Search Results (44)

Eurasian aspen (Populus tremula L.) is a tree species with recognised ecological and economic importance for both natural and plantation forests. For the fast cloning of selected aspen genotypes, the method of plant propagation through in vitro culture (micropropagation) is often recommended. The efficiency of this method is related to the use of shoot-inducing chemical growth regulators, among which cytokinins, a type of plant hormone, dominate. Although cytokinins can inhibit rooting, this effect is avoided by using cytokinin-free media. This study sought to identify concentrations and combinations of growth regulators that would stimulate one type of P. tremula organogenesis (either shoot or root formation) without inhibiting the other. The investigated growth regulators included cytokinin 6-benzylaminopurine (BAP), auxin transport inhibitor 2,3,5-triiodobenzoic acid (TIBA), auxins indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA), gibberellin biosynthesis inhibitor paclobutrazol (PBZ), and a gibberellin mixture (GA₄/₇). Both BAP and TIBA increased shoot number per P. tremula explant and decreased the number of adventitious roots, but TIBA, in contrast to BAP, did not inhibit lateral root formation. However, for the maintenance of both adventitious shoot and root formation above the control level, the combination of PBZ and GA_4/7 was shown to be especially promising. Full article

Toxoplasma gondii is an Apicomplexan parasite that possesses a well-developed system of scavengers of reactive oxygen species (ROS). Among its components, T. gondii mitochondrial superoxide dismutase (TgSOD2) is essential, as predicted by the CRISPR phenotype index and evidenced by the non-viability of its constitutive knockouts. As an obligate intracellular parasite, TgSOD2 is upregulated during extracellular stages. Herein, we generated a viable TgSOD2 knockdown mutant using an inducible auxin–degron system to explore the biological role of TgSOD2 in T. gondii. Depletion of TgSOD2 led to impaired parasite growth and replication, reduced mitochondrial membrane potential (MMP), abnormalities in the distribution of ATP synthase within its mitochondrial electron transport chain (mETC), and increased susceptibility to mETC inhibitors. Through a proximal biotinylation approach, we identified the interactions of TgSOD2 with complexes IV and V of its mETC, suggesting that these sites are sensitive to ROS. Our study provides the first insights into the role of TgSOD2 in maintaining its mitochondrial redox homeostasis and subsequent parasite replication fitness. Significance: Toxoplasma gondii infects nearly a third of the world population and can cause fetal miscarriages or life-threatening complications in vulnerable patients. Current therapies do not eradicate the parasite from the human hosts, rendering them at risk of recurrence during their lifetimes. T. gondii has a single mitochondrion, which is well-known for its susceptibility to oxidative damage that leads to T. gondii’s death. Therefore, targeting T. gondii mitochondrion remains an attractive therapeutic strategy for drug development. T. gondii’s mitochondrial superoxide dismutase is an antioxidant protein in the parasite mitochondrion and is essential for its survival. Understanding its biological role could reveal mitochondrial vulnerabilities in T. gondii and provide new leads for the development of effective treatments for T. gondii infections. Full article

Grafting is a prevalent horticultural technique that enhances crop yields and stress resilience; nevertheless, compatibility issues frequently constrain its efficacy. This research examined the physiological, hormonal, and transcriptional factors regulating compatibility between the litchi (Litchi chinensis Sonn.) cultivars Feizixiao (FZX) and Ziniangxi (ZNX). The anatomical and growth investigations demonstrated significant disparities between compatible (FZX as scion and ZNX as rootstock) and incompatible (ZNX as scion and FZX as rootstock) grafts, with the latter showing reduced levels of indole acetic acid (IAA). Exogenous 1-naphthalene acetic acid (NAA) application markedly improved the graft survival, shoot development, and hormonal synergy, whereas the auxin inhibitor tri-iodobenzoic acid (TIBA) diminished these parameters. The incompatible grafts showed downregulation of auxin transporter genes, including ATP-binding cassette (ABC) transporter, AUXIN1/LIKE AUX1 (AUX/LAX), and PIN-FORMED (PIN) genes, suggesting impaired vascular tissue growth. Metabolomic profiling revealed dynamic interactions between auxin, salicylic acid, and jasmonic acid, with NAA-treated grafts exhibiting enhanced levels of stress-responsive metabolites. Transcriptome sequencing identified differentially expressed genes (DEGs) linked to auxin signaling (ARF, GH3), seven additional phytohormones, secondary metabolism (terpenoids, anthocyanins, and phenylpropanoids), and ABC transporters. Gene ontology and KEGG analyses highlighted the significance of hormone interactions and the biosynthesis of secondary metabolites in successful grafting. qRT-PCR validation substantiated the veracity of the transcriptome data, emphasizing the significance of auxin transport and signaling in effective graft development. This study provides an in-depth review of the molecular and physiological factors influencing litchi grafting. These findings provide critical insights for enhancing graft success rates in agricultural operations via targeted hormonal and genetic approaches. Full article

The initiation and specification of germline cells are crucial for plant reproduction and the continuity of species. In Arabidopsis thaliana, auxin plays a vital role in guiding the transition of somatic cells into germline fate, orchestrating the specification of both male archesporial cells and female megaspore mother cells. This process is regulated through interaction with the transcription factor Sporocyteless/Nozzle, which forms a feedback mechanism that modulates germ cell specialization. Auxin biosynthesis, polar transport, and signal transduction pathways collectively ensure the accurate determination of germ cell fate. Furthermore, the coordination of auxin signaling with epigenetic regulation and miRNA-mediated control fine-tunes the differentiation between germline and somatic cells. This review discusses the mechanisms underlying auxin-guided germ cell specification. It proposes future research directions, including studies on PIN-FORMED-mediated polar transport, the role of the YUCCA family in auxin biosynthesis, and the involvement of the Transport Inhibitors Response 1/Auxn Signaling F-Box-Auxin Response Factor (TIR1/AFB-ARF) signaling pathway in germ cell fate determination. These insights will enhance our understanding of plant reproductive biology and provide new strategies for crop breeding. Full article

The gravitropic growth of roots is crucial for plants to adapt to terrestrial environments and acquire nutrients from the soil. Tomatoes are a vital economic crop that requires abundant water and nutrients for growth and development. However, there are few reports on the regulatory mechanisms of tomato root gravitropism, particularly auxin-mediated root gravitropic growth. Here, we revealed the signaling pathway of auxin regulating tomato root gravity response through exogenous auxin and auxin inhibitor treatment combined with transcriptome profiling. Our data underscore the necessity of auxin biosynthesis, transport, and optimal levels for the gravitropic growth of tomato roots. Treatment with exogenous auxin or auxin biosynthesis inhibitors diminished gravitropic response in tomato roots. Conversely, treatment with an auxin transport inhibitor led to a robust agravitropic response. Furthermore, we observed that auxin controls root gravitropic growth by establishing concentration gradients and influences root perception of gravity signals by positively regulating starch granule accumulation. Treatment with the exogenous auxin NAA heightened starch synthesis, while exogenous application of the auxin biosynthesis inhibitor yucasin dampened starch synthesis in tomato roots. Our study observed a slow gravitropic response in cultivated cherry tomato (Aisheng) roots. Time series analysis showed that tomato roots bend toward gravity at a slower rate. Transcriptome analysis revealed that many (2770) differentially expressed genes (DEGs) were identified in roots following 36 h of gravity stimulation. In contrast, only 58 DEGs were detected after 3 h of gravity stimulation, further supporting the slow gravitropic response phenotype of tomato roots. GO and KEGG analysis highlighted auxin response, starch and sugar metabolism, and cell wall modification as the major regulatory pathways involved in the gravitropic response and growth of tomato roots. Our results indicate that auxin mediates root sensing of gravity signals through feedback regulation of starch accumulation and controls root gravitropic bending by regulating the expression of cell wall modification-related genes. Full article

As an endogenous hormone, auxin plays a crucial role in regulating plants’ growth and development, and also in the responses to abiotic stresses. However, the effects and mechanism of auxin and its inhibitors on plant growth and mineral nutrient absorption in citrus have not been thoroughly studied. Therefore, we used trifoliate orange (citrus’s rootstock, Poncirus trifoliata) as the experimental material to supplement the research content in this area. The trifoliate orange seedlings were treated with exogenous auxin (indolebutyric acid, IBA) and auxin inhibitor (2-naphthoxyacetic acid, 2-NOA) in a sand culture system. The results showed that compared to the control, exogenous auxin (1.0 µmol L⁻¹ IBA) significantly enhanced the taproot length, lateral root length, and lateral root number by 17.56%, 123.07%, and 88.89%, respectively, while also markedly elevating the levels of nitrogen (N), phosphorus (P), potassium (K), copper (Cu), and zinc (Zn) by 14.29%, 45.61%, 23.28%, 42.86%, and 59.80%, respectively. Again compared to the control, the auxin inhibitor (50.0 µmol L⁻¹ 2-NOA) dramatically reduced the taproot length, lateral root length, and lateral root number by 21.37%, 10.25%, and 43.33%, respectively, while also markedly decreasing the levels of N, magnesium (Mg), iron (Fe), Cu, and Zn by 7.94%, 10.42%, 24.65%, 39.25%, and 18.76%, respectively. Furthermore, IBA increased auxin accumulation in the root hair, stele, and epidermal tissues of citrus taproots, and promoted the up-regulation of auxin synthesis genes (TAR2, YUC3, YUC4, YUC6, YUC8) and transport genes (ABCB1, ABCB19, AUX1, LAX1, LAX2, PIN1, PIN3, PIN4). In contrast, 2-NOA decreased auxin levels in the root hair, stele, and epidermal tissues of citrus taproots, and was involved in the down-regulation of auxin synthesis genes (TAR2, YUC3, YUC4, YUC6) and transport genes (ABCB1, AUX1, LAX1, LAX2, LAX3, PIN3). Interestingly, 2-NOA dramatically elevated auxin level specifically in the root tip of citrus taproot. Therefore, 2-NOA disrupts auxin reflux from the root tip to root hair and epidermal tissues in citrus taproot through down-regulation of auxin transport genes, thereby creating localized (i.e., root hair zone and epidermal tissues) auxin deficiencies that compromise root system architecture and nutrient acquisition capacity. According to the results of this study, exogenous auxin analogs could regulate citrus growth and mineral nutrient absorption through the auxin synthesis and transport pathways. Full article

Auxins play crucial regulatory roles in plants coping with cadmium (Cd) stress. However, the regulatory mechanism by which auxins alleviate Cd toxicity in tomato seedlings remains unclear. Here, we demonstrate that exposure to Cd stress leads to dynamic changes in the auxin response in tomato roots, characterized by an initial increase followed by a subsequent weakening. Under Cd stress, tomato seedlings show primary root- and hypocotyl-growth inhibition, accompanied by the accumulation of Cd and reactive oxygen species (ROS) in the roots. The exogenous application of 1-naphthylacetic acid (NAA) does not mitigate the inhibitory effect of Cd toxicity on primary root growth, but it does significantly enhance lateral root development under Cd stress. Auxin transport inhibitors, such as 1-N-naphthylphthalamic acid (NPA) and 2,3,5-triiodobenoic acid (TIBA), aggravate the growth inhibition of primary roots caused by Cd stress. Additionally, lateral root development was inhibited by NPA. However, applying auxin synthesis inhibitors L-kynurenine (kyn) and yucasin alleviated the tomato root growth inhibition caused by Cd stress; between them, the effect of yucasin was more pronounced. Yucasin mitigates Cd toxicity in tomato seedlings by reducing Cd²⁺ absorption and auxin accumulation, strengthening ROS scavenging, and reducing cell death in roots. These observations suggest that yucasin potentially mitigates Cd toxicity and improves the tolerance of tomato seedlings to Cd stress. Full article

The optimization of the sink-source relationship is of great importance for crop yield regulation. Cucumber is a typical raffinose family oligosaccharide (RFO)-transporting crop. DNA methylation is a common epigenetic modification in plants, but its role in sink-source regulation has not been demonstrated in RFO-translocating species. Here, whole-genome bisulfite sequencing (WGBS-seq) was conducted to compare the nonfruiting-node leaves (NFNLs) and leaves of fruit setting (FNLs) at the 12th node by removing all female flowers in other nodes of the two treatments. We found considerable differentially methylated genes enriched in photosynthesis and carbohydrate metabolic processes. Comparative transcriptome analysis between FNLs and NFNLs indicated that many differentially expressed genes (DEGs) with differentially methylated regions were involved in auxin, ethylene and brassinolide metabolism; sucrose metabolism; and RFO synthesis pathways related to sink-source regulation. Moreover, DNA methylation levels of six sink-source-related genes in the pathways mentioned above decreased in leaves after 5-aza-dC-2′-deoxycytidine (5-Aza-dC, a DNA methyltransferase inhibitor) treatment on FNLs, and stachyose synthase (CsSTS) gene expression, enzyme activity and stachyose content in RFO synthesis pathway were upregulated, thereby increasing fruit length and dry weight. Taken together, our findings proposed an up-to-date inference for the potential role of DNA methylation in the sink-source relationship, which will provide important references for further exploring the molecular mechanism of DNA methylation in improving the yield of RFO transport plants. Full article

The precise control of free auxin (indole-3-acetic acid, IAA) gradient, which is orchestrated by biosynthesis, conjugation, degradation, hydrolyzation, and transport, is critical for all aspects of plant growth and development. Of these, the GRETCHEN HAGEN 3 (GH3) acyl acid amido synthetase family, pivotal in conjugating IAA with amino acids, has garnered significant interest. Recent advances in understanding GH3-dependent IAA conjugation have positioned GH3 functional elucidation as a hot topic of research. This review aims to consolidate and discuss recent findings on (i) the enzymatic mechanisms driving GH3 activity, (ii) the influence of chemical inhibitor on GH3 function, and (iii) the transcriptional regulation of GH3 and its impact on plant development and stress response. Additionally, we explore the distinct biological functions attributed to IAA-amino acid conjugates. Full article

Growth and development of the Ceratopteris hermaphroditic gametophytes are dependent on cell proliferation in the marginal meristem, which when destroyed will regenerate at a new location on the body margin. We established a laser ablation method to destroy a single initial cell in the meristem. Ablation caused the cessation of cell proliferation accompanied by the disappearance of the expression of an auxin synthesis gene (CrTAA2) and a cell proliferation marker gene (CrWOXB). New meristem regeneration occurred within a predictable distance from the original two days post-ablation, signified by cell proliferation and the expression of CrTAA2. Treatment with the naturally occurring auxin indole-3-acetic acid (IAA), synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4-D), or the transport inhibitor naphthylphthalamic acid (NPA) altered positioning of the original marginal meristem toward the apex of the gametophyte. IAA altered positioning of the regenerated meristem after damaging the original meristem. A model of auxin involvement in the positioning of the marginal meristem in Ceratopteris is presented to encompass these results. Full article

The plant architecture of higher plants is regulated through environmental and genetic factors, as well as phytohormones. Phytohormones play a critical role in regulating shoot branching. We determined the branching phenotype of D16 and N99-6, the content of strigolactones, the genetic expression level, and the interaction between auxin and strigolactones. We found that the branching development of the two soybean varieties under shading was significantly slower than that under normal light. The average branch length of N99-6 decreased by 40.9% after shading; however, the branch length of D16 was not significantly affected. Meanwhile, the branch formation rate in D16 was significantly higher than in N99-6. In addition, after shading treatment, the content of strigolactones in D16 and N99-6 axillary buds increased significantly, and the expression of phytochrome genes, PhyA and PhyB, showed opposite changes. However, strigolactone synthesis gene GmMAX4 and signal transduction gene GmMAX2 expression levels of D16 were lower than those of N99-6 after 24 h of shading. In addition, the application of strigolactone inhibitor TIS108 and auxin inhibitor NPA to soybean had no significant effect on the branch phenotype. The expression of the GmMAX2 gene was significantly up-regulated after the external application of the auxin analog, and the expression of auxin transporter gene GmPINI was significantly down-regulated after external application of the strigolactone analog under shade. In this study, we investigated the adverse effect of shade on soybean branching development, which may be due to the interaction of strigolactones with auxins. Full article

The establishment of plant architecture requires coordination of distinct processes including shoot branching and apical dominance (AD). AD involves the bud apical shoot, mainly through indole-3-acetic acid (IAA) synthetized by the cells of the meristem and young leaves. The rootward flow generates an auxin gradient in the stem and buds, regulating lateral bud (LB) outgrowth. Phytochromes and AD are involved in the shade-avoidance syndrome in woody plants. However, the underlying mechanisms remain poorly understood. The aim of this study was to evaluate the sensitivity of cherry rootstocks to light, mediated by the photoreceptor phytochrome, and its effect on the role of auxin in driving branching by AD. Pharmacological treatments using transport inhibitors and a competitor of IAA were applied to transgenic lines of Colt cherry rootstock, which showed different sensitivities to light because of the ectopic expression of a rice phyA gene. Results showed different physiological behaviours among the transgenic lines and between themselves and the Colt-wt line. Exogenous IBA inhibited Colt-wt LB outgrowth, and this inhibition was less intense in transgenic lines. The IAA-inhibitors and IAA-competitor promoted branching. In in vitro phyA-transgenic plantlets, the ectopic gene induced greater branching and a higher number of buds developed in new shoots. This work confirms a positive action of phytochrome on lateral branching in cherry rootstock, playing a role in the regulation of AD. Moreover, we suggest that the confined in vitro system might now be used as a phenotyping screening to test the plasticity of the response, highlighting the behaviour of modified genotypes due to an ectopic insertion event by simple and rapid procedures. Full article

The mechanisms underlying the de novo regeneration of adventitious roots are still poorly understood, particularly in trees. We developed a system for studying adventitious rooting (AR) at physiological and molecular levels using leaves excised from chestnut microshoots of the same genotype but with two distinct ontogenetic origins that differ in rooting competence. Leaves were treated with auxin and N-1-naphthyl-phthalamic acid (NPA), an inhibitor of auxin polar transport (PAT). The physiological effects were investigated by recording rooting rates and the number and quality of the roots. Molecular responses were examined by localizing and monitoring the changes in the expression of CsSCL1, an auxin-inducible gene in juvenile and mature shoots during AR. The rooting response of leaves was ontogenetic-stage dependent and similar to that of the donor microshoots. Initiation of root primordia and root development were inhibited by application of NPA, although its effect depended on the timing of application. CsSCL1 was upregulated by auxin only in rooting-competent leaves during the novo root organogenesis, and the expression was reduced by NPA. The inhibitory effect on gene expression was detected during the reprograming of rooting competent cells towards root initials in response to auxin, indicating that PAT-mediated upregulation of CsSCL1 is required in the initial steps of AR in chestnut leaves. The localized expression of CsSCL1 in the quiescent center (QC) also suggests a role for this gene in the maintenance of meristematic competence and root radial patterning. Full article

Fruit softening is an important characteristic of peach fruit ripening. The auxin receptor TIR1 (Transport Inhibitor Response 1) plays an important role in plant growth and fruit maturation. Still, little research has been conducted on the relation of TIR1 to the softening of peach fruits. In this study, the hardness of isolated peach fruits was reduced under exogenous NAA treatment at low concentrations. At the same time, the low concentration of NAA treatment reduced the transcription level of PpPG and Ppβ-GAL genes related to cell wall softening and PpACS1 genes related to ethylene synthesis. The transient overexpression of the PpTIR1 gene in peach fruit blocks caused significant down-regulation of the expression of early auxin-responsive genes, ethylene synthesis, and cell wall metabolic genes related to fruit firmness. Through yeast two-hybrid technology, bimolecular fluorescence complementary technology, and a firefly luciferase complementation imaging assay, we were able to unveil an interaction between PpTIR1 and PpIAA1/3/5/9/27 proteins. Furthermore, it was determined that the interaction depended on auxin and its type and concentration. These results show that the PpTIR1-Aux/IAA module has a possible regulatory effect on fruit ripening and softening. Full article

MicroRNAs (miRNAs) belong to non-coding small RNAs which have been shown to take a regulatory function at the posttranscriptional level in plant growth development and response to abiotic stress. Hemerocallis fulva is an herbaceous perennial plant with fleshy roots, wide distribution, and strong adaptability. However, salt stress is one of the most serious abiotic stresses to limit the growth and production of Hemerocallis fulva. To identify the miRNAs and their targets involved in the salt stress resistance, the salt-tolerant H. fulva with and without NaCl treatment were used as materials, and the expression differences of miRNAs–mRNAs related to salt-tolerance were explored and the cleavage sites between miRNAs and targets were also identified by using degradome sequencing technology. In this study, twenty and three significantly differential expression miRNAs (p-value < 0.05) were identified in the roots and leaves of H. fulva separately. Additionally, 12,691 and 1538 differentially expressed genes (DEGs) were also obtained, respectively, in roots and leaves. Moreover, 222 target genes of 61 family miRNAs were validated by degradome sequencing. Among the DE miRNAs, 29 pairs of miRNA targets displayed negatively correlated expression profiles. The qRT-PCR results also showed that the trends of miRNA and DEG expression were consistent with those of RNA-seq. A gene ontology (GO) enrichment analysis of these targets revealed that the calcium ion pathway, oxidative defense response, microtubule cytoskeleton organization, and DNA binding transcription factor responded to NaCl stress. Five miRNAs, miR156, miR160, miR393, miR166, and miR396, and several hub genes, squamosa promoter-binding-like protein (SPL), auxin response factor 12 (ARF), transport inhibitor response 1-like protein (TIR1), calmodulin-like proteins (CML), and growth-regulating factor 4 (GRF4), might play central roles in the regulation of NaCl-responsive genes. These results indicate that non-coding small RNAs and their target genes that are related to phytohormone signaling, Ca²⁺ signaling, and oxidative defense signaling pathways are involved in H. fulva’s response to NaCl stress. Full article