Light perception and signalling by phytochrome A.

In etiolated seedlings, phytochrome A (phyA) mediates very-low-fluence responses (VLFRs), which initiate de-etiolation at the interphase between the soil and above-ground environments, and high-irradiance responses (HIR), which complete de-etiolation under dense canopies and require more sustained activation with far-red light. Light-activated phyA is transported to the nucleus by FAR-RED ELONGATED HYPOCOTYL1 (FHY1). The nuclear pool of active phyA increases under prolonged far-red light of relatively high fluence rates. This condition maximizes the rate of FHY1-phyA complex assembly and disassembly, allowing FHY1 to return to the cytoplasm to translocate further phyA to the nucleus, to replace phyA degraded in the proteasome. The core signalling pathways downstream of nuclear phyA involve the negative regulation of CONSTITUTIVE PHOTOMORPHOGENIC 1, which targets for degradation transcription factors required for photomorphogenesis, and PHYTOCHROME-INTERACTING FACTORs, which are transcription factors that repress photomorphogenesis. Under sustained far-red light activation, released FHY1 can also be recruited with active phyA to target gene promoters as a transcriptional activator, and nuclear phyA signalling activates a positive regulatory loop involving BELL-LIKE HOMEODOMAIN 1 that reinforces the HIR.

Conjugated linoleic acid (CLA) and polyunsaturated fatty acids in muscle lipids of lambs from the Patagonian area of Argentina.

The concentrations of fatty acids were measured in total lipids, triacyglycerol and phospholipid fractions of intramuscular fat (IMF) from the Longissimus dorsi (LD) muscle of 10 lambs reared to approximately 30kg live weight on natural pasture with their dams. Fatty acid composition was also measured in 25 (five of each) Semitendinosus (ST), Semimembranosus (SM), Rectus femoris (RF), Gluteus (GLU) and Tensor fascia latea (TFL) muscles. Intramuscular fat percentages were similar for all muscles. Aspects of the fatty-acid patterns of relevance to human nutrition tended to favor the leg muscles with lower saturated fatty acids (SFA %), n-6/n-3 fatty acid ratios (p<0.01) and higher concentrations of the conjugated linoleic acid (CLA) (p<0.05). The estimated fatty acid concentrations (mg/100g of meat) showed higher contribution of arachidonic (C20:4 n-6), eicosapentanoic (C20:5 n-3), docosapentanoic (C22:5 n-3) and docosahexanoic (C22:6 n-3) acids in leg compared to LD lipids.

Beef lipids in relation to animal breed and nutrition in Argentina.

Fatty acid (FA) composition of intramuscular fat (IMF) in M. Longissimus dorsi (LD) was measured in 72 steers from Angus (A), Charolais×Angus (CHA×A) and Holstein Argentine (HA) breeds. The steers were allotted to four dietary treatments of six animals each: T1, steers grazed on pasture; T2, steers supplemented with cracked corn grain (0.7% of live-weight) daily and free access to pasture; T3, steers supplemented with cracked corn grain (1% of live-weight) daily and free access to pasture; and T4, feedlot (concentrate based on corn, alfalfa hay and soybean meal without access to pasture). At slaughter weight, samples of LD at the 11th rib were used for intramuscular lipid analysis. The diet was shown to be more important than breed in determining FA composition. Pasture beef had higher percentages of saturated fatty acids (SFA), n-3 polyunsaturated fatty acids (PUFA) and conjugated linoleic acid (CLA) and lower percentages of IMF, monounsaturated fatty acids (MUFA), n-6 PUFA and n-6/n-3 ratios than feedlot beef. HA beef presented lower percentages of SFA and more MUFA with a higher n-6/n-3 ratio than A and CHA×A. Comparing grass and feedlot beef the amounts of FA in muscle (mg/100g) were, respectively 18:3 n-3 (44 vs. 11mg), CLA (20 vs. 12mg), 20:5 n-3 (20 vs. 11mg), 22:5 n-5 (20 vs. 11mg), 22:6 n-3 (12 vs. 6mg) and n-3 PUFA (84 vs. 32mg). Feedlot beef has more SFA (1372 vs. 1081mg), MUFA (1574 vs. 1078mg), PUFA (350 vs. 227mg) and n-6 PUFA (318 vs.143mg).

A quadruple photoreceptor mutant still keeps track of time.

Time measurement and light detection are inextricably linked. Cryptochromes, the blue-light photoreceptors shared between plants and animals, are critical for circadian rhythms in flies and mice [1-3]. WC-1, a putative blue-light photoreceptor, is also essential for the maintenance of circadian rhythms in Neurospora [4]. In contrast, we report here that in Arabidopsis thaliana the double mutant lacking the cryptochromes cry1 and cry2, and even a quadruple mutant lacking the red/ far-red photoreceptor phytochromes phyA and phyB as well as cry1 and cry2, retain robust circadian rhythmicity. Interestingly, the quadruple mutant was nearly blind for developmental responses but perceived a light cue for entraining the circadian clock. These results indicate that cryptochromes and phytochromes are not essential components of the central oscillator in Arabidopsis and suggest that plants could possess specific photosensory mechanisms for temporal orientation, in addition to cryptochromes and phytochromes, which are used for both spatial and temporal adaptation.

Phytochrome A Mediates the Promotion of Seed Germination by Very Low Fluences of Light and Canopy Shade Light in Arabidopsis.

Seeds of the wild type (WT) and of the phyA and phyB mutants of Arabidopsis thaliana were exposed to single red light (R)/far-red light (FR) pulses predicted to establish a series of calculated phytochrome photoequilibria (Pfr/P). WT and phyB seeds showed biphasic responses to Pfr/P. The first phase, i.e. the very-low-fluence response (VLFR), occurred below Pfr/P = 10-1%. The second phase, i.e. the low-fluence response, occurred above Pfr/P = 3%. The VLFR was similarly induced by either a FR pulse saturating photoconversion or a subsaturating R pulse predicted to establish the same Pfr/P. The VLFR was absent in phyA seeds, which showed a strong low-fluence response. In the field, even brief exposures to the very low fluences of canopy shade light (R/FR ratio < 0.05) promoted germination above dark controls in WT and phyB seeds but not in the phyA mutant. Seeds of the phyA mutant germinated normally under canopies providing higher R/FR ratios or under deep canopy shade light supplemented with R from light-emitting diodes. We propose that phytochrome A mediates VLFR of A. thaliana seeds.

Resetting of the circadian clock by phytochromes and cryptochromes in Arabidopsis.

The authors sought to investigate the role of phytochromes A and B (phyA and phyB) and cryptochromes 1 and 2 (cryl and cry2) in the synchronization of the leaf position rhythm in Arabidopsis thaliana. The seedlings were transferred from white light-dark cycles to free-running conditions with or without exposure to a light treatment during the final hours of the last dark period. The phase advance caused by a far-red light treatment was absent in the phyA mutant, deficient in the fhy1 and fhy3 mutants involved in phyA signaling, and normal in the cryl and cryl cry2 mutants. The phase shift caused by blue light was normal in the cry2 mutant; reduced in the phyA, cryl, phyA cry1, and cry1 cry2 mutants; and abolished in the phyA cryl cry2 triple mutant. The phase shift caused by red light was partially retained by the phyA phyB double mutant. The authors conclude that cryl and cry2 participate as photoreceptors in the blue light input to the clock but are not required for the phyA-mediated effects on the phase of the circadian rhythm of leaf position. The signaling proteins FHY1 and FHY3 are shared by phyA-mediated photomorphogenesis and phyA input to the clock.

Phytochromes, cryptochromes, phototropin: photoreceptor interactions in plants.

In higher plants, natural radiation simultaneously activates more than one photoreceptor. Five phytochromes (phyA through phyD), two cryptochromes (cry1, cry2) and phototropin have been identified in the model species Arabidopsis thaliana. There is light-dependent epistasis among certain photoreceptor genes because the action of one pigment can be affected by the activity of others. Under red light, phyA and phyB are antagonistic, but under far-red light, followed by brief red light, phyA and phyB are synergistic in the control of seedling morphology and the expression of some genes during de-etiolation. Under short photoperiods of red and blue light, cry1 and phyB are synergistic, but under continuous exposure to the same light field the actions of phyB and cry1 become independent and additive. Phototropic bending of the shoot toward unilateral blue light is mediated by phototropin, but cry1, cry2, phyA and phyB positively regulate the response. Finally, cry2 and phyB are antagonistic in the induction of flowering. At least some of these interactions are likely to result from cross talk of the photoreceptor signaling pathways and uncover new avenues to approach signal transduction. Experiments under natural radiation are beginning to show that the interactions create a phototransduction network with emergent properties. This provides a more robust system for light perception in plants.

Two photobiological pathways of phytochrome A activity, only one of which shows dominant negative suppression by phytochrome B.

The plant receptor phytochrome A (phyA) mediates responses like hypocotyl growth inhibition and cotyledon unfolding that require continuous far-red (FR) light for maximum expression (high-irradiance responses, HIR), and responses like seed germination that can be induced by a single pulse of FR (very-low-fluence responses, VLFR). It is not known whether this duality results from either phyA interaction with different end-point processes or from the intrinsic properties of phyA activity. Etiolated seedlings of Arabidopsis thaliana were exposed to pulses of FR (3 min) separated by dark intervals of different duration. Hypocotyl-growth inhibition and cotyledon unfolding showed two phases. The first phase (VLFR) between 0.17 and 0.5 pulses.h-1, a plateau between 0.5 and 2 pulses.h-1 and a second phase (HIR) at higher frequencies. Reciprocity between fluence rate and duration of FR was observed within phases, not between phases. The fluence rate for half the maximum effect was 0.1 and 3 mumol.m-2.s-1 for hourly pulses of FR (VLFR) and continuous FR (HIR), respectively. Overexpression of phytochrome B caused dominant negative suppression under continuous but not under hourly FR. We conclude that phyA is intrinsically able to initiate two discrete photoresponses even when a single end-point process is considered.

Effect of body fat distribution on the fatty acid composition of the depot fat of cattle.

The influence of liveweight, total carcass fat and the amount of fat at five specific anatomical locations on the fatty acid composition of the depot fat of adult ruminants was examined. The total carcassfat and the amount offat at specific locations seem to affect the fatty acid composition significantly. No important significant correlation between liveweight and fatty acid composition was found. There was an increase in odd, even and branched chain fatty acids, and a decrease in linoleic acid, with increase in animal age.

A comparison of distribution and composition of intramuscular fat in Duroc Jersey and Hampshire pigs at 100 kg liveweight.

The intramuscular fat content, its fatty acid composition and that from the triglyceride fraction, were determined in several muscles from the right sides of four Duroc Jersey and four Hampshire castrated male pigs slaughtered at 100 kg liveweight. The Hampshires had heavier muscles than the Duroc Jersey pigs, but did not show significant differences in the percentages of intramuscular fat. The concentrations of linoleic acid were, in general, higher in the Hampshire than in the Duroc Jersey pigs and in many muscles the differences were highly significant (p < 0·05) The concentrations of linoleic acid (18:2) in the triglyceride fraction did not differ among muscles within breeds but were higher in Hampshire than in Duroc Jersey pigs.

Effect of breed-type on the relationships between intramuscular and total body fat in steers.

The partitioning of total dissectible body fat and the amounts of intramuscular fat in Psoas major, Semitendinosus and Biceps brachii muscles were determined in two groups of A. Angus and AA × Nelore steers with similar averages of total dissectible fat (27·7 kg). In addition, the fatty acid composition of total fat and the triglyceride fraction from dissectible and intramuscular fats were determined. The AA × Nelore steers have higher levels of subcutaneous fat and lower levels of intermuscular fat than the A. Angus but contain lower levels of intramuscular fat in the three muscles. The allometric regressions varied according to the muscle and breed type. The fatty acid composition of subcutaneous and kidney fats were similar but differences in the percentages of 14:0, 18:0, 18:2 and 20:4 fatty acids in intramuscular fats between the two genetic groups were detected.

Butterfat in rations for pigs fed skimmed whey: Growth rate, feed conversion, economics, carcass evaluation and fat quality.

An experiment was carried out, using 81 castrated male pigs, to estimate the effect of adding butterfat (70% fat, 30% buttermilk), as an energy supplement, to the rations of pigs fed skimmed whey ad libitum. The rations were fed daily in a restricted amount and butterfat added to rations at two levels: T-2 and T-3, 140 and 280 g/day/animal, respectively. In treatment T-1, no butterfat was added. The pigs were weighed every 21 days until they reached average liveweights of 100 (group 1) or 120 kg (group 2), when they were killed for carcass studies. There were significant differences in liveweight gains and feed conversion for the pigs supplemented with butterfat. A negative result, from an economical point of view, was obtained due to the relatively high price of the butterfat. A higher yield of carcass weight on field liveweight was obtained in treatments T-2 and T-3, reflecting the higher degree of fatness. The relative carcass length, muscle depth and percentage of defatted ham were higher in T-1. Conversely, the relative depth of fat was more marked in the groups and treatments that consumed butterfat with their rations. Both groups of T-1 yielded the highest percentage of muscle and the lowest of fat in their carcasses, the differences from T-2 and T-3 being significant. T-2 and T-3 did not differ significantly from one another. There were significant differences in the fatty acid composition due to sampling location. The differences due to treatment were statistically significant in respect of the concentrations of 14:0, 16:1, 17:0, 17:1, 18:0 and 18:1 fatty acids. The unusually low concentration of 18:2 in the dissected fat tissues produces high quality porcine fats, for use in manifactured and prepared foods.

Phytochrome A enhances the promotion of hypocotyl growth caused by reductions in levels of phytochrome B in its far-red-light-absorbing form in light-grown Arabidopsis thaliana.

We sought to determine if phytochrome B (phyB)-mediated responses to the red light (R)/far-red light (FR) ratio are affected by phytochrome A (phyA) activity in light-grown seedlings of Arabidopsis thaliana. Pulses of FR delayed into the dark period were less effective than end-of-day (EOD) FR in promoting hypocotyl growth over a given period in darkness. White light minus blue light interposed instead of darkness between the end of the white-light photoperiod and the FR pulse was sufficient to maintain responsivity to the decrease in phyB in FR-light-absorbing form in wild-type (WT) seedlings, but not in the phyA mutant. Compared with EOD R, hourly R+FR pulses provided throughout the night caused a stronger promotion of stem growth than a single EOD R+FR pulse in WT Arabidopsis, cucumber, mustard, sunflower, tobacco, and tomato, but not in phyA Arabidopsis or in the aurea mutant of tomato. WT seedlings of Arabidopsis responded to a range of high EOD R/FR ratios, whereas the phyA mutant required stronger reductions in the EOD R/FR ratio. In sunlight, phyA seedlings of Arabidopsis showed no response to the "early warning" signals of neighboring vegetation, and hypocotyl-growth promotion occurred at higher plant densities than in the WT. Thus, under a series of light conditions, the sensitivity or responsivity to reductions in the R/FR ratio were larger in WT than in phyA seedlings. A product of phyA is therefore proposed to enhance the hypocotyl-growth response to decreases in phyB in FR-light-absorbing form in light grown seedlings.

Coupling of phytochrome B to the control of hypocotyl growth in Arabidopsis.

Etiolated seedlings of the wild-type (WT) and of the phyB-1 mutant of Arabidopsis thaliana (L.) Heynh. were exposed to red-light (R) and far-red light (FR) treatments to characterize the action of phytochrome B on hypocotyl extension growth. A single R or FR pulse had no detectable effects on hypocotyl growth. After 24-h pre-treatment with continuous FR (FRc) a single R, compared to FR pulse inhibited (more than 70%) subsequent hypocotyl growth in the WT but not in the phyB-1 mutant. This effect of FRc was fluence-rate dependent and more efficient than continuous R (Rc) or hourly FR pulses of equal total fluence. Hypocotyl growth inhibition by Rc was larger in WT than phyB-1 seedlings when chlorophyll screening was reduced either by using broad-band Rc (maximum emission 610 nm) or by using narrow-band Rc (658 nm) over short periods (24 h) or with seedlings bleached with Norflurazon. Hourly R or R+FR pulses had similar effects in WT and phyB-1 mutant etiolated seedlings. It is concluded that phytochrome B is not the only photoreceptor of Rc and that the action of phytochrome B is enhanced by a FRc high-irradiance reaction. Complementary experiments with the phyA-201 mutant indicate that this promotion of a phytochrome B-mediated response occurs via co-action with phytochrome A.

Co-action between phytochrome B and HY4 in Arabidopsis thaliana.

A combination of physiological and genetic approaches was used to investigate whether phytochromes and blue light (BL) photoreceptors act in a fully independent manner during photomorphogenesis of Arabidopsis thaliana (L.) Heynh. Wild-type seedlings and phyA, phyB and hy4 mutants were daily exposed to 3 h BL terminated with either a red light (R) or a far-red light (FR) pulse. In wild-type and phyA-mutant seedlings, BL followed by an R pulse inhibited hypocotyl growth and promoted cotyledon unfolding. The effects of BL were reduced if exposure to BL was followed by an FR pulse driving phytochrome to the R-absorbing form (Pr). In the wild type, the effects of R versus FR pulses were small in seedlings not exposed to BL. Thus, maximal responses depended on the presence of both BL and the FR-absorbing form of phytochrome (Pfr) in the subsequent dark period. Impaired responses to BL and to R versus FR pulses were observed in phyB and hy4 mutants. Simultaneous irradiation with orange light indicated that BL, perceived by specific BL photoreceptors (i.e. not by phytochromes), required phytochrome B to display a full effect. These results indicate interdependent co-action between phytochrome B and BL photoreceptors, particularly the HY4 gene product. No synergism between phytochrome A (activated by continuous or pulsed FR) and BL photoreceptors was observed.

Conditional synergism between cryptochrome 1 and phytochrome B is shown by the analysis of phyA, phyB, and hy4 simple, double, and triple mutants in Arabidopsis.

Wild-type or phyA, phyB, or hy4 mutant Arabidopsis seedlings lacking phytochrome A (phyA), phytochrome B (phyB), or cryptochrome 1 (cry1), respectively, and the double and triple mutants were used in combination with blue-light treatments given simultaneously with red or far-red light. We investigated the interaction between phytochromes and cry1 in the control of hypocotyl growth and cotyledon unfolding. Under conditions deficient for cry1 (short exposures to blue light) or phyB (far-red background), these photoreceptors acted synergistically: Under short exposures to blue light (3 h/d) added to a red-light background, cry1 activity required phyB (e.g. the hy4 mutant was taller than the wild type but the phyBhy4 mutant was not taller than the phyB mutant). Under prolonged exposures to blue light (24 h/d) added to a far-red light background, phyB activity required cry1 (e.g. the phyAphyB mutant was taller than the phyA mutant but the phyAphyBhy4 mutant was not taller than the phyAhy4 mutant). Under more favorable light inputs, i.e. prolonged exposures to blue light added to a red-light background, the effects of cry1 and phyB were independent. Thus, the synergism between phyB and cry1 is conditional. The effect of cry1 was not reduced by the phyA mutation under any tested light condition. Under continuous blue light the triple mutant phyAphyBhy4 showed reduced hypocotyl growth inhibition and cotyledon unfolding compared with the phyAphyB mutant. The action of cry1 in the phyAphyB double mutant was higher under the red-light than the far-red-light background, indicating a synergistic interaction between cry1 and phytochromes C, D, or E; however, a residual action of cry1 independent of any phytochrome is likely to occur.

The loci of perception for phytochrome control of internode growth in light-grown mustard: Promotion by low phytochrome photoequilibria in the internode is enhanced by blue light perceived by the leaves.

Under continuous white light (WL), extension growth of the first internode in Sinapis alba L. was promoted by low red (R): far-red (FR) ratios reaching the stem and-or the leaves. Conversely, the growth promotion by end-of-day light treatments was only triggered by FR perceived by the leaves and cotyledons, while FR given to the growning internode alone was tatally ineffective. Continuous WL+FR given to the internode was also in-effective if the rest of the shoot remained in darkness. Both the background stem growth, and the growth promotion caused by either an end-of-day FR pulse or continuous WL+FR given to the internode, increased with increasing fluence rates of WL given to the rest of the shoot. The increase by WL of the growth-stimulatory effect of low phytochrome photoequilibria in the internode appears to be mediated by a specific blue-light-absorbing photoreceptor, as blue-deficient light from sodium-discharge lamps, or from filtered fluorescent tubes, promoted background stem growth similarly to WL but did not amplify the response to the R:FR ratio in the internode. Supplementing the blue-deficient light (94 µmol·m(-2)·s(-1)) with low fluence rates of blue (<9 µmol·m(-2)·s(-1)) restored the promotive effect of low R:FR reaching the internode.

Persistent effects of changes in phytochrome status on internode growth in light-grown mustard: Occurrence, kinetics and locus of perception.

Extension growth of the first internode in fully de-etiolated mustard (Sinapis alba L.) seedlings (11-12.5 d old) is under the control of both the current phytochrome photoequilibrium (Pfr/P, ratio of the far-red-absorbing form of phytochrome to total phytochrome) and that established by short (<12 h) pretreatments. Plants were pretreated with either light pulses providing different calculated Pfr/P followed by dark incubations of different durations (a), or with a 12-h period of white light establishing different Pfr/P (b). After the pretreatments, the plants received either light pulses providing different Pfr/P, followed by dark incubations (c), or continuous white light with or without addtional far-red light (d). Thus, four experimental approaches were followed: (a)→(c); (a)→(d); (b)→(c) and (b)→(d). Extension growth during the second period (c or d) was not only affected by the current phytochrome status, but also by that established during the pretreatment period (a or b). The results show the existence of a long-term promotion of stem growth which persists after the end of the low Pfr/P pretreatment. This effect is different from the previously reported rapid effect of far-red light added to background white light as follows: (i) the duration of low Pfr/P required to effect a full response is longer (2.5 h); (ii) the duration of the promotion after returning to high Pfr/P is longer (approx. 24 h) and (iii) the locus of perception is mainly in the leaves, rather than the growing internode.

Temperature-dependent internode elongation in vegetative plants of Arabidopsis thaliana lacking phytochrome B and cryptochrome 1.

Vegetative plants of Arabidopsis thaliana (L.) Heynh. form a compact rosette of leaves in which internode growth is virtually arrested. Rapid extension of the internodes occurs after flower buds are present in the reproductive apex. Under natural radiation, continuous light from fluorescent lamps, or short photoperiods of light from fluorescent lamps, plants of the phyB cry1 double mutant (lacking both phytochrome B and cryptochrome 1) did not form normal rosettes because all the internodes showed some degree of elongation. Internode elongation was weak in thephyB single mutant and absent in the cry1 mutant, indicating redundancy between phytochrome B and cryptochrome 1. The absence of phytochrome A caused no effects. The failure to form normal rosettes was conditional because internode elongation was arrested at low temperatures in all the mutant combinations. In contrast, the temperature dependence of phytochrome B and cryptochrome 1 effects on hypocotyl growth was weak. The elongation of the internodes in phyB cry1 was not accompanied by early flowering as showed by the lack of effects on the final number of leaves. Apex dissection indicated that in phyB cry1 double mutants internode elongation anticipated the transition from the vegetative to the reproductive stage. Thus, stem growth in Arabidopsis thaliana is not fully dependent on the program of reproductive development.

Effects of Light Quality on Tiller Production in Lolium spp.

Variations in phytochrome status in grasses were provoked by end of day red and far-red treatments and above canopy irradiations. Lolium perenne L. and Lolium multiflorum Lam. plants developed more tillers when illuminated with higher red/far-red ratios. These results show that branching of grasses is controlled by phytochrome activity in a way similar to that in dicotyledon plants.