GA Synthesis Research Articles

Arabidopsis thaliana E3 ligase AIRP4 is involved in GA synthesis

Arabidopsis abscisic acid ABA-Insensitive RING Proteins (AtAIRP1–4) are RING E3s that play significant roles in ABA-signaling pathways. However, it is still unclear whether they have other functions. Here, AtAIRP4 was determined to play a role in response to gibberellin A3 (GA3) in Arabidopsis thaliana. After proAtAIRP4::GUS transgenic lines were treated with GA3, the GUS activity decreased in hypocotyls. Increased hypocotyl elongation in response to GA3 seen in WT was not observed in the AtAIRP4-overexpression lines, whereas AtAIRP4-overexpression lines were hypersensitive to Paclobutrazol (PAC, an inhibitor of GA biosynthesis) during the seed germination stage. Additionally, AtAIRP4-overexpressing lines showed the lowest level of primary root elongation in the presence of GA3. The levels of endogenous GA3 in 35S::AtAIRP4 lines were lower than those in wild-type. In addition, among the plants, the mRNA levels of the GA synthetic gene GIBBERELLIN 20-OXIDASE1 (GA20ox1) was the lowest in overexpressing line. However, the expression of the response gene DELLA RGA-LIKE3 (RGL3) was the highest in overexpressing lines after treatment with GA3. Thus, AtAIRP4 plays a negative role in GA-mediated hypocotyl elongation and root growth, and it inhibits the synthesis of endogenous biologically active GA3 to some extent.

Three multi-enzyme cascade pathways for conversion of C1 to C2/C4 compounds

A feasible and promising carbon reduction strategy is to convert CO 2 into C1 compounds through renewable energy-driven first and then use a multi-enzyme cascade to obtain compounds with higher carbon numbers and market value. In this study, three multi-enzyme cascade pathways for preparing important C2/C4 compounds (ethylene glycol [EG], glycolic acid [GA], and d -erythrose) from methanol were proposed. All pathways use glycolaldehyde as the key intermediate, which can be obtained from methanol through glycolaldehyde synthase and alcohol oxidase cascade. Then, the target compounds can be acquired by adding different enzymes. As a result, 0.90 g/L EG, 0.33 g/L GA, and 53.65 mg/L d -erythrose were produced from methanol. Compared with the titers of EG (6.6 mM) and GA (1.2 g/L) synthesis routes from formaldehyde reported in the literature, the titers of this study (27.58 mM EG and 1.59 g/L GA from formaldehyde) were significantly improved. • Glycolic acid and d -erythrose pathways from CH 3 OH were proposed for the first time • DERA were found to have the function of condensing glycolaldehyde to d -erythrose • GOX were used to synthesize ethylene glycol from glycolaldehyde • ALDH were used to oxidize glycolaldehyde to glycolic acid Under the background of CO 2 is one of the main gases leading to the greenhouse effect and the inexpensive and widespread feedstock. Coupling the renewable energy-driven CO 2 capture technology with multi-enzyme cascade catalytic conversion of C1 compounds to multi-carbon compounds is a promising method for CO 2 utilization. In this case, we successfully constructed three multi-enzyme cascade pathways to synthesize important C2/C4 compounds (ethylene glycol, glycolic acid, and d -erythrose) from C1 compounds. The pathways proposed herein could contribute to realizing the recycling of carbon resources and solving the problems of plastic pollution and energy shortage. They could also provide a new impetus for the design of chemo-bio routes for the efficient conversion of CO 2 into compounds with higher carbon numbers and market value. Junhui et al. successfully constructed three multi-enzyme pathways for synthesizing C2/C4 compounds (ethylene glycol, glycolic acid, and d -erythrose) from C1 compounds. Glycolic acid or d -erythrose pathways were proposed for the first time. Three new kinds of enzymes were found to catalyze intermediate glycolaldehyde to synthesize such C2/C4 compounds.

MiR3633a-GA3ox2 Module Conducts Grape Seed-Embryo Abortion in Response to Gibberellin.

Seedlessness is one of the important quality and economic traits favored by grapevine consumers, which are mainly affected by phytohormones, especially gibberellin (GA). GA is widely utilized in seedless berry production and could effectively induce grape seed embryo abortion. However, the molecular mechanism underlying this process, like the role of RNA silencing in the biosynthesis pathway of GA remains elusive. Here, Gibberellin 3-β dioxygenase2 (GA3ox2) as the last key enzyme in GA biosynthesis was predicated as a potential target gene for miR3633a, and two of them were identified as a GA response in grape berries. We also analyzed the promoter regions of genes encoding GA biosynthesis and found the hormone-responsive elements to regulate grape growth and development. The cleavage interaction between VvmiR3633a and VvGA3ox2 was validated by RLM-RACE and the transient co-transformation technique in tobacco in vivo. Interestingly, during GA-induced grape seed embryo abortion, exogenous GA promoted the expression of VvmiR3633a, thereby mainly repressing the level of VvGA3ox2 in seed embryos. We also observed a negative correlation between down-regulated VvGA20ox2/VvGA3ox2 and up-regulated VvGA2ox3/VvGA2ox1, of which GA inactivation was greater than GA synthesis, inhibited active GA content, accompanied by the reduction of VvSOD and VvCAT expression levels and enzymatic activities. These series of changes might be the main causes of grape seed embryo abortion. In conclusion, we have preliminarily drawn a schematic mode of GA-mediated VvmiR3633a and related genes regulatory network during grape seed abortion induced by exogenous GA. Our findings provide novel insights into the GA-responsive roles of the VvmiR3633a-VvGA3ox2 module in the modulation of grape seed-embryo abortion, which has implications for the molecular breeding of high-quality seedless grape berries.

The mechanism of exogenous gibberellin A3 protecting sorghum shoots from S-metolachlor Phytotoxicity.

S-metolachlor (MET) was used to prevent weed infestation in sorghum fields, but inappropriate application could result in phytotoxicity on sorghum. Exogenous gibberellin A3 (GA3 ) has been applied for alleviating the phytotoxicity of MET. However, its detoxification mechanism is still not well known. Leaf deformity of sorghum caused by 200 mg/L MET was alleviated by treating sorghum shoots with 800 mg/L GA3 , and the injury recovery rate of growth index was over 73%. More importantly, GA3 could not accelerate the metabolic rate of MET in sorghum. The result of phytohormone metabolomics showed that endogenous GA3 content in sorghum decreased by 78.10% with MET treatment, while abscisic acid (ABA) content increased by 120.2%, resulting in 10.3-fold increase of ABA/GA3 ratio. Content of ABA and GA3 increased by 11.9- and 21.1-fold with MET and GA3 treatment, respectively, leading to ABA/GA3 ratio restoration. Moreover, MET inhibited the expression of genes encoding key enzymes related to GA synthesis including CPS1, KO2, KAO, GA20ox1D and ABA8ox gene related to ABA metabolism. The transcription levels of GA metabolism-related genes CYP714D1 and GA2ox were up-regulated by 11.2- and 7.2-fold, while ABA synthesis-related genes NCED and ZEP were up-regulated by 8.0- and 3.0-fold, respectively, with MET and GA3 treatment. In this study, exogenous GA3 protecting sorghum shoots from MET phytotoxicity was due to supplement the MET-induced GA3 deficiency by absorbing exogenous GA3 , and restore homeostasis of ABA and GA3 by promoting ABA synthesis, which provides novel insights for mechanism of GA3 alleviating MET phytotoxicity. © 2022 Society of Chemical Industry.

Evolutionary and Integrative Analysis of Gibberellin-Dioxygenase Gene Family and Their Expression Profile in Three Rosaceae Genomes (F. vesca, P. mume, and P. avium) Under Phytohormone Stress.

The gibberellin-dioxygenase (GAox) gene family plays a crucial role in regulating plant growth and development. GAoxs, which are encoded by many gene subfamilies, are extremely critical in regulating bioactive GA levels by catalyzing the subsequent stages in the biosynthesis process. Moreover, GAoxs are important enzymes in the GA synthesis pathway, and the GAox gene family has not yet been identified in Rosaceae species (Prunus avium L., F. vesca, and P. mume), especially in response to gibberellin and PCa (prohexadione calcium; reduce biologically active GAs). In the current investigation, 399 GAox members were identified in sweet cherry, Japanese apricot, and strawberry. Moreover, they were further classified into six (A-F) subgroups based on phylogeny. According to motif analysis and gene structure, the majority of the PavGAox genes have a remarkably well-maintained exon–intron and motif arrangement within the same subgroup, which may lead to functional divergence. In the systematic investigation, PavGAox genes have several duplication events, but segmental duplication occurs frequently. A calculative analysis of orthologous gene pairs in Prunus avium L., F. vesca, and P. mume revealed that GAox genes are subjected to purifying selection during the evolutionary process, resulting in functional divergence. The analysis of cis-regulatory elements in the upstream region of the 140 PavGAox members suggests a possible relationship between genes and specific functions of hormone response-related elements. Moreover, the PavGAox genes display a variety of tissue expression patterns in diverse tissues, with most of the PavGAox genes displaying tissue-specific expression patterns. Furthermore, most of the PavGAox genes express significant expression in buds under phytohormonal stresses. Phytohormones stress analysis demonstrated that some of PavGAox genes are responsible for maintaining the GA level in plant-like Pav co4017001.1 g010.1.br, Pav sc0000024.1 g340.1.br, and Pav sc0000024.1 g270.1.mk. The subcellular localization of PavGAox protein utilizing a tobacco transient transformation system into the tobacco epidermal cells predicted that GFP signals were mostly found in the cytoplasm. These findings will contribute to a better understanding of the GAox gene family’s interaction with prohexadione calcium and GA, as well as provide a strong framework for future functional characterization of GAox genes in sweet cherry.

Identification of BcARR Genes and CTK Effects on Stalk Development of Flowering Chinese Cabbage.

Flowering Chinese cabbage (Brassica campestris L. ssp. Chinensis var. utilis Tsen et Lee) is an important and extensively cultivated vegetable in south China, whose major food product is the stalk. In the process of stalk formation, its initiation and development are regulated by a series of hormonal signals, such as cytokinin and gibberellin. In this study, we analyzed the effects of zeatin (ZT) and gibberellin A3 (GA3), and their interaction, on the bolting of flowering Chinese cabbage. The results indicated that the three-true-leaf spraying of ZT and GA synthesis inhibitor (PAC) inhibited plant height but increased stem diameter. Cytokinin (CTK) synthesis inhibitor (YZJ) and GA3 treatment increased plant height and decreased stem diameter. In addition, ZT and GA3 co-treated plants displayed antagonistic effect. Further, 19 type-B authentic response regulators (ARR-Bs), the positive regulators of cytokinin signal transduction were identified from flowering Chinese cabbage. Comprehensive analysis of phylogeny showed BcARR-Bs clustered into three subfamilies with 10 conserved motifs. Analysis of their expression patterns in different tissues and at various growth stage, and their response to hormone treatment suggest that ARR1-b localized in the nucleus displayed unique highest expression patterns in stem tips, are responsive both to ZT and GA, suggesting a significant role in mediating the crosstalk of ZT and GA in the bolting of flowering Chinese cabbage.

Transcriptome Analysis Reveals the Regulatory Networks of Cytokinin in Promoting Floral Feminization in Castanea henryi.

Castanea henryi is a monoecious plant with a low female-to-male ratio, which limits its yield. The phytohormone cytokinin (CK) plays a crucial role in flower development, especially gynoecium development. Here, the feminizing effect of CK on the development of C. henryi was confirmed by the exogenous spraying of N-(2-chloro-4-pyridyl)-N’-phenylurea (CPPU). Spraying CPPU at 125 mg·L−1 thrice changed the male catkin into a pure female catkin, whereas at 5 mg·L−1 and 25 mg·L−1, only a part of the male catkin was transformed into a female catkin. A comparative transcriptome analysis of male catkins subjected to CPPU was performed to study the mechanism of the role of CKs in sex differentiation. Using Pearson’s correlation analysis between hormone content and hormone synthesis gene expression, four key genes, LOG1, LOG3, LOG7 and KO, were identified in the CK and GA synthesis pathways. Moreover, a hub gene in the crosstalk between JA and the other hormone signaling pathways, MYC2, was identified, and 15 flowering-related genes were significantly differentially expressed after CPPU treatment. These results suggest that CK interacts with other phytohormones to determine the sex of C. henryi, and CK may directly target floral organ recognition genes to control flower sex.

Unveiling Molecular Mechanisms of Nitric Oxide-Induced Low-Temperature Tolerance in Cucumber by Transcriptome Profiling.

Cucumber (Cucumis sativus L.) is one of the most popular cultivated vegetable crops but it is intrinsically sensitive to cold stress due to its thermophilic nature. To explore the molecular mechanism of plant response to low temperature (LT) and the mitigation effect of exogenous nitric oxide (NO) on LT stress in cucumber, transcriptome changes in cucumber leaves were compared. The results showed that LT stress regulated the transcript level of genes related to the cell cycle, photosynthesis, flavonoid accumulation, lignin synthesis, active gibberellin (GA), phenylalanine metabolism, phytohormone ethylene and salicylic acid (SA) signaling in cucumber seedlings. Exogenous NO improved the LT tolerance of cucumber as reflected by increased maximum photochemical efficiency (Fv/Fm) and decreased chilling damage index (CI), electrolyte leakage and malondialdehyde (MDA) content, and altered transcript levels of genes related to phenylalanine metabolism, lignin synthesis, plant hormone (SA and ethylene) signal transduction, and cell cycle. In addition, we found four differentially expressed transcription factors (MYB63, WRKY21, HD-ZIP, and b-ZIP) and their target genes such as the light-harvesting complex I chlorophyll a/b binding protein 1 gene (LHCA1), light-harvesting complex II chlorophyll a/b binding protein 1, 3, and 5 genes (LHCB1, LHCB3, and LHCB5), chalcone synthase gene (CSH), ethylene-insensitive protein 3 gene (EIN3), peroxidase, phenylalanine ammonia-lyase gene (PAL), DNA replication licensing factor gene (MCM5 and MCM6), gibberellin 3 beta-dioxygenase gene (GA3ox), and regulatory protein gene (NPRI), which are potentially associated with plant responses to NO and LT stress. Notably, HD-ZIP and b-ZIP specifically responded to exogenous NO under LT stress. Taken together, these results demonstrate that cucumber seedlings respond to LT stress and exogenous NO by modulating the transcription of some key transcription factors and their downstream genes, thereby regulating photosynthesis, lignin synthesis, plant hormone signal transduction, phenylalanine metabolism, cell cycle, and GA synthesis. Our study unveiled potential molecular mechanisms of plant response to LT stress and indicated the possibility of NO application in cucumber production under LT stress, particularly in winter and early spring.

14-3-3 gene of Zostera japonica ZjGRF1 participates in gibberellin signaling pathway.

Zostera japonica is a unique seagrass species in Asia with important ecological value. Gibberellin (GA) is a plant hormone crucial in the regulation of plant growth and development, including seed longevity, seed germination, anti-aging, hypocotyl elongation, leaf development, reproductive organ development, and abiotic stress responses. However, the role of 14-3-3 gene of Z. japonica in GA signaling pathway remains unclear. Herein, gibberellin content and expression of GA synthesis genes were lower in Arabidopsis overexpressing ZjGRF1, 14-3-3 gene of Z. japonica, than in wild type (WT). Moreover, the expression level of GA receptors was lower in transgenic ZjGRF1 Arabidopsis than in WT. The expression level of GA response gene expansin8 (EXP8) was lower in transgenic ZjGRF1 Arabidopsis than in WT. In contrast, the expression levels of PACLOBUTRAZOL RESISTANT1 (PRE1), PRE5 and SCARECROW-LIKE 3 (SCL3) were higher in the transgenic ZjGRF1 Arabidopsis than in WT. Transgenic ZjGRF1 Arabidopsis has decreased sensitivity to paclobutrazol, an inhibitor of GA synthesis. The expression level of flowering regulatory genes was lower in transgenic ZjGRF1 Arabidopsis than in WT. This is the first study to report the function of 14-3-3 gene family in seagrass. These findings can be used in future studies on the regulation role of 14-3-3 gene on plant development.

Daminozide enhances the vigor and steviol glycoside yield of stevia (Stevia rebaudiana Bert.) propagated in temporary immersion bioreactors

Stevia (Stevia rebaudiana Bertoni) produces steviol glycosides (SG), a group of diterpenoid secondary metabolites widely used as natural sweeteners. SGs share the same precursor with gibberellins (GAs), ent-kaurenoic acid. In this study, we attempted to increase the SG content in stevia propagated in Temporary Immersion Bioreactors (TIBs) by blocking ent-kaurenoic acid entry to the GA synthesis pathway using Daminozide (0, 10, 20 ppm), a known GA inhibitor. The maximum increase in biomass weight and SG content (up to 40% for stevioside and rebaudioside A) was observed following 10 ppm Daminozide treatment. The treatment also increased transcript accumulation for genes involved in SG biosynthesis, such as SrKA13H, SrUGT85C2, and SrUGT76G1, indicating that the SG pathway became more active due to GA pathway inhibition. Furthermore, GA inhibition following inhibitor exposure was also indicated by the upregulated expression of a GA biosynthesis gene (GA3ox) due to feedback regulation, and the downregulated expression of a GA catabolism gene (GA2ox2) as a result of feed-forward regulation.

Overexpression of ZmSPL12 confers enhanced lodging resistance through transcriptional regulation of D1 in maize.

Overexpression of ZmSPL12 confers enhanced lodging resistance through transcriptional regulation of D1 in maize.

ScGAIL, a sugarcane N-terminal truncated DELLA-like protein, participates in gibberellin signaling in Arabidopsis.

The hormone gibberellin (GA) is crucial for internode elongation in sugarcane. DELLA proteins are critical negative regulators of the GA signaling pathway. ScGAI encodes a DELLA protein that was previously implicated in the regulation of sugarcane culm development. Here, we characterized ScGAI-like (ScGAIL) in sugarcane, which lacked the N-terminal region but was otherwise homologous to ScGAI. ScGAIL differed from ScGAI in its chromosomal location, expression patterns, and cellular localization. Although transgenic Arabidopsis overexpressing ScGAIL were insensitive to GAs, GA synthesis was affected in these plants, suggesting that ScGAIL disrupted the GA signaling pathway. After GA treatment, the expression patterns of GA-associated genes differed between ScGAIL-overexpressing and wild-type Arabidopsis, and the degradation of AtDELLA proteins in transgenic lines was significantly inhibited compared with wild-type lines. A sugarcane GID1 gene (ScGID1) encoding a putative GA receptor was isolated and interacted with ScGAIL in a GA-independent manner. Five ScGAIL-interacting proteins were verified by yeast two-hybrid assays, and only one interacted with ScGAI. Therefore, ScGAIL may inhibit plant growth by modulating the GA signaling pathway.

Exogenous Gibberellin Treatment Enhances Melatonin Synthesis for Melatonin-Enriched Rice Production.

Melatonin production is induced by many abiotic and biotic stressors; it modulates the levels of many plant hormones and their signaling pathways. This study investigated the effects of plant hormones on melatonin synthesis. Melatonin synthesis in rice seedlings was significantly induced upon exogenous gibberellin 3 (GA3) treatment, while it was severely decreased by GA synthesis inhibitor paclobutrazol. In contrast, abscisic acid (ABA) strongly inhibited melatonin synthesis, whereas its inhibitor norflurazon (NF) induced melatonin synthesis. The observed GA-mediated increase in melatonin was closely associated with elevated expression levels of melatonin biosynthetic genes such as TDC3, T5H, and ASMT1; it was also associated with reduced expression levels of catabolic genes ASDAC and M2H. In a paddy field, the treatment of immature rice seeds with exogenous GA led to enhanced melatonin production in rice seeds; various transgenic rice plants downregulating a GA biosynthesis gene (GA3ox2) and a signaling gene (Gα) showed severely decreased melatonin levels, providing in vivo genetic evidence that GA has a positive effect on melatonin synthesis. This is the first study to report that GA is positively involved in melatonin synthesis in plants; GA treatment can be used to produce melatonin-rich seeds, vegetables, and fruits, which are beneficial for human health.

Integrative signaling of hydrogen peroxide and gibberellin on Zn-mediated alleviation of thermodormancy in sorghum seeds.

Increasing global temperatures could result in decreasing crop production by decreasing seed germination in the field due to thermodormancy acquisition. Certain metals appear to modulate seed thermodormancy, although the exact mechanisms of that effect have not yet been elucidated. We report here the effects of Zn on the thermodormancy of sorghum seeds. Seeds treated with 0 or 200 mg Zn L-1 were germinated at optimal (30°C) and supra-optimal (40°C) temperatures and their germinability and oxidative stress markers were evaluated. The integrative effects of Zn, abscisic acid (ABA), gibberellin (GA), and H2 O2 on the physiology of seed thermodormancy were examined. The supra-optimal germination temperature (40°C) induced seed thermodormancy, which was, however, alleviated by treatment with 200 mg Zn L-1 . Thermodormancy acquired at supra-optimal temperatures in sorghum seeds must reflect de novo synthesis and accumulation of ABA. Although Zn treatment did not prevent ABA accumulation, it increased the activities of mitochondrial ETC enzymes and decreased the antioxidant enzymes' activity, leading to the accumulation of H2 O2 . By increasing mitochondria activity and H2 O2 production, Zn may induce GA synthesis and alleviate thermodormancy in sorghum seeds. The pretreatment of sorghum seeds with Zn may therefore improve seed germination and assure increased crop performance under normal (30°C) or rising (up to 40°C) temperatures.

Inhibition of gibberellin accumulation by water deficiency promotes fast and long-term 'drought avoidance' responses in tomato.

Plants reduce transpiration to avoid dehydration during drought episodes by stomatal closure and inhibition of canopy growth. Previous studies have suggested that low gibberellin (GA) activity promotes these 'drought avoidance' responses. Using genome editing, molecular, physiological and hormone analyses, we examined if drought regulates GA metabolism in tomato (Solanum lycopersicum) guard cells and leaves, and studied how this affects water loss. Water deficiency inhibited the expression of the GA biosynthesis genes GA20 oxidase1 (GA20ox1) and GA20ox2 and induced the GA deactivating gene GA2ox7 in guard cells and leaf tissue, resulting in reduced levels of bioactive GAs. These effects were mediated by abscisic acid-dependent and abscisic acid-independent pathways, and by the transcription factor TINY1. The loss of GA2ox7 attenuated stomatal response to water deficiency and during soil dehydration, ga2ox7 plants closed their stomata later, and wilted faster than wild-type (WT) M82 cv. Mutations in GA20ox1 and GA20ox2, had no effect on stomatal closure, but reduced water loss due to the mutants' smaller canopy areas. The results suggested that drought-induced GA deactivation in guard cells, contributes to stomatal closure at the early stages of soil dehydration, whereas inhibition of GA synthesis in leaves suppresses canopy growth and restricts transpiration area.

Delayed germination of Brassica parachinensis seeds by coumarin involves decreased GA4 production and a consequent reduction of ROS accumulation

AbstractThe plant allelochemical coumarin effectively inhibits the germination of Brassica parachinensis (B. parachinensis) seeds. Quantification of endogenous phytohormones showed that contents of abscisic acid (ABA), ABA glucose ester, gibberellin A20 (GA20), GA3, GA15, GA24, GA9 and GA4 were higher in germinating seeds than in seedlings. Moreover, the presence of coumarin significantly reduced the content of bioactive GA4 which is thought to positively regulate seed germination. Histochemical staining and spectrophotometry of reactive oxygen species (ROS) revealed that exogenous GA3 and GA4+7 could effectively promote the production of endogenous ROS during germination and that the GA synthesis inhibitor paclobutrazol could effectively inhibit production of ROS. Coumarin significantly inhibited the accumulation of ROS, especially superoxide anion radical (${\rm O}_2^{{\cdot}{-}} $). This inhibitory effect could be restored by the addition of exogenous GA3 and GA4+7. Coumarin also inhibited the activity of the ROS-degrading enzymes such as superoxide dismutase, catalase and peroxidase as well as β-amylase in seeds and seedlings. Taken together, we propose a model for the regulation of seed germination in B. parachinensis by coumarin, Gas and ROS, in which coumarin may delay seed germination by reducing endogenous GA4, thus decreasing the accumulation of ROS.

Chilling Requirement Validation and Physiological and Molecular Responses of the Bud Endodormancy Release in Paeonia lactiflora 'Meiju'.

The introduction of herbaceous peony (Paeonia lactiflora Pall.) in low-latitude areas is of great significance to expand the landscape application of this world-famous ornamental. With the hazards of climate warming, warm winters occurs frequently, which makes many excellent northern herbaceous peony cultivars unable to meet their chilling requirements (CR) and leads to their poor growth and flowering in southern China. Exploring the endodormancy release mechanism of underground buds is crucial for improving low-CR cultivar screening and breeding. A systematic study was conducted on P. lactiflora ‘Meiju’, a screened cultivar with a typical low-CR trait introduced from northern China, at the morphological, physiological and molecular levels. The CR value of ‘Meiju’ was further verified as 677.5 CUs based on the UT model and morphological observation. As a kind of signal transducer, reactive oxygen species (ROS) released a signal to enter dormancy, which led to corresponding changes in carbohydrate and hormone metabolism in buds, thus promoting underground buds to acquire strong cold resistance and enter endodormancy. The expression of important genes related to ABA metabolism, such as NCED3, PP2C, CBF4 and ABF2, reached peaks at the critical stage of endodormancy release (9 January) and then decreased rapidly; the expression of the GA2ox8 gene related to GA synthesis increased significantly in the early stage of endodormancy release and decreased rapidly after the release of ecodormancy (23 January). Cytological observation showed that the period when the sugar and starch contents decreased and the ABA/GA ratio decreased was when ‘Meiju’ bud endodormancy was released. This study reveals the endodormancy regulation mechanism of ‘Meiju’ buds with the low-CR trait, which lays a theoretical foundation for breeding new herbaceous peony cultivars with the low-CR trait.

Installation and Optimization of 68Ga Synthesis Module for Clinical Use: An Institutional Experience.

Introduction:With advent of gallium-labeling somatostatin analogs and its evaluation under positron emission tomography–computed tomography, there has been a tremendous surge in its application. Gallium 68 can be made available either from onsite cyclotron production or in the form of ready-to-use 68Ge/68Ga generators. Wherein setting up and running of cyclotron amounts to huge investment and dedicated team, the 68Ge/68Ga generator has proved to be a better option and viable project. Moreover, due to long half-life of 68Ge, i.e. 271 days, it enables the usage of generator for several months. The preparation of gallium-labeled peptides is much simpler in comparison to 18F radiochemistry, but the radiation exposure has always been an area of concern owing to high-energy annihilation photon of 511 keV.Materials and Methods:In this study, we share our experience of self-installation of 68Ge/68Ga generator during lockdown and the various steps involved in installation of fully automated peptide-labeling system in customized mini hot cell module, synthesis steps, and quality control steps of gallium-based radiopharmaceutical.Results:The installation was successfully completed with online assistance during the pandemic situation. The average elution yield met company specification (>80%), and 68Ga-labeled peptides were prepared with high radiochemical purity (>95%). The overall exposure in single batch of production and quality control never exceeded 3 μSv as shielding was well-taken care of with customized mini hot cell.Conclusion:With the described experience and validation process, one can easily think of making an installation at his/her center and cater to the needs of society with a new radiopharmaceutical.

Genome-wide identification and expression analysis of the GA2ox gene family in maize (Zea mays L.) under various abiotic stress conditions

GA 2-oxidases (GA2oxs) are a class of enzymes that inhibit the biosynthesis of bioactive GAs in plants. Although GA2oxs have clear roles in the development and defence responses in Arabidopsis, rice, and wheat, their potential effects on maize remain unclear. This study identified thirteen ZmGA2ox genes in maize and further characterized them using phylogenetic, gene structure, genomic locus, expression pattern analyses and GA content determination. Phylogenetic relationship analysis clearly divided the ZmGA2ox family into three groups—seven in C19-GA2ox class I, three in C19-GA2ox class II, and three in C20-GA2ox class. Evolutionary analysis suggested that ZmGA2ox1;1 and ZmGA2ox1;2, ZmGA2ox3;1 and ZmGA2ox3;2, and ZmGA2ox7;1 and ZmGA2ox7;2 are three pairs of segmental duplicated genes. Prediction of cis-regulatory elements in promoters suggested that ZmGA2ox genes were mainly associated with growth, development, hormones, and biotic/abiotic stress. Therefore, their spatial and temporal expression patterns and responses to various stress treatments were analysed on the basis of published RNA-seq data. Moreover, the changes of ZmGA2ox expression in leaves and roots of maize seedlings was detected under salt, alkali, dehydration, and cold stresses by qRT-PCR. The ZmGA2oxs exhibited obvious expression tendencies or characteristics in various organs under different abiotic stresses. The variations in the expression of three ZmGA2ox genes in the C20-GA2ox class in maize seedling roots showed significant regularity and a clear negative correlation with bioactive GA contents under cold and drought conditions, indicating that these three genes might exert key effects on the regulation of GA synthesis and the response to drought and cold stress. Taken together, this study is useful for further dissection of the effect of ZmGA2oxs on abiotic stress responses and might provide potential targets for the genetic improvement of maize.

OsGRF6 interacts with SLR1 to regulate OsGA2ox1 expression for coordinating chilling tolerance and growth in rice

Low temperature is one of the abiotic stressors that affect growth and productivity of rice. The plant hormone gibberellin not only regulates growth and development but is also involved in stress defense. Our rice seedling experiments demonstrated that overexpression of SLR1, a gene that encodes the rice DELLA protein, enhanced chilling tolerance. In contrast, overexpression of the active GA synthesis gene OsGA20ox1 reduced chilling tolerance, indicating that weakening GA signaling promoted plant defense against cold stress. CoIP-MS and BiFC assays showed that SLR1 physically interacted with OsGRF6. After cold treatment and recovery, the survival rates of OsGRF6-overexpression lines and an osgrf6 mutant and its complementary lines indicated that OsGRF6 is a negative regulator of chilling tolerance in rice. The yeast one-hybrid, qRT-PCR, and transactivation assays showed that both SLR1 and OsGRF6 can bind to the promoter of the active GA catabolic gene OsGA2ox1, where SLR1 promoted and OsGRF6 suppressed OsGA2ox1 expression. At normal temperature, OsGRF6 was responsible for maintaining active GA levels by inhibiting OsGA2ox1. When rice seedlings were subjected to chilling stress, the repressive effect of OsGRF6 on OsGA2ox1 was released by cold-induced SLR1, which activated OsGA2ox1 expression to decrease the active GA levels, enhancing chilling tolerance. These results suggest that OsGRF6 is an important regulator in the balance between growth and chilling tolerance in rice.