Petal Movement Research Articles

Behavioural Thermoregulation of Flowers via Petal Movement.

Widely documented in animals, behavioural thermoregulation mitigates negative impacts of climate change. Plants experience especially strong thermal variability but evidence for plant behavioural thermoregulation is limited. Along a montane elevation gradient, Argentina anserina flowers warm more in alpine populations than at lower elevation. We linked floral temperature with phenotypes to identify warming mechanisms and documented petal movement and pollinator visitation using time-lapse cameras. High elevation flowers were more cupped, focused light deeper within flowers and were more responsive to air temperature than low; cupping when cold and flattening when warm. At high elevation, a 20° increase in petal angle resulted in a 0.46°C increase in warming. Warming increased pollinator visitation, especially under cooler high elevation temperatures. A plasticity study revealed constitutive elevational differences in petal cupping and stronger temperature-induced floral plasticity in high elevation populations. Thus, plant populations have evolved different behavioural responses to temperature driving differences in thermoregulatory capacity.

Autogamy ensures reproductive success in the bromeliad Dyckia dissitiflora Schult.f., endemic to the Brazilian caatinga domain

AbstractThe genus Dyckia (Bromeliaceae) is endemic to South America, and its species show important ecological roles in xeric environments. The flowering phenology as well as floral and reproductive biology were monitored monthly in Dyckia dissitiflora at two sites (Bela Vista: 11°42′03.4″ S 39°05′21.7″ W and Barra do Vento: 11°37′58.1″ S 39°03′33.7″ W) in the municipality of Serrinha, Bahia, Brazil. Flowering occurs in the early austral summer with a peculiar time of beginning of anthesis (10:00–11:00), offering resources 18 h after the first petal movements. Pollen viability was 98%, with a high production of pollen grains. Stigma receptivity is continuous from flower opening to senescence, and nectar production initiates at 04:00 and extends until 18:00. The species is self‐compatible and autogamous, but not agamospermic. Pollen tubes developed and reached the micropyle within 24 h in all of the reproductive tests. The hummingbird Chlorostilbon lucidus was the only floral visitor and pollinator (frequency = 0.82 ± 0.02). Fruiting occurs 3 days after anthesis, and the seeds are wind‐dispersed. Self‐compatibility and autogamy are essential strategies for the reproductive success of D. dissitiflora in light of the low number and visit frequency of its pollinators, and it is favored by slow floral movements and the position of the stigmas in relation to stamens.

Floral biology and pollinator captivating semiochemicals in Goniothalamus wynaadensis (Bedd.) Bedd. (Annonaceae) an endemic tree species of the Western Ghats

Goniothalamus wynaadensis (Bedd.) Bedd. (Family Annonaceae) is an endemic tree species growing in the evergreen forests of the Western Ghats up to 900m asl. The present study aims at understanding the functional floral traits and semiochemicals found in G. wynaadensis which support plant-pollinator interactions. The study revealed G. wynaadensis exhibits abbreviated anthesis (c.28hrs). Goniothalamus flowers exhibit unique features of cantharophily. The study indicated that outcrossing is promoted by protogyny, and pollination efficiency is enhanced by pollen aggregation, anthesis duration, and pollinator trapping which involves a close alignment between petal movements and the circadian rhythm of pollinators. Curculionidae and Nitidulidae beetles are effective pollinators of the plant. In beetle-pollinated flowers, floral scent is a crucial component for pollinator attraction. G. wynaadensis has a strong fruity aroma. The floral scent analysis performed using headspace solid- phase microextraction combined with GC-MS detected various semiochemicals that support plant-pollinator interactions. Ethyl butyrate, isobutyl acetate, 3 hydroxy 3 methyl 2 butanol, isopentyl acetate, and 3 methyl 1 butanol dominate the floral scent of G. wynaadensis.

The ARF2-MYB6 module mediates auxin-regulated petal expansion in rose.

In cut rose (Rosa hybrida), the flower-opening process is closely associated with vase life. Auxin induces the expression of transcription factor genes that function in petal growth via cell expansion. However, the molecular mechanisms underlying the auxin effect during flower opening are not well understood. Here, we identified the auxin-inducible transcription factor gene RhMYB6, whose expression level is high during the early stages of flower opening. Silencing of RhMYB6 delayed flower opening by controlling petal cell expansion through down-regulation of cell expansion-related genes. Furthermore, we demonstrated that the auxin response factor RhARF2 directly interacts with the promoter of RhMYB6 and represses its transcription. Silencing of RhARF2 resulted in larger petal size and delayed petal movement. We also showed that the expression of genes related to ethylene and petal movement showed substantial differences in RhARF2-silenced petals. Our results indicate that auxin-regulated RhARF2 is a critical player that controls flower opening by governing RhMYB6 expression and mediating the crosstalk between auxin and ethylene signaling.

Circadian autonomy and rhythmic precision of the Arabidopsis female reproductive organ.

The plant circadian clock regulates essential biological processes including flowering time or petal movement. However, little is known about how the clock functions in flowers. Here, we identified the circadian components and transcriptional networks contributing to the generation of rhythms in pistils, the female reproductive organ. When detached from the rest of the flower, pistils sustain highly precise rhythms, indicating organ-specific circadian autonomy. Analyses of clock mutants and chromatin immunoprecipitation assays showed distinct expression patterns and specific regulatory functions for clock activators and repressors in pistils. Genetic interaction studies also suggested a hierarchy of the repressing activities that provide robustness and precision to the pistil clock. Globally, the circadian function in pistils primarily governs responses to environmental stimuli and photosynthesis and controls pistil growth and seed weight and production. Understanding the circadian intricacies in reproductive organs may prove useful for optimizing plant reproduction and productivity.

Better time of flower opening, a good time to improve the efficiency of hybrid seed production

The proper time of flower opening, which ensures successful pollination by exposure of stamens and gynoecia via petal movement, is locally adapted to diverse environmental changes among different plants (Cheng et al., 2021). Consequently, flower opening time may differ among divergent populations as a result of long-term adaptation, thereby promoting prezygotic reproductive isolation by preventing mating and hybridization.

The JA pathway is rapidly down-regulated in petal abscission zones prior to flower opening and affects petal abscission in fragrant roses during natural and ethylene-induced petal abscission

Organ abscission is a developmental process regulated through changes in the abscission zone (AZ) and governed primarily by ethylene. In roses, the opening of flowers is also an ethylene-sensitive process possibly requiring changes at the AZ to facilitate petal movement. A study of the rose petal AZ transcriptome of the early-abscising fragrant rose, R. bourboniana, had indicated suppression of the jasmonic acid (JA) pathway in the AZ before abscission. Detailed investigation of 24 genes representing JA biosynthesis/perception/signalling revealed a rapid and substantial down-regulation of the entire JA pathway in the AZ prior to flower opening and abscission. Transcript levels of 17 genes encoding JA biosynthesis members of the PLD/LOX/AOS/AOC/OPR families and the receptor/signalling components of COI1/JAZ/MYC2/NINJA families decreased by 50–90% in petal AZ within 4 h of ethylene treatment as well as during natural field abscission (ethylene untreated). In contrast, R. hybrida flowers (which show delayed opening and abscission even after ethylene treatment) were much less affected in 10/24 genes while transcript levels of 9/24 JA pathway genes actually increased several fold during or just prior to abscission. JA treatment delayed flower opening as well as petal abscission by more than 24 h in R. bourboniana and also increased transcription of the JA pathway genes, unlike ethylene. The strong and rapid reduction of the entire JA pathway in the AZ in R. bourboniana flowers, prior to flower opening and petal abscission, suggests a negative role for JA in these processes with JA acting opposite to ethylene and possibly reducing ethylene sensitivity.

Cherry Blossom Forecast Based on Transcriptome of Floral Organs Approaching Blooming in the Flowering Cherry (Cerasus × yedoensis) Cultivar 'Somei-Yoshino'.

To gain insights into the genetic mechanisms underlying blooming and petal movement in flowering cherry (Cerasus × yedoensis), we performed time-course RNA-seq analysis of the floral buds and open-flowers of the most popular flowering cherry cultivar, ‘Somei-Yoshino.’ Independent biological duplicate samples of floral buds and open-flowers were collected from ‘Somei-Yoshino’ trees grown at three different locations in Japan. RNA-seq reads obtained from floral bud and open-flower samples collected in the current study (in 2019) and in a previous study (in 2017) were aligned against the genome sequence of ‘Somei-Yoshino’ to quantify gene transcript levels. Clustering analysis of RNA-seq reads revealed dynamic changes in the transcriptome, with genes in seven modules predominantly expressed at specific time points, ranging from 5 weeks before flowering to 2 weeks after flowering. Based on the identified gene modules and Gene Ontology (GO) terms enriched at different floral stages, we speculate that the genetic mechanisms underlying petal movement and flower opening in cherry involve the processes of development, cell wall organization, reproduction, and metabolism, which are executed by genes encoding transcription factors, phytohormones, transporters, and polysaccharide metabolic enzymes. Furthermore, we established a statistical model for cherry bloom forecasting, based on gene expression levels as RNA markers at different time points before flowering.

Why flowers close at noon? A case study of an alpine species Gentianopsis paludosa (Gentianaceae).

Repeatable floral closure with diurnal rhythms, that is, flower opening in the morning and closing in the evening, was widely reported. However, the rhythm of flower opening in the morning but closing in the midday received much less attention. Gentianopsis paludosa, Gentianaceae, has an obvious petal movement rhythm opening in the morning and closing at noon at northeast of the Qinghai‐Tibetan Plateau. In this study, we examined the effects of temperature (T), relative humidity (RH), and illumination intensity (II) on G. paludosa's flower closure. Furthermore, we monitored the environmental changes inside and outside of the flowers, aiming to test the effect of floral closure on the stability of microenvironment inside the flower. Finally, we artificially interrupted temporal petal closure and investigated its effects on reproductive fitness. The results showed that high/low temperature contributed more to the flower closure than low RH, while illumination intensity had no significant effect on it. The medium temperature, relative humidity and illumination intensity (environmental conditions at 10:00) did not delay flower closure when flowers at pre‐closing period or stimulate reopen when flowers full closed. Floral closure provided a stable temperature condition and a higher RH condition inside the flower. Meanwhile, compulsive opening and delayed closure of flowers decreased the seed‐set ratio while no effect was found when flowers were forced to close. We conclude that endogenous rhythm regulates floral closure. The rhythm of petal movement providing a stable microenvironment for G. paludosa, increasing the seed production and saving energy from flower opening maintenance, which might be an adaptive strategy to against unfavorable environmental conditions.

Pollination adaptations of group-by-group stamen movement in a meadow plant with temporal floral closure

Floral sexual organ (stamen and pistil) movements are selective adaptations that have different functions in male-female reproduction and the evolution of flowering plants. However, the significance of stamen movements in the spatial–temporal function and separation of male and female organs has not been experimentally determined in species exhibiting floral temporal closure. The current study investigated the role of slow stamen (group-by-group) movement in male-female sexual function, and the effect of stamen movement on pollen removal, male-male and male-female interference, and mating patterns of Geranium pratense, a plant with temporal floral closure. This species uses stamen group-by-group movement and therefore anther-stigma spatial–temporal separation. Spatial separation (two whorls of stamen and pistil length) was shown to be stronger than temporal separation. We found that stamen movements to the center of the flower increase pollen removal, and the most common pollinators visited more frequently and for longer durations during the male floral stage than during the female floral stage. Petal movements increased both self-pollen deposition rate and sexual interference in G. pratense. The fruit and seed set of naturally and outcrossed pollinated flowers were more prolific than those of self-pollinated flowers. Group-by-group stamen movement, dehiscence of stamens, pistil movement, and male-female spatial–temporal functional separation of G. pratense before floral temporal closure may prevent male-female and stamen-stamen interference and pollen discounting, and may increase pollen removal and cross-pollination.

Ethylene-regulated asymmetric growth of the petal base promotes flower opening in rose (Rosa hybrida).

Flowers are the core reproductive structures and key distinguishing features of angiosperms. Flower opening to expose stamens and gynoecia is important in cases where pollinators much be attracted to promote cross-pollination, which can enhance reproductive success and species preservation. The floral opening process is accompanied by the coordinated movement of various floral organs, particularly petals. However, the mechanisms underlying petal movement and flower opening are not well understood. Here, we integrated anatomical, physiological, and molecular approaches to determine the petal movement regulatory network using rose (Rosa hybrida) as a model. We found that PETAL MOVEMENT-RELATED PROTEIN1 (RhPMP1), a homeodomain transcription factor (TF) gene, is a direct target of ETHYLENE INSENSITIVE3, a TF that functions downstream of ethylene signaling. RhPMP1 expression was upregulated by ethylene and specifically activated endoreduplication of parenchyma cells on the adaxial side of the petal (ADSP) base by inducing the expression of RhAPC3b, a gene encoding the core subunit of the Anaphase-Promoting Complex. Cell expansion of the parenchyma on the ADSP base was subsequently enhanced, thus resulting in asymmetric growth of the petal base, leading to the typical epinastic movement of petals and flower opening. These findings provide insights into the pathway regulating petal movement and associated flower-opening mechanisms.�.

The evolution of key functional floral traits in the early divergent angiosperm family Annonaceae

AbstractPotential key functional floral traits are assessed in the species‐rich early divergent angiosperm family Annonaceae. Pollinators (generally beetles) are attracted by various cues (particularly visual, olfactory, and thermogenic), with pollinators rewarded by nectar (generally as stigmatic exudate), heat, and protection within the partially enclosed floral chamber. Petals sometimes function as pollinator brood sites, although this could be deceptive. Annonaceae species are self‐compatible, with outcrossing promoted by a combination of protogyny, herkogamy, floral synchrony, and dicliny. Pollination efficiency is enhanced by pollen aggregation, changes in anthesis duration, and pollinator trapping involving a close alignment between petal movements and the circadian rhythms of pollinators. Most Annonaceae flowers are apocarpous, with syncarpy restricted to very few lineages; fertilization is therefore optimized by intercarpellary growth of pollen tubes, either by stigmatic exudate (suprastylar extragynoecial compitum) or possibly the floral receptacle (infrastylar extragynoecial compitum). Although Annonaceae lack a distinct style, the stigmas in several lineages are elongated to form “pseudostyles” that are hypothesized to function as sites for pollen competition. Flowers can be regarded as immature fruits in which the ovules are yet to be fertilized, with floral traits that may have little selective advantage during anthesis theoretically promoting fruit and seed dispersal. The plesiomorphic apocarpous trait may have been perpetuated in Annonaceae flowers as it promotes the independent dispersal of fruit monocarps (derived from separate carpels), thereby maximizing the spatial/temporal distance between seedlings. This might compensate for the lack of genetic diversity among seeds within fruits arising from the limited diversity of pollen donors.

Stimuli-responsive composite biopolymer actuators with selective spatial deformation behavior

Bioinspired actuators with stimuli-responsive and deformable properties are being pursued in fields such as artificial tissues, medical devices and diagnostics, and intelligent biosensors. These applications require that actuator systems have biocompatibility, controlled deformability, biodegradability, mechanical durability, and stable reversibility. Herein, we report a bionic actuator system consisting of stimuli-responsive genetically engineered silk-elastin-like protein (SELP) hydrogels and wood-derived cellulose nanofibers (CNFs), which respond to temperature and ionic strength underwater by ecofriendly methods. Programmed site-selective actuation can be predicted and folded into three-dimensional (3D) origami-like shapes. The reversible deformation performance of the SELP/CNF actuators was quantified, and complex spatial transformations of multilayer actuators were demonstrated, including a biomimetic flower design with selective petal movements. Such actuators consisting entirely of biocompatible and biodegradable materials will offer an option toward constructing stimuli-responsive systems for in vivo biomedicine soft robotics and bionic research.

The Plant Circadian Oscillator.

It has been nearly 300 years since the first scientific demonstration of a self-sustaining circadian clock in plants. It has become clear that plants are richly rhythmic, and many aspects of plant biology, including photosynthetic light harvesting and carbon assimilation, resistance to abiotic stresses, pathogens, and pests, photoperiodic flower induction, petal movement, and floral fragrance emission, exhibit circadian rhythmicity in one or more plant species. Much experimental effort, primarily, but not exclusively in Arabidopsis thaliana, has been expended to characterize and understand the plant circadian oscillator, which has been revealed to be a highly complex network of interlocked transcriptional feedback loops. In addition, the plant circadian oscillator has employed a panoply of post-transcriptional regulatory mechanisms, including alternative splicing, adjustable rates of translation, and regulated protein activity and stability. This review focuses on our present understanding of the regulatory network that comprises the plant circadian oscillator. The complexity of this oscillatory network facilitates the maintenance of robust rhythmicity in response to environmental extremes and permits nuanced control of multiple clock outputs. Consistent with this view, the clock is emerging as a target of domestication and presents multiple targets for targeted breeding to improve crop performance.

A new pollination system in non-cleistogamous species of Viola results from nyctinastic (night-closing) petal movements – A mixed outcrossing-selfing strategy

Viola banksii of the eastern Australian and Tasmanian sect. Erpetion produces exclusively chasmogamous flowers, unlike most other temperate taxa of Viola which additionally produce obligatory self-pollinated cleistogamous flowers. This study explored flower structure and nyctinastic flower movements (temporal flower closure), the correlation of the timing of nyctinastic movements with stigma receptivity, and self-compatibility of flowers. Petal movement in day/night cycles was documented by two cameras. Floral morphology, anther protuberance anatomy, stigma receptivity, and pollen tube growth were examined using scanning electron microscopy, fluorescent microscopy, histochemical and sectioning techniques. Diurnal petal movements that cause flowers to be open during the day and closed during the night were documented in V. banksii. Chasmogamous flower colour, fragrance, anterior petal venation, and the indurated green area at the base of the anterior petal are all likely to play roles for pollinator attraction. Unlike most other Viola species, the anther protuberances do not function as nectaries. The flowers offer pollinators no nectar reward. The short time of stigma receptivity (2–3 days) of individual flowers limits opportunities for insect visitation and cross-pollination. Night-closed flowers of V. banksii appear to facilitate self-pollination. Self-compatibility was confirmed by tracking the pollen tube growth from the stigma to the ovule in spontaneous and hand self-pollinated flowers. Over the floral phenological cycle, nyctinasty coincides in time with stigma receptivity and, hence, the ability of self-pollination. We discovered that the same chasmogamous flower of V. banksii can cross-pollinate while open during the day, but self-pollinate while closed during the night (i.e. acting as a cleistogamous-like flower). Several floral characters indicate that V. banksii exhibits a pollination deceit strategy. Such a pollination system has not been described elsewhere in Viola.

Comprehensive characteristics and genetic diversity of the endemic Australian Viola banksii (section Erpetion, Violaceae)

Viola banksii, the type species of section Erpetion, is endemic in eastern mainland Australia. In this paper we characterise morphological and anatomical features and assess genome size and genetic diversity in combination with the breeding system. V. banksii develops exclusively chasmogamous flowers. Ovules are anatropous, crassinucellate and bitegmic, the female gametophyte is of the Polygonum type, and the embryo is of Asterad type surrounded by nuclear endosperm. Pollen is non-heteromorphic, 3-aperturate, and highly viable. V. banksii grows in shade on moist, well drained, often sandy soils, and this is reflected in the anatomy of its organs, which includes a lack of subepidermal collenchyma in aerial parts, large leaf epidermal cells with thin cell walls, a narrow cuticle layer, and vascular bundles with xylem that are not rich in vessels. V. banksii is tolerant to zinc and lead based on phytotoxicity test. The high chromosome number (2n = 10x = 50) does not correspond to a small genome size (2C DNA = 1.27 pg). Low mean intra-populational gene diversity (HS = 0.077) detected by ISSR markers confirms the strong influence of selfing and clonal propagation by pseudostolons. Unique morphological traits of V. banksii include nyctinastic petal movement, the lack of a floral spur, the presence of gland-like protuberances on two stamens, and the presence of pseudostolons, which could be a synapomorphy for the whole section.

Effect of change in floral openness with floral age on floral display and reproduction in Gentiana

Some plant species have flowers that open widely and then close slowly over time, resulting in changes in the width of the corolla opening. We examined the adaptive significance of retaining flowers with relatively closed corollas using Gentiana triflora var. japonica, a perennial species with petal movement, in Hakkoda, northern Japan. Under open pollination, floral openness changed with floral age. It increased from days 1–3 after the flower initially opened and decreased thereafter until floral senescence. Hand-pollinated flowers showed a significant reduction in floral openness compared with unpollinated flowers, suggesting that a floral openness reflects its demand for pollination. Pollinators visited inflorescences with many flowers irrespective of their openness, suggesting that the reduction in floral openness does not affect the attractiveness of the inflorescences to pollinators. Within inflorescences, pollinators visited less-pollinated flowers with wider floral openness more frequently and more-pollinated flowers with lower floral openness less frequently, possibly because of the time cost of entering flowers with low openness. This strategy appears adaptive because if all flowers maintain wide floral openness irrespective of their pollination demand, pollinators may visit more-pollinated flowers in an inflorescence and may leave the inflorescence without visiting less-pollinated flowers. Reduction of openness of more-pollinated flowers fits a strategy of enhancing effective pollination within inflorescences.

Differential expression of several xyloglucan endotransglucosylase/hydrolase genes regulates flower opening and petal abscission in roses

Flower opening is a process that requires movement of petals from a closed position to a horizontal open position, while petal abscission requires cell-wall disassembly. Both processes are controlled by ethylene and require cell-wall modification at the junction (abscission zone) of the petal and thalamus to facilitate the movement or separ- ationofpetals.Inthepresentstudy,afamilyofxyloglucanendotransglucosylase/hydrolase(XTH)geneswasstudiedto understand their role in petal abscission in flowers of Rosa bourboniana (ethylene sensitive, early abscising) and Rosa hybrida(lessethylenesensitive,lateabscising).TranscriptomesequencingofpetalabscissionzonecDNAwasperformed atdifferenttimepoints(ethylenetreatedanduntreated)andscreenedforXTHgenes.ThestudyidentifiedninenewXTH genes that showed differential changes in gene expression during flower opening and abscission. Of these, RbXTH3, RbXTH5, RbXTH6 and RbXTH12 were rapidly induced by ethylene within 1-4 h of ethylene treatment, corresponding to the period of flower opening. These genes also showed an early up-regulation during flower opening under ethylene-untreated (field abscission) conditions, indicating a possible role in anthesis and petal movement during flower opening. Other genes such as RbXTH4 and RbXTH9 were up-regulated later at 8-12 h after ethylene treatment and at 24-36 h under natural abscission conditions, indicating a possible role in abscission. Treatment with a higher ethylenedose(15 m LL 21 ethylene)acceleratedabscission,leadingtohighersteady-statelevelsofXTHgenetranscripts atanearliertimepointcomparedwith0.5 m LL 21 ethylene.Incontrast,transcriptaccumulationofmostoftheXTHswas considerablydelayedin thelate-abscising rose,R.hybrida, inkeeping with the slowerfloweropeningand delayedpetal abscission. The results suggest coordinated action of different XTHs in cell-wall modification of xyloglucan moieties during flower opening as well as cell separation during abscission.

Expression profiles of aquaporin homologues and petal movement during petal development in Tulipagesneriana

Previously, we have characterized two tonoplast intrinsic proteins (TIPs) and four plasma membrane intrinsic proteins (PIPs) from the 2-day-old petals of tulip (Tulipa gesneriana). In this study, we analyzed the development of tulip petals and stems, temperature-dependent petal movement, the amount of ³H₂O transported into petals and stems during petal movement, and the transcript levels of two TIP (TgTIP1;1 and TgTIP1;2) and four TgPIP genes in petals and stems, from the first day of petal opening to day 12. The development of the petals and stems was completed by days 6 and 9, respectively, after the first day of petal opening. Temperature-dependent petal movement and the amount of ³H₂O that was transported into petals could be detected at significant levels up to day 6 with petal movement reaching a peak at day 3. Real-time reverse transcription-polymerase chain reaction analysis revealed that TgTIP1;1 and TgTIP1;2 were expressed ubiquitously in petals, stems, leaves, bulbs and roots. However, the expression level of TgTIP1;2 was very low in bulbs. The expression of both TgTIP1 genes was upregulated in close association with the development of petals but not with that of the stem. The four TgPIP genes were expressed at almost the same level during the development of the petals and the stem. However, the levels of the TgTIP1 and TgPIP transcripts in petals decreased during the course of petal wilting from day 9 onwards. These results suggest that TgTIP1;1 and TgTIP1;2 may contribute to petal development.

Characterization of Four Plasma Membrane Aquaporins in Tulip Petals: A Putative Homolog is Regulated by Phosphorylation

We suggested previously that temperature-dependent tulip (Tulipa gesneriana) petal movement that is concomitant with water transport is regulated by reversible phosphorylation of an unidentified plasma membrane intrinsic protein (PIP). In this study, four full-length cDNAs of PIPs from tulip petals were identified and cloned. Two PIPs, namely TgPIP1;1 and TgPIP1;2, are members of the PIP1 subfamily, and the remaining two PIPs, namely TgPIP2;1 and TgPIP2;2, belong to the PIP2 subfamily of aquaporins and were named according to the nomenclature of PIP genes in plants. Of these four homologs, only TgPIP2;2 displayed significant water channel activity in the heterologous expression assay using Xenopus laevis oocytes. The water channel activity of this functional isoform was abolished by mercury and was affected by inhibitors of protein kinase and protein phosphatase. Using a site-directed mutagenesis approach to substitute several serine residues with alanine, and assessing water channel activity using the methylotrophic yeast Pichia pastoris expression assay, we showed that Ser35, Ser116 and Ser274 are the putative phosphorylation sites of TgPIP2;2. Real-time reverse transcription-PCR analysis revealed that the transcript levels of TgPIP1;1 and TgPIP1;2 in tulip petals, stems, leaves, bulbs and roots are very low when compared with those of TgPIP2;1 and TgPIP2;2. The transcript level of TgPIP2;1 is negligible in roots, and TgPIP2;2 is ubiquitously expressed in all organs with significant transcript levels. From the data reported herein, we suggest that TgPIP2;2 might be modulated by phosphorylation and dephosphorylation for regulating water channel activity, and may play a role in transcellular water transport in all tulip organs.