Possible evidence for morning and evening oscillators in Drosophila melanogaster populations selected for early and late adult emergence

In this paper, we report the results of our study aimed at a systematic analysis of the circadian phenotypes of fruit flies Drosophila melanogaster selected for early and late adult emergence, in light of the "morning and evening oscillator" (M and E) model for circadian clocks. We monitored adult emergence and activity/rest rhythms in these flies under light/dark (LD) cycles with short (8:16 h), normal (12:12 h) and long (16:8 h) photoperiods, as well as under constant darkness (DD). Across all the three LD cycles, the early populations displayed a morning phenotype with peak of emergence and activity occurring earlier than the controls and greater anticipation to "lights-on" and weak anticipation to "lights-off", while the late populations showed an evening phenotype with peak of emergence and activity occurring later than the controls and greater anticipation to lights-off and weak anticipation to lights-on. The gate of adult emergence and duration of activity in the early populations was narrower than the controls, while those of the late populations were wider than the controls. In addition, the circadian periodicities of adult emergence and activity/rest rhythms of the early flies were significantly shorter than the controls, while those of the late flies were significantly longer than the controls. In summary, the circadian phenotypes indicate that the early populations have evolved a dominant M oscillator, while the late populations have evolved a dominant E oscillator, thus providing an empirical support for the M and E model in Drosophila.     

Entrainment analysis of the circadian locomotor activity rhythm in specimens of the crayfish, Faxonella clypeata, having activity peaks at different times of the solar day.

Locomotor activity rhythms in the crayfish, Faxonella clypeata, were recorded under conditions of controlled light and temperature. In LD 12:12, dark active rhythms with a major activity onset at lights-off, and bimodally active rhythms with onsets at both lights-on and lights-off were recorded. In DD, most of the LD dark active rhythms retained the lights-off activity onset. However, among the majority of the LD bimodally active crayfish, only the lights-on activity peak persisted in DD. A small number of the LD bimodal rhythms remained bimodal in DD. DD records revealed free-running period lengths both greater and less than 24 hrs. An hypothesis stating that the two recorded LD rhythms represent two basically different rhythmic types is presented. The dark active crayfish appear to entrain to the lights-off position, but the bimodally rhythmic crayfish appear to entrain to the lights-on position.     

Dopaminergic Regulation of Circadian Food Anticipatory Activity Rhythms in the Rat

Circadian activity rhythms are jointly controlled by a master pacemaker in the hypothalamic suprachiasmatic nuclei (SCN) and by food-entrainable circadian oscillators (FEOs) located elsewhere. The SCN mediates synchrony to daily light-dark cycles, whereas FEOs generate activity rhythms synchronized with regular daily mealtimes. The location of FEOs generating food anticipation rhythms, and the pathways that entrain these FEOs, remain to be clarified. To gain insight into entrainment pathways, we developed a protocol for measuring phase shifts of anticipatory activity rhythms in response to pharmacological probes. We used this protocol to examine a role for dopamine signaling in the timing of circadian food anticipation. To generate a stable food anticipation rhythm, rats were fed 3h/day beginning 6-h after lights-on or in constant light for at least 3 weeks. Rats then received the D2 agonist quinpirole (1 mg/kg IP) alone or after pretreatment with the dopamine synthesis inhibitor α-methylparatyrosine (AMPT). By comparison with vehicle injections, quinpirole administered 1-h before lights-off (19h before mealtime) induced a phase delay of activity onset prior to the next meal. Delay shifts were larger in rats pretreated with AMPT, and smaller following quinpirole administered 4-h after lights-on. A significant shift was not observed in response to the D1 agonist SKF81297. These results provide evidence that signaling at D2 receptors is involved in phase control of FEOs responsible for circadian food anticipatory rhythms in rats.     

Cultivar selection prior to introduction may increase invasiveness: evidence from Ardisia crenata

Ardisia crenata (Myrsinaceae), an evergreen shrub with attractive red fruits introduced from Japan to the USA for ornamental purpose, invades the understory of mesic hardwood forests, forming dense patches (up to 300 stems per m²), and competitively displaces native understory plants by creating dense local shade. Comparison of the wild genotype that grows in mature evergreen broadleaf forests in central Kyushu, Japan, with the ecotype invading north central Florida revealed how selection for desirable cultivars might have inadvertently selected for traits that enhance the invasive potential of the species. In Japanese wild populations in deeply shaded evergreen forests, natural selection apparently maintained efficient architecture with a low degree of self-shading and large seed mass to enhance seedling shade tolerance. Cultivar selection for showy appearance can explain the greater fecundity but smaller seed size observed in the Florida populations compared to the Japanese population. Artificial selection for densely foliated appearance can also explain the greater degree of self-shading and less-efficient light use in the Florida genotype compared to the Japanese wild type grown under a common environment. Furthermore, the Florida ecotype allocated more biomass to root carbohydrate storage. These trait modifications resulted in slower growth rates, but greater competitive ability to cast shade upon neighbors and higher resprouting potential in the Florida populations. How traits are modified through the processes of artificial selection and cultivation must be taken into consideration in the evolutionary ecology of many other invasive plants introduced as ornamental plants.     

A role of melatonin in the initial stage of photoperiodism in the Japanese quail

To estimate whether melatonin is involved in gonadal activity in the male quail, the dynamics of plasma melatonin at an early stage of the photoperiodic response were investigated. Nocturnal levels of melatonin were manipulated by treatment with anti-melatonin (anti-M). By means of 4 additional hours of photic stimulation of the brain (provided by a red light-emitting diode inserted through the back of the head) after the environmental lights (8L:16D, lights-on, 1000 h) were turned off, the elevation of levels of melatonin after lights-off was significantly suppressed on Days 1 and 2 (p less than 0.01); after 5 days of brain-lighting, gonadal growth first became noticeable. However, 4 h of brain-lighting before lights-on elicited no change in levels of melatonin or in gonadal growth. The injections of anti-M just before lights-off (at 1800 h) for the first 3 days caused significant gonadal growth (p less than 0.01), whereas injections at 2200, 0200, or 0600 h were without effect. In addition, 4 h of brain-lighting before lights-on became gonadostimulatory (p less than 0.01) when it was accompanied by the injection of anti-M at 1800 h, but remained without effect when anti-M was injected at 0600 h. These results suggest that melatonin is involved in the initial stage of photoperiodism in birds, and the timing of suppression of the elevation of melatonin levels is critical in gonadal development.     

古老铅锌矿区生态型东南景天对锌耐性及超积累特性的研究

 植物长期生长在重金属污染的生境中,逐渐进化成不同的生态型。通过调查中国东南部古老Pb/zn矿和非矿山生境中的植物种群,发现生长在古老Pb／Zn矿的东南景天(Sedum alfredii Hance)是一种新的Zn超积累植物。在自然和控制条件下,古老Pb／Zn矿生态型比非矿山生态型植株的茎粗、叶片大、植株高。在矿山土壤Zn有效含量为105.5～325.4mg·kg-1时,矿山生态型东南景天植株地上部Zn含量为4134～5000mg·kg-1;当营养液中Zn浓度为1223.6μmol时,其Zn含量高达2％。在相同Zn浓度下,矿山生态型地上部Zn含量比非矿山生态型高30倍左右。两种生态型体内Zn分布也不同,古老铅锌矿山生态型的不同器官中Zn含量以茎>叶片>根系,而非矿山生态型则以根系>茎>叶片。古老铅锌矿山生态型地上部积累的Zn占植株总积累量的90％以上,其中叶片和茎分别占41.66％±5.46％和54.75％±5.87％;非矿山生态型各器官中积累的Zn远远低于古老铅锌矿山生态型,各器官中积累的Zn以茎>根系>叶片。本研究表明,这两种生态型东南景天的发现,为今后探讨植物耐高Zn胁迫和超积累Zn的微进化过程提供了非常有价值的材料,也为Zn污染土壤的植物修复提供了一种很有潜力的候选材料。     

Diurnal variation of arachidonoylethanolamine, palmitoylethanolamide and oleoylethanolamide in the brain of the rat

The diurnal variations of the endocannabinoid arachidonoylethanolamine (anandamide, ANA) as well as palmitoylethanolamide (PEA) and oleoylethanolamide (OEA) were detected and quantified in cerebrospinal fluid (CSF), pons, hippocampus, and hypothalamus in the rat over 24 h using HPLC/MS. In CSF, the 3 compounds presented an increase in their concentration during the lights-on period and a remarkable decrease in their values during the lights-off period. In the pons, ANA, PEA and OEA showed the maximum values during the dark phase. On the other hand, we found that in the hippocampus, ANA increased its concentration during the lights-off period and PEA showed the highest peak at the beginning of the same period. OEA concentration showed no diurnal variations in the hippocampus. Finally, in the hypothalamus, ANA rose during the lights-on period whereas PEA and OEA presented the highest concentration at the end of the lights-off period. We postulate that all compounds are likely to be accumulated in parenchyma during the lights-off period (when animal is awake) and then, released into the CSF in order to reach target regions in turn to modulate diverse behaviors, such as feeding and sleep.     

Immigrant and extrinsic hybrid seed inviability contribute to reproductive isolation between forest and dune ecotypes of Epipactis helleborine (Orchidaceae)

Reproductive isolation caused by divergent natural selection arising from differences between ecological environments or ecological interactions represents a major mechanism contributing to speciation, but its relative importance is poorly known. In this study, controlled reciprocal crossings and seed germination experiments were combined with genetic and morphometric analyses to test the hypothesis that previously described differences in mycorrhizal communities between forest and dune ecotypes of Epipactis helleborine were sufficiently strong to create complete reproductive isolation between the two ecotypes. Molecular analyses using 770 SNP markers showed that the two ecotypes were genetically distinct and that populations of the dune ecotype were genetically impoverished compared to populations of the forest ecotype. Morphologically, the two ecotypes were also significantly different, with plants of the dune ecotype generally being smaller than plants of the forest ecotype. The results further showed that immigrant seeds had a significantly lower probability of protocorm formation than native seeds, indicating strong immigrant inviability. Although both ecotypes were able to cross easily and to produce a large number of viable seeds, hybrid seeds showed significantly lower protocorm formation than pure seeds, further contributing to reproductive isolation. Overall, these results indicate that interfertile populations of a widespread orchid adapting to contrasting environments diverge as a consequence of concurrent selection acting against immigrants and hybrids and suggest that mycorrhizal fungi can play a role in the early stages of plant speciation.     

Wood vessel diameter is related to elevation and genotype in the Hawaiian tree Metrosideros polymorpha (Myrtaceae)

We tested the hypothesis that trees growing at high elevations with occasional freezing temperatures have smaller diameter xylem vessels than trees of the same species growing at lower and warmer elevations. The young branch wood of the wide-ranging Hawaiian tree species Metrosideros polymorpha (Myrtaceae) was examined in three natural field populations (high, middle, and low elevations: 2469, 1280, and 107 m a.s.l., respectively) and contrasted with seedlings from these populations that were grown in a common garden at middle elevation (1190 m). Previous studies showed that these populations have some genetic differences and have distinctive leaf structure and ecophysiological traits. Vessel diameter was significantly smaller in the high elevation field and common garden plants than in middle elevation plants. However, high elevation vessels were wider in common garden plants compared to field plants, indicating that vessel diameter is determined both by genotype (parental populations) and environment (growing conditions different from those of parents). Reduced vessel diameter has implications for resistance to cavitation induced by freezing and/or drought in plants growing near tree line in Hawaii.     

草原地区不同生态类型的植物生理特性的比较研究

比较研究了4种不同水分生态型植物在不同水分胁迫下的光合作用、叶片含水量和气孔阻力等生理指标的反应.结果表明,不同水分生态型植物抵御干旱的机制是不同的.中生植物主要是通过增加气孔阻力限制蒸腾失水,而旱生植物则依靠高浓度的细胞原生质减少水分的散失,后者保水效率远高于前者.植物从中生种到旱生种,生理特性亦显示出规律性的种间差异,叶片含水量和气孔阻力水平降低,而单位叶面积的净光合速率增加.     

The relative Zeitgeber strength of lights-on and lights-off is changed in old mice

The daily activity pattern of old mice is characterized by a decreased amplitude, a phase advance, and less stable relationship between lights-off and the onset of the main activity maximum. When analyzing the possible causes of these changes, it must be remembered that the activity rhythm of laboratory mice is bimodal, with a main peak in the first half of the dark time and a secondary one shortly after lights-on. Thus it seems to be controlled by at least two circadian oscillators--an "evening oscillator" coupled more strongly to lights-off and a "morning oscillator" coupled to lights-on--though both oscillators are also coupled to each other. The objective of the present paper was to investigate the putative changes in the strength of these couplings in HaZ:ICR mice of different ages (adult animals of 20 weeks, n = 12; old mice of 72 and 91 weeks of age, n = 6 each) and kept in a 24 h LD-cycle with a gradually reduced light:dark ratio. In adult mice, lengthening the dark time caused the onset of the main maximum of activity to be delayed in relation to the time of lights-off, while the morning maximum of activity was advanced in relation to lights-on. On average, the sizes of the advance and the delay were equal. As a consequence, the activity pattern did not shift in relation to the middle of the dark time. Lengthening the dark time resulted in a bigger (on average, 1.5 h) difference between the evening and morning activity onsets. Under short photoperiods (< or = 2 h of light) the activity rhythm started to free run, and the difference between evening and morning activity onsets decreased again. The changes obtained in senile mice were similar. However, the limits of entrainment were reached with longer photoperiods compared to adult animals. Also, the phase delay of the activity onset in the evening was much less, nearly zero. As a consequence, the activity pattern as whole phase-advanced in relation to the middle of the dark time. A model was proposed in which lights-off triggers advances of the "evening oscillator," lights-on delays the "morning oscillator," and the two oscillators are coupled with each other. Though it was probably the case, decreased coupling strengths could not be shown with the present experimental approach. However, it was clearly evident that, with increasing age, the advancing effect of lights-off exceeded the delaying effect of lights-on.     

The Relative Zeitgeber Strength of Lights-On and Lights-Off Is Changed in Old Mice

The daily activity pattern of old mice is characterized by a decreased amplitude, a phase advance, and less stable relationship between lights-off and the onset of the main activity maximum. When analyzing the possible causes of these changes, it must be remembered that the activity rhythm of laboratory mice is bimodal, with a main peak in the first half of the dark time and a secondary one shortly after lights-on. Thus it seems to be controlled by at least two circadian oscillators—an “evening oscillator” coupled more strongly to lights-off and a “morning oscillator” coupled to lights-on—though both oscillators are also coupled to each other. The objective of the present paper was to investigate the putative changes in the strength of these couplings in HaZ:ICR mice of different ages (adult animals of 20 weeks, n=12; old mice of 72 and 91 weeks of age, n=6 each) and kept in a 24h LD-cycle with a gradually reduced light:dark ratio.

In adult mice, lengthening the dark time caused the onset of the main maximum of activity to be delayed in relation to the time of lights-off, while the morning maximum of activity was advanced in relation to lights-on. On average, the sizes of the advance and the delay were equal. As a consequence, the activity pattern did not shift in relation to the middle of the dark time. Lengthening the dark time resulted in a bigger (on average, 1.5h) difference between the evening and morning activity onsets. Under short photoperiods (≤2h of light) the activity rhythm started to free run, and the difference between evening and morning activity onsets decreased again. The changes obtained in senile mice were similar. However, the limits of entrainment were reached with longer photoperiods compared to adult animals. Also, the phase delay of the activity onset in the evening was much less, nearly zero. As a consequence, the activity pattern as a whole phase-advanced in relation to the middle of the dark time.

A model was proposed in which lights-off triggers advances of the “evening oscillator,” lights-on delays the “morning oscillator,” and the two oscillators are coupled with each other. Though it was probably the case, decreased coupling strengths could not be shown with the present experimental approach. However, it was clearly evident that, with increasing age, the advancing effect of lights-off exceeded the delaying effect of lights-on.     

Limited field establishment of a weed biocontrol agent, Floracarus perrepae (Acariformes: Eriophyidae), against Old World climbing fern in Florida--a possible role of mite resistant plant genotypes

The leaflet galling mite Floracarus perrepae Knihinicki & Boczek was released on Lygodium microphyllum (Cav.) in 63 plots in Florida from 2008 to 2009. Mites transferred onto field plants in 34 plots, but failed to establish populations in the majority of plots. Leaflet galls were observed in only six plots, and in only two plots did mite populations persist for >12 mo. Rates of mite transfer onto field plants were similar for methods using direct transfer of galls versus approaches using passive transfer of mites from infested plants. Often leaflets on some L. microphyllum plants were heavily galled by F. perrepae, whereas leaflets on intertwined stems of other L. microphyllum plants were ungalled but exhibited a characteristic browning and scorching of the leaflet tips. Living mites were consistently present on the undersurface of scorched leaflet tips on ungalled plants, suggesting that this damage might be caused by mite feeding on L. microphyllum genotypes that did not support induction of leaflet galls. Plant nutritional status did not account for differences in galling response, because there were no differences in leaflet nitrogen between galled and ungalled stems. We review those factors known to affect the colonization of biological control agents, and discuss how they may have contributed to the lower than expected rate of F. perrepae establishment.     

Leaf structural dynamics associated with adaptation of two Ebenus cretica ecotypes

Morphological and anatomical features of Ebenus cretica leaflet, such as lanceolate shape, reduced size, dense cover with non-glandular hairs, epidermis of small cells, compact mesophyll, amphipleurous presence of palisade parenchyma, thick cuticle, development of numerous mesophyll phenol-storing cells and the amphistomatic type, disclose the xeromorphic character of the plant. In the island of Crete two ecotypes of E. cretica, ecotype A and ecotype C, are greatly extended. In ecotype A leaflets, the above features are more prominent than in ecotype C. This fact accomplished by physiological data favours the suggestion that plants of ecotype A are better adapted to the xerothermic environment of the island of Crete. This may be the reason that ecotype A occupies the major portion of the island and is predominant in the western and central regions. The distinction of ecotypes A and C, by evaluating the strategies these plants used in order to better adapt and the characteristics of their inflorescences may be used as a criterion for the selection of the most appropriate ecotype for application in floriculture and ornamental horticulture.     

Life history variation in the heavy metal tolerant plant Thlaspi caerulescens growing in a network of contaminated and noncontaminated sites in southern France: role of gene flow, selection and phenotypic plasticity

* Here we explore life history differences in a set of neighbouring metallicolous and nonmetallicolous populations of the heavy metal tolerant plant Thlaspi caerulescens. * We contrasted data from field observations and from a common garden experiment, in which soil zinc (Zn) concentration and light availability were manipulated, and data on microsatellite molecular variation. * The two ecotypes showed few differences in life history in the field, but large differences in their response to Zn concentration in the common garden. Soil toxicity affected most characters in nonmetallicolous plants, while it had no effect on metallicolous plants. The two ecotypes responded similarly to light. Genetic differentiation for quantitative characters between ecotypes contrasted with the absence of differentiation for microsatellites. Conversely, populations of the same ecotype showed similar responses to Zn, despite their high differentiation for molecular markers. * We conclude that divergent selection related to soil toxicity has had a predominant role in shaping life history differences between ecotypes, gene flow weakly opposing local adaptation despite geographical proximity.     

The Effect of Age, Pre-conditioning, and Water Stress on the Transpiration Rates of Douglas-Fir (Pseudotsuga menziesii) Seedlings of Several Ecotypes

Seedlings of Douglas-fir from seed of a number of mesic and xeric origins were grown in growth chambers and a nursery to various ages up to 16 weeks. Measurements were made to determine the effect of seedling age, growth chamber and nursery pre-conditioning, and seed source on transpiration rates under closely controlled laboratory conditions. Additional experiments were conducted on seedlings of two contrasting ecotypes to determine the effect of different pre-conditioning combinations of plant and soil water potential on seedling transpiration rates. Results show that well-watered seedlings of two mesic ecotypes show no decline in transpiration rates per unit leaf area up to 16 weeks of age while corresponding seedlings of three exeric ecotypes do decline. The growth chamber pre-conditioning results in lower seedling transpiration rates and more decline in seedling transpiration rates with increasing plant water stress than for nursery pre-conditioning. In a similar way, the xeric ecotype seedlings have more decline in transpiration rates with increasing plant water stress than do the mesic ecotype seedlings. Soil water potential influences transpiration rates through pre-conditioning effects. Seedlings which have experienced prior soil moisture stress decrease transpiration more in response to low plant water potentials than do plants which have experienced no soil moisture stress. These behavioral characteristics illustrate adaptive means by which seedlings conserve water through the interaction of genetic and preconditioning mechanisms.     

The daily rhythms of melatonin and free fatty acids in goats under varying photoperiods and constant darkness

The purpose of the study was to explore parallel and divergent features of the daily rhythms of melatonin and plasma free fatty acids (FFA) in goats exposed to different lighting conditions. From these features, we attempted to analyze whether the endogenous melatonin rhythm plays any role in the maintenance of the FFA rhythm. Seven Finnish landrace goats were kept under artificial lighting that simulated the annual changes of photoperiod at 60°N (longest photoperiod, 18 h; shortest, 6 h). The ambient temperature and feeding regimen were kept constant. Blood samples were collected 6 times a year at 2 h intervals for 2 d, first in the prevailing light-dark (LD) conditions and then after 3 d in constant darkness (DD). In LD conditions, the melatonin levels always increased immediately after lights-off and declined around lights-on, except in winter (18 h darkness), when the low daytime levels were restored clearly before lights-on. The FFA levels also displayed a consistent rhythmicity, with low levels at night and a transient peak around lights-on. In DD conditions, the melatonin profiles were very similar to those found in the habitual LD conditions, but the rhythm tended to advance. The FFA rhythm persisted also in DD, and the morning peak tended to advance. There was an overall parallelism between the two rhythms, with one significant exception. In winter in LD conditions, the morning rise in FFA levels coincided with lights-on and not with the declining phase of melatonin, whereas in DD conditions, the FFA peak advanced several hours and coincided with the declining phase of melatonin. From this finding and comparisons of the calculated rhythm characteristics, i.e., phase-shifts, phase differences, and correlations, we conclude that the daily rhythm of FFA levels is most probably generated by an endogenous oscillator, primarily adjusted by dawn, whereas the melatonin rhythm in this species is regulated by an oscillator primarily adjusted by dusk. The results did not exclude a modulatory effect of melatonin on the daily FFA profiles, but melatonin secretion, alone, does not explain the patterns sufficiently.     

Genotype by environment interactions for fitness in hybrid genotypes of Avena barbata

We examined genotype (G) by environment (E) interactions for fitness in mesic and xeric ecotypes of the self-fertilizing annual grass, Avena barbata and their recombinant inbred hybrid progeny. Fitness was assayed (1) in experimental water and nutrient treatments in the greenhouse and (2) in common gardens in each ecotype's native habitat. G x E interactions were significant in the greenhouse. Nevertheless, the same recombinant genotypes tended to have high fitness across all water and nutrient treatments. G x E interactions were less pronounced in the field, and were driven by the contrast between the uniformly low survivorship at the mesic site in 2004 and genetic variation in fitness at the other years/site combinations. Moreover, the mesic ecotype consistently outperformed the xeric in both field and greenhouse. Several of the recombinant genotypes outperformed the parents in the novel greenhouse treatments, but these genotypes did not outperform the mesic parent in field trials. Indeed, it is only in the comparison between field and greenhouse environments that there was a noticeable change in the identity of the most-fit genotype. The results provide evidence that hybridization can create genotypes that are better adapted to newer environments such as those imposed in our greenhouse experiments.     

Phenotypic plasticity of sun and shade ecotypes of Stellaria longipes in response to light quality signaling: Cytokinins

From two distinct ecotypes of Stellaria longipes, one genotype was chosen from each of two very different locations, an alpine (sun) and a prairie (shade) habitat. Plants were clonally propagated and grown in controlled environment chambers under low and moderate red to far-red (R/FR) ratios. The prairie ecotype plants exhibited increased stem elongation, leaf expansion and flowering (6-fold) in response to a low R/FR ratio, relative to plants grown under the moderate R/FR ratio. In contrast, plants of the alpine ecotype showed no increased growth in response to a low R/FR ratio and their flowering was reduced, all relative to the plants grown under the moderate R/FR ratio. These different phenotypic responses to the reduction in R/FR ratio were associated with very different profiles and concentrations of endogenous cytokinins (CKs) assessed in growing tissues of the upper shoots. Specifically, increased total CKs were associated with the rapid growth of plants of the prairie ecotype under a low R/FR ratio. In particular, concentrations of bioactive trans-zeatin and dihydrozeatin, were increased during the period of most rapid shoot growth by 2- to 4- fold for these prairie ecotype plants grown under the low R/FR ratio treatment. In contrast, changes in CK levels for the alpine ecotype plants grown under low R/FR ratios were muted. Of especial interest, plants of the alpine ecotype had a predominance of cis-pathway CKs, whereas the low elevation, prairie ecotype plants accumulated predominantly trans-pathway CKs. Speculatively, the pattern emphasizing trans-pathway CKs may be explained by increased LONELY GUY enzyme activity. This enzyme converts and activates nucleotide CKs to free base CKs (bypassing riboside CKs). It could thus explain, in part, the prairie ecotype's ability to respond to shade light with such a high degree of plasticity if one assumes that high trans-CKs levels are causal for the increased shoot growth seen under a low R/FR ratio.