axillary bud outgrowth by using physiological, biochemical and genetic approaches. GSNOR negatively regulated NO homeostasis. Suppression of GSNOR promoted axillary bud outgrowth by inhibiting the expression of FZY in both apical and axillary buds. Meanwhile, AUX1 and PIN1 were down-regulated in apical buds but up-regulated in axillary buds in GSNOR-suppressed plants. Thus, reduced IAA accumulation was shown in both apical buds and axillary buds of GSNOR-suppressed plants. GSNOR-mediated changes of NO and auxin affected cytokinin biosynthesis, transport, and signaling. And a decreased ratio of auxin: cytokinin was shown in axillary buds of GSNOR-suppressed plants, leading to bud dormancy breaking. We also found that the original NO signaling was generated by nitrate reductase (NR) catalyzing nitrate as substrate. NR-mediated NO reduced the GSNOR activity through S-nitrosylation of Cys-10, then induced a further NO burst, which played the above roles to promote axillary buds outgrowth. Together, GSNOR-mediated NO played important roles in controlling axillary buds outgrowth by altering the homeostasis and signaling of auxin and cytokinin in tomato plants.

The ectopic expression of Arabidopsis glucosyltransferase UGT74D1 affects leaf positioning through modulating indole-3-acetic acid homeostasis

Leaf angle is an important agronomic trait affecting photosynthesis efficiency and crop yield. Although the mechanisms involved in the leaf angle control are intensively studied in monocots, factors contribute to the leaf angle in dicots are largely unknown. In this article, we explored the physiological roles of an Arabidopsis glucosyltransferase, UGT74D1, which have been proved to be indole-3-acetic acid (IAA) glucosyltransferase in vitro. We found that UGT74D1 possessed the enzymatic activity toward IAA glucosylation in vivo and its expression was induced by auxins. The ectopically expressed UGT74D1 obviously reduced the leaf angle with an altered IAA level, auxin distribution and cell size in leaf tissues. The expression of several key genes involved in the leaf shaping and leaf positioning, including PHYTOCHROME KINASE SUBSTRATE (PKS) genes and TEOSINTE BRANCHED1, CYCLOIDEA, and PCF (TCP) genes, were dramatically changed by ectopic expression of UGT74D1. In addition, clear transcription changes of YUCCA genes and other auxin related genes can be observed in overexpression lines. Taken together, our data indicate that glucosyltransferase UGT74D1 could affect leaf positioning through modulating auxin homeostasis and regulating transcription of PKS and TCP genes, suggesting a potential new role of UGT74D1 in regulation of leaf angle in dicot Arabidopsis.

Comparative transcriptome analysis reveals molecular regulators underlying pluripotent cell induction and callus formation in Anthurium andraeanum “Alabama”

Shoot regeneration from pluripotent cell masses is an important process for genetic improvement of Anthurium andraeanum through Agrobacterium-mediated transformation. However, the mechanisms underlying pluripotent cell induction are not well understood. Here, three half-leaf explant samples collected after 0-, 2-, and 30-d culture on callus-inducing medium were used for RNA isolation and transcriptome profiling. In total, 225,752 non-redundant unigenes were assembled with a mean length of 1299 bp. Differentially expressed genes among the three samples were identified by comparative transcriptome analysis. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed the greatest number of differentially expressed genes in the comparison between 0- and 2-d cultures were related to the “ribosome.” Many differentially expressed genes in the comparison between 2- and 30-d cultures were involved in “plant hormone signal transduction,” “starch and sucrose metabolism,” and “phenylpropanoid biosynthesis.” Further investigation was focused on genes related to transcription factors, epigenetic regulation, nitrous oxide metabolism, and homeostasis of plant hormones auxin, cytokinin, brassinolide, and strigolactone. Experiments were designed to confirm the effects of nitrous oxide and strigolactone on callus formation in Anthurium andraeanum. Callus formation was enhanced by exogenous application of sodium nitroprusside, which produces nitrous oxide, but was inhibited under rac-GR24 (a strigolactone analogue) treatment. Our comparative transcriptome analysis revealed that the signal molecule nitrous oxide and plant hormones auxin, cytokinin, and brassinolide act as positive regulators in promoting callus formation, while strigolactone may inhibit callus formation. Transcriptional and epigenetic regulations also play important roles in pluripotent cell induction in Anthurium andraeanum.

Ethylene-auxin crosstalk regulates postharvest fruit ripening process in apple

The ripening of climacteric fruits, such as apple, is represented by a series of genetically programmed events orchestrated by the action of several hormones. In this study, we investigated the existence of a hormonal crosstalk between ethylene and auxin during the post-harvest ripening of three internationally known apple cultivars: 'Golden Delicious', 'Granny Smith' and 'Fuji'. The normal climacteric ripening was impaired by the exogenous application of 1-methylcyclopropene (1-MCP) that affected the production of ethylene and the physiological behaviour of specific ethylene-related quality traits, such as fruit texture and the production of volatile organic compounds. The application of 1-MCP induced, moreover, a de-novo accumulation of auxin. The RNA-Seq wide-transcriptome analysis evidenced as the competition at the level of the ethylene receptors induced a cultivar-dependent transcriptional re-programming. The DEGs annotation carried out through the KEGG database identified as most genes were assigned to the plant hormone signaling transduction category, and specifically related to auxin and ethylene. The interplay between these two hormones was further assessed through a candidate gene analysis that highlighted a specific activation of GH3 and ILL genes, encoding key steps in the process of the auxin homeostasis mechanism. Our results showed that a compromised ethylene metabolism at the onset of the climacteric ripening in apple can stimulate, in a cultivar-dependent fashion, an initial de-novo synthesis and de-conjugation of auxin as a tentative to restore a normal ripening progression.