Induction and promotion of flowering by heat-treated catecholamines in Lemna paucicostata

Alkali- and heat-treated norepinephrine, a catecholamine, induced flowering of short-day (SD) plant Lemna paucicostata 151 even under long-day (LD) conditions. Flowering induced with a lower concentration of heat-treated norepinephrine was promoted under SD conditions but inhibited by a night break. The related compounds L-dopa and dopamine also promoted flowering under SD conditions when they were heat-treated.     

A Comparative Study on the Short-Day- and the Benzoic Acid-Induced Flowering in Lemna paucicostata

The addition of a high concentration of FeCl₃ to the medium(1/10 strength M medium) slightly inhibited the benzoic acid(BA)-induced flowering of Lemna paucicostata 151 in continuouslight, although it promoted the flowering induced by short-day(SD) conditions. SD treatment had no significant effect on BA-inducedflowering in the medium with a standard concentration of iron,in which this plant hardly responds to SD, but greatly promotedit in the medium containing iron at a high concentration, inwhich this plant clearly responds to SD. The effect of BA seemsto be independent of but additive to the photoperiodic stimulus. In photosensitive strains 6746 and 441, a low concentrationof BA slightly lengthened the critical photoperiod but had noflower-inducing effect under continuous light. Since an optimalconcentration of BA induced flowering even under continuouslight in these strains, this was considered to be due to photoperiod-independentpromotion of flowering rather than shortening of the time-measuringprocess in the photoperiodic reaction. (Received August 19, 1986; Accepted February 21, 1987)     

A genetic and physiological analysis of late flowering mutants in Arabidopsis thaliana

Summary Monogenic mutants of the early ecotype Landsberg erecta were selected on the basis of late flowering under long day (LD) conditions after treatment with ethyl methanesulphonate or irradiation. In addition to later flowering the number of rosette and cauline leaves is proportionally higher in all mutants, although the correlation coefficient between the two parameters is not the same for all genotypes. Forty-two independently induced mutants were found to represent mutations at 11 loci. The mutations were either recessive, intermediate (co locus) or almost completely dominant (fwa locus). The loci are located at distinct positions on four of the five Arabidopsis chromosomes. Recombinants carrying mutations at different loci flower later than or as late as the later parental mutant. This distinction led to the assignment of eight of the loci to three epistatic groups. In wild type, vernalization promotes flowering to a small extent. For mutants at the loci fca, fve, fy and fpa, vernalization has a large effect both under LD and short day (SD) conditions, whereas co, gi, fd and fwa mutants are almost completely insensitive to this treatment. SD induces later flowering except for mutants at the co and gi loci, which flower with the same number of leaves under LD and SD conditions. This differential response of the mutants to environmental factors and their subdivision into epistatic groups is discussed in relation to a causal model for floral initiation in Arabidopsis thaliana.     

Flowering and dwarfism induced by DNA demethylation in Pharbitis nil

Flowering and dwarfism induced by 5‐azacytidine and zebularine, which both cause DNA demethylation, were studied in a short‐day (SD) plant Pharbitis nil (synonym Ipomoea nil), var. Violet whose photoinduced flowering state does not last for a long period of time. The DNA demethylating reagents induced flowering under non‐inductive long‐day (LD) conditions. The flower‐inducing effect of 5‐azacytidine did not last for a long period of time, and the plants reverted to vegetative growth. The progeny of the plants that were induced to flower by DNA demethylation did not flower under the non‐inductive photoperiodic conditions. These results suggest that the flowering‐related genes were activated by DNA demethylation and then remethylated again in the progeny. The DNA demethylation also induced dwarfism. The dwarfism did not last for a long period of time, was not heritable and was overcome by gibberellin A₃ but not by t‐zeatin or kinetin. The change in the genome‐wide methylation state was examined by methylation‐sensitive amplified fragment length polymorphism (MS‐AFLP) analysis. The analysis detected many more polymorphic fragments between the DNA samples isolated from the cotyledons treated with SD than from the cotyledons under LD conditions, indicating that the DNA methylation state was altered by photoperiodic conditions. Seven LD‐specific fragments were extracted from the gel of the MS‐AFLP and were sequenced. One of these fragments was highly homologous with the genes encoding ribosomal proteins.     

CONSTANS delays flowering and affects tuber yield in potato

CONSTANS (CO) has a central role in the photoperiodic regulation of flowering in Arabidopsis thaliana. We show here that potato (Solanum tuberosum ssp. andigena) plants constitutively expressing Arabidopsis CO (pACo plants) flower late under all photoperiodic conditions tested. Exogenous application of gibberellic acid to pACo plants corrected their short stem phenotype but not their late flowering. To further understand the effect of CO in potato, we used three photoperiodic conditions: short days (SD), which strongly induce tuberisation of wild type plants, SD supplemented with a night break (SD+NB), which are moderately inductive, and tuberisation-inhibiting long days. Tuberisation of pACo plants was delayed under SD and very strongly delayed or completely inhibited under SD+NB, suggesting that CO affects an autonomous pathway controlling potato tuberisation. In addition, tuber yield, a trait of high agronomic relevance, was significantly increased in pACo plants expressing moderate CO levels. Our results indicate that CO affects flowering and stem elongation through distinct mechanisms and suggest that its effects on flowering and tuberisation in potato are photoperiod-independent.     

CONSTANS delays Arabidopsis flowering under short days

Long days (LD) promote flowering of Arabidopsis thaliana compared with short days (SD) by activating the photoperiodic pathway. Here we show that growth under very‐SD (3 h) or darkness (on sucrose) also accelerates flowering on a biological scale, indicating that SD actively repress flowering compared with very‐SD. CONSTANS (CO) repressed flowering under SD, and the early flowering of co under SD required FLOWERING LOCUS T (FT). FT was expressed at a basal level in the leaves under SD, but these levels were not enhanced in co. This indicates that the action of CO in A. thaliana is not the mirror image of the action of its homologue in rice. In the apex, CO enhanced the expression of TERMINAL FLOWER 1 (TFL1) around the time when FT expression is important to promote flowering. Under SD, the tfl1 mutation was epistatic to co and in turn ft was epistatic to tfl1. These observations are consistent with the long‐standing but not demonstrated model where CO can inhibit FT induction of flowering by affecting TFL1 expression.     

The alleles at the E1 locus impact the expression pattern of two soybean FT-like genes shown to induce flowering in Arabidopsis

A small gene family of phosphatidyl ethanolamine-binding proteins (PEBP) has been shown to function as key regulators in flowering; in Arabidopsis thaliana the FT protein promotes flowering whilst the closely related TFL1 protein represses flowering. Control of flowering time in soybean [Glycine max (L.) Merrill] is important for geographic adaptation and maximizing yield. Soybean breeders have identified a series of loci, the E-genes, that control photoperiod-mediated flowering time, yet how these loci control flowering is poorly understood. The objectives of this study were to evaluate the expression of GmFT-like genes in the E1 near-isogenic line (NIL) background. Of the 20 closely related PEBP proteins in the soybean genome, ten are similar to the Arabidopsis FT protein. Expression analysis of these ten GmFT-like genes confirmed that only two are detectable in the conditions tested. Further analysis of these two genes in the E1 NILs grown under short-day (SD) and long-day (LD) conditions showed a diurnal expression and tissue specificity expression commensurate with soybean flowering time under SD and LD conditions, suggesting that these were good candidates for flowering induction in soybean. Arabidopsis ft mutant lines flowered early when transformed with the two soybean genes, suggesting that the soybean genes can complement the Arabidopsis FT function. Flowering time in E1 NILs is consistent with the differential expression of the two GmFT-like genes under SD and LD conditions, suggesting that the E1 locus, at least in part, impacts time to flowering through the regulation of soybean FT expression.     

Mutagenesis of GmFT2a and GmFT5a mediated by CRISPR/Cas9 contributes for expanding the regional adaptability of soybean

Flowering time is a key agronomic trait that directly influences the successful adaptation of soybean (Glycine max) to diverse latitudes and farming systems. GmFT2a and GmFT5a have been extensively identified as flowering activators and integrators in soybean. Here, we identified two quantitative trait loci (QTLs) regions harbouring GmFT2a and GmFT5a, respectively, associated with different genetic effects on flowering under different photoperiods. We analysed the flowering time of transgenic plants overexpressing GmFT2a or GmFT5a, ft2a mutants, ft5a mutants and ft2aft5a double mutants under long‐day (LD) and short‐day (SD) conditions. We confirmed that GmFT2a and GmFT5a are not redundant, they collectively regulate flowering time, and the effect of GmFT2a is more prominent than that of GmFT5a under SD conditions whereas GmFT5a has more significant effects than GmFT2a under LD conditions. GmFT5a, not GmFT2a, was essential for soybean to adapt to high latitude regions. The ft2aft5a double mutants showed late flowering by about 31.3 days under SD conditions and produced significantly increased numbers of pods and seeds per plant compared to the wild type. We speculate that these mutants may have enormous yield potential for the tropics. In addition, we examined the sequences of these two loci in 202 soybean accessions and investigated the flowering phenotypes, geographical distributions and maturity groups within major haplotypes. These results will contribute to soybean breeding and regional adaptability.     

Natural variation and CRISPR/Cas9‐mediated mutation in GmPRR37 affect photoperiodic flowering and contribute to regional adaptation of soybean

Flowering time is a critical determinant of the geographic distribution and regional adaptability of soybean (Glycine max) and is strongly regulated by photoperiod and temperature. In this study, quantitative trait locus (QTL) mapping and subsequent candidate gene analysis revealed that GmPRR37, encoding a pseudo‐response regulator protein, is responsible for the major QTL qFT12‐2, which was identified from a population of 308 recombinant inbred lines (RILs) derived from a cross between a very late‐flowering soybean cultivar, ‘Zigongdongdou (ZGDD)’, and an extremely early‐flowering cultivar, ‘Heihe27 (HH27)’, in multiple environments. Comparative analysis of parental sequencing data confirmed that HH27 contains a non‐sense mutation that causes the loss of the CCT domain in the GmPRR37 protein. CRISPR/Cas9‐induced Gmprr37‐ZGDD mutants in soybean exhibited early flowering under natural long‐day (NLD) conditions. Overexpression of GmPRR37 significantly delayed the flowering of transgenic soybean plants compared with wild‐type under long photoperiod conditions. In addition, both the knockout and overexpression of GmPRR37 in soybean showed no significant phenotypic alterations in flowering time under short‐day (SD) conditions. Furthermore, GmPRR37 down‐regulated the expression of the flowering‐promoting FT homologues GmFT2a and GmFT5a, and up‐regulated flowering‐inhibiting FT homologue GmFT1a expression under long‐day (LD) conditions. We analysed haplotypes of GmPRR37 among 180 cultivars collected across China and found natural Gmprr37 mutants flower earlier and enable soybean to be cultivated at higher latitudes. This study demonstrates that GmPRR37 controls soybean photoperiodic flowering and provides opportunities to breed optimized cultivars with adaptation to specific regions and farming systems.     

Effect of Heat-Treated Noradrenaline on Flowering in Lemna

In a previous study, heat-treated noradrenaline induced flowering of the short-day plant Lemna paucicostata Hegelmaier 151. In the present study, we found that heat-treated noradrenaline also had flower-inducing activity in short-day L. paucicostata strains 441 and 6746 and in long-day L. gibba strain G3. The flower-inducing activity in these plants was enhanced by water homogenates of eggplant (Solanum melongena L.).     

Flower formation in the short-day plant Kalanchoë by grafting with a long-day and a short-long-day Echeveria

Flower formation was induced in the shortday plant Kalanchoë blossfeldiana Poellnitz under long-day conditions by grafting with flowering shoots of the short-long-day plant Echeveria harmsii (Rose) MacBr. and of the long-day plant Echeveria pulvoliver (E. pulvinata Rose x E. harmsii). Vegetative shoots from induced Echeveria plants failed to cause a flowering response in Kalanchoë. The presence of flowering and vegetative shoots side by side on induced Echeveria plants provides evidence for physiological chimeras in this genus.Abbreviations LD long day(s) - SD short day(s) - LDP long-day plant(s) - LSDP long-short-day plant(s) - SDP short-day plant(s) - SLDP short-long-day plant(s)     

Nitric Oxide Induces Flowering in the Duckweed Lemna aequinoctialis Welw. (Syn. L. paucicostata Hegelm.) Under Noninductive Conditions

Nitric oxide (NO) plays diverse roles in the growth and development of plants and in their responses to various abiotic and biotic stresses. It has also been reported to repress flowering in Arabidopsis thaliana. In the present study, NO donors sodium nitroprusside (SNP), S-nitroso-N-acetyl penicillamine (SNAP), and 3-morpholinosydnonimine (SIN-1) induced flowering in Lemna aequinoctialis 6746 (a short-day strain) and in L. aequinoctialis LP₆ (a photoperiod-insensitive strain) under noninductive conditions. Nitrate and nitrite, two stable metabolites of NO, did not induce flowering. On the other hand, cyanide donors potassium ferricyanide {K₃[Fe(CN)₆]} and potassium cyanide (KCN) induced flowering in both strains under noninductive conditions. The flowering induced under a 8-h daily photoperiod regime in the short-day strain L. aequinoctialis 6746 was inhibited by NO and cyanide donors. Vegetative multiplication of both strains was adversely affected by NO and cyanide donors, irrespective of the photoperiod conditions. The observed effects of NO donors on flowering were substantially negated by NO scavengers c-PTIO [2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide] and methylene blue. This confirmed the role of NO in induction of flowering. The inductive effect of CN⁻ also appeared to be partly mediated through NO as NO scavengers partially negated the effect of CN⁻.     

Obligatory short‐day plant,Perilla frutescens var. crispa can flower in response to low‐intensity light stress under long‐day conditions

An obligatory short‐day plant, Perilla frutescens var. crispa was induced to flower under long‐day conditions when grown under low‐intensity light (30 µmol m^?2 s^?1). Plant size was smaller under lower light intensity, indicating that the low‐intensity light acted as a stress factor. The phenomenon is categorized as stress‐induced flowering. Low‐intensity light treatment for 4 weeks induced 100% flowering. The plants responded to low‐intensity light immediately after the cotyledons expanded, and the flowering response decreased with increasing plant age. The induced plants produced fertile seeds, and the progeny developed normally. The plants that flowered under low‐intensity light had greener leaves. This greening was because of the decrease in anthocyanin content, and there was a negative correlation between the anthocyanin content and percent flowering. Treatment with L‐2‐aminooxy‐3‐phenylpropionic acid, an inhibitor of phenylalanine ammonia‐lyase (PAL), did not induce flowering under non‐inductive light conditions and inhibited flowering under inductive low‐intensity light conditions. The metabolic pathway regulated by PAL may be involved in the flowering induced by low‐intensity light.     

Expression of <Emphasis Type="Italic">FLOWERING LOCUS T</Emphasis> from <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> induces precocious flowering in soybean irrespective of maturity group and stem growth habit

The flowering integrator gene FLOWERING LOCUS T (FT) in Arabidopsis thaliana is conserved between diverse plant species and is thought to be the flowering signal ‘‘florigen’’, a universal long-distance mobile signal. In soybean, two FT homologs having a function to induce flowering in Arabidopsis have been identified. In this study, we showed that the expression of FT from Arabidopsis by the Apple latent spherical virus (ALSV) vector promoted precocious flowering and terminated vegetative growth in a wide range of genotypes of soybean, without using a short-day treatment. Four determinate and two indeterminate cultivars, infected with ALSV expressing FT (FT-ALSV), set flower buds on shoot apices and terminated vegetative growth at the fourth- to seventh-node stages under long-day conditions. In contrast, non-infected, healthy plants did not set flower buds on shoot apices at the same stage under the same conditions. After flowering, soybean cultivars infected with FT-ALSV, belonging to different maturity groups and stem growth habits, matured and produced seeds. The results suggest that the basic flowering pathway controlled by FT in A. thaliana might also be conserved in soybean. A system for precocious flowering and shortening of generation time using FT-ALSV would be a useful and novel technology for efficient soybean breeding.     

Stress enhances the gene expression and enzyme activity of phenylalanine ammonia-lyase and the endogenous content of salicylic acid to induce flowering in pharbitis

The involvement of salicylic acid (SA) in the regulation of stress-induced flowering in the short-day plant pharbitis (also called Japanese morning glory) Ipomoea nil (formerly Pharbitis nil) was studied. Pharbitis cv. Violet was induced to flower when grown in 1/100-strength mineral nutrient solution under non-inductive long-day conditions. All fully expanded true leaves were removed from seedlings, leaving only the cotyledons, and flowering was induced under poor-nutrition stress conditions. This indicates that cotyledons can play a role in the regulation of poor-nutrition stress-induced flowering. The expression of the pharbitis homolog of PHENYLALANINE AMMONIA-LYASE, the enzyme activity of phenylalanine ammonia-lyase (PAL; E.C. 4.3.1.5) and the content of SA in the cotyledons were all up-regulated by the stress treatment. The Violet was also induced to flower by low-temperature stress, DNA demethylation and short-day treatment. Low-temperature stress enhanced PAL activity, whereas non-stress factors such as DNA demethylation and short-day treatment decreased the activity. The PAL enzyme activity was also examined in another cultivar, Tendan, obtaining similar results to Violet. The exogenously applied SA did not induce flowering under non-stress conditions but did promote flowering under weak stress conditions in both cultivars. These results suggest that stress-induced flowering in pharbitis is induced, at least partly, by SA, and the synthesis of SA is promoted by PAL.     

The promotive effect of applying mixtures of (S)-(+)-abscisic acid and gibberellic acid on flowering in long-day plants

Application of mixtures of natural type abscisic acid,(S)-(+)-abscisic acid (SABA), and gibberellic acid (GA₃)promoted floral-bud initiation and flowering in the long-day plants, spinach,pansy, primrose and petunia, even under short-day conditions. The effectiveconcentrations for spray application of SABA/GA₃ were restrictedwithin the limits of 10–1 respectively. Applications ofSABAand/or GA₃ as high as 50 induced no flowering.Flowering in short-day plants, dahlia, morning-glory and Christmas-cactus wasnot promoted by the mixtures.