Gibberellic Acid Biosynthesis Research Articles

Ameliorative Effects of Paclobutrazol via Physio-Biochemical and Molecular Manifestation in Rice under Water Deficit Stress

We investigated the effects of an optimized dose (100 mg/kg) of paclobutrazol (PBZ) on various physio-biochemical and molecular parameters in three rice varieties (N22, IR64, and IR64 DTY1.1) under water deficit stress (WDS). Concurrently, a dose of 100 mg/kg of PBZ was applied through drenching. Samples were collected at both mild stress [75%‒80% relative water content (RWC)] and severe stress (60%‒65% RWC) conditions. Physio-biochemical parameters, including dry matter, RWC, membrane stability index (MSI), grain yield, total chlorophyll content, chlorophyll stability index, drought susceptibility index, drought tolerance efficiency, total antioxidant activity, abscisic acid (ABA) content, and gibberellic acid (GA) content, were measured. The results showed that PBZ treatment positively influenced the physio-biochemical parameters, significantly increasing dry matter (16.27%‒ 61.91%), RWC (6.48%‒16.34%), MSI (4.37%‒10.35%), and total chlorophyll content (8.97%‒29.09%) in the rice varieties under both mild and severe WDS. Moreover, PBZ treatment reduced drought susceptibility (0.83‒0.95) and enhanced drought tolerance efficiency (60.92%‒86.78%), indicating its potential as a stress-mitigating agent. This study also explored the molecular responses of the rice varieties to PBZ treatment under WDS. Global methylation analysis revealed changes in DNA methylation patterns, indicating the regulatory influence of PBZ on gene expression. The expression analysis of genes involved in the diversification of geranylgeranyl pyrophosphate towards the biosynthesis of ABA, GA, and chlorophyll showed alterations in their expression levels, suggesting the involvement of PBZ in the isoprenoid pathway. Overall, this study provides valuable insights into the potential mechanisms by which PBZ modulates physiological and molecular responses in rice plants under WDS. The findings highlight the importance of PBZ as a promising tool for enhancing drought tolerance in rice and offer valuable information for future research in crop stress management.

Functional analysis of a wheat class III peroxidase gene, TaPer12-3A, in seed dormancy and germination

BackgroundClass III peroxidases (PODs) perform crucial functions in various developmental processes and responses to biotic and abiotic stresses. However, their roles in wheat seed dormancy (SD) and germination remain elusive.ResultsHere, we identified a wheat class III POD gene, named TaPer12-3A, based on transcriptome data and expression analysis. TaPer12-3A showed decreasing and increasing expression trends with SD acquisition and release, respectively. It was highly expressed in wheat seeds and localized in the endoplasmic reticulum and cytoplasm. Germination tests were performed using the transgenic Arabidopsis and rice lines as well as wheat mutant mutagenized with ethyl methane sulfonate (EMS) in Jing 411 (J411) background. These results indicated that TaPer12-3A negatively regulated SD and positively mediated germination. Further studies showed that TaPer12-3A maintained H2O2 homeostasis by scavenging excess H2O2 and participated in the biosynthesis and catabolism pathways of gibberellic acid and abscisic acid to regulate SD and germination.ConclusionThese findings not only provide new insights for future functional analysis of TaPer12-3A in regulating wheat SD and germination but also provide a target gene for breeding wheat varieties with high pre-harvest sprouting resistance by gene editing technology.

Differential expression of antimicrobial metabolites, phenylpropanoid and phytohormone metabolic pathway genes determines resistance or susceptibility to Ascochyta rabiei in chickpea

AbstractBlight caused by Ascochyta rabiei is a major constraint in the productivity of chickpea (Cicer arietinum). The mechanisms governing resistance/susceptibility to blight in chickpea are poorly understood. We used a blight‐resistant (HC1) and a blight‐susceptible (GPF2) genotype of chickpea and genes of pathogenesis‐related proteins (PRPs), phenylpropanoid pathway metabolites, abscisic acid (ABA), gibberellic acid (GA) and jasmonic acid (JA) to understand the role of these in A. rabiei resistance/susceptibility. The JA, ABA and GA biosynthesis genes of chickpea were retrieved, characterized and gene‐specific primers were used for transcriptional studies. Gene expression revealed that chickpea activated its defences rather quickly and well before initiation of spore germination. In resistant HC1, the majority of the JA, GA and phenylpropanoid pathway genes had peak maxima at 2 h post‐inoculation (hpi) whereas PRPs/defence genes had peak maxima at 24/36 hpi implying that defence to A. rabiei in chickpea is composed of a two‐tier system separated by time: immediately after spore attachment and at or just prior to host penetration. Unlike HC1, susceptible GPF2 was late in activation of defence responses or did not activate them. Another striking difference between HC1 and GPF2 was up‐regulation of ABA biosynthesis genes in inoculated GPF2 and down‐regulation in HC1. This study revealed that phenylpropanoids, PRPs, JA, 8‐(1R,2R)‐3‐oxo‐2‐(Z)‐pent‐2‐enyl cyclopentyl octanoate, (15Z)‐12‐oxophyto‐10,15‐dienoic acid and methyl‐jasmonate govern resistance to A. rabiei in chickpea whereas ABA governs susceptibility.

GRAS gene family in rye (Secale cereale L.): genome-wide identification, phylogeny, evolutionary expansion and expression analyses

BackgroundThe GRAS transcription factor family plays a crucial role in various biological processes in different plants, such as tissue development, fruit maturation, and environmental stress. However, the GRAS family in rye has not been systematically analyzed yet.ResultsIn this study, 67 GRAS genes in S. cereale were identified and named based on the chromosomal location. The gene structures, conserved motifs, cis-acting elements, gene replications, and expression patterns were further analyzed. These 67 ScGRAS members are divided into 13 subfamilies. All members include the LHR I, VHIID, LHR II, PFYRE, and SAW domains, and some nonpolar hydrophobic amino acid residues may undergo cross-substitution in the VHIID region. Interested, tandem duplications may have a more important contribution, which distinguishes them from other monocotyledonous plants. To further investigate the evolutionary relationship of the GRAS family, we constructed six comparative genomic maps of homologous genes between rye and different representative monocotyledonous and dicotyledonous plants. The response characteristics of 19 ScGRAS members from different subfamilies to different tissues, grains at filling stages, and different abiotic stresses of rye were systematically analyzed. Paclobutrazol, a triazole-based plant growth regulator, controls plant tissue and grain development by inhibiting gibberellic acid (GA) biosynthesis through the regulation of DELLA proteins. Exogenous spraying of paclobutrazol significantly reduced the plant height but was beneficial for increasing the weight of 1000 grains of rye. Treatment with paclobutrazol, significantly reduced gibberellin levels in grain in the filling period, caused significant alteration in the expression of the DELLA subfamily gene members. Furthermore, our findings with respect to genes, ScGRAS46 and ScGRAS60, suggest that these two family members could be further used for functional characterization studies in basic research and in breeding programmes for crop improvement.ConclusionsWe identified 67 ScGRAS genes in rye and further analysed the evolution and expression patterns of the encoded proteins. This study will be helpful for further analysing the functional characteristics of ScGRAS genes.

Process parameters optimization for biosynthesis of gibberellic acid using orange waste as an alternative substrate

Gibberellic acid (GA3) is the most important gibberellin that affects various plant developmental processes. As such, it has an international market value of $106 million dollars. However, it is costly in the market because of high downstream processing cost that is linked to its current mode of production via submerged fermentation. Solid-state fermentation, in contrast, minimizes this limitation of cost when optimal process parameters are used. Hence, this study utilized orange waste as a cheap substrate in the bio-production of gibberellic acid through the optimization of three process parameters (incubation temperature, pH and substrate concentration). The study also investigated the interactive effects of these parameters on the yield of gibberellic acid. Using Design Expert® software, a 19-run experiment, varied at five levels, was generated. Accordingly, each fermentation reaction that had been supplemented with 0.03% FeSO4.7H2O and 0.01% (NH4)2SO4 was inoculated with a constant fungal inoculum. The highest yield of gibberellic acid was with a combination of 19.5 g of substrate set at 30 °C incubation temperature with a pH of 5.5. The interaction between the three factors was a linear relationship. The bio-production of gibberellic acid using orange waste as an alternative substrate suggests the possibility of a further reduction in cost of production of high-end value metabolites when proper optimization is carried out.

Meiotic instability and irregular chromosome pairing underpin heat-induced infertility in bread wheat carrying the Rht-B1b or Rht-D1b Green Revolution genes.

Mutations in the Rht-B1a and Rht-D1a genes of wheat (Triticum aestivum; resulting in Rht-B1b and Rht-D1b alleles) cause gibberellin-insensitive dwarfism and are one of the most important elements of increased yield introduced during the 'Green Revolution'. We measured the effects of a short period of heat imposed during the early reproductive stage on near-isogenic lines carrying Rht-B1b or Rht-D1b alleles, with respect to the wild-type (WT). The temperature shift caused a significant fertility loss within the ears of Rht-B1b and Rht-D1b wheats, greater than that observed for the WT. Defects in chromosome synapsis, reduced homologous recombination and a high frequency of chromosome mis-segregation were associated with reduced fertility. The transcription of TaGA3ox gene involved in the final stage of gibberellic acid (GA) biosynthesis was activated and ultra-performance liquid chromatography-tandem mass spectrometry identified GA1 as the dominant bioactive GA in developing ears, but levels were unaffected by the elevated temperature. Rht-B1b and Rht-D1b mutants were inclined to meiotic errors under optimal temperatures and showed a higher susceptibility to heat than their tall counterparts. Identification and introduction of new dwarfing alleles into modern breeding programmes is invaluable in the development of climate-resilient wheat varieties.

Apple MdbHLH4 promotes the flowering transition through interactions with FLOWERING LOCUS C and transcriptional activation of FLOWERING LOCUS T

Apple (Malus domestica) is one of the most widely cultivated fruit trees in the world and plays a central role in the agricultural industry and the economy. Flowering is a multi-path synergistic process that helps plants achieve high quality and yield in a multilateral environment. Here, overexpressing MdbHLH4 promoted the flowering transition in Arabidopsis, while myc70, a mutant of the MdbHLH4 Arabidopsis homolog, only exhibited a late flowering phenotype under short days. Moreover, the late flowering myc70 mutant phenotype was attenuated by ectopic expression of MdbHLH4. The myc70 mutant recovered from late flowering under short days by exogenous application of gibberellic acid (GA)4+7. The expression of GA biosynthesis-related genes was upregulated and that of GA catabolic genes was downregulated in the apical meristems of the Arabidopsis MdbHLH4 transgenic lines compared with the wild-type, and the opposite was observed in the myc70 mutant. Furthermore, yeast-two-hybrid (Y2H) assays revealed that MdbHLH4 and DELLA family proteins interacted with the C-terminal domain of MdFLC (FLOWERING LOCUS C) protein, suggesting potential competition between MdbHLH4 and MdDELLAs to interact with MdFLC in apples. Additionally, MdbHLH4 directly bound to the MdFT (FLOWERING LOCUS T) promoter and activated MdFT expression; thus promoting the floral transition. Our results suggest that MdbHLH4 is a key gene involved in plant flowering under short day conditions by regulating GA biosynthesis and signaling as well as the expression of MdFT, which provides new insight into the apple flowering mechanism.

Sweet cherry PavGA20ox-2 positive regulation of plant growth, flowering time, and seed germination

GA20 oxidase (GA20ox) is an enzyme that regulates GA (Gibberellic acid) biosynthesis and is essential in maintaining the dynamic balance of gibberellin in plants. We identified the PavGA20ox-2 gene from sweet cherry. The expression level of PavGA20ox-2 increased from winter to spring and significantly decreased after GA4+7 treatment. Surprisingly, the expression level of PavGA20ox-2 in sweet cherry flower buds with transient infection of overexpression PavWRKY31 was significantly higher than that in the control group. Besides, PavWRKY31 is directly bound to the W-box motif in the promoter of PavGA20ox-2 to activate the transcriptional level of PavGA20ox-2. The plant height was also increased, and the flowering time was advanced when PavGA20ox-2 was overexpressed in Arabidopsis. Moreover, the seed germination rate of Arabidopsis thaliana was higher than that of the wild type. In conclusion, PavGA20ox-2 could promote plant growth, accelerate flowering time, and increase seed germination. Thus, PavGA20ox-2 is considered involved in regulating multiple processes of plant growth and development by affecting GA biosynthesis. Our results will contribute to a better understanding of the molecular mechanism of GA20ox-mediated fruit tree development.

Transcriptome analysis during vernalization in wheat (Triticum aestivum L.)

BackgroundVernalization, as a vital process in the life cycle of winter cereal, has important effects on floral organ formation and flowering time. Many morphological changes together with molecular changes occur during the vernalization period. Here, we used transcriptome sequencing to analyze the transcriptomic changes in wheat leaves before, during and after vernalization using the winter wheat cultivar ‘Shiluan02-1’.ResultsA total of 16,370 differentially expressed genes were obtained across different vernalization periods. Gene Ontology enrichment analysis revealed that photoperiodism, photoprotection, photosynthesis, lipid transport and biosynthetic process, and chlorophyll metabolic process were closely related to vernalization. In addition, AP2/ERF, C2H2, bHLH, WRKY, MYB, MYB-related, and NAC transcription factors were significantly enriched during vernalization, and the transcription factor expression patterns suggested the intricate regulation of transcription factor modules in plant vernalization pathways. Analysis of gene expression patterns of the MADS-box transcription factor genes showed different expression patterns during vernalization phases, among which VERNALIZATION1 (VRN1) genes were found to gradually increase during vernalization periods from V0 to V35, while decline in the V42 phase, then increase after vernalization. The Tavrt-2 gene cooperated with Tavrn1 to regulate flowering induced by vernalization, and its expression level was rapidly increased by vernalization but declined in the V42 phase and then increased after vernalization. Some genes from the ICE-CBF-COR pathway were also identified, and additional analysis indicated that some key genes related to phytohormone biosynthesis and signal transduction were enriched during the vernalization period, such as gibberellic acid, ethylene, abscisic acid and jasmonic acid biosynthesis and signaling pathway genes.ConclusionsOur study provides valuable molecular information for future studies on wheat vernalization regulation and also serves as an excellent reference for future wheat breeding.

A large-scale whole-exome sequencing mutant resource for functional genomics in wheat.

Hexaploid wheat (Triticum aestivum), a major staple crop, has a remarkably large genome of ~14.4 Gb (containing 106 913 high-confidence [HC] and 159 840 low-confidence [LC] genes in the Chinese Spring v2.1 reference genome), which poses a major challenge for functional genomics studies. To overcome this hurdle, we performed whole-exome sequencing to generate a nearly saturated wheat mutant database containing 18 025 209 mutations induced by ethyl methanesulfonate (EMS), carbon (C)-ion beams, or γ-ray mutagenesis. This database contains an average of 47.1 mutations per kb in each gene-coding sequence: the potential functional mutations were predicted to cover 96.7% of HC genes and 70.5% of LC genes. Comparative analysis of mutations induced by EMS, γ-rays, or C-ion beam irradiation revealed that γ-ray and C-ion beam mutagenesis induced a more diverse array of variations than EMS, including large-fragment deletions, small insertions/deletions, and various non-synonymous single nucleotide polymorphisms. As a test case, we combined mutation analysis with phenotypic screening and rapidly mapped the candidate gene responsible for the phenotype of a yellow-green leaf mutant to a 2.8-Mb chromosomal region. Furthermore, a proof-of-concept reverse genetics study revealed that mutations in gibberellic acid biosynthesis and signalling genes could be associated with negative impacts on plant height. Finally, we built a publically available database of these mutations with the corresponding germplasm (seed stock) repository to facilitate advanced functional genomics studies in wheat for the broad plant research community.

Arabidopsis-expressing lysine-null SUMO1 reveals a non-essential role for secondary SUMO modifications in plants.

The reversible conjugation of small ubiquitin-like modifier (SUMO) to other proteins has pervasive roles in various aspects of plant development and stress defense through its selective attachment to numerous intracellular substrates. An intriguing aspect of SUMO is that it can be further modified by SUMOylation and ubiquitylation, which isopeptide-link either or both polypeptides to internal lysines within previously bound SUMOs. Although detectable by mass spectrometry, the functions of these secondary modifications remain obscure. Here, we generated transgenic Arabidopsis that replaced the two related and essential SUMO isoforms (SUMO1 and SUMO2) with a lysine-null SUMO1 variant (K0) immune to further SUMOylation/ubiquitylation at these residues. Remarkably, homozygous SUMO1(K0) sumo1 sumo2 plants developed normally, were not hypersensitive to heat stress, and have nearly unaltered SUMOylation profiles during heat shock. However, subtle changes in tolerance to salt, paraquat, and the DNA-damaging agents bleomycin and methane methylsulfonate were evident, as were increased sensitivities to ABA and the gibberellic acid biosynthesis inhibitor paclobutrazol, suggesting roles for these secondary modifications in stress defense, DNA repair, and hormone signaling. We also generated viable sumo1 sumo2 lines expressing a SUMO1(K0) variant specifically designed to help isolate SUMO conjugates and map SUMOylation sites, thus offering a new tool for investigating SUMO in planta.

PROTEIN PHOSPHATASE 2C08, a Negative Regulator of Abscisic Acid Signaling, Promotes Internode Elongation in Rice

Clade A protein phosphatase 2Cs (PP2CAs) negatively regulate abscisic acid (ABA) signaling. Here, we investigated the functions of OsPP2CAs and their crosstalk with ABA and gibberellic acid (GA) signaling pathways in rice (Oryza sativa). Among the nine OsPP2CAs, OsPP2C08 had the highest amino acid sequence similarity with OsPP2C51, which positively regulates GA signaling in rice seed germination. However, OsPP2C08 was expressed in different tissues (internodes, sheaths, and flowers) compared to OsPP2C51, which was specifically expressed in seeds, and showed much stronger induction under abiotic stress than OsPP2C51. Transgenic rice lines overexpressing OsPP2C08 (OsPP2C08-OX) had a typical ABA-insensitive phenotype in a post-germination assay, indicating that OsPP2C08, as with other OsPP2CAs, negatively regulates ABA signaling. Furthermore, OsPP2C08-OX lines had longer stems than wild-type (WT) plants due to longer internodes, especially between the second and third nodes. Internode cells were also longer in OsPP2C08-OX lines than in the WT. As GA positively regulates plant growth, these results suggest that OsPP2C08 might positively regulate GA biosynthesis. Indeed, the expression levels of GA biosynthetic genes including gibberellin 20-oxidase (OsGA20ox4) and Ent-kaurenoic acid oxidase (OsKAO) were increased in OsPP2C08-OX lines, and we observed that GIBBERELLIN 2-OXIDASE 4 (OsGA2ox4), encoding an oxidase that catalyzes the 2-beta-hydroxylation of several biologically active GAs, was repressed in the OsPP2C08-OX lines based on a transcriptome deep sequencing and RT-qPCR analysis. Furthermore, we compared the accumulation of SLENDER RICE 1 (SLR1), a DELLA protein involved in GA signaling, in OsPP2C08-OX and WT plants, and observed lower levels of SLR1 in the OsPP2C08-OX lines than in the WT. Taken together, our results reveal that OsPP2C08 negatively regulates ABA signaling and positively regulates GA signaling in rice. Our study provides valuable insight into the molecular mechanisms underlying the crosstalk between GA and ABA signaling in rice.

Regulation of reactive oxygen species and phytohormones in osmotic stress tolerance during seed germination in indica rice.

Climate change due to global warming is now affecting agricultural production worldwide. In rice, one of the most important crops, water limitation due to irregular rainfall in rainfed lowlands during crop growth limits yield. Dry direct-sowing has been proposed as a water-efficient approach to cope with water stress during rice growth, but poor seedling establishment due to drought during germination and emergence is a problem. Here, we germinated indica rice cultivars Rc348 (drought tolerant) and Rc10 (drought sensitive) under osmotic stress induced by PEG to elucidate mechanisms of germination under drought. Rc348 had higher germination rate and germination index under severe osmotic stress of -1.5 MPa, above those of Rc10. Rc348 showed up-regulated GA biosynthesis, down-regulated ABA catabolism, and up-regulated α-amylase gene expression in imbibed seeds under PEG treatment compared to that of Rc10. During germination, reactive oxygen species (ROS) play important roles in antagonism between gibberellic acid (GA) and abscisic acid (ABA). Embryo of Rc348 treated with PEG had significantly greater expression of NADPH oxidase genes and higher endogenous ROS levels, together with significantly increased endogenous GA1, GA4 and ABA contents compared to that of Rc10. In aleurone layers treated with exogenous GA, expression of α-amylase genes was higher in Rc348 than in Rc10, and expression of NADPH oxidase genes was enhanced with significantly higher ROS content in Rc348, suggesting higher sensitivity of GA to ROS production and starch degradation in aleurone cells of Rc348. These results suggest that the osmotic stress tolerance of Rc348 is due to enhancement of ROS production, GA biosynthesis, and GA sensitivity, resulting in a higher germination rate under osmotic stress.

Phytochrome-interacting factor PIF3 integrates phytochrome B and UVB signaling pathways to regulate gibberellin- and auxin-dependent growth in cucumber hypocotyls.

In Arabidopsis, the photoreceptors phytochrome B- (PhyB) and UVB resistance 8- (UVR8) mediated light responses play a major role in regulating photomorphogenic hypocotyl growth, but how they crosstalk to coordinate this process is not well understood. Here we report map-based cloning and functional characterization of a UVB-insensitive, long-hypocotyl mutant, lh1, and a wild-type-like lh2 mutant in cucumber, Cucumis sativus, which encodes a defective CsPhyB and a key gibberellic acid (GA) biosynthesis enzyme CsGA20ox-2, respectively. The lh2 mutation was epistatic to lh1 and partly suppressed long-hypocotyl phenotype in the lhl1lh2 double mutant. We identified phytochrome interacting factor (PIF) CsPIF3 that played a critical role in integrating the red/far-red and UVB light responses for hypocotyl growth. We show that two modules, CsPhyB-CsPIF3-CsGA20ox-2 (GA oxidase2)-DELLA and CsPIF3-CsARF18 (auxin response factor 18) mediate CsPhyB-regulated hypocotyl elongation through GA and auxin pathways, respectively, in which CsPIF3 binds to the G-/E-box motifs in the promoters of CsGA20ox-2 and CsARF18 to regulate their expression. We also identified a new physical interaction between CsPIF3 and CsUVR8 mediating CsPhyB-dependent, UVB-induced hypocotyl growth inhibition. Our work suggests that hypocotyl growth in cucumber involves a complex interplay of multiple photoreceptor- and phytohormone-mediated signaling pathways that show both conservation with and divergence from those in Arabidopsis.

Artificial selection of the Green Revolution gene Semidwarf 1 is implicated in upland rice breeding

Semidwarf breeding has boosted crop production and is a well-known outcome from the first Green Revolution. The Green Revolution gene Semidwarf 1 (SD1), which modulates gibberellic acid (GA) biosynthesis, plays a principal role in determining rice plant height. Mutations in SD1 reduce rice plant height and promote lodging resistance and fertilizer tolerance to increase grain production. The plant height mediated by SD1 also favors grain yield under certain conditions. However, it is not yet known whether the function of SD1 in upland rice promotes adaptation and grain production. In this study, the plant height and grain yield of irrigated and upland rice were comparatively analyzed under paddy and dryland conditions. In response to dryland environments, rice requires a reduction in plant height to cope with water deficits. Upland rice accessions had greater plant heights than their irrigated counterparts under both paddy and dryland conditions, and appropriately reducing plant height could improve adaptability to dryland environments and maintain high grain yield formation. Moreover, upland rice cultivars with thicker stem diameters had stronger lodging resistance, which addresses the lodging problem. Knockout of SD1 in the upland rice cultivar IRAT104 reduced the plant height and grain yield, demonstrating that the adjustment of plant height mediated by SD1 could increase grain production in dryland fields. In addition, an SD1 genetic diversity analysis verified that haplotype variation causes phenotypic variation in plant height. During the breeding history of rice, SD1 allelic mutations were selected from landraces to improve the grain yield of irrigated rice cultivars, and this selection was accompanied by a reduction in plant height. Thus, five known mutant alleles were analyzed to verify that functional SD1 is required for upland rice production. All these results suggest that SD1 might have undergone artificial positive selection in upland rice, which provides further insights concerning greater plant height in upland rice breeding.

Vernalization Promotes GA-Mediated Bolting Initiation via the Inhibition of ABA and JA Biosynthesis

To understand the relationship between vernalization and endogenous phytohormones on bolting, phytohormone levels and transcriptome changes were measured by LC-MS/MS and RNA sequencing before vernalization (CK), at the end of 16 weeks of vernalization (Vel) and at 5 days after vernalization (Re). A total of 32 phytohormone compounds significantly changed after vernalization; especially, the content of abscisic acid (ABA) and jasmonic acid (JA) was dramatically decreased more than sixteen and three times in the Vel and Re samples. In addition, GA19 accumulated after vernalization, while the content of GA53 and GA20 decreased. A total of 7471 differentially expressed genes (DEGs) were identified in response to vernalization. These DEGs were enriched with GO terms including response to stimulus (GO:0050896), response to hormone (GO:0009725) and shoot system development (GO:0048367). KEGG mapping indicated that 16, 13 and 11 DEGs were involved in gibberellic acid (GA), ABA and JA biosynthesis, suggesting a negative role of vernalization in phytohormone biosynthesis. Vernalization also repressed the expression of BvABFs and BvMYC2s, implying the inhibition of ABA and JA signalling. Additionally, vernalization plays a positive role in GA signalling, owing to the down-regulation of BvDELLAs. We also found that GA3-induced bolting could be retarded by exogenous ABA and methyl jasmonate (MeJA). In total, our results suggest that vernalization can promote GA-induced bolting by decreasing BvDELLA repressors of GA signalling and eliminating the antagonistic effects of ABA and JA.

SlZF3 regulates tomato plant height by directly repressing SlGA20ox4 in the gibberellic acid biosynthesis pathway.

Plant height is an important target trait for crop genetic improvement. Our previous work has identified a salt-tolerant C2H2 zinc finger, SlZF3, and its overexpression lines also showed a semi-dwarf phenotype, but the molecular mechanism remains to be elucidated. Here, we characterized the dwarf phenotype in detail. The dwarfism is caused by a decrease in stem internode cell elongation and deficiency of bioactive gibberellic acids (GAs), and can be rescued by exogenous GA3 treatment. Gene expression assays detected reduced expression of genes in the GA biosynthesis pathway of the overexpression lines, including SlGA20ox4. Several protein-DNA interaction methods confirmed that SlZF3 can directly bind to the SlGA20ox4 promoter and inhibit its expression, and the interaction can also occur for SlKS and SlKO. Overexpression of SlGA20ox4 in the SlZF3-overexpressing line can recover the dwarf phenotype. Therefore, SlZF3 regulates plant height by directly repressing genes in the tomato GA biosynthesis pathway.

Isolation and characterization of full-length coding sequences of gibberellic acid biosynthetic genes in coconut (Cocos nucifera L.) and their expression analysis

Gibberellic acid (GA) is intricately associated with plant growth and developmental pathways. To elucidate GA metabolism in coconut, we cloned full-length coding regions of the seven GA biosynthetic pathway genes (CPS, KS, KO, KAO, GA20ox, GA3ox and GA2ox) using a comparative genomics approach, using sequences from oil palm and date palm. The predicted amino acid sequences of the seven coconut GA biosynthetic genes shared high identities with oil palm and date palm GA biosynthetic genes. In silico structural and functional analyses were performed, and secondary and tertiary structures were predicted. Compared to the other two palms, the evolutionary analysis of the seven genes in coconut revealed that these genes may not be evolving rapidly enough to keep pace with occurring mutations. Additionally, we performed expression profiling of these seven genes during germination of mature zygotic embryos under in vitro conditions in two phenotypically contrasting coconut accessions (West Coast Tall and Chowghat Green Dwarf) and in response to exogenously supplemented GA3 and paclobutrazol (PBZ) in the embryo culture medium. While PBZ had an antagonistic effect on the GA biosynthesis pathway, GA3 enhanced the overall GA production. These results provide the first insights into GA biosynthesis in coconut, laying the foundation for future studies to decipher the genetics of dwarfism in coconut.

Gibberellic acid sensitive dwarf encodes an ARPC2 subunit that mediates gibberellic acid biosynthesis, effects to grain yield in rice.

The plant hormone gibberellic acid (GA) is important for plant growth and productivity. Actin-related proteins (ARPs) also play central roles in plant growth, including cell elongation and development. However, the relationships between ARPs and GA signaling and biosynthesis are not fully understood. Here, we isolated OsGASD, encoding an ARP subunit from rice (Oryza sativa), using the Ac/Ds knockout system. The osgasd knockout (Ko) mutation reduced GA3 content in shoots as well as plant growth and height. However, GA application restored the plant height of the osgasd Ko mutant to a height similar to that of the wild type (WT). Rice plants overexpressing OsGASD (Ox) showed increased plant height and grain yield compared to the WT. Transcriptome analysis of flag leaves of OsGASD Ox and osgasd Ko plants revealed that OsGASD regulates cell development and the expression of elongation-related genes. These observations suggest that OsGASD is involved in maintaining GA homeostasis to regulate plant development, thereby affecting rice growth and productivity.

Ectopic expression of GmNF-YA8 in Arabidopsis delays flowering via modulating the expression of gibberellic acid biosynthesis- and flowering-related genes and promotes lateral root emergence in low phosphorus conditions.

NUCLEAR FACTOR Y subunit alpha (NF-YA), together with NF-YB and NF-YC, regulates plant growth and development, as well as plant responses to biotic and abiotic stresses. Although extensive studies have examined the functions of NF-YAs in Arabidopsis thaliana, the roles of NF- YAs in Glycinme max are poorly understood. In this study, we identified a phosphorus (P) starvation-responsive NF-YA8 in soybean. The expression of GmNF-YA8 is induced by low P or low nitrogen in leaves, but not by potassium or iron starvation, respectively. GmNF-YA8 is localized in the nucleus and plasma membrane. Ectopic expression of GmNF-YA8 inhibits plant growth and delayed flowering in Arabidopsis. Exogenous application of gibberellic acid (GA) rescues the delayed flowering phenotype in Arabidopsis overexpressing GmNF-YA8 lines GmNF-YA8OE-05 and GmNF-YA8OE-20. Moreover, quantitative real time PCR (qRT-PCR) verified that overexpression of GmNF-YA8 downregulates GA20ox2 and GA3ox2 expression, but upregulates GA2ox2 and GA2ox3 that encode enzymes, which inactive bioactive GAs. Consistent with the late flowering phenotype of Arabidopsis trangenic lines that overexpress GmNF-YA8, the transcript levels of flowering-promoting genes AP1, CO, LFY, and SOC1 are reduced. In addition, overexpression of GmNF-YA8 promotes the emergence of lateral root (LR) primordium from epidermis rather than the initiation of LR in low P, and increases the LR density in low nitrogen. Our results provide insights into the roles of GmNF-YA8.