Gibberellin Activity Research Articles

Identification and characterization of the Chenopodium quinoa gibberellin oxidase gene family and its role in seed germination

Pre-harvest sprouting (PHS) is an urgent problem in the quinoa breeding process. Gibberellin oxidases are key enzymes in the biosynthesis and degradation of gibberellins, playing a significant role in the regulation of active gibberellins (GAs). In this study, 18 CqGAox genes were identified in quinoa and characterized according to phylogenetic relationships, gene structure, conserved motifs, codon use pattern, and expression patterns. Based on their phylogenetic relationships, the CqGAox genes were classified into four groups: C19CqGA2ox, C20CqGA2ox, CqGA3ox, and CqGA20ox. Evolutionary analyses showed that homologous GAox genes from Arabidopsis, rice, and quinoa have been subject to purifying selection during evolution. According to the codon use patterns, mutational stress may cause codon bias in CqGAox genes. Prediction of promoter cis-regulatory elements suggested that the CqGAox genes contained multiple elements that responded to phytohormones and stress. We further evaluated the transcriptional responses of CqGAox genes during quinoa seed germination and under low-temperature stress treatments of seedlings. Moreover, qRT-PCR was used to confirm the changes in CqGAox gene expression during quinoa seed germination under GA3 and paclobutrazol (PAC) treatments. Compared with the control, expression of CqGAox genes was significantly altered by treatment with GA3 or PAC. These findings indicated that the CqGAox genes are essential for regulating GA biosynthesis and degradation. This study lays the foundation for further investigations of the roles of CqGAox genes in GA-regulated quinoa seed germination and provides potential targets for addressing the challenge of quinoa PHS.

Study on the physiological mechanism and transcriptional regulatory network of early fruit development in Gleditsia sinensis Lam. (Fabaceae)

BackgroundGleditsia sinensis Lam. (Fabaceae) is a medicinal legume characterized by its spines and pods, which are rich in saponins, polysaccharides, and various specialized metabolites with potential medicinal and industrial applications. The low fruit set rate in artificially cultivated economic forests significantly impedes its development and utilization. A comprehensive understanding of the cellular events, physiological and biochemical processes, and molecular regulatory mechanisms underlying fruit initiation and early fruit development is essential for enhancing yield. However, such information for G. sinensis remains largely unexplored.ResultsIn this study, we identified that the early fruit development process in G. sinensis can be categorized into three distinct stages: pollination, the critical period of fertilization, and the initial fruit development followed by subsequent growth. The dynamic changes in non-structural carbohydrates and endogenous plant hormones within the ovary were found to play a significant role during fruit set and the early stages of fruit development. Additionally, the high activity of gibberellin, cytokinin, and sucrose-metabolizing enzymes in the ovary was conducive to early fruit development. Furthermore, we generated high-resolution spatiotemporal gene expression profiles in the ovary from the stage of efflorescence to early fruit development. Comparative transcriptomics and weighted gene co-expression network analysis revealed specific genes and gene modules predominant at distinct developmental stages, thereby highlighting unique genetic programming. Overall, we identified the potential regulatory network governing fruit initiation and subsequent development, as well as the sets of candidate genes involved, based on the aforementioned results.ConclusionsThe results offer a valuable reference and resource for the application of exogenous substances, such as hormones and sugars, during critical fruit development periods, and for the development of molecular tools aimed at improving yield.

ZmGDIα-hel counters the RBSDV-induced reduction of active gibberellins to alleviate maize rough dwarf virus disease

Maize rough dwarf disease (MRDD) threatens maize production globally. The P7-1 effector of the rice black-streaked dwarf virus (RBSDV) targets maize Rab GDP dissociation inhibitor alpha (ZmGDIα) to cause MRDD. However, P7-1 has difficulty recruiting a ZmGDIα variant with an alternative helitron-derived exon 10 (ZmGDIα-hel), resulting in recessive resistance. Here, we demonstrate that P7-1 can recruit another maize protein, gibberellin 2-oxidase 13 (ZmGA2ox7.3), which also exhibits tighter binding affinity for ZmGDIα than ZmGDIα-hel. The oligomerization of ZmGA2ox7.3 is vital for its function in converting bioactive gibberellins into inactive forms. Moreover, the enzymatic activity of ZmGA2ox7.3 oligomers increases when forming hetero-oligomers with P7-1/ZmGDIα, but decreases when ZmGDIα-hel replaces ZmGDIα. Viral infection significantly promotes ZmGA2ox7.3 expression and oligomerization in ZmGDIα-containing susceptible maize, resulting in reduced bioactive GA1/GA4 levels. This causes an auxin/cytokinin imbalance and ultimately manifests as MRDD syndrome. Conversely, in resistant maize, ZmGDIα-hel counters these virus-induced changes, thereby mitigating MRDD severity.

MeGT2.6 increases cellulose synthesis and active gibberellin content to promote cell enlargement in cassava.

Cassava, a pivotal tropical crop, exhibits rapid growth and possesses a substantial biomass. Its stem is rich in cellulose and serves as a crucial carbohydrate storage organ. The height and strength of stems restrict the mechanised operation and propagation of cassava. In this study, the triple helix transcription factor MeGT2.6 was identified through yeast one-hybrid assay using MeCesA1pro as bait, which is critical for cellulose synthesis. Over-expression and loss-of-function lines were generated, and results revealed that MeGT2.6 could promote a significant increase in the plant height, stem diameter, cell size and thickness of SCW of cassava plant. Specifically, MeGT2.6 upregulated the transcription activity of MeGA20ox1 and downregulated the expression level of MeGA2ox1, thereby enhancing the content of active GA3, resulting in a large cell size, high plant height and long stem diameter in cassava. Moreover, MeGT2.6 upregulated the transcription activity of MeCesA1, which promoted the synthesis of cellulose and hemicellulose and produced a thick secondary cell wall. Finally, MeGT2.6 could help supply additional substrates for the synthesis of cellulose and hemicellulose by upregulating the invertase genes (MeNINV1/6). Thus, MeGT2.6 was found to be a multiple regulator; it was involved in GA metabolism and sucrose decomposition and the synthesis of cellulose and hemicellulose.

JrGA20ox1-transformed rootstocks deliver drought response signals to wild-type scions in grafted walnut.

Targeted regulation using transgrafting technology has become a trend. However, the mechanisms of transgene-derived signal communication between rootstocks and scions remain unclear in woody plants. Here, we grafted wild-type (WT) walnut (Juglans regia L.) on WT (WT/WT), JrGA20ox1 (encodes a gibberellin 20-oxidase)-overexpressing (WT/OE), and JrGA20ox1-RNAi transformation (WT/RNAi) walnut in vitro. We aimed to elucidate the mechanisms of JrGA20ox1-derived signal communication under PEG-simulated drought stress between rootstocks and scions in walnut. We demonstrated that JrGA20ox1-OE and JrGA20ox1-RNAi rootstocks could transport active gibberellins (GAs) and JrGA20ox1-RNAi vector-produced sRNAs to WT scions under PEG-simulated drought stress, respectively. The movement of sRNAs further led to a successive decline in JrGA20ox1 expression and active GA content. Meanwhile, unknown mobile signals may move between rootstocks and scions. These mobile signals reduced the expression of a series of GA-responsive and GA-non-responsive genes, and induced ROS production in guard cells and an increase in ABA content, which may contribute to the drought tolerance of WT/RNAi, while the opposite occurred in WT/OE. The findings suggest that JrGA20ox1-derived rootstock-to-scion movement of signals is involved in drought tolerance of scions. Our research will provide a feasible approach for studying signal communication in woody plants.

Overexpression of MsDREB1C Modulates Growth and Improves Forage Quality in Tetraploid Alfalfa (Medicago sativa L.).

DREB has been reported to be involved in plant growth and response to environmental factors. However, the function of DREB in growth and development has not been elucidated in alfalfa (Medicago sativa L.), a perennial tetraploid forage cultivated worldwide. In this study, an ortholog of MtDREB1C was characterized from alfalfa and named MsDREB1C accordingly. MsDREB1C was significantly induced by abiotic stress. The transcription factor MsDREB1C resided in the nucleus and had self-transactivation activity. The MsDREB1C overexpression (OE) alfalfa displayed growth retardation under both long-day and short-day conditions, which was supported by decreased MsGA20ox and upregulated MsGA2ox in the OE lines. Consistently, a decrease in active gibberellin (GA) was detected, suggesting a negative effect of MsDREB1C on GA accumulation in alfalfa. Interestingly, the forage quality of the OE lines was better than that of WT lines, with higher crude protein and lower lignin content, which was supported by an increase in the leaf-stem ratio (LSR) and repression of several lignin-synthesis genes (MsNST, MsPAL1, MsC4H, and Ms4CL). Therefore, this study revealed the effects of MsDREB1C overexpression on growth and forage quality via modifying GA accumulation and lignin synthesis, respectively. Our findings provide a valuable candidate for improving the critical agronomic traits of alfalfa, such as overwintering and feeding value of the forage.

Highlights in gibberellin research: A tale of the dwarf and the slender.

It has been almost a century since biologically active gibberellin (GA) was isolated. Here, we give a historical overview of the early efforts in establishing the GA biosynthesis and catabolism pathway, characterizing the enzymes for GA metabolism, and elucidating their corresponding genes. We then highlight more recent studies that have identified the GA receptors and early GA signaling components (DELLA repressors and F-box activators), determined the molecular mechanism of DELLA-mediated transcription reprograming, and revealed how DELLAs integrate multiple signaling pathways to regulate plant vegetative and reproductive development in response to internal and external cues. Finally, we discuss the GA transporters and their roles in GA-mediated plant development.

Application of In Silico Analysis to Determine Morphogenesis in Plant Tissue Culture

The work proposed and tested a new approach to optimizing biotechnological processes, including the process of microclonal propagation. The proposed method is based on constructing a map of the similarity of the structures of molecules of secondary metabolites of plant extracts and molecules that regulate the processes of plant morphogenesis (primarily phytohormones) with subsequent prediction of the action of the extract. Lichen extract of Cetraria islandica (L.) Ach. (Parmeliaceae)was used as an example, for which the range of secondary metabolites contained is well known. The structural similarity of aliphatic secondary compounds of lichen (protolichesteric and lichesteric acids) with strigolactones (to a greater extent), as well as with gibberellins and brassinosteroids, was revealed. Based on the analysis of the results obtained, a prediction was made about the dose-dependent effect of the lichen extract of C. islandica on growth processes and rhizogenesis of microshootsin vitro. This hypothesis was experimentally tested in experiments with microclonal propagation of higher plants Lonicera caerulea L. and Populus tremula L. As a result of the work carried out, it was established that the addition of extract from C. islandicaat a concentration of 10–50 mg/L in the nutrient medium increased the multiplication rate of L. caerulea (by 31%) and P. tremula (by 8%). The rhizogenic activity of the lichen extract at the same concentrations (10–50 mg/L of medium), similar to the activity of strigolactones and gibberellins, has been experimentally proven. The extract has also been shown to have a positive effect of C. islandica (50 mg/L) on elongation of microshoots of both cultures and hemogenesisof P. tremula.The proposed approach allows for optimizing studies aimed at identifying the effect of various extracts on plant morphogenesis in vitroby preliminary constructing a map of the similarity of secondary metabolites contained in extracts (including according to literature data) and known growth regulators (including phytohormones) with subsequent prediction of the effect of the extract.

Growth Responses to two Commercial Plant Growth Retardants with Different Concentration in Potted Geranium

Background and objective: The aim of this study was to elucidate the effect of varying treatment concentrations of two different plant growth retardants, acting at different inhibition positions during the biosynthetic process, on the growth characteristics of geranium (<i>Pelargonium</i> × <i>hortorum</i>) and then to provide essential data to improve the quality of pot plant production.Methods: Aqueous solutions of diniconazole (Binnari) and daminozide (B-9) at concentrations of 0 (distilled water), 50, 100, 200, and 400 mg⋅L<sup>-1</sup> were prepared for the geranium 'Ringo 2000TM Deep Red'. These solutions were applied by foliar spraying twice at 10-day intervals. Growth characteristics such as plant height, leaf size and relative chlorophyll content (SPAD) were then assessed.Results: When Binnari was applied at concentrations of 0, 50, 100, 200, and 400 mg⋅L<sup>-1</sup> , the corresponding plant heights were 29.5, 23.8, 24.1, 23.0, and 18.4 cm, respectively. There was a statistically significant decrease in plant height with increasing concentration. Conversely, for B-9, plant height was significantly reduced to 26.7 cm only at the 400 mg⋅L<sup>-1</sup> concentration, indicating a less pronounced inhibitory effect compared to Binnari. Leaf width showed a significant decreasing trend with increasing Binnari application concentrations and was 4.6, 4.7, 4.1, 4.2, and 3.9 cm respectively. However, there was no significant difference due to the B-9 treatment. Chlorophyll content increased with increasing concentrations of both retardants.Conclusion: In conclusion, Binnari showed a greater inhibitory effect on geranium growth compared to B-9. The application of Binnari at 400 mg⋅L<sup>-1</sup> resulted in the greatest increase in the ornamental value of the geranium. Furthermore, diniconazole acts during the early stage before the splitting of the GA<sub>1</sub> and GA<sub>4</sub> pathways, thus inhibiting the biosynthesis of both active gibberellins. In contrast, daminozide inhibits the step just before GA<sub>1</sub> biosynthesis. It is concluded that GA<sub>4</sub> serves as the dominant active gibberellin in geranium plants.

Manifestation of Triploid Heterosis in the Root System after Crossing Diploid and Autotetraploid Energy Willow Plants.

Successful use of woody species in reducing climatic and environmental risks of energy shortage and spreading pollution requires deeper understanding of the physiological functions controlling biomass productivity and phytoremediation efficiency. Targets in the breeding of energy willow include the size and the functionality of the root system. For the combination of polyploidy and heterosis, we have generated triploid hybrids (THs) of energy willow by crossing autotetraploid willow plants with leading cultivars (Tordis and Inger). These novel Salix genotypes (TH3/12, TH17/17, TH21/2) have provided a unique experimental material for characterization of Mid-Parent Heterosis (MPH) in various root traits. Using a root phenotyping platform, we detected heterosis (TH3/12: MPH 43.99%; TH21/2: MPH 26.93%) in the size of the root system in soil. Triploid heterosis was also recorded in the fresh root weights, but it was less pronounced (MPH%: 9.63-19.31). In agreement with root growth characteristics in soil, the TH3/12 hybrids showed considerable heterosis (MPH: 70.08%) under in vitro conditions. Confocal microscopy-based imaging and quantitative analysis of root parenchyma cells at the division-elongation transition zone showed increased average cell diameter as a sign of cellular heterosis in plants from TH17/17 and TH21/2 triploid lines. Analysis of the hormonal background revealed that the auxin level was seven times higher than the total cytokinin contents in root tips of parental Tordis plants. In triploid hybrids, the auxin-cytokinin ratios were considerably reduced in TH3/12 and TH17/17 roots. In particular, the contents of cytokinin precursor, such as isopentenyl adenosine monophosphate, were elevated in all three triploid hybrids. Heterosis was also recorded in the amounts of active gibberellin precursor, GA19, in roots of TH3/12 plants. The presented experimental findings highlight the physiological basics of triploid heterosis in energy willow roots.

Screening Candidate Genes at the Co Locus Conferring to the Columnar Growth Habit in Apple (Malus × Domestica Borkh.).

The columnar growth trait of apple (Malus × domestica Borkh.) is genetically controlled by the Columnar (Co) locus on 10 chromosomes, including several candidate genes. Except for MdCo31, other candidate genes at the Co locus are less elucidated. In this study, a strategy of step-by-step screening was adopted to select 11 candidate genes by experimental cloning, transient expression, and genetic transformation. There existed several SNPs in four genes by sequence alignment in columnar and non-columnar apples. Two genes were detected in the nucleus and three genes in the cell membrane, other genes were located in multiple cellular structures by subcellular location. Ectopic expression demonstrated that more branching occurred in MdCo38-OE by upregulating NtPIN1 and NtGA2ox and enlarged leaves in MdCo41-OE tobaccos by upregulating NtCCDs. Transcripts of MdCo38 and MdCo41 were associated with the Co genotypes in apples. The results indicate that MdCo38 and MdCo41 are involved in the columnar growth phenotype in apple, probably through altering polar auxin transport, active gibberellin levels, and strigolactone biosynthesis.

Penicillium citrinum Provides Transkingdom Growth Benefits in Choy Sum (Brassica rapa var. parachinensis).

Soil-borne beneficial microbes establish symbioses with plant hosts and play key roles during growth and development therein. In this study, two fungal strains, FLP7 and B9, were isolated from the rhizosphere microbiome associated with Choy Sum (Brassica rapa var. parachinensis) and barley (Hordeum vulgare), respectively. Sequence analyses of the internal transcribed spacer and 18S ribosomal RNA genes combined with colony and conidial morphology identified FLP7 and B9 to be Penicillium citrinum strains/isolates. Plant-fungus interaction assays revealed that isolate B9 showed significant growth promotion effects in Choy Sum plants cultivated in normal soil, as well as under phosphate-limiting conditions. In comparison to the mock control, B9-inoculated plants showed a 34% increase in growth in aerial parts and an 85% upsurge in the fresh weight of roots when cultivated in sterilized soil. The dry biomass of such fungus-inoculated Choy Sum increased by 39% and 74% for the shoots and roots, respectively. Root colonization assays showed that P. citrinum associates directly with the root surface but does not enter or invade the root cortex of the inoculated Choy Sum plants. Preliminary results also indicated that P. citrinum can promote growth in Choy Sum via volatile metabolites too. Interestingly, we detected relatively higher amounts of gibberellins and cytokinins in axenic P. citrinum culture filtrates through liquid chromatography-mass spectrometry analyses. This could plausibly explain the overall growth induction in P. citrinum-inoculated Choy Sum plants. Furthermore, the phenotypic growth defects associated with the Arabidopsis ga1 mutant could be chemically complemented by the exogenous application of P. citrinum culture filtrate, which also showed accumulation of fungus-derived active gibberellins. Our study underscores the importance of transkingdom beneficial effects of such mycobiome-assisted nutrient assimilation and beneficial fungus-derived phytohormone-like metabolites in the induction of robust growth in urban farmed crops.

Base Editing of EUI1 Improves the Elongation of the Uppermost Internode in Two-Line Male Sterile Rice Lines

The use of male sterile lines (MSLs) of rice is essential for heterosis utilization. However, MSLs have a common defect in the elongation of the uppermost internode, eventually leading to incomplete panicle exsertion, blocking pollination, and reducing the hybrid rice seed yield. Previously, the elongated uppermost internode 1 (EUI1) was identified as an active gibberellin-deactivating enzyme that plays a key role in panicle exsertion from the flag leaf sheath in rice (Oryza sativa L.). We used an adenine base editor to edit EUI1 and obtained two types of homozygous transgenic plants (eui1-1 and eui1-2). The transcription and translation levels of EUI1 in the two mutants were significantly lower than in the wild type, as was the oxidation activity of EUI1 to active gibberellins (GAs), which also decreased. The contents of the plant hormones GA1, GA3, and GA4 in eui1-1 (1.64, 1.55, and 0.92 ng/g) and eui1-2 (0.85, 0.64, and 0.65 ng/g) panicles were significantly higher than the wild type (0.70, 0.57, and 0.42 ng/g). The uppermost internode lengths of the mutant were 26.5 and 23.6 cm, which were significantly longer than that of the wild type (18.0 cm), and the cell lengths of the mutant were 161.10 and 157.19 μm, which were longer than that of the wild type (89.28 μm). Our results indicate that the adenine base editing system could increase the content of endogenous bioactive GAs in young panicles by fine-tuning EUI1 activity, reduce the defect of panicle enclosure in MSLs and increase the yield of hybrid rice seed production.

PsRGL1 negatively regulates chilling- and gibberellin-induced dormancy release by PsF-box1-mediated targeting for proteolytic degradation in tree peony.

Tree peony bud endodormancy is a common survival strategy similar to many perennial woody plants in winter, and the activation of the GA signaling pathway is the key to breaking endodormancy. GA signal transduction is involved in many physiological processes. Although the GA-GID1-DELLA regulatory module is conserved in many plants, it has a set of specific components that add complexity to the GA response mechanism. DELLA proteins are key switches in GA signaling. Therefore, there is an urgent need to identify the key DELLA proteins involved in tree peony bud dormancy release. In this study, the prolonged chilling increased the content of endogenously active gibberellins. PsRGL1 among three DELLA proteins was significantly downregulated during chilling- and exogenous GA3-induced bud dormancy release by cell-free degradation assay, and a high level of polyubiquitination was detected. Silencing PsRGL1 accelerated bud dormancy release by increasing the expression of the genes associated with dormancy release, including PsCYCD, PsEBB1, PsEBB3, PsBG6, and PsBG9. Three F-box protein family members responded to chilling and GA3 treatments, resulting in PsF-box1 induction. Yeast two-hybrid and BiFC assays indicated that only PsF-box1 could bind to PsRGL1, and the binding site was in the C-terminal domain. PsF-box1 overexpression promoted dormancy release and upregulated the expression of the dormancy-related genes. In addition, yeast two-hybrid and pull-down assays showed that PsF-box1 also interacted with PsSKP1 to form an E3 ubiquitin ligase. These findings enriched the molecular mechanism of the GA signaling pathway during dormancy release, and enhanced the understanding of tree peony bud endodormancy.

Brassinosteroids Mediate Endogenous Phytohormone Metabolism to Alleviate High Temperature Injury at Panicle Initiation Stage in Rice

High temperatures cause physiological and biochemical changes and significantly affect young panicle development of rice (Oryza sativa L.). Brassinosteroids play important roles in enhancing crop stress resistance. In this study, we subjected rice cultivars Huanghuazhan (heat-resistant) and IR36 (heat-sensitive) to high temperature (HT, 40 ºC) or normal temperature (NT, 33 ºC) for 7 d at the panicle initiation stage, in conjunction with application of 24-epibrassinolide [EBR, a synthetic brassinolide (BR)] or brassinazole (BRZ, a BR biosynthesis inhibitor) at the beginning of the treatments. HT exacerbated spikelet degeneration and inhibited young panicle growth, which were partially prevented by EBR application, while BRZ application aggravated the reduction in spikelet number. HT decreased the contents of BR, active cytokinins (aCTK), active gibberellins (aGA) and indole-3-acetic acid (IAA), but increased the content of abscisic acid (ABA) in young panicles. The activities of key enzymes involved in sucrose hydrolysis, glycolysis and the tricarboxylic acid cycle in young panicles were decreased with the change of endogenous hormone levels under HT. In addition, the contents of H2O2 and malondialdehyde (MDA) were increased and the activities of antioxidant enzymes were decreased in young panicles. Exogenous application of EBR induced the expression of phytohormone biosynthesis-related genes and down-regulated the expression of phytohormone catabolism-related genes to increase the contents of endogenous BR, aCTK, aGA and ABA, thus promoting the decomposition and utilization of sucrose in young panicles, enhancing the activities of superoxide dismutase, catalase and peroxidase, and reducing the accumulation of H2O2 and MDA in young panicles, whereas application of BRZ had the opposite physiological effects. These results showed that brassinosteroids mediate endogenous phytohormone metabolism to alleviate HT injury at the panicle initiation stage in rice.

Study on the flowering in rice plant (Oryza sativa cv. OM5451)

Introduction: The consumption of rice is rising in response to worldwide population development. The purpose of this study was to determine the stages of plant development, specifically the transition stage from panicle initiation to the flowering stage, which determines the number of spikelets per branch. In addition, this research studied the changes in physiology and morphology during the flowering development of rice in a garden. Methods: Plant height, number of leaves, number of tillers, and flowering time were all measured. Plant growth regulators such as cytokinin, auxin, gibberellic acid (GA), and acid abscisic (ABA) in the apical shoot at different stages of flower development were analyzed. Results: The activities of indole acetic acid (IAA), zeatin, and gibberellins in the apical shoot of the stem elongation stage are higher than those in plants of the panicle initiation stage. At the flowering stage, the rates of photosynthesis and respiration of the leaves at positions 1, 2, and 3 were higher than those of the other leaves. In addition, photosynthetic pigments at leaf positions 2, 3, and 4 were higher than those at positions 1 and 3. Conclusion: The results show that the relationship between changes in morphological, physiological, and biochemical indicators during the rice growth stages was discussed.

OsHLS1 regulates plant height and development by controlling active gibberellin accumulation in rice (Oryza sativa L.)

In this study, we identified a gene related to plant height, leaf, and premature senescence in rice, and named it OsHLS1. Through bioinformatics analysis, it was found that this gene belongs to a new gene family-HLS family, and this gene family exists widely in higher plants. Expression of OsHLS1 was significantly brought about by gibberellin (GA). Subcellular localization showed that OsHLS1 was located in the nucleus. oshls1–3 displayed a GA-deficient phenotype, with dwarf plants. In addition, oshls1–3 also showed premature senescence, shorter and narrower leaves, and pollen abortion. Exogenous GA3 can restore the plant height of oshls1–3. Histomorphological analysis showed that the gene affected the progress of internode cells in the first and third nodes under the rice panicle. Through the verification of the homologous gene AT4G25690 in Arabidopsis, it was found that the mutant at4g25690 lines also showed plant dwarfing, premature senescence, and shortening and narrowing of leaves and pollen abortion. OsHLS1 affected the expression levels of genes involved in the GA metabolic pathway and affected the content of active GA, thereby regulating plant height development in rice. In conclusion, we suggest that OsHLS1 regulates plant height and development by controlling the accumulation of active gibberellins in rice.

The Interaction between Hydromulching and Arbuscular Mycorrhiza Improves Escarole Growth and Productivity by Regulating Nutrient Uptake and Hormonal Balance.

To improve water and nutrient use efficiencies some strategies have been proposed, such as the use of mulching techniques or arbuscular mycorrhizal fungi (AMF) inoculation. To gain insights into the interaction between the use of hydromulch and AMF inoculation on plant growth and productivity, escarole plants (Cichorium endivia, L.) were inoculated with the AMF Rhizophagus irregularis and grown with non-inoculated plants under different soil cover treatments: ecological hydromulching based on the substrate of mushroom cultivation (MS), low-density black polyethylene (PE), and non-covered soil (BS). AMF inoculation or the use of mulching alone, but especially their interaction, increased the plant growth. The growth improvement observed in AMF-inoculated escarole plants grown under hydromulching conditions was mainly associated with the upgrading of nitrogen and phosphorous use efficiency through the regulation of the hormonal balance. Both hydromulching and AMF inoculation were found to increase the active gibberellins (GAs) and cytokinins (CKs), resulting in a positive correlation between these hormones and the growth-related parameters. In contrast, the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) and abscisic acid (ABA) decreased in AMF-inoculated plants and especially in those grown with the MS treatment. This study demonstrates that there exists a positive interaction between AMF and hydromulching which enhances the growth of escarole plants by improving nutrient use efficiency and hormonal balance.

MdCo31 interacts with an RNA polymerase II transcription subunit 32 to regulate dwarf growth with short internodes in columnar apple

The dominant Co locus controls the columnar growth phenotype of apple (Malus × domestica) trees. Candidate gene MdCo31, encoding 2-oxoglutarate-dependent dioxygenase, causes dwarf growth with short internodes in transgenic plants by reducing the abundance of biologically active gibberellin. However, the pathway regulating MdCo31 in the dwarfism of apple trees remains unclear. In this study, expression of MdCo31 was proved to be negatively correlated with internode length in F1 populations created by crossing columnar parents, and with dwarfism in transgenic apple plantlets. Yeast (Saccharomyces cerevisiae) two-hybrid screening identified the RNA polymerase II transcription subunit MdMED32 as putative interactor of MdCo31. Bimolecular fluorescence complementation, co-immunoprecipitation, and dual-luciferase reporter assays confirmed this interaction both in vivo and in vitro. Ectopic expression of MdMED32 in Nicotiana tabacum led to a dwarf phenotype, similar to that of MdCo31 transgenic apple plants. Expression of GA2ox1 and GA20ox1, encoding key enzymes of gibberellin metabolism, was upregulated in transgenic plants. Transient transcriptional activity demonstrated that MdMED32 functioned as an activator, promoting expression of MdGA2ox1 and MdGA20ox1. These findings indicate that the interaction between MdCo31 and MdMED32 functions in the regulation of internode length in columnar apple.

Cloning and Functional Identification of Gibberellin Receptor SvGID1s Gene of Salix viminalis.

Gibberellins (GAs) play a key role in the transition from vegetative growth to flowering and the GA receptor GID1 (GIBBERELLIN INSENSITIVE DWARF1) is the central part of GA-signaling. The differential expression of SvGID1 was found in the transcriptome sequencing in our previous study, which was further verified at different stages of flowering of Salix viminalis. In order to reveal the function GID1 of S. viminalis, two genes of SvGID1b and SvGID1c were cloned and transformed into Arabidopsis thaliana, respectively. The results showed that the full ORF length of SvGID1b and SvGID1c genes were both 1035bp, encoding 344 amino acids, which were typical globular proteins. The peptide chain contained more α-helix structure, and had 99% similarity with GID1b and GID1c amino acid sequences of Salix suchowensis. Phylogenetic analysis showed that SvGID1s had close genetic relationship with woody plants such as Populus alba and Populus tomentosa, and had far genetic relationship with rice. After overexpression in A. thaliana, the total gibberellin, active gibberellin content and the expression level of GA3ox1, the key gene for GA4 synthesis, were not significantly different from those in the wild-type, while the expression levels of FUL, SOC1 and FT, the key genes for flowering in plants, were increased, and the expression levels of FLC and GAI were decreased. The ectopic expression of SvGID1s increased the sensitivity of plants to gibberellin and enhanced gibberellin effect, caused early bolting, budding and flowering, led to higher plant, longer hypocotyl and other phenomena. The results provide a theoretical basis for clarifying the regulation of gibberellin on flower bud differentiation of flowering plants.