Effects of Gibberellin Pre-Treatment on Seed Germination and Seedling Physiology Characteristics in Industrial Hemp under Drought Stress Condition.

The present study aimed to explore the effects of exogenous gibberellins (GAs) on seed germination and subsequent seedling growth of hemp (Cannabis sativa L.) under drought stress. Seeds of two industrial hemp cultivars i.e., 'Yunma 1', (YM) and 'Bamahuoma', (BM) were treated with different concentrations of GA3 solution (0, 200, 400, 600, 800 mg/L) at 20 °C for 8 h. The effect of pre-treatment was assessed on germination characteristics and physiological indexes on subsequent exposure to drought stress using 20% (m/v) polyethylene glycol (PEG) for 7 days. The results revealed that seed germination in hemp was sensitive to drought stress, as the germination indexes (germination rate and germination potential) decreased significantly, and seedling growth (hypocotyl length and radicle length) was impeded under 20% PEG-6000 condition. GA3 pre-treatment affected germination rate, germination potential, hypocotyl length and radicle length. With increasing GA3 concentration, these indexes first increased and then decreased. For seedling physiology characteristics in hemp, GA3-pretreatment remarkedly increased the osmotic regulating substances (soluble sugar and soluble protein contents) and the activities of antioxidant enzymes (SOD, superoxide dismutase and POD, peroxidase), while sharply decreased the lipid peroxidation (malondialdehyde, MDA) in seedlings grown under PEG-6000 induced drought stress. These results suggested that seeds pre-treated with GA3 could enhance the drought tolerance of hempseeds, and the optimal effect of GA3 for seed pre-treatment of YM and BM could be obtained when the concentration of GA3 solution reached 400 mg/L and 600 mg/L, respectively.

Integrative Application of Foliar Yeast Extract and Gibberellic Acid Improves Morpho-Physiological Responses and Nutrient Uptake of Solidago virgaurea Plant in Alkaline Soil.

Alkaline soils have fertility issues due to poor physical qualities, which have a negative impact on crop growth and output. Solidago is used in flower arrangements, bouquet filler, and traditional medicine. The possible biological fertilizers' eco-friendly and cost-effective nature favours farmers because of the vital role in soil productivity and environmental sustainability. A field experiment was performed during two successive seasons to explore the effect of applying yeast extract (YE) at (0, 0.5, 1.0, and 1.5 g/L) and/or gibberellic acid (GA3) at (control, 100, 200, and 300 ppm) on the morpho-physiological parameters, macronutrients, and biochemical constituents of Solidago virgaurea. The results emphasize that YE (1.5 g/L) and/or GA3 (300 ppm) treatments show the highest significant increase in plant growth (i.e., plant height, no. of branches, fresh and dry weight of shoots); photosynthetic efficiency (i.e., chlorophyll (a), chlorophyll (b) and total carotenoids); macronutrient content (i.e., N, P, and K); and biochemical constituents (i.e., total soluble sugars, total phenolic, total flavonoids, and total glycosides). The study results recommend using YE and GA3 in combination at concentrations of 1.5 g/L and 300 ppm, respectively, to improve Solidago production sustainability under alkaline soil conditions.

Comparative proteomic analysis between mature and germinating seeds in Paris polyphylla var. yunnanensis.

The long dormancy period of Paris polyphylla var. yunnanensis seeds affects the supply of this scarce plant, which is used as an important traditional Chinese medicine. Mature seeds with a globular embryo and germinating seeds with developed embryo were used to explore the mechanisms of seed germination in this species. The protein profiles between the mature and germinating seeds were compared using the isobaric tags for relative and absolute quantification (iTRAQ) approach. Of the 4,488 proteins identified, a total of 1,305 differentially expressed proteins (DEPs) were detected. A Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of these DEPs indicated that metabolic pathways and the biosynthesis of secondary metabolites were the two top pathways. Additionally, phytohormone quantification shows that the abscisic acid (ABA) level significantly decreased, whereas the GA3 level dramatically increased among nine endogenous gibberellins (GAs), resulting in a significant increase of the GA3/ABA ratio in germinating seeds. The biosynthesis pathways of carotenoid as a precursor for ABA production and GA were further analyzed, and showed that proteinic expressions of the candidate genes in the two pathways did not correlate with the transcriptional abundances. However, 9-cis-epoxycarotenoid dioxygenase (NCED), a rate limited enzyme for ABA biosynthesis, was significantly decreased in mRNA levels in germinating seeds. By contrast, gibberellin 20-oxidase (GA20ox), a key enzyme GA biosynthesis, exhibited the major increase in one copy and a slight decrease in three others at the protentional level in germinating seeds. Gibberellin 2-oxidase (GA2ox), an inactivate enzyme in bioactive GAs, has the tendency to down-regulate in mRNA or at the proteinic level in germinating seeds. Altogether, these results suggested that the analyses of ABA and GA levels, the GA3/ABA ratio, and the expressional patterns of their regulatory genes may provide a novel mechanistic understanding of how phytohormones regulate seed germination in P. polyphylla var. yunnanensis.

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.

Gibberellin Signaling Promotes the Secondary Growth of Storage Roots in Panax ginseng.

Gibberellins (GAs) are an important group of phytohormones associated with diverse growth and developmental processes, including cell elongation, seed germination, and secondary growth. Recent genomic and genetic analyses have advanced our knowledge of GA signaling pathways and related genes in model plant species. However, functional genomics analyses of GA signaling pathways in Panax ginseng, a perennial herb, have rarely been carried out, despite its well-known economical and medicinal importance. Here, we conducted functional characterization of GA receptors and investigated their physiological roles in the secondary growth of P. ginseng storage roots. We found that the physiological and genetic functions of P. ginseng gibberellin-insensitive dwarf1s (PgGID1s) have been evolutionarily conserved. Additionally, the essential domains and residues in the primary protein structure for interaction with active GAs and DELLA proteins are well-conserved. Overexpression of PgGID1s in Arabidopsis completely restored the GA deficient phenotype of the Arabidopsis gid1a gid1c (atgid1a/c) double mutant. Exogenous GA treatment greatly enhanced the secondary growth of tap roots; however, paclobutrazol (PCZ), a GA biosynthetic inhibitor, reduced root growth in P. ginseng. Transcriptome profiling of P. ginseng roots revealed that GA-induced root secondary growth is closely associated with cell wall biogenesis, the cell cycle, the jasmonic acid (JA) response, and nitrate assimilation, suggesting that a transcriptional network regulate root secondary growth in P. ginseng. These results provide novel insights into the mechanism controlling secondary root growth in P. ginseng.

Plant Growth Hormones Increase the Stimulation Efficiency of Seedlings Development for Spring Wheat Seeds upon Pre-sowing Treatment.

The influence of pre-sowing treatment of spring wheat seeds with combined use of plant growth hormones and sorption preparations based on bentonite-humate mixtures on seeds germination and their development in soils was studied. In some cases, the combined use of plant growth hormones and the sorption preparation (CB-H-BYA) that can decrease the intake of allelotoxins from soil to seeds allows noticeably increasing the efficiency of plant growth hormones used for pre-sowing treatment. The inclusion of cytokinins (6-benzylaminopurine, kinetin, and forchlorophenuron) into the sorption preparation (CB-H-BYA) had markedly different effects on seeds germination. The addition of Polysorbate 20 to the sorption preparation (CB-H-BYA) leads to an increase in the effectiveness of its action on seed germination.

Comprehensive Insight into Gibberellin- and Jasmonate-Mediated Stamen Development.

In flowering plants, proper development of male generative organs is required for successful sexual reproduction. Stamen primordia arise in the third whorl of floral organs and subsequently differentiate into filaments and anthers. The early phase of stamen development, in which meiosis occurs, is followed by a late developmental phase, which consists of filament elongation coordinated with pollen maturation, anther dehiscence and finally viable pollen grain release. Stamen development and function are modulated by phytohormones, with a key role of gibberellins (GAs) and jasmonates (JAs). Long-term, extensive investigations, mainly involving GA/JA-deficient and GA/JA-response mutants, have led to a better understanding of the hormone-dependent molecular mechanisms of stamen development. In several species, the principal functions of GAs are to stimulate filament elongation through increased cell elongation and to promote anther locule opening. In the GA-dependent regulation of early stamen development, both the tapetum and developing pollen were identified as major targets. JAs mainly control the late stages of stamen development, such as filament elongation, viable pollen formation and anther dehiscence. A hierarchical relationship between GAs and JAs was recognized mainly in the control of late stamen development. By repressing DELLA proteins, GAs modulate the transcriptional activity of JA biosynthesis genes to promote JA production. A high level of JAs induces a complex of transcription factors crucial for normal stamen development.

Effects of High Temperature on Embryological Development and Hormone Profile in Flowers and Leaves of Common Buckwheat (Fagopyrum esculentum Moench).

Common buckwheat is a valuable crop, mainly due to the beneficial chemical composition of its seeds. However, buckwheat cultivation is limited because of unstable seed yield. The most important reasons for the low yield include embryo and flower abortion. The aim of this work is to verify whether high temperature affects embryological development in this plant species. The experiment was conducted on plants of a Polish cultivar 'Panda' and strain PA15, in which the percentage of degenerating embryo sacs was previously determined and amounted to 32% and 10%, respectively. The plants were cultivated in phytotronic conditions at 20 °C (control), and 30 °C (thermal stress). The embryological processes and hormonal profiles in flowers at various developmental stages (buds, open flowers, and wilted flowers) and in donor leaves were analyzed in two-month-old plants. Significant effects of thermal stress on the defective development of female gametophytes and hormone content in flowers and leaves were observed. Ovules were much more sensitive to high temperature than pollen grains in both genotypes. Pollen viability remained unaffected at 30 °C in both genotypes. The effect of temperature on female gametophyte development was visible in cv. Panda but not in PA15 buds. A drastic reduction in the number of properly developed embryo sacs was clear in open flowers at 30 °C in both genotypes. A considerable increase in abscisic acid in open flowers ready for fertilization may serve as a signal inducing flower senescence observed in the next few days. Based on embryological analyses and hormone profiles in flowers, we conclude that cv. 'Panda' is more sensitive to thermal stress than strain PA15, mainly due to a much earlier response to thermal stress involving impairment of embryological processes already in the flower buds.

One-pot sample preparation approach for profiling spatial distribution of gibberellins in a single shoot of germinating cereal seeds.

Sample preparation remains a bottleneck in the rapid and reliable quantification of gibberellins (GAs) for obtaining an insight into the physiological processes mediated by GAs. The challenges arise from not only the extremely low content of GAs in complex plant matrices, but the poor detectability of GAs by mass spectrometry (MS) in negative ion mode. In an effort to solve these urgent difficulties, we present a spatial-resolved analysis method to investigate the distribution of GAs in tiny plant tissues based on a simplified one-pot sample preparation approach coupled with ultrahigh-performance liquid chromatography-tandem MS. By integrating extraction and derivatization into one step, target GAs were effectively extracted from plant materials and simultaneously reacted with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide, the sample preparation time was largely shortened, the probability of sample loss was minimized and the detection sensitivity of MS was also greatly improved compared with underivatized GAs. Under optimal conditions, the method was validated from the quantification linearity, limits of detection and limits of quantification in the presence of plant matrices, recoveries, and precision. With the proposed method, 15 endogenous GAs were detected and, among these, 11 GAs could be quantified in 0.50 mg fresh weight (FW) wheat shoot samples, and five GAs were quantified in only 0.15 mg FW developing seed samples of Arabidopsis thaliana. The distribution patterns of GAs along both the non-13-hydroxylation pathway and the early 13-hydroxylation pathway in a single shoot of germinating wheat, rice and maize seeds were finally profiled with a spatial resolution down to approximately 1 mm2 .

Core features of the hormonal status in in vitro grown potato plants.

Some time ago, potato transformants expressing Agrobacterium-derived auxin synthesis gene tms1 were generated. These tms1-transgenic plants, showing enhanced productivity, were studied for their hormonal status, turnover and responses in comparison with control plants. For this purpose, contents of phytohormones belonging to six different classes (auxins, cytokinins, gibberellins, abscisic, jasmonic and salicylic acids) were determined by a sensitive UPLC-MS/MS method in tubers and shoots of in vitro grown plants. To date, this study represents the most comprehensive analysis of the potato hormonal system. On the basis of obtained results, several new generalizations concerning potato hormonal status were drawn. Overall, these data can serve as a framework for forthcoming integrative studies of the hormonal system in potato plants.

Molecular mechanism of seed dormancy release induced by fluridone compared with cod stratification in Notopterygium incisum.

BACKGROUND: Notopterygium incisum is an important Chinese medicinal plant. Its mature seeds have underdeveloped embryos and are physiological dormant. We found the seeds with full developed embryos can germinate after treated by fluridone (FL), an inhibitor of abscisic acid (ABA). In order to understand the molecular mechanisms underlying seed dormancy release by FL, we compared the transcriptomic changes in dormancy release induced by two different methods, FL and cold stratification (CS) in N. incisum. We further analyzed the gene expression patterns involved in seed germination and dormancy using quantitative reverse-transcription PCR. RESULTS: RNA-sequence analysis revealed more dramatic changes in the transcriptomes of FL than those in CS, particularly for genes involved in the biosynthesis and regulation of gibberellins (GAs) and ABA. The down-regulation of ABA biosynthesis genes and the dramatic up-regulation of NiCYP707As, an ABA catabolic gene, contributed to the reduced ABA levels in FL. The increased GA3 levels in CS-treated seeds were due to the up-regulation of NiGA3OX. Both NiABI5 (a positive ABA regulator) and NiGAI (a negative regulator of GA) were down-regulated in FL and CS. The upregulation of strigolactones (SLs; the metabolites with the same precursor as ABA) biosynthesis and regulatory genes in both FL- and CS-treated seeds indicates that SLs contribute positively to seed dormancy release in N. incisum. CONCLUSIONS: Our results indicated that FL- and CS-seed dormancy release possibly depends on two totally different mechanisms: alleviation of the effects of ABA and potentiation of the effects of GA, respectively. However, NiABI5 and NiGAI probably function as common factors integrating the effects of ABA and GA on seed dormancy release.

Gene mapping and transcriptome profiling of a practical photo-thermo-sensitive rice male sterile line with seedling-specific green-revertible albino leaf.

Abnormal environment weather can cause rice photoperiod-thermo-sensitive genic male sterile (PTGMS) lines fertile or partially fertile and thus cause the mixture of true hybrids with selfing seeds. Seedling-specific green-revertible albino leaf color mutant can be used to distinguish the real hybrids. Besides, it can also be used as an ideal material to research the development of chloroplast and biosynthesis of chlorophyll. The phenotype of leaf color mutants includes light green, yellowing, albino, green-revertible albino. Gene mutations affecting the synthesis and degradation of photosynthetic pigments, lycopene and heme, the differentiation and development of chloroplast, gibberellins (GAs) biosynthesis, can change the leaf color. We have created a PTGMS line with seedling-specific green-revertible albino leaf named W01S. The leaf phenotype, pollen sterility and fertility, agronomic traits, heredity, gene mapping and RNA-Seq of the differentially expressed genes between albino and green-revertible leaves were investigated. The results showed that W01S is a practical PTGMS line as Pei'ai 64S. The mutation of candidate gene Os03g0594100 (ent-isokaurene C2-hydroxylase-like) in W01S can be related to the biosynthesis of GAs, indole acetic acids, ethylene.

Differential expression of gibberellin- and abscisic acid-related genes implies their roles in the bud activity-dormancy transition of tea plants.

KEY MESSAGE: Thirty genes involved in GA and ABA metabolism and signalling were identified, and the expression profiles indicated that they play crucial roles in the bud activity-dormancy transition in tea plants. Gibberellin (GA) and abscisic acid (ABA) are fundamental phytohormones that extensively regulate plant growth and development, especially bud dormancy and sprouting transition in perennial plants. However, there is little information on GA- and ABA-related genes and their expression profiles during the activity-dormancy transition in tea plants. In the present study, 30 genes involved in the metabolism and signalling pathways of GA and ABA were first identified, and their expression patterns in different tissues were assessed. Further evaluation of the expression patterns of selected genes in response to GA3 and ABA application showed that CsGA3ox, CsGA20ox, CsGA2ox, CsZEP and CsNCED transcripts were differentially expressed after exogenous treatment. The expression profiles of the studied genes during winter dormancy and spring sprouting were investigated, and somewhat diverse expression patterns were found for GA- and ABA-related genes. This diversity was associated with the bud activity-dormancy cycle of tea plants. These results indicate that the genes involved in the metabolism and signalling of GA and ABA are important for regulating the bud activity-dormancy transition in tea plants.

Biogenic Synthesis and Spatial Distribution of Endogenous Phytohormones and Ginsenosides Provide Insights on Their Intrinsic Relevance in Panax ginseng.

Ginseng, the root and rhizome of Panax ginseng C. A. Mey., is a well-known and valuable traditional Chinese medicine. The pharmacological activities of ginseng are mainly attributed to the presence of ginsenosides, which are considered to be critical biomarkers for evaluating ginseng quality. The biosynthesis of triterpenes, which serve defensive functions in plants, is regulated by endogenous phytohormones that play key roles in growth and defense of plant populations. However, the role of major hormones that are closely related to secondary metabolism pathways in P. ginseng is poorly understood. To gain insight into their potential correlation, we performed a spatial synthesis analysis and studied the distribution of endogenous phytohormones and ginsenosides in different tissue regions of the entire P. ginseng plant. Gibberellins are growth hormones that accumulate in the fiber root. In contrast, abscisic acid (ABA), salicylic acid (SA), and jasmonic acid (JA), which are considered stress hormones, were predominantly found in the leaf and leaf peduncle. We observed a tissue-specific distribution of phytohormones consistent with the expression of genes involved in hormone biosynthesis that influenced ginsenoside synthesis and distribution. The aim of this study was to investigate the role of different endogenous phytohormones on triterpene metabolites in ginseng innate immunity.

Developmental landmarks during floral ontogeny of jalapeño chili pepper (Capsicum annuum L.) and the effect of gibberellin on ovary growth.

Pepper (Capsicum annuum L.) is an important horticultural crop in many regions of the world. The final shape and size of the fruit are known to be determined at a very early step of flower development. During flower development hormonal treatments using gibberellins seem to promote growth resulting in higher yield and fruit quality. However, the morphological changes that occur in the pepper flowers after these treatments are largely unknown. In the present study, we provide a description of floral development landmarks of jalapeño chili pepper (cultivar Huichol), divided in nine representative stages from its initiation until the opening of the bud. We established a correlation among external flower development and the time and pattern of reproductive organogenesis. Male and female gametogenesis progression was used to define specific landmarks during flower maturation. The pattern of expression of key genes involved in gibberellin metabolism and response was also evaluated in the nine flower stages. The proposed development framework was used to analyze the effect of gibberellin treatments in the development of the flower. We observed both an effect of the treatment in the histology of the ovary tissue and an increase in the level of expression of CaGA2ox1 and CaGID1b genes. The developmental stages we defined for this species are very useful to analyze the molecular and morphological changes after hormonal treatments.

Enhancing crop yield with the use of N-based fertilizers co-applied with plant hormones or growth regulators.

Crop yield, vegetative or reproductive, depends on access to an adequate supply of essential mineral nutrients. At the same time, a crop plant's growth and development, and thus yield, also depend on in situ production of plant hormones. Thus optimizing mineral nutrition and providing supplemental hormones are two mechanisms for gaining appreciable yield increases. Optimizing the mineral nutrient supply is a common and accepted agricultural practice, but the co-application of nitrogen-based fertilizers with plant hormones or plant growth regulators is relatively uncommon. Our review discusses possible uses of plant hormones (gibberellins, auxins, cytokinins, abscisic acid and ethylene) and specific growth regulators (glycine betaine and polyamines) to enhance and optimize crop yield when co-applied with nitrogen-based fertilizers. We conclude that use of growth-active gibberellins, together with a nitrogen-based fertilizer, can result in appreciable and significant additive increases in shoot dry biomass of crops, including forage crops growing under low-temperature conditions. There may also be a potential for use of an auxin or cytokinin, together with a nitrogen-based fertilizer, for obtaining additive increases in dry shoot biomass and/or reproductive yield. Further research, though, is needed to determine the potential of co-application of nitrogen-based fertilizers with abscisic acid, ethylene and other growth regulators.

Down regulation of StGA3ox genes in potato results in altered GA content and affect plant and tuber growth characteristics.

GA biosynthesis and catabolism has been shown to play an important role in regulating tuberization in potato. Active GAs are inactivated in the stolon tips shortly after induction to tuberization. Overexpression of a GA inactivation gene results in an earlier tuberization phenotype, while reducing expression of the same gene results in delayed tuberization. In addition, overexpression of genes involved in GA biosynthesis results in delayed tuberization, while decreased expression of those genes results in earlied tuberization. The final step in GA biosynthesis is catalysed by StGA3ox1 and StGA3ox2 activity, that convert inactive forms of GA into active GA1 and GA4. In this study we cloned StGA3ox2 gene in an RNAi construct and used this construct to transform potato plants. The StGA3ox2 silenced plants were smaller and had shorter internodes. In addition, we assayed the concentrations of various GAs in the transgenic plants and showed an altered GA content. No difference was observed on the time point of tuber initiation. However, the transgenic clones had increased number of tubers with the same yield, resulting in smaller average tuber weight. In addition, we cloned the promoter of StGA3ox2 to direct expression of the GUS reporter gene to visualize the sites of GA biosynthesis in the potato plant. Finally, we discuss how changes of several GA levels can have an impact on shoot, stolon and tuber development, as well as the possible mechanisms that mediate feed-forward and feed-back regulation loops in the GA biosynthetic pathway in potato.