Crosstalk in the darkness: bulb vernalization activates meristem transition via circadian rhythm and photoperiodic pathway.

BACKGROUND: Geophytes possess specialized storage organs - bulbs, tubers, corms or rhizomes, which allow their survival during unfovarable periods and provide energy support for sprouting and sexual and vegetative reproduction. Bulbing and flowering of the geophyte depend on the combined effects of the internal and external factors, especially temperature and photoperiod. Many geophytes are extensively used in agriculture, but mechanisms of regulation of their flowering and bulbing are still unclear. RESULTS: Comparative morpho-physiological and transcriptome analyses and quantitative validation of gene expression shed light on the molecular regulation of the responses to vernalization in garlic, a typical bulbous plant. Long dark cold exposure of bulbs is a major cue for flowering and bulbing, and its interactions with the genetic makeup of the individual plant dictate the phenotypic expression during growth stage. Photoperiod signal is not involved in the initial nuclear and metabolic processes, but might play role in the later stages of development, flower stem elongation and bulbing. Vernalization for 12 weeks at 4 °C and planting in November resulted in flower initiation under short photoperiod in December-January, and early blooming and bulbing. In contrast, non-vernalized plants did not undergo meristem transition. Comparisons between vernalized and non-vernalized bulbs revealed ~ 14,000 differentially expressed genes. CONCLUSIONS: Low temperatures stimulate a large cascades of molecular mechanisms in garlic, and a variety of flowering pathways operate together for the benefit of meristem transition, annual life cycle and viable reproduction results.The circadian clock appears to play a central role in the transition of the meristem from vegetative to reproductive stage in bulbous plant, serving as integrator of the low-temperature signals and the expression of the genes associated with vernalization, photoperiod and meristem transition. The reserved photoperiodic pathway is integrated at an upstream point, possibly by the same receptors. Therefore, in bulb, low temperatures stimulate cascades of developmental mechanisms, and several genetic flowering pathways intermix to achieve successful sexual and vegetative reproduction.

Storage temperature controls the timing of garlic bulb formation via shoot apical meristem termination.

MAIN CONCLUSION: Timing of bulb formation and floral stem induction in garlic is controlled by preplanting storage temperature and shoot apical meristem termination, probably via FLOWERING LOCUS T (FT) genes. Garlic is planted in the winter, undergoes a vegetative stage, then forms bulbs in response to increasing temperature and lengthening photoperiod. Herein, the storage conditions for propagation bulbs are shown to potentially affect future vegetative-stage length and timing of bulb formation. Storage temperatures of 2 or 33 °C inhibited internal bud growth. Levels of endogenous abscisic acid (ABA) and its inactive isomer trans-ABA were significantly higher in the internal bud of cloves stored at 33 vs. 2 °C, and exogenous ABA treatment before planting confirmed its inhibitory effect on foliage leaf development. Bulb formation started 30 and 60 days after planting of cloves stored at 2 and 33 °C, respectively. Warm storage temperature induced the formation of multiple leaves and cloves after planting. Plants from cloves stored at warm temperature developed a floral stem, whereas those from cold storage did not. Allium sativum FLOWERING LOCUS T1 (AsFT1) was upregulated 2.5- and 4.5-fold in the internal bud and storage leaf, respectively, after 90 and 150 days of cold vs. warm storage. Expression of AsFT4, expected to be antagonist to AsFT1, was 2- to 3-fold lower in the internal bud from cold storage. Expression of AsFT2, associated with floral termination, was 2- to 3- and 10- to 12-fold higher for cold vs. warm storage temperatures, in the internal bud and storage leaf, respectively. Early bulb formation, induced by cold storage, is suggested to inhibit normal foliage leaf development and transition of the shoot apical meristem to reproductive meristem, through regulation of FT genes.

Effects of different temperature regimes on flower development, microsporogenesis and fertility in bolting garlic (Allium sativum).

Garlic (Allium sativum L.) cultivars do not develop fertile flowers and seeds. Therefore, garlic production and improvement depend exclusively on vegetative propagation. Recent advances in garlic research have enabled fertility restoration and the discovery of fertile and male-sterile genotypes; however, the environmental regulation of the reproductive process is still not clear. Garlic seeds are successfully produced in the Mediterrenean region, where the photoperiod is relatively short, whereas spring and summer temperatures are high. We hypothesise that, in bolting garlic, various stages of florogenesis are differentially regulated by temperature and that high temperatures might obstruct pollen production. The effects of eight combinations of controlled growth temperatures on fertile and male-sterile garlic clones were studied. In both genotypes, a gradual temperature increase before and during anthesis favoured intact flower development. Surprisingly, continuous exposure to moderate temperatures during the entire growth period resulted in poor flowering, anther abortion and reduced pollen production. In the male-sterile genotype, no growth regime improved pollen production, which is controlled by genetic mechanisms. In the male-fertile genotype, gradual temperature increase supported pollen production but a sharp transition to high temperatures resulted in rapid flower senescence and pollen abortion, thus supporting our research hypothesis. In both fertile and male-sterile plants, the most vulnerable phase of microsporogenesis is the unicellular microspore stage. Tapetal malformation is the major cause for malnutrition of the microspores, with consequent production of nonviable pollen grains.