Two native pools of phytochrome A in monocots: Evidence from fluorescence investigations of phytochrome mutants of rice

Fluorescence investigations of phytochrome (phy) in rice (Oryza sativa L. cv. Nipponbare) mutants deficient in phyA, phyB and phyA plus phyB were performed. Total content of the pigment (P(tot)) and its spectroscopic and photochemical characteristics were determined in different parts of the dark-grown and far-red light (FR)-grown coleoptiles. Spectroscopically, phyA in the phyB mutant was identical to phyA in the wild-type (WT) and the extent of the conversion from Pr to lumi-R at 85 K was the same for phyA in both lines and varied similarly, depending on the part of the coleoptile used. The latter finding proved that phyA in rice is heterogeneous and comprises two phyA populations, phyA' and phyA". Functional properties of phyA were also determined. In the dark the phyB mutant had a higher content of phyA, inactive protochlorophyllide (Pchlide633) and active protochlorophyllide (Pchlide655) than WT and its coleoptile was longer, indicating that phyB may affect the development of WT seedlings in the dark. Constant FR drastically reduced the content of phyA, Pchlide633 and Pchlide655 and brought about coleoptile shortening and appearance of the first leaf, whereas pulsed FR of equal fluence was less effective. This suggested that the reactions were primarily of the high irradiance responses type, which are likely to be mediated by phyA'. The effects on protochlorophyllide biosynthesis and growth responses type were more pronounced in the phyB mutant than in the WT seedlings, which can be connected with the higher phyA' content in the phyB mutant and/or phyB interference with its action in WT seedlings. In the phyA mutant induction of Pchlide633 and Pchlide655 biosynthesis was observed under constant FR, indicating that phyC may be responsible for this effect.     

RHYTHMIC OSCILLATION OF CYCLIC 3',5'AMP AND -GMP CONCENTRATION AND STIMULATION OF FLOWERING BY CYCLIC 3',5'-GMP IN Lemna paucicostata 381

Abstract— Rhythmic oscillation of the concentration of cyclic 3',5'-AMP and -GMP in a short day plant, Lemna paucicostata 381 in continuous darkness was detected after 2 cycles of 12 h dark and 12 h light entrainment. Cyclic 3',5'-AMP and -GMP, extracted from whole plant showed parallel oscillations in their concentrations for initial 36 h in continuous darkness and the oscillation in the concentration of cyclic 3',5'-AMP was roughly circadian. Their concentrations decreased during the initial 12 h (subjective night) and increased during 12 to 28 h. Exogenous addition of 2 μ.M of cyclic 3',5'-GMP or the dibutyryl derivative of it stimulated floral induction by 20 to 30%, when the plants were grown under 12 h light and 12 h dark regime. Cyclic 3',5'-AMP or the dibutyryl derivative of it showed little effect on flowering.     

光周期敏感不育水稻中激素的二阶导数卷积伏安法研究

二阶导数卷积伏安法应用于水稻中痛量雌雄激素的检测，具有快速，灵敏，干扰少的特点。本文以光周期敏感核不育水稻农垦５８Ｓ为材料，以农垦５８作对照，了叶焉和穗中雌，雄性激素的含量，研究了不同光周期条件下两类激素的变化。     

Phytochrome Control of Anthocyanin Biosynthesis in Tomato Seedlings: Analysis Using Photomorphogenic Mutants

Anthocyanin biosynthesis has been studied in hypocotyls and whole seedlings of tomato (Lycoperskon esculentum Mill.) wild types (WTs) and photomorphogenic mutants. In white light (WL)/dark (D) cycles the fri¹ mutant, deficient in phytochrome A (phyA), shows an enhancement of anthocyanin accumulation, whereas the tri¹ mutant, deficient in phytochrome Bl (phyBl) has a WT level of anthocyanin. Under pulses of red light (R) or R followed by far-red light (FR) given every 4 h, phyA is responsible for the non-R/FR reversible response, whereas phyBl is partially responsible for the R/FR reversible response. From R and blue light (B) pretreatment studies, B is most effective in increasing phytochrome responsiveness, whereas under R itself it appears to be dependent on the presence of phyBl. Anthocyanin biosynthesis during a 24 h period of monochromatic irradiation at different flu-ence rates of 4 day-old D-grown seedlings has been studied. At 660 nm the fluence rate-response relationships for induction of anthocyanin in the WT are similar, yet complex, showing a low fluence rate response (LFRR) and a fluence rate-dependent high irradiance response (HIR). The high-pigment-1 (hp-1) mutant exhibits a strong amplification of both the LFRR and HIR. The fri¹ mutant lacks the LFRR while retaining a normal HIR. In contrast, a transgenic tomato line overexpressing the oat PHYA3 gene shows a dramatic amplification of the LFRR. The tri¹ mutant, retains the LFRR but lacks the HIR, whereas the fri¹, tri¹ double mutant lacks both components. Only an LFRR is seen at 729 nm in WT; however, an appreciable HIR is observed at 704 nm, which is retained in the tri¹ mutant and is absent in the fri¹ mutant, indicating the labile phyA pool regulates this response component.     

KINETIC AND SPECTROSCOPIC EFFECTS OF PROTEIN-CHROMOPHORE ELECTROSTATIC INTERACTIONS IN BACTERIORHODOPSIN

Abstract— Available pH effects on the absorption spectrum and on the rates of dark adaptation (all- trans → 13 -cis interconversion) of bacteriorhodopsin are analyzed in terms of a model based on protein-chromophore electrostatic interactions. Plausible locations of three protein acid-base groups controlling the spectrum and the activation energy of the thermal trans → 13- cis process are suggested. The importance of the protein charges in controlling the cycles of bacteriorhodopsin is indicated.     

CONTROL BY PHYTOCHROME OF EXTENSION GROWTH and POLAROTROPISM IN CHLORONEMATA OF Funaria hygrometrica

Abstract— The photocontrol of extension growth and polarotropism has been investigated in chloronemata of the moss Funaria hygrometrica Hedw. When grown on a 12/12 h light/dark cycle, chloronemata show light-stimulated diurnal variations in elongation rate with no evidence of a circadian rhythm. Regulation of elongation by a low energy, photoreversible phytochrome mechanism was demonstrated by brief red (R) and far red (FR) irradiations given either at the end of the day (EOD) or 6 h later as a night break (NB). Night break irradiation with polarised R caused polarotropic reorientations of chloronemal growth also through a FR-reversible, low energy phytochrome mechanism. Such transformations of P_r to P_fr were accompanied by a 90° shift in the orientation of the phytochrome chromophore. Microbeam irradiation indicated that the phytochrome chromophores involved in both responses were predominantly, if not entirely, located in the tip of the apical chloronemal cell.     

Kinetics of Stem Elongation in Light-Grown Tomato Plants. Responses to Different Photosynthetically Active Radiation Levels by Wild-Type and aurea Mutant Plants

Abstract— In this research, we measured the short- and long-term, stem elongation responses of wild-type and aurea(au) mutant tomato plants to different photosynthetically active radiation (PAR) levels by using linear voltage transducers. Stem elongation was continuously measured in green tomato plants over 2.75 days, under 12 h light/12 h dark photoperiods or in darkness after a 6 h irradiation period. There is no significant difference in stem elongation between wild-type plants pregrown at either LOO or 400 μmol m^?2 s^?1 and then exposed to 12 h photoperiods. However, in the au mutant there is a very large difference between plants pregrown under 100 or 400 umol m ^?2 s^?1 and then exposed either to 12 h photoperiods or to continuous darkness. Total stem elongation of the wild type appears to be maximal at 100 umol m^?2 s^?1, while that of the au mutant appears to be maximal with PAR 400 umol m^?2 s^?1. Wild-type plants displayed PAR-dependent (in the range 100-800 umol m^?2 s^?1) inhibition of growth both during the day and during the night. In contrast, the au mutant showed a fluence-rate-dependent promotion of growth during the dark periods in the range of 10-400 umol m^?2 s^?1. Large, fast and opposite changes in stem elongation rate at the light/dark and dark/light transitions were present in both genotypes. Internode elongation rate in the first half of the night was always modest in wild-type tomato, whereas it increased rapidly in the au mutant. Stem elongation rate of wild type starts to increase after about 6 h in darkness, showing the typical time course of escape from Pfr-mediated inhibition of elongation by an end-of-day response. The role of phytochrome level and type in sensing light quantity is discussed.     

Phytochrome-mediated inhibition of coleoptile growth in rice: age-dependency and action spectra

Phytochrome has been shown to be the major photoreceptor involved in the photo-inhibition of coleoptile growth in Japonica-type rice (Oryza sativa L.). We have characterized this typical photomorphogenetic response of rice using mutants deficient in phytochrome A (phyA) and phytochrome B (phyB) and with respect to age-dependency and action spectra. Seedlings were irradiated with a pulse of light 40 h or 80 h after germination (i.e. at an early or late developmental stage) and the final coleoptile length of these seedlings was determined. A saturating pulse of red light (R) had a stronger effect when it was given in the late stage than in the early stage. It was found that the photoinhibition is mediated by both the phyA and the phyB in the late stage but predominantly by phyB in the early stage. Consistent with many other reported responses, the photo-inhibition in the phyA mutant, which was observed in the early and late developmental stages and is thought to be mediated mainly by phyB, occurred in the low-fluence range (10(1)-10(3) micromol m(-2)) of R and was far-red-light (FR)-reversible; the photo-inhibition in the phyB mutant, which was observed in the late developmental stage and is thought to be mediated mainly by phyA, occurred in the very-low-fluence range (10(-2)-10(0) micromol m(-2)) and was FR-irreversible. The action spectra (350-800 nm at 50 nm intervals) obtained at the two developmental stages using phyA and phyB mutants indicated that both the phyB-mediated low-fluence response and the phyA-mediated very-low-fluence response have a major peak at 650 nm and a minor peak at 400 nm.     

EXTENSION GROWTH AND ANTHOCYANIN RESPONSES OF PHOTOMORPHOGENIC TOMATO MUTANTS TO CHANGES IN THE PHYTOCHROME: PHOTOEQUILIBRIUM DURING THE DAILY PHOTOPERIOD

Four genotypes of tomato (Lycopersicon esculentum Mill.) in the genetic background ‘Aha Craig’ were used: an aurea (au) mutant, deficient in the bulk light-labile phytochrome pool; a high pigment (hp) mutant, showing exaggerated phytochrome responses at the time of de-etiolation; the au,hp double mutant and the isogenic wild type (WT). A dramatic increase in plant height resulting from an increase in the length of all internodes for each of the genotypes studied was observed upon reduction of the red light: far-red light photon ratio (R:FR) from 6.90 to 0.13 by addition of FR for the whole photoperiod. A concomitant increase in leaf length was also observed. Since au and au,hp mutants, deficient in the bulk light-labile phytochrome pool, respond to this reduction in the R:FR these data demonstrate that the phytochrome pool that mediates this response is present and fully functional. Anthocyanin was detectable in the comparably developed young growing leaves of the WT and hp mutant under the high R:FR, but not in the au and au,hp mutants, suggesting that the potential for anthocyanin synthesis is correlated with the presence of the bulk light-labile phytochrome pool. The kinetics of anthocyanin decrease in the young growing leaves were investigated in the hp mutant and the results suggest a very rapid cessation of flavonoid biosynthesis upon reduction of the R:FR. The functions of different phytochrome types are discussed.     

INHIBITORY EFFECT OF CONTINUOUS FAR-RED PREIRRADIATION ON CHLOROPHYLL ACCUMULATION OF PHARBITIS NIL COTYLEDONS

Abstract The effect of continuous far-red (FR) preirradiation on the accumulation of chlorophyll (Chi) during a white light (WL; 500 lx) period was examined using Pharbitis nil cotyledons. The saturation level of accumulated Chi attained after prolonged exposure to WL was always lowered by continuous FR irradiation preceding the WL. The rate of Chi accumulation during the rapid increase phase (operationally defined as the amount of Chi accumulated during a 24-h WL period) was enhanced by preirradiation with up to 36 h of FR. However, when the FR preirradiation lasted longer, the rate was reduced below the dark control level. Even FR preirradiation of up to 36 h fully reduced the rate of Chi accumulation under WL when 36 h or longer darkness was spaced between the FR and the WL period.     

Isovitexin-2'-O-beta-[6-O-E-p-coumaroylglucopyranoside] from UV-B irradiated leaves of rice, Oryza sativa L. inhibits fertility of Helicoverpa armigera

UV-B irradiated rice leaves (Oryza sativa L.) contained four closely related flavonoids, with either an isoorientin or isovitexin aglycone. These flavonoids have previously been purified and characterized, and were added to artificial diets of the African bollworm (Helicoverpa armigera Hübner) at 0.1x concentration found in irradiated rice leaves. Consumption of different diets had relatively small effects on laval, pupal and adult duration, weight and survival, indicating the insects lived near normal life cycles on all diets. However, one of the compounds, flavonoid IIa, isovitexin-2'-O-beta-[6-O-E-p-coumaroylglucopyranoside], dramatically reduced the number of fertile eggs laid to 7% of control insects (P<0.001) when added to insect diets at 18 nmol gFW(-1) (14 ppm). A similar antifertility effect was observed when only the male partner consumed diet containing flavonoid IIa, indicating that the reduced fertility may be male specific. In contrast, the fecundity and fertility of insects eating diets containing the closely related flavonoids, isoorientin-2'-O-beta-[6-O-E-p-coumaroylglucopyranoside] or isoorientin-2'-O-beta-[6-O-E-p-feruloylglucopyranoside], were not significantly different to control diets.     

Effects of Light Environment During Culture on UV-lnduced Cyclobutyl Pyrimidine Dimers and Their Photorepair in Rice (Oryza sativa L.)

Abstract— We examined the effects of a light environment during culture of rice plants ( Oryza sativa ) on the steady-state cyclobutyl pyrimidine dimer (CPD) level, CPD induction by challenge UVB exposure and the ability to photorepair CPD. The steady-state CPD level in plants grown under visible radiation with supplemental UVB radiation in a growth chamber was several times higher than in plants grown without supplemental UVB radiation, whereas in outdoor-grown plants, it was not enhanced by supplemental UVB radiation. The susceptibility to CPD induction by challenge UVB exposure was highest in dark-grown plants and decreased with increasing irradiance of visible radiation at low and high levels and outdoors. Chronic UVB radiation reduced the susceptibility to UV-induced CPD in plants grown both indoors and outdoors. There was a significant negative correlation between CPD levels induced by challenge UVB exposure and the content of UV-absorbing compounds. The UV-induced CPD could be reduced by subsequent blue radiation in all samples except in dark-grown seedlings. The higher the irradiance of visible radiation in the culture, the greater the ability to photorepair CPD. Chronic UVB radiation did not increase the ability to photorepair CPD.     

The Heterotrimeric G-protein Complex Modulates Light Sensitivity in Arabidopsis thaliana Seed Germination

Release of dormancy and induction of seed germination are complex traits finely regulated by hormonal signals and environmental cues such as temperature and light. The Red (R):Far-Red (FR) phytochrome photoreceptors mediate light regulation of seed germination. We investigated the possible involvement of heterotrimeric G-protein complex in the phytochrome signaling pathways of Arabidopsis thaliana seed germination. Germination rates of null mutants of the alpha (Gα) and beta (Gβ) subunits of the G-protein (At gpa1-4 and agb1-2 , respectively) and the double mutant ( agb1-2/gpa1-4 ) are lower than the wildtype (WT) under continuous or pulsed light. The Gα and Gβ subunits play a role in seed germination under hourly pulses of R lower than 0.1 μmol m⁻² s⁻¹ whereas the Gβ subunit plays a role in higher R fluences. The germination of double mutants of G-protein subunits with phyA-211 and phyB-9 suggests that AtGPA1 seems to act as a positive regulator of phyA and probably phyB signaling pathways, while the role of AGB1 is ambiguous. The imbibition of seeds at 4°C and 35°C alters the R and FR light responsiveness of WT and G-protein mutants to a similar magnitude. Thus, Gα and Gβ subunits of the heterotrimeric G-protein complex modulate light induction of seed germination by phytochromes and are dispensable for the control of dormancy by low and high temperatures prior to irradiation. We discuss the possible indirect role of the G-protein complex on the phytochrome-regulated germination through hormonal signaling pathways.     

LASER PHOTOLYSIS OF 3-METHYLLUMIFLAVIN

Abstract— Using high-intensity actinic light, the chlorophyll a fluorescence transient from HCO^-₃-depleted chloroplasts shows a rapid initial rise (O → I) followed by a slow phase (I → P). In the presence of HCO^-₃, the O → I rise is delayed but the I → P phase is much more rapid. Using low-intensity actinic light, the chlorophyll a fluorescence transient from 3-(3,4-dichlorophenyl)-1,1 dimethylurea (DCMU)-treated chloroplasts is delayed in the presence of HCO^-₃. Bicarbonate increases the amount of delayed light emission from chloroplasts given 10 s illumination with weak blue light (0·4 W/m²). DCMU greatly increases the amount of delayed light seen in the presence of HCO^-₃ under these conditions but decreases the amount seen in the absence of HCO^-₃. It is suggested that HCO^-₃ may somehow form or stabilize, in the dark, a number of reaction centers corresponding to the S₁ state in the model of B. Forbush, B. Kok and M. McGloin ( Photochem. Photobiol. 14, 307–321, 1971).     

PHOTOTACTIC BEHAVIOR OF INDIVIDUAL CELLS OF CRYPTOMONAS SP. IN RESPONSE TO CONTINUOUS AND INTERMITTENT LIGHT STIMULI

Abstract— The phototactic response of cells of Cryptomonas sp. to stimulation with continuous or intermittent lateral light was determined by an individual cell method using photomicrography and videomicrography. The cells showed positive phototaxis under the conditions studied. The phototactic orientation of individual cells was induced most effectively by irradiation with light of 570 nm; blue light was less effective, and no orientation was found in red light. An intermittent stimulus regime with a long dark interval (250 ms) elicited a weaker phototactic orientation than did a regime with a short dark interval (63 ms) irrespective of the duration of light pulses (16, 250 and 1000 ms). The swimming rate was ca. 240 ums ^-1 and the rotation period ca. 450 ms in the dark, neither of which was greatly affected by stimulation with continuous or intermittent light. Neither step-up nor step-down photophobic responses were observed at the time of onset or removal of the light stimulus under the experimental conditions. The swimming direction of individual cells became gradually oriented toward the light source. Phototactic response was detectable within 4 s after the onset of light stimulation, reaching a saturation level after more than 30 s.     

Identification of a light-regulated protein kinase activity from seedlings of Arabidopsis thaliana

Protein kinase transduction pathways are thought to be involved in light signaling in plants, but other than the photoreceptors, no protein kinase activity has been shown to be light-regulated in vivo. Using an in-gel protein kinase assay technique with histone H III SS as an exogenous substrate, we identified a light-regulated protein kinase activity with an apparent molecular weight ca 50 kDa. The kinase activity increased transiently after irradiation of dark-grown seedlings with continuous far red light (FR) and blue light (B) and decreased after irradiation with red light (R). The maximal activation was achieved after 30 min to 1 h with FR or B. After irradiation times longer than 2 h, the kinase activity decreased to below the sensitivity level of the assay. In Arabidopsis mutants lacking either the photoreceptors phytochrome A, phytochrome B or the blue-light receptor cryptochrome 1, kinase activity was undetectable, whereas in the photomorphogenic mutants cop1 and det1 the kinase activity was also observed in the absence of light signals, though still stimulated by B and FR. Interestingly, the R inhibition of the kinase activity was lost in the mutant hy5. Pretreatment with cycloheximide blocked the kinase activity.     

Light effects in yeast: persisting oscillations in cell division activity and amino acid transport in cultures of Saccharomyces cerevisiae entrained by light-dark cycles.

Abstract— In addition to the direct inhibitory effects of visible light (cool-white fluorescent, < 3500 lux) on growth and amino acid transport previously reported for cultures of the yeast Saccharomyces cereui-siae (strain Y185 rho⁺) grown at 12°C in medium containing glucose, yeast carbon base, KH₂PO₄, ammonium ion and proline, we have found evidence for endogenous, light-entrainable, self-sustaining circadian and ultradian oscillators underlying both cell division and transport activity. Diurnal light (? 3000 lux) cycles (LD), imposed on cultures previously grown in the dark, phased or synchronized cell number increase to a 24-h period with bud release being confined primarily to the dark intervals (although not necessarily every cell divided during any given division ‘burst’). The observed division or budding rhythm freeran with a circadian period (?26h) only approximating 24 h for a number of days in constant darkness (DD) following prior entrainment by LD, thereby implicating an endogenous circadian clock. Further, a similar light-entrainable circadian rhythm in the uptake of “C-histidine or ¹4C-lysine occurred in nondividing (or very slowly dividing) cultures during the “stationary” (infradian) phase of growth synchronized by a 24-h LD cycle and then released into DD for as long as 10 days. Some experiments revealed a bimodal (ultradian) periodicity in both LD and DD with secondary peaks or shoulders occurring at intervals of ?12h, corresponding approximately to subjective ‘dawn’ and ‘dusk’. Transport in cultures of the Y185 rho^- petite mutant, which lacks cytochromes a/a₃, b and c_l, could not be synchronized by LD cycles, a finding that is consistent with the hypothesis that these chromophores may be primary photoreceptors for synchronization of rhythms in this microorganism.     

SPECIFIC PROTEIN OCCURRENCE RELATED WITH PHOTOPERIODIC FLOWER INDUCTION IN THE COTYLEDONS OF Pharbitis nil CV. VIOLET

In our experiment, three groups of seedlings of SDP Pharbitis nil cv. violet were sepa-rately treated with three different photoperiods (1,16 h dark period--SD; 2, continuous illumi-nation--CL; 3, 16 h dark treatment with 10 min white light in the middle of the dark period--NB). By analysing proteins in the cotyledons from three groups with 2-D PAGE, we found nodifference in protein pattern between the three groups at 0 or 8 h after photoperiodic treatments.At 24 h after the treatments, a specific protein(MW:19 kD; pI: 4.5)appeared only in the cotyledonsof the seedlings which endured SD. This protein disappeared at 72 h after SD. ActinomycinD could inhibit flowering and the specific protein occurrence when applied to cotyledonsprior to SD, but it had no inhibition effect on flowering as well as the specific proteinoccurrence when applied to cotyledons after SD. Chloroamphenicol, a protein synthesisinhibitor, inhibited flowering when applied to cotyledons prior to or immediately after SD,but it did not i     

PHOTORESPONSES OF Arabidopsis SEEDLINGS EXPRESSING AN INTRODUCED OAT phyA cDNA: PERSISTENCE OF ETIOLATED PLANT TYPE RESPONSES IN LIGHT-GROWN PLANTS

The photocontrol of hypocotyl elongation has been studied in etiolated and light-grown wild type (WT) Arabidopsis thaliana (L. Heynh) seedlings, and in two homozygous isogenic lines that have been transformed with the oat phy A gene coding sequence under the control of the cauliflower mosaic virus (CaMV) 35S promoter. For etiolated seedlings the inhibition of hypocotyl elongation by continuous broad band far-red light (FR) is saturated at much lower photon fluence rates in the transgenic seedlings compared with WT seedlings. Furthermore, whereas de-etiolation of WT seedlings leads to loss of responsiveness of the hypocotyls to prolonged FR, de-etiolated transgenic seedlings continue to show a pronounced FR-mediated inhibition of elongation. This may reflect the persistence of a FR-high irradiance response (HIR) mediated by the introduced oat phytochrome A. Although the hypocotyls of light-grown transgenic seedlings display a qualitatively normal end-of-day FR growth promotion, such seedlings display an aberrant shade-avoidance response to reduced red:far-red ratio (R:FR). These results are discussed in relation to the proposal that the constitutive expression of phytochrome A leads to the persistence of photoresponse modes normally restricted to etiolated plants.     

MULCH SURFACE COLOR AFFECTS ACCUMULATION OF EPICUTICULAR WAX ON DEVELOPING LEAVES

Abstract— Leaf waxes are important to plant growth because they impede water loss and may influence entry of light. Leaf wax quantity and composition were studied in Capsicum annuum plants in trickle-irrigated field plots covered with white, black or red plastic mulches. The quantity of reflected blue light (BL) was greatest over white and about the same over black versus red surfaces; and reflected far-red to red ratios (FR/R) were about the same over white versus black and higher over red. The greatest quantity of total epicuticular wax developed on leaves of plants grown over white mulch (443 μg/cm²) while plants grown over black and red mulches had 229 and 227 μg/cm², respectively. When individual lipid classes were expressed as percentages of the totals, esters constituted the highest percentages over white and black surfaces while secondary free fatty alcohols dominated over red. The higher ester content of epicuticular wax on plants grown over black (versus red) was almost exactly equalled by increased contents of free fatty alcohols and free fatty acids in the epicuticular wax of plants grown over red mulch. We conclude that the total wax concentration on leaves of field-grown plants was influenced by quantity of BL, and the percentage of individual components was influenced by the FR/R ratio.