Flower Induction Research Articles

Cloning and function analysis of DlWRKY9 gene in longan (Dimocarpus longan)

WRKY is one of the largest plant transcription factors (TFs) which is widely involved in plant growth, development, and responses to stresses. In the present study, a WRKY TF DlWRKY9 was cloned from longan (Dimocarpus longan). The coding sequence (CDS) of DlWRKY9 is 762 bp in length and encodes 253 amino acids. It has a typical WRKY domain and zinc finger structure which belongs to type IIa WRKY protein. The molecular weight of DlWRKY9 protein was 30.27kda and the theoretical isoelectric point (PI) was 5.24. It is an unstable hydrophilic protein. The secondary structure of DlWRKY9 protein consists of helical structure (17.39%), extended chain (8.70%) and other structures (turn and random coil) (73.91%). The amino acid sequence of DlWRKY9 protein had the highest similarity with DlWRKY9 (xp_006450293.1) of citrus Clementina. DlWRKY9 gene promoter elements contain light, abscisic acid, gibberellin, jasmonic acid and other response elements. The results of qRT-PCR showed that the relative expression level of DlWRKY9 gene in pericarp was higher, followed by young fruits and floral organs. Meanwhile, DlWRKY9 gene specifically down-regulated in the early stage of flower induction in ‘Sijimi’ (SJ) longan. The results of transient expression of Arabidopsis protoplasts showed that the fluorescence signal was mainly concentrated in the nucleus. Moreover, overexpression of DlWRKY9 in Arabidopsis promoted early flowering. These results provide useful information for revealing the biological roles of DlWRKY9 in longan and increase our understanding of the WRKY family in fruit trees.

Genome-wide characterization and expression profiling of GASA gene family in Triticum turgidum ssp. durum (desf.) husn. (Durum wheat) unveils its involvement in environmental stress responses

Family members within the plant-specific gibberellic acid-stimulated Arabidopsis (GASA) gene serve a crucial role in plant growth and development, particularly in flower induction and seed development. Through a genome-wide analysis of Triticum turgidum ssp. Durum (durum wheat), we identified 19 GASA genes, designated as TdGASA1‒19. Moreover, the chromosomal locations, exon–intron distribution and the physiochemical properties of these genes were determined and the subcellular localization of their encoded proteins was estimated. Analyses of their domain structure, motif arrangements, and phylogeny revealed four distinct groups that share a conserved GASA domain. Additionally, a real-time q-PCR analysis revealed differential expression patterns of TdGASA genes in various tissues (including leaves, roots, stems, and seeds) and in response to salinity, osmotic stress, and treatment with exogenous phytohormones (abscisic and gibberellic acid), implying that these genes may play a role in the growth, development, and stress responses of Triticum turgidum. Heterologous expression of TdGASA1, TdGASA4, TdGASA14, and TdGASA19 in Saccharomyces cerevisiae improved its tolerance to salt, osmotic, oxidative, and heat stresses, which suggests the involvement of these genes in abiotic stress tolerance mechanisms. The present study is the first to identify and analyze the expression profile of T. turgidum GASA genes, therefore offering novel insights for their further functional characterization, which may serve as a novel resource for molecular breeding of durum wheat.

Sugar metabolism mediates temperature-dependent flowering induction in saffron (Crocus sativus L.)

Saffron, which comprises the stigma of the Crocus sativus flower, is an important culinary and medicinal product. Saffron productivity depends on its flower formation, which is regulated by temperature. Inappropriate temperatures at the flowering stages result in flower atrophy and reduced or no flowering. The effect of temperature on saffron flowering was investigated using morphological studies coupled with transcriptomics and metabolic studies of meristem-enriched tissue collected from corms stored and grown at two different temperatures [high (25°C) and low (8°C)] during flowering. Morphological and histological studies suggested that high temperature, during the resting phase of the saffron corm life cycle, promotes flowering, whereas low temperature suppresses it. Low temperatures suppress flowering by inhibiting flower induction. Transcriptome analysis during the flower induction stage indicated that sugar metabolism genes were differentially regulated at different temperatures. Genes coding for starch and sucrose metabolism were enriched and differentially regulated at 25°C vs 8°C. High temperature promoted starch breakdown and soluble sugar formation, whereas low temperature had a vice-versa effect. Further, we identified that low-temperature mediated suppression of flowering involves the downregulation of the floral integrator gene CsatFT3. Exogenous sugar feeding, mainly sucrose, can rescue the low-temperature suppression of flower induction via upregulation of the CsatFT3 gene. Overall the results suggest that environmental temperatures alter sugar metabolism, which mediates temperature-dependent flower induction in saffron corms via regulation of the FT3 gene. These findings will aid in identifying temperature driven molecular factors controlling flowering in saffron.

Effect of 6-Benzylaminopurine (BA) and Paclobutrazol (PBZ) with Light Intensity Variations for Cherry Tomatoes In vitro Flowering

Utilization of Solanum lycopersicum L. and its varieties contribute to plant biotechnology to study the effect of genes and flower transition events. This study aims to optimize media and light intensity for in vitro flowering of a cherry tomato variety. Induction of flowering was carried out using seven types of media combined with 3 light intensities, followed by feeding by adding liquid medium MS+PBZ 1 mg/L. A positive response is shown at the light intensity of 2,600 lux; there are 5 explants that have flower buds. The percentage of flower buds for each treatment combination was 50% from two replications. The first shoots occurred at 142 days after planting (DAP) on explants grown on media containing 0.5 mg/L BA, 0.75 mg/L PBZ, with feeding treatment. It can also be assumed that the emergence of flower buds occurred not because of the PGR induction treatment, but because at that time the explants had entered the generative phase, considering that the plantlets had been growing for a long time. Although the effects of BA and PBZ have not been confirmed, the use of 2,600 lux light appears to be the best condition for cherry tomato in vitro flowering induction.

Overexpression of two CONSTANS-like 2 (MiCOL2) genes from mango delays flowering and enhances tolerance to abiotic stress in transgenic Arabidopsis

The CO/COL gene family plays an important role in regulating photoperiod-dependent flowering time in plants. In this study, two COL2 gene homologs, MiCOL2A and MiCOL2B, were isolated from ‘SiJiMi’ mango, and their expression patterns and functions were characterized. The MiCOL2A and MiCOL2B genes both belonged to the group Ⅰ of CO/COL gene family. MiCOL2A and MiCOL2B exhibited distinct circadian rhythms and were highly expressed in leaves during the flowering induction period. Subcellular localization analysis revealed that MiCOL2A and MiCOL2B are localized in the nucleus. The overexpression of MiCOL2A and MiCOL2B significantly delayed flowering time in Arabidopsis under both long-day (LD) and short-day (SD) conditions. The MiCOL2A and MiCOL2B overexpression Arabidopsis plants exhibited more tolerance to slat and drought stress after abiotic stress treatments, with greater ROS scavenging capacity and protective enzyme activity, less cell damage and death and higher expression of stress response genes than wild type plants. Bimolecular fluorescence complementation (BiFC) analysis showed that MiCOL2A and MiCOL2B interacted with several stress-related proteins, including zinc finger protein 4 (MiZFP4), MYB30-INTERACTING E3 LIGASE 1 (MiMIEL1) and RING zinc finger protein 34 (MiRZFP34). The results indicate that MiCOL2A and MiCOL2B are not only involved in flowering time but also play a positive role in abiotic stress responses in plants.

Effects of hormone and fertilizers on early flower induction of Dendrobium anosmum hybrid seedlings under ex vitro condition

Early flowering of new orchids is important to save time for selecting valuable flowers and artificial induction of flowering is a critical consideration in the orchid production industry. In this study, a new Dendrobium anosmum hybrid was generated by cross-breeding between D. anosmum ‘Chau Nhu’ and D. anosmum ‘Di Linh’. The ancestors and hybrid seedlings from in-vitro culture were trained in the net house and their growth and flowering were evaluated under ex vivo conditions with specific fertilizers and hormones. The results suggest that the hybrid plants grew better than their parents in terms of stem height, stem diameter, and leaf number. Growth hormones were applied to stimulate early flowering in matured hybrids and it was discovered that ‘Keiki pro’, a commercial hormone product, produced the best results, with a flowering rate of 66.67% after two applications. Hybrid flowers varied in width from 36.36% (3.0-6.0 cm) to 63.64 % (more than 6.0 cm) from ancestral width in medium-sized and large-sized flowers, respectively. Also, the hybrid flower colours was mostly a combination of pink/violet (75C) and purple/pink (68A), which is different from their parents. Importantly, the dorsal sepal, petal colours, and shape of hybrid flowers varied significantly among individual hybrids, between hybrids and their progenitors. Some mutations in the lips and columns of the novel hybrid flowers were also visualized. Hence, the D. anosmum hybrid seedlings successfully induced flowers after a year of culture under optimal hormones and fertilizers conditions. The results can serve as a critical reference for the early flowering of the orchid seedlings.

Creating optimal nutrient conditions in soil and plants during the flower induction process of sweet lime (Citrus limettioides) under calcareous soil conditions to increase the yield

A study was conducted on 6-year-old sweet lime trees on the Mexican lime (Citrus aurantifolia Swingle) rootstock in a randomized complete block design with ten treatments and three replications in southern Fars to determine the appropriate time of plant supplementary feeding to increase flower production and the yield in calcareous soils. In 2016 and 2017, 40 trees that had the same planting and growing conditions, were selected to determine the flower induction time. Ten times (22nd of October, 1st, 11th, and 20th November, December, and January), four branches that had a length of 120 cm and a diameter of half a centimeter were selected on each tree in four geographical directions. At each time, the branches of three trees were ringed at a distance of 120 cm from the tip of the branch, and all the leaves were removed. At the time of flower emergence, the number of flowers was counted. The results showed that the flower induction in these trees was in early December. Based on the results of this experiment and previous research in this field, in September 2017, the 7-year-old trees grafted on the Mexican lime rootstock were treated with the foliar application of nitrogen as urea [CO(NH2)2] and zinc as zinc sulfate (ZnSO4) at three concentrations including 0, 3, and 5 g∙L–1, alone and by combination in two separate pieces in a randomized complete-block factorial design including nine treatments and three replications to increase the yield using elements involved in the flowering. At the harvest time, fruit juice, TSS, TA, vitamin C, average fruit weight, and single tree yield were measured. According to the results, the combination of urea and zinc sulfate at a concentration of 5 g∙L–1 led to an increase in the average fruit weight (58 g) and the yield (68 kg) compared to the control.

Effect of Salicylic Acid and Pre-Cold Treatment on Flower Induction in Saffron.

Saffron is an important flowering plant, generally known as a golden condiment. The present study was performed to find the influence of different levels of SA and pre-cold treatment in the dormancy period of saffron and their effects on content enzyme activity. The results indicated that the SA2%, SA1%, and, pre-cold treatments took the shortest day to flowering. PAL enzyme activity was highest in pre-cold treatment. The higher total amount of protein was measured in the control, SA1% and SA2%. The highest amount of starch content and total soluble sugar was detected in pre-cold, SA2%, and control, respectively. No significant differences between treatments were present for CAT, PPO, GPX, and APX enzymes activity. There was a negative significant correlation between flowering time and some studied traits, i.e., starch and PAL activity. Applying SA and pre-cold treatment can induce saffron flowering and effect on pal enzyme activity and corm total protein, sugar, and starch content accordingly.

Repeated runner removal after harvest and floating row cover during fall affect carbohydrate status and yield potential of strawberry cvs. Polka and Wendy in the Northern climate

We investigated the impact of repeated runner removal after harvest and row cover during fall on strawberry (Fragaria × ananassa) cvs. Polka and Wendy cropping potential in the joint project of the University of Helsinki and the Rural Advisory Services Finland. Field trials were carried out in 2017 to 2019 on strawberry farms in Southern Ostrobothnia, Finland, at 62°49′N. The treatment effects on the crown carbohydrate reserves, flowering, and yield were recorded. Runner removal three times in three-week intervals after harvest decreased the crown starch reserves but increased the length of the apical inflorescence initials as observed through flower mapping in November, and the numbers of inflorescences and flowers in cv. Polka. Floating row cover after flower induction in September to October hastened floral development in the fall and increased the numbers of inflorescences, flowers, and fruit in cv. Wendy. Although the effects may depend on a cultivar and weather conditions during fall, repeated removal of runners after harvest and covering plants with a floating row cover after flower induction are recommended to increase strawberry yield potential in the Northern climate.

A comparative proteomic analysis provides insight into the molecular mechanism of bud break in longan

BackgroundThe timing of bud break is very important for the flowering and fruiting of longan. To obtain new insights into the underlying regulatory mechanism of bud break in longan, a comparative analysis was conducted in three flower induction stages of two longan varieties with opposite flowering phenotypes by using isobaric tags for relative and absolute quantification (iTRAQ).ResultsIn total, 3180 unique proteins were identified in 18 samples, and 1101 differentially abundant proteins (DAPs) were identified. “SX” (“Shixia”), a common longan cultivated variety that needs an appropriate period of low temperatures to accumulate energy and nutrients for flower induction, had a strong primary inflorescence, had a strong axillary inflorescence, and contained high contents of sugars, and most DAPs during the bud break process were enriched in assimilates and energy metabolism. Combined with our previous transcriptome data, it was observed that sucrose synthase 6 (SS6) and granule-bound starch synthase 1 (GBSSI) might be the key DAPs for “SX” bud break. Compared to those of “SX”, the primary inflorescence, axillary inflorescence, floral primordium, bract, and prophyll of “SJ” (“Sijimi”) were weaker. In addition, light, rather than a high sugar content or chilling duration, might act as the key signal for triggering bud break. In addition, catalase isozyme 1, an important enzyme in the redox cycle, and RuBisCO, a key enzyme in the Calvin cycle of photosynthetic carbon assimilation, might be the key DAPs for SJ bud break.ConclusionOur results present a dynamic picture of the bud break of longan, not only revealing the temporal specific expression of key candidate genes and proteins but also providing a scientific basis for the genetic improvement of this fruit tree species.

Drought and low temperature-induced NF-YA1 activates FT expression to promote citrus flowering.

Flower induction in adult citrus is mainly regulated by drought and low temperatures. However, the mechanism of FLOWERING LOCUS T regulation of citrus flowering (CiFT) under two flower-inductive stimuli remains largely unclear. In this study, a citrus transcription factor, nuclear factor YA (CiNF-YA1), was found to specifically bind to the CiFT promoter by forming a complex with CiNF-YB2 and CiNF-YC2 to activate CiFT expression. CiNF-YA1 was induced in juvenile citrus by low temperature and drought treatments. Overexpression of CiNF-YA1 increased drought susceptibility in transgenic citrus, whereas suppression of CiNF-YA1 enhanced drought tolerance in silenced citrus plants. Furthermore, a GOLDEN2 - LIKE protein (CiFE) that interacts with CiFT protein was also isolated. Further experimental evidence showed that CiFE binds to the citrus LEAFY (CiLFY) promoter and activates its expression. In addition, the expressions of CiNF-YA1 and CiFE showed a seasonal increase during the floral induction period and were induced by artificial drought and low-temperature treatments at which floral induction occurred. These results indicate that CiNF-YA1 may activate CiFT expression in response to drought and low temperatures by binding to the CiFT promoter. CiFT then forms a complex with CiFE to activate CiLFY, thereby promoting the flowering of adult citrus.

Effect of Pre-Harvest Foliar Calcium and Silicon Fertilization on Pineapple Quality and Fruit Collapse Incidence

Pineapple can be affected by fruit collapse, a disease caused by the bacterium Dickeya zeae. However, adequate fertilization can increase the fruit’s resistance to this illness. Therefore, the impact of preharvest foliar calcium and silicon fertilization on pineapple quality and fruit collapse incidence was assessed in this study. The experiment implemented a split-plot design with two factors. The first factor has two terms of inoculation (flower induction and before harvest). The second factor uses a control with three foliar fertilization treatments, A (control: No foliar fertilizers applied), B (Ca from 13 to 11 weeks before harvest/from 6 weeks to harvest), C (Si from 13 to 11 weeks before harvest/from 6 weeks to harvest), and D (Ca + Si from 13 to 11 weeks before harvest/from 6 weeks to harvest). Treatment D gave the best response. It had the lowest fruit collapse incidence (21.70%), highest ascorbic acid (71.64 mg/kg), elevated β-carotene (4.87 mg/kg) and mineral content (Ca: 1851.10 mg/kg, Si: 1164.87 mg/kg), essentially under the before harvest term of inoculation, which was more harmful for the fruit. In conclusion, mixed foliar calcium and silicon fertilization manage to improve the tolerance to fruit collapse incidence, impacting the pineapple quality positively

Long-day photoperiod and cool temperature induce flowering in cassava: Expression of signaling genes.

Cassava is a staple food crop in the tropics, and is of particular importance in Africa. Recent development of genomic selection technology have improved the speed of cassava breeding; however, cassava flower initiation and development remains a bottleneck. The objectives of the current studies were to elucidate the effect of photoperiod, temperature and their interactions on the time of flowering and flower development in controlled environments, and to use RNA-sequencing to identify transcriptome expression underlying these environmental responses. Compared to a normal tropical day-length of 12 h, increasing the photoperiod by 4 h or decreasing the air temperature from 34/31 to 22°/19°C (day/night) substantially hastened the time to flowering. For both photoperiod and temperature, the environment most favorable for flowering was opposite the one for storage root harvest index. There was a pronounced treatment interaction: at warm day-time temperatures, percent flowering was low, and photoperiod had little effect. In contrast, at cooler temperatures, percent flowering increased, and long-day (LD) photoperiod had a strong effect in hastening flowering. In response to temperature, many differentially expressed genes in the sugar, phase-change, and flowering-time-integrator pathways had expression/flowering patterns in the same direction as in Arabidopsis (positive or negative) even though the effect of temperature on flowering operates in the reverse direction in cassava compared to Arabidopsis. Three trehalose-6-phosphate-synthase-1 (TPS1) genes and four members of the SPL gene family had significantly increased expression at cool temperature, suggesting sugar signaling roles in flower induction. In response to LD photoperiod, regulatory genes were expressed as in Arabidopsis and other LD flowering plants. Several hormone-related genes were expressed in response to both photoperiod and temperature. In summary, these findings provide insight on photoperiod and temperature responses and underlying gene expression that may assist breeding programs to manipulate flowering for more rapid crop improvement.

Determining Reproductive Parameters, which Contribute to Variation in Yield of Olive Trees from Different Cultivars, Irrigation Regimes, Age and Location.

Olive (Olea europaea L.) trees can reach a very old age and still bear fruit. Although traditional groves are planted at low density and are rainfed, many newer groves are planted at higher densities and irrigated. As expected, initial yields per area are larger in high density plantations, yet some farmers claim they experience a reduction in productivity with grove age, even in well maintained trees. In order to test the accuracy of this claim and its underlying cause, we measured several productivity parameters in selected branches of trees in seven sites differing in cultivar (‘Barnea’ or ‘Souri’), location and irrigation regime (rainfed or irrigated) for two consecutive years. For each site (cultivar/location/regime), we compared neighboring groves of different ages, altogether 14 groves. There was no consistent reduction in productivity in older groves. Differences in productivity between irrigated cultivars were mostly due to variation in the percentage of inflorescences that formed fruit. Several parameters were higher in irrigated, compared to rainfed ‘Souri’. Differences in productivity between years within the same grove was mostly due to variation in the percentage of nodes forming inflorescences. We studied the expression of OeFT2 encoding a FLOWERING LOCUS T protein involved in olive flower induction in leaves of trees of different ages, including juvenile seedlings. Expression increased during winter in mature trees and correlated with the percentage of inflorescences formed. The leaves of juvenile seedlings expressed higher levels of two genes encoding APETALA2-like proteins, potential inhibitors of OeFT2 expression. The buds of juvenile seedlings expressed higher levels of OeTFL1, encoding a TERMINAL FLOWER 1 protein, a potential inhibitor of OeFT2 function in the meristem. Our results suggest that olives, once past the juvenile phase, can retain a similar level of productivity even in densely planted well maintained groves.

Genome-Wide Analysis of the SPL Gene Family and Expression Analysis during Flowering Induction in Prunus × yedoensis 'Somei-yoshino'.

SQUAMOSA Promoter-Binding Protein-Like (SPL) genes encode plant-specific transcription factors which bind to the SQUAMOSA promoter of the MADS-box genes to regulate its expression. It plays important regulatory roles in floral induction and development, fertility, light signals and hormonal transduction, and stress response in plants. In this study, 32 PySPL genes with complete SBP (squamosa promoter binding protein) conserved domain were identified from the genome of Prunus × yedoensis ‘Somei-yoshino’ and analyzed by bioinformatics. 32 PySPLs were distributed on 13 chromosomes, encoding 32 PySPL proteins with different physical and chemical properties. The phylogenetic tree constructed with Arabidopsis thaliana and Oryza sativa can be divided into 10 subtribes, indicating PySPLs of different clusters have different biological functions. The conserved motif prediction showed that the number and distribution of motifs on each PySPL is varied. The gene structure analysis revealed that PySPLs harbored exons ranging from 2 to 10. The predictive analysis of acting elements showed that the promoter of PySPLs contain a large number of light-responsive elements, as well as response elements related to hormone response, growth and development and stress response. The analysis of the PySPLs expressions in flower induction and flower organs based on qRT-PCR showed that PySPL06/22 may be the key genes of flower development, PySPL01/06 and PySPL22 may play a role in the development of sepal and pistil, respectively. The results provide a foundation for the study of SPL transcription factors of Prunus × yedoensis ‘Somei-yoshino’ and provide more reference information of the function of SPL gene in flowering.

Application Paclobutrazol and Duration of Drought Stress to Flowering Induction in Chokun Orange

Induction of flowering is one of the efforts that can extend the production period of Chokun oranges. This study aims to determine the effect of the dose of paclobutrazol (PBZ), duration of drought stress and the combination of treatments that gives the best results on the Chokun orange (<em>Citrus</em> sp.). The research design used was randomized complete block design (RCBD), consisting of 3 replications. The treatment in this study was a combination of the dose of PBZ (control, active ingredient 0.75 g plant<sup>-1</sup> and active ingredient 1.5 g plant<sup>-1</sup>) and duration of drought stress (control, 1, 2 and 3 weeks). The results show that the application of PBZ and duration of drought stress can induce the flowering of citrus plants as seen from the generative shoot variables with a quadratic model on the equation y = -17.778x<sup>2</sup> + 31.556x + 26.667 at the optimum dose of 0.89 g plant<sup>-1</sup> of active ingredient and 1 week of drought. The dry period of 3 weeks gives the best results seen from variables of the number of flowers and number of fruits. In general, the results suggest that the application of PBZ and duration of drought stress can transfer from the vegetative phase to the generative phase which in turn could induce the flowering of citrus plants.

Transcriptome analysis reveals dual action of salicylic acid application in the induction of flowering in Malus domestica

In the apple tree, insufficient flower bud production is an intractable challenge, and very little information is available in this field due to the fact that research done in this sector is very rare owing to its extended life cycles and low rate of genetic transformation. Here we display novel changes and events in spur buds of Malus × domestica trees after they were exposed to salicylic acid (SA) treatment during the flower induction period. We found a significant increase in morphological indexes, followed by a wider and well-defined shoot apical meristem in SA-treated spur buds. Additionally, we observed increased oxidative stress markers and enzymatic antioxidants in control-treated buds during the flower induction period, while non-enzymatic antioxidants were recorded higher in SA-treated buds. Maximum flowering was observed in SA-treated trees in the next year. Furthermore, ultra-high-performance liquid chromatography (u-HPLC) analysis displays that SA treatment enhances SA and indole acetic acid (IAA), while having an antagonistic effect on gibberellin (GA). At different time points, transcriptome analysis was conducted to analyze the transcriptional response of CK and SA treated buds. Pathway enrichment was detected in differentially expressed genes (DEGs). Agamous (AGL) and SQUAMOSA-promoter binding protein-like (SPL) family related flowering genes display a positive signal for the increased flowering in SA-treated trees, which confirms our findings. As far as we know, there is no report available on the response of spur buds to SA treatment during the flower induction period. This data provides a new theoretical reference for the management of apple tree flowering and also provides an essential basis for future analysis of the regulation and control of flowering in M. domestica.

FLOWERING LOCUS M isoforms differentially affect the subcellular localization and stability of SHORT VEGETATIVE PHASE to regulate temperature-responsive flowering in Arabidopsis

Temperature is an important environmental cue that affects flowering time in plants. The MADS-box transcription factor FLOWERING LOCUS M (FLM) forms a heterodimeric complex with SHORT VEGETATIVE PHASE (SVP) and controls ambient temperature-responsive flowering in Arabidopsis. FLM-β and FLM-δ, two major splice variants produced from the FLM locus, exert opposite effects on flowering, but the molecular mechanism by which the interaction between FLM isoforms and SVP affects temperature-responsive flowering remains poorly understood. Here, we show that FLM-β and FLM-δ play important roles in modulating the temperature-dependent behavior, conformation, and stability of SVP. Nuclear localization of SVP decreases as temperature increases. FLM-β is required for SVP nuclear translocation at low temperature, whereas SVP interacts with FLM-δ mainly in the cytoplasm at high temperature. SVP preferentially binds to FLM-β at low temperature in tobacco leaf cells. SVP shows high binding affinity to FLM-β at low temperature and to FLM-δ at high temperature. SVP undergoes similar structural changes in the interactions with FLM-β and FLM-δ; however, FLM-δ likely causes more pronounced conformational changes in the SVP structure. FLM-δ causes rapid degradation of SVP at high temperature, compared with FLM-β, possibly via ubiquitination. Mutation of lysine 53 or lysine 165 in SVP causes increased abundance of SVP due to reduced ubiquitination of SVP and thus delays flowering at high temperature. Our findings suggest that temperature-dependent differential interactions between SVP and FLM isoforms modulate the temperature-responsive induction of flowering in Arabidopsis.

Role of FaSOC1 and FaCO in the seasonal control of reproductive and vegetative development in the perennial crop Fragaria × ananassa.

The diploid woodland strawberry (F. vesca) represents an important model for the genus Fragaria. Significant advances in the understanding of the molecular mechanisms regulating seasonal alternance of flower induction and vegetative reproduction has been made in this species. However, this research area has received little attention on the cultivated octoploid strawberry (F. × ananassa) despite its enormous agronomical and economic importance. To advance in the characterization of this intricated molecular network, expression analysis of key flowering time genes was performed both in short and long days and in cultivars with seasonal and perpetual flowering. Analysis of overexpression of FaCO and FaSOC1 in the seasonal flowering ‘Camarosa’ allowed functional validation of a number of responses already observed in F. vesca while uncovered differences related to the regulation of FaFTs expression and gibberellins (GAs) biosynthesis. While FvCO has been shown to promote flowering and inhibit runner development in the perpetual flowering H4 accession of F. vesca, our study showed that FaCO responds to LD photoperiods as in F. vesca but delayed flowering to some extent, possibly by induction of the strong FaTFL1 repressor in crowns. A contrasting effect on runnering was observed in FaCO transgenic plants, some lines showing reduced runner number whereas in others runnering was slightly accelerated. We demonstrate that the role of the MADS-box transcription factor FaSOC1 as a strong repressor of flowering and promoter of vegetative growth is conserved in woodland and cultivated strawberry. Our study further indicates an important role of FaSOC1 in the photoperiodic repression of FLOWERING LOCUS T (FT) genes FaFT2 and FaFT3 while FaTFL1 upregulation was less prominent than that observed in F. vesca. In our experimental conditions, FaSOC1 promotion of vegetative growth do not require induction of GA biosynthesis, despite GA biosynthesis genes showed a marked photoperiodic upregulation in response to long days, supporting GA requirement for the promotion of vegetative growth. Our results also provided insights into additional factors, such as FaTEM, associated with the vegetative developmental phase that deserve further characterization in the future.

Expression Patterns of Key Genes in the Photoperiod and Vernalization Flowering Pathways in Lilium longiflorum with Different Bulb Sizes.

Lilium longiflorum is a wild Lilium, and its flowering transition requires a long period of cold exposure to meet the demand of vernalization. The responses of different sized bulbs to cold exposure and photoperiod are different, and the floral transition pathways of small and large bulbs are different. In this study, small and large bulbs were placed in cold storage for different weeks and then cultured at a constant ambient temperature of 25 °C under long day (LD) and short day (SD) conditions. Then, the flowering characteristics and expression patterns of key genes related to the vernalization and photoperiod pathways in different groups were calculated and analyzed. The results showed that the floral transition of Lilium longiflorum was influenced by both vernalization and photoperiod, that vernalization and LD conditions can significantly improve the flowering rate of Lilium longiflorum, and that the time from planting to visible flowering buds’ appearance was decreased. The flowering time and rate of large bulbs were greatly influenced by cold exposure, and the vernalization pathway acted more actively at the floral transition stage. The floral transition of small bulbs was affected more by the photoperiod pathway. Moreover, it was speculated that cold exposure may promote greater sensitivity of the small bulbs to LD conditions. In addition, the expression of LlVRN1, LlFKF1, LlGI, LlCO5, LlCO7, LlCO16, LlFT1, LlFT3 and LlSOC1 was high during the process of floral transition, and LlCO13, LlCO14 and LlCO15 were highly expressed in the vegetative stage. The expression of LlCO13 and LlCO14 was different under different lighting conditions, and the flowering induction function of LlCO9 and LlFT3 was related to vernalization. Moreover, LlFKF1, LlGI, LlCO5, LlCO16, LlSOC1 and LlFT2 were involved in the entire growth process of plants, while LlCO6, LlCO16 and LlFT1 are involved in the differentiation and formation of small bulblets of plants after the inflorescence stage, and this process is also closely related to LD conditions. This study has great significance for understanding the molecular mechanisms of the vernalization and photoperiod flowering pathways of Lilium longiflorum.