Etiolated Tissues Research Articles

Iron uptake of etioplasts is independent from photosynthesis but applies the reduction-based strategy.

Iron (Fe) is one of themost important cofactors in the photosynthetic apparatus, and its uptake by chloroplasts has also been associated with the operation of the photosynthetic electron transport chain during reduction-based plastidial Fe uptake. Therefore, plastidial Fe uptake was considered not to be operational in the absence of the photosynthetic activity. Nevertheless, Fe is also required for enzymatic functions unrelated to photosynthesis, highlighting the importance of Fe acquisition by non-photosynthetic plastids. Yet, it remains unclear how these plastids acquire Fe in the absence of photosynthetic function. Furthermore, plastids of etiolated tissues should already possess the ability to acquire Fe, since the biosynthesis of thylakoid membrane complexes requires a massive amount of readily available Fe. Thus, we aimed to investigate whether the reduction-based plastidial Fe uptake solely relies on the functioning photosynthetic apparatus. In our combined structure, iron content and transcript amount analysis studies, we used Savoy cabbage plant as a model, which develops natural etiolation in the inner leaves of the heads due to the shading of the outer leaf layers. Foliar and plastidial Fe content of Savoy cabbage leaves decreased towards the inner leaf layers. The leaves of the innermost leaf layers proved to be etiolated, containing etioplasts that lacked the photosynthetic machinery and thus were photosynthetically inactive. However, we discovered that these etioplasts contained, and were able to take up, Fe. Although the relative transcript abundance of genes associated with plastidial Fe uptake and homeostasis decreased towards the inner leaf layers, both ferric chelate reductase FRO7 transcripts and activity were detected in the innermost leaf layer. Additionally, a significant NADP(H) pool and NAD(P)H dehydrogenase activity was detected in the etioplasts of the innermost leaf layer, indicating the presence of the reducing capacity that likely supports the reduction-based Fe uptake of etioplasts. Based on these findings, the reduction-based plastidial Fe acquisition should not be considered exclusively dependent on the photosynthetic functions.

Antibacterial and hypoglycemic activity of flavonoids in etiolated tissues from fresh-cut Chinese water-chestnuts (Eleocharis tuberosa)

Chinese water chestnut (Eleocharis tuberosa) is prone to etiolation after fresh-cut treatment. Though etiolation caused undesired yellow color, it comes with accumulation of bioactive flavonoids. In this work, flavonoids were extracted from etiolated fresh-cut Chinese water chestnut. Their species and content were investigated. The antibacterial and hypoglycemic activities of crude extract were also characterized in vitro. The results showed that four main flavonoids naringenin, eriodictyol, quercetin and myricetin were identified from fresh-cut Chinese water chestnut and the content of total flavonoids greatly increased with the acceleration of etiolation. The antibacterial test showed that the flavonoid extract had significant inhibitory effect on Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Salmonella with the minimum inhibitory concentration of 2.5, 10, 2.5 and 5 mg/mL, respectively. The hypoglycemic kinetic analysis suggested that the flavonoid extracts exhibited competitive inhibition to both of α-glucosidase and α-amylase. The increased total flavonoids in extracts brought higher antibacterial and hypoglycemic ability during etiolation. These results revealed that the etiolated fresh-cut Chinese water chestnut can be developed and utilized for antibacterial and hypoglycemia activity.

AUXIN-BINDING-PROTEIN1 (ABP1) in phytochrome-B-controlled responses

The auxin receptor ABP1 directly regulates plasma membrane activities including the number of PIN-formed (PIN) proteins and auxin efflux transport. Red light (R) mediated by phytochromes regulates the steady-state level of ABP1 and auxin-inducible growth capacity in etiolated tissues but, until now, there has been no genetic proof that ABP1 and phytochrome regulation of elongation share a common mechanism for organ elongation. In far red (FR)-enriched light, hypocotyl lengths were larger in the abp1-5 and abp1/ABP1 mutants, but not in tir1-1, a null mutant of the TRANSPORT-INHIBITOR-RESPONSE1 auxin receptor. The polar auxin transport inhibitor naphthylphthalamic acid (NPA) decreased elongation in the low R:FR light-enriched white light (WL) condition more strongly than in the high red:FR light-enriched condition WL suggesting that auxin transport is an important condition for FR-induced elongation. The addition of NPA to hypocotyls grown in R- and FR-enriched light inhibited hypocotyl gravitropism to a greater extent in both abp1 mutants and in phyB-9 and phyA-211 than the wild-type hypocotyl, arguing for decreased phytochrome action in conjunction with auxin transport in abp1 mutants. Transcription of FR-enriched light-induced genes, including several genes regulated by auxin and shade, was reduced 3-5-fold in abp1-5 compared with Col and was very low in abp1/ABP1. In the phyB-9 mutant the expression of these reporter genes was 5–15-fold lower than in Col. In tir1-1 and the phyA-211 mutants shade-induced gene expression was greatly attenuated. Thus, ABP1 directly or indirectly participates in auxin and light signalling.

A thylakoid-localised FK506-binding protein in wheat may be linked to chloroplast biogenesis

Plant chloroplasts contain a large proportion of immunophilins, comprising the FK506-binding proteins (FKBPs) and cyclophilins (CYPs), which are members of the peptidyl-prolyl cis/trans isomerase (PPIase) family of proline-folding enzymes. Some of the chloroplastic immunophilins are known to chaperone certain photosynthetic proteins, however the functions of a majority of these proteins are unknown. This work focussed on characterisation of genes encoding the chloroplast-localised FKBP16-1 from wheat and its progenitor species, and identification of its putative promoters, as well as investigations into the effects of light regulation and plant development on its expression. The work identified several alternatively spliced FKBP16-1 transcripts, indicating expression of FKBP16-1 may be post-transcriptionally regulated. FKBP16-1 was expressed in both green and etiolated tissues, and highest levels were detected in developing tissues, indicating a role in chloroplast biogenesis. We also report a novel transcription module, designated 'chloroplast biogenesis module' (CBM) in the FKBP16-1 promoter of cereals that also appears to be involved in the regulation of additional genes involved in chloroplast biogenesis or other aspects of plant development. The results point to considerable potential for a role for FKBP16-1 in early chloroplast development, architecture of photosynthetic apparatus and plant development.

Quantitative correlation between the concentration of photoreactive phytochrome and light-induced formation of adventitious shoots in horse-radish hairy roots

The hairy roots of horse-radish (Armoracia rusticana Gaert., Mey. et Scherb.) were found to contain spectrophotometrically active phytochrome. The absorption maxima of the red-absorbing and the far red-absorbing forms of phytochrome were the same as those of phytochrome I in etiolated tissues and of purified phytochrome A. Red light dramatically decreased the concentration of phytochrome in the hairy roots. The phytochrome was concentrated at the proximal ends of hairy roots and the concentration of phytochrome at each position in hairy roots was quantitatively correlated with the frequency of the light-induced formation of adventitious shoots at the same position. The level of phytochrome reflected the duration of culture in darkness. The increase in the concentration of phytochrome with time in darkness was positively correlated with the frequency of light-induced formation of adventitious shoots. The involvement of phytochromes A and B in the induction by light of budding of adventitious shoots from horse-radish hairy roots is discussed on the basis of the newly revealed correlations and earlier results.

Changes in the Pools of Carotenoids and Protochlorophyll(ide) in Etiolated Cucumber (Cucumis Sativus) Cotyledons Treated with Norflurazon and KC 6361

Changes in the pools of carotenoids and protochlorophyll(ide) were investigated in etiolated cucumber cotyledons treated with norflurazon (NF) and an experimental herbicide KC 6361 (KC). Both the NF- and the KC-treated tissues considerably accumulated the colourless carotenes phytoene and phytofluene with a concomitant depletion of the coloured carotenoids lutein and β-carotene in darkness. However, the profiles of changes in chlorophylls (Chls) and carotenoids were different for the two herbicides. The plants were also influenced by the photosynthetic photon flux densities (PPFD's), with a more pronounced decline of Chl under high PPFD than under low PPFD. The ratios of protochlorophyll (PChl)/protochlorophyllide (PChlide) were greatly altered due to a decrease and an increase of PChl in the NF- and the KC-treated etiolated tissues, respectively, whereas the PChlide content was not significantly influenced by the inhibitors. Large increase of PChls in the KC-treated tissues seems to derive from the binding of accumulated geranylgeraniol (GG) equivalents, through carotenogenic inhibition, to PChlide. Therefore, the alterations of PChl and PChlide occurring under disturbed carotenogenesis may suggest an interaction between the biosynthetic pathways of Chls and carotenoids. In addition, the great proportion of PChl GG and PChl dihydro-GG in the KC-treated tissues implies that PChl formation is regulated at the level of hydrogenation.

Cloning of Novel Genes From Sorghum bicolor

Two novel cDNAs from Sorghum bicolor have been cloned and sequenced. The sequence analysis revealed that one of the cDNAs, pSbaNS5, is a partial clone which may encode a putative transcription regulator, and its transcript was markedly upregulated by white light. The second cDNA, pSbaNS4, may encode a putative glycoprotein of 307 amino acids. Its transcript was present in etiolated tissues and was not significantly altered by white light.

Twilight-zone and canopy shade induction of the Athb-2 homeobox gene in green plants.

We present evidence that a novel phytochrome (other than phytochromes A and B, PHYA and PHYB) operative in green plants regulates the "twilight-inducible" expression of a plant homeobox gene (Athb-2). Light regulation of the Athb-2 gene is unique in that it is not induced by red (R)-rich daylight or by the light-dark transition but is instead induced by changes in the ratio of R to far-red (FR) light. These changes, which normally occur at dawn and dusk (end-of-day FR), also occur during the daytime under the canopy (shade avoidance). By using pure light sources and phyA/phyB null mutants, we demonstrated that the induction of Athb-2 by changes in the R/FR ratio is mediated for the most part by a novel phytochrome operative in green plants. Furthermore, PHYB plays a negative role in repressing the accumulation of Athb-2 mRNA in the dark and a minor role in the FR response. The strict correlation of Athb-2 expression with FR-induced growth phenomena suggests a role for the Athb-2 gene in mediating cell elongation. This interpretation is supported by the finding that the Athb-2 gene is expressed at high levels in rapidly elongating etiolated seedlings. Furthermore, as either R or FR light inhibits cell elongation in etiolated tissues, they also down-regulate the expression of Athb-2 mRNA. Thus, these data support the notion that changes in light quality perceived by a novel phytochrome regulate plant development through the action of the Athb-2 homeobox gene.

Separation of Mono- and Divinyl Chlorophyll Species by High-Performance Liquid Chromatography Using an Octadecyl Polyvinyl Alcohol Polymer Column

Most of the chlorophylls and their related compounds from nonesterified chlorophyllide to esterified pheophytin were separated by high-performance liquid chromatography using a wide pore, C 18 polyvinyl alcohol polymer column with an elution using a binary gradient starting from a buffered mobile phase. The high selectivity of this system enabled not only the separation of common chlorophyll species but also the resolution of structurally similar compounds such as mono- and divinyl chlorophyllide and pheophorbide species which usually coelute on monomeric bonded phases. The method is successfully applied to analyses of the pigments extracted from green and etiolated tissues of higher plants and photosynthetic bacterial cells.

Cloning and developmental expression of pea ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit N-methyltransferase.

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) large subunit (LS) N-methyltransferase (protein methylase III, Rubisco LSMT, EC 2.1.1.43) catalyzes methylation of the epsilon-amino group of Lys-14 in the LS of Rubisco. With limited internal amino acid sequence information obtained from HPLC-purified peptic polypeptides from Rubisco LSMT, a full-length cDNA clone was isolated utilizing polymerase chain reaction-based technology and conventional bacteriophage library screening. The 1802 bp cDNA of Rubisco LSMT encodes a 489 amino acid polypeptide with a predicted molecular mass of ca. 55 kDa. A derived N-terminal amino acid sequence with features common to chloroplast transit peptides was identified. The deduced sequence of Rubisco LSMT did not exhibit regions of significant homology with other protein methyltransferases. Southern blot analysis of pea genomic DNA indicated a low gene copy number of Rubisco LSMT in pea. Northern analysis revealed a single mRNA species of about 1.8 kb encoding for Rubisco LSMT which was predominately located in leaf tissue. Illumination of etiolated pea seedlings showed that the accumulation of Rubisco LSMT mRNA is light-dependent. Maximum accumulation of Rubisco LSMT transcripts occurred during the initial phase of light-induced leaf development which preceded the maximum accumulation of rbcS and rbcL mRNA. Transcript levels of Rubisco LSMT in mature light-grown tissue were similar to transcript levels in etiolated tissues indicating that the light-dependent accumulation of Rubisco LSMT mRNA is transient. This is the first reported DNA and amino acid sequence for a protein methylase III enzyme.

The effect of ultraviolet‐B radiation on gene expression and pigment composition in etiolated and green pea leaf tissue: UV‐B‐induced changes are gene‐specific and dependent upon the developmental stage

ABSTRACTThe effect of ultraviolet‐B radiation (UV‐B: 280–320nm) on gene expression and pigment composition has been investigated in pea tissue at different stages of development. Pea (Pisum sativum L., cv. Feltham First) seedlings were grown for 17d and then exposed to supplementary UV‐B radiation. Chlorophyll a per unit fresh weight decreased by more than 20% compared with control levels after exposure to UV‐B radiation for 7d. In contrast, chlorophyll b content remained the same or increased slightly. Leaf protein biosynthesis, as determined by 35S‐methionine incorporation, was rapidly inhibited by UV‐B radiation, although the steady‐state levels of proteins were either unchanged or only slightly altered. RNA transcripts for the chlorophyll a/b binding protein (cab) were also rapidly reduced to low or even undetectable levels in the expanded third leaf or younger leaf bud tissue after exposure to UV‐B radiation. In contrast, cab RNA transcripts were either low or undetectable in etiolated pea tissue, but increased substantially in light and during exposure to UV‐B radiation. The cab RNA transcripts were still present at control levels in pea plants after 7d of greening under supplementary UV‐B radiation or UV‐B alone. The protein composition changed significantly over the 7d of greening, but no differences could be detected between the light treatments. The increase in chlorophyll content was slightly greater during de‐etiolation under supplementary UV‐B radiation than under control irradiance. Under UV‐B radiation alone, chlorophyll was synthesized at a greatly reduced rate. Changes in protective pigments were also determined. Anthocyanins did not change in either etiolated or green tissue exposed to UV‐B radiation. However, other flavonoids increased substantially in either tissue during exposure to light and UV‐B radiation. The RNA levels for chalcone synthase were measured in green and etiolated tissue exposed to UV‐B radiation. The chs RNA transcripts were present in low or undetectable amounts in control tissues. In green leaf tissue exposed to supplementary UV‐B, a transient increase was detected. The transcripts for chs reached a maximum level after approximately 8 h UV‐B exposure, and then declined to lower levels over subsequent days of diurnal photoperiods. However, a constant increase in chs was found after continuous exposure to UV‐B for up to 30 h. In etiolated tissue, either white‐light, supplementary UV‐B or UV‐B alone gave small increases in chs, only 8 h of UV‐B radiation alone gave any substantial increase in chs expression. Overall, these results clearly demonstrate that the response to increased levels of UV‐B radiation is dependent upon the developmental stage of the tissue and involves complex changes in gene expression.

Monovinyl and Divinyl Protochlorophyllide Pools in Etiolated Tissues of Higher Plants

The composition of chlorophyll-precursor pigments, particularly the contents of monovinyl (MV) and divinyl (DV) protochlorophyllides (Pchlides), in etiolated tissues of higher plants were determined by polyethylene-column HPLC (Y. Shioi, S. I. Beale [1987] Anal Biochem 162: 493-499), which enables the complete separation of these pigments. DV-Pchlide was ubiquitous in etiolated tissue of higher plants. From the analyses of 24 plant species belonging to 17 different families, it was shown that the concentration of DV-Pchlide was strongly dependent on the plant species and the age of the plants. The ratio of DV-Pchlide to MV-Pchlide in high DV-Pchlide plants such as cucumber and leaf mustard decreased sharply with increasing age. Levels of DV-Pchlide in Gramineae plants were considerably lower at all ages compared with those of other plants. Etiolated tissues of higher plants such as barley and corn were, therefore, good sources of MV-Pchlide. Absorption spectra of the purified MV- and DV-Pchlides in ether are presented and compared.

Genetic Regulation of Development in Sorghum bicolor

Phytochrome content of three near-isogenic genotypes of Sorghum bicolor was analyzed using immunological and spectrophotometric means. Seedlings of the photoperiodically sensitive genotypes 90M (Ma(1)Ma(1), Ma(2)Ma(2), ma(3)ma(3)) and 100M (Ma(1)Ma(1), Ma(2)Ma(2), Ma(3)Ma(3)) contain 126- and 123-kilodalton phytochromes. The 126-kilodalton protein is immunostained by antibodies Oat-16 and Pea-25. The 123-kilodalton phytochrome is immunostained by antibodies Pea-25 and Green-Oat-7. Seedlings of the photoperiodically insensitive genotype 58M (Ma(1)Ma(1), Ma(2)Ma(2), ma(3) (r)ma(3) (r)) contain only the 126-kilodalton phytochrome. In 58M seedlings, 123-kilodalton phytochrome is not detected by either Pea-25 or Green-Oat-7. Deetiolation by white light causes the 126-kilodalton phytochrome to disappear but does not greatly affect the abundance of the 123-kilodalton phytochrome. In 58M, 90M, and 100M seedlings, the 126-kilodalton phytochrome is the most abundant in etiolated tissue, whereas the 123-kilodalton phytochrome of 90M and 100M seedlings predominates in green tissue. Spectrophotometric assays show that the bulk phytochrome of etiolated tissues of all three genotypes degrades similarly upon exposure to light. At least two phytochromes are detected in sorghum: a light-labile 126-kilodalton phytochrome that predominates in etiolated tissue and a 123-kilodalton phytochrome that predominates in green tissue. Photoperiodic control of flowering in sorghum is correlated with the presence of the 123-kilodalton phytochrome.

Variation of the Polypeptide Composition of Mitochondria Isolated from Different Potato Tissues

The protein contents of mitochondria from different potato (Solanum tuberosum L.) tissues (tubers, dark-grown shoots, and green leaves) grown in a greenhouse or in vitro were compared by two-dimensional polyacrylamide gel electrophoresis. Two different methods were used: using the method that gave the highest resolution, an average number of 360 polypeptides was revealed on the mitochondrial patterns after silver staining. The mitochondrial protein patterns of etiolated tissues (tubers, dark-grown shoots) are roughly similar but distinct from those of green leaves. The four subunits of the glycine decarboxylase complex (involved in photorespiration) and a few other polypeptides are very abundant in green tissues, compared with nonphotosynthetic tissues. Conversely, some other polypeptides that are abundant in tubers and dark-grown shoots are hardly detectable in green leaf mitochondria. A rabbit antiserum was raised against a 40 kilodalton polypeptide that is among the most characteristic of these nonphotosynthetic tissue-specific polypeptides, and the N-terminal sequence of this polypeptide was determined. No effect of in vitro culture was observed on the protein composition of mitochondria isolated from differentiated tissues. However, the protein patterns of callus and cell suspension mitochondria are distinct from those of any differentiated tissues, although their basic pattern is clearly mitochondrial.

Retention of Phytochrome-Mediated Shade Avoidance Responses in Phytochrome-Deficient Mutants of Arabidopsis, Cucumber and Tomato

Summary Phytochrome-mediated shade avoidance responses have been studied in light-grown wild type and phytochrome-deficient mutant seedlings of Arabidopsis thaliana , tomato and cucumber. Light-grown seedlings of the long-hypocotyl hy -1, hy -2 and hy -6 mutants of A. thaliana retain a qualitatively normal shade avoidance response to growth under a low red: far-red (R:FR) ratio, namely an increase in elongation growth and a acceleration of flowering. This indicates that, despite the fact that etiolated tissues of these mutants are severely deficient in spectrophotometrically-detectable phytochrome, light-grown seedlings of the mutants possess some functional phytochrome. The hy -3 mutant, which is deficient in seed phytochrome, retains a similar, but less dramatic, shade avoidance response. The aurea ( au ) mutant of tomato, in which etiolated tissues are deficient in spectrophotometrically- and immunochemically-detectable phytochrome, also retains a qualitatively normal shade avoidance response to reduced R:FR ratio. This is consistent with the hypothesis that au seedlings are deficient only in a light-labile phytochrome species that normally accumulates in etiolated tissues and is involved in the deetiolation process. Light-grown seedlings of the long-hypocotyl ( lh ) mutant of cucumber, a mutation that appears to lead to a deficiency in a light-stable phytochrome species, retain a small, but significant, phytochrome-mediated increase in elongation growth in response to reduced R:FR ratio.

Changes to the Stoichiometry of Glycine Decarboxylase Subunits during Wheat (Triticum aestivum L.) and Pea (Pisum sativum L.) Leaf Development

Changes in the levels of the four subunits of the mitochondrial enzyme glycine decarboxylase (EC 2.1.2.10) have been investigated during development in the 8 day old primary leaf of wheat (Triticum aestivum L.). Proteins were extracted from wheat leaf sections between the basal meristem and 8.5 centimeters. The individual glycine decarboxylase subunits were detected by Western blotting, using subunit-specific polyclonal antibodies, and quantified by laser densitometry. P, T, and H subunits showed similar developmental patterns along the leaf. All were below the level of detection up to 1.5 centimeters from the meristem, but then increased over the leaf length examined. In contrast, the increase in the L protein (lipoamide dehydrogenase) was more gradual, and levels in the youngest regions of the leaf were maintained at approximately 14% of those at 8.5 centimeters. In a complementary study, levels of the four subunits in light-grown leaf tissues were compared to those in etiolated leaves from wheat and pea (Pisum sativum L.), using the activity of the mitochondrial marker enzyme fumarase as the basis for comparison. For both wheat and pea, levels of P, T, and H proteins in etiolated tissues were between 25 and 30% of those in lightgrown tissue. However, in etiolated tissues L protein was present at levels of 60 to 70% of that in light-grown tissues. The results indicate that discrete mechanisms may control the synthesis of L, as compared to P, T, and H proteins.

A simplified method for determining 1-aminocyclopropane-1-carboxylic acid (ACC) in plant tissues using a mass selective detector

A sensitive and specific method is described for the routine assay of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) in 100–200 mg fresh weight samples of green or etiolated tissue. The method involves high performance liquid chromatography (HPLC) and gas chromatography linked to mass spectrometry (GCMS) and uses 14C-labelled ACC as an internal standard, N-benzoyl n-propyl ACC as an easily prepared derivative for HPLC and GCMS, and N-benzoyl isobutyl ACC as an internal standard for GCMS. The procedure is faster and safer than an existing GCMS method and more specific and reliable than indirect assays widely in use. The method has been used to measure ACC in maize roots, young leaves of cucumber, and aerobic or anaerobic seedlings of rice.

Demonstration of ATP-Dependent, Ubiquitin-Conjugating Activities in Higher Plants

Ubiquitin is a highly conserved, 76-amino acid polypeptide with several important regulatory functions in both plants and animals that all arise from its covalent ligation to other cellular proteins. Here, we demonstrate that higher plants have the capacity to conjugate ubiquitin to other plant proteins in vitro. Using (125)I-labeled human ubiquitin as a substrate, conjugating activities were observed in crude etiolated tissue extracts from all species tested, including oats, rye, barley, corn, zucchini squash, pea, soybean, and sunflower. The reaction has a soluble distribution, is specific for ATP, and requires the protease inhibitor, leupeptin, to protect ubiquitin from inactivation during the assay. Conjugation is inhibited by N-ethylmaleimide and high concentrations of 2-mercaptoethanol suggesting that the mechanism of ubiquitin ligation in plants involves a similar thiolester intermediate to that found in the mammalian pathway. The conjugating activity in etiolated oat extracts is extremely labile with a half-life of about 20 minutes at 30 degrees C. Detectable but low ATP-stimulated, conjugating activities were also observed in extracts from dry seeds and green leaves of oats. In addition to this conjugating activity, crude plant extracts have the capacity to degrade ubiquitin-protein conjugates formed in vitro. These results demonstrate that higher plants contain several of the enzymic activities necessary for ubiquitin's functions and provide a method for assaying ubiquitin conjugation in vitro.

FMC 57020 Effects on Chloroplast Development in Pitted Morningglory (Ipomoea lacunosa) Cotyledons

The effect of FMC 57020 [2-(2-chlorophenyl) methyl-4,4-dimethyl-3-isoxalidinone] on chloroplast development was examined in the cotyledons of 5-day-old, etiolated pitted morningglory (Ipomoea lacunosaL. ♯ IPOLA) seedlings grown from seeds inbibed for 24 h in water or 0.5 mM FMC 57020. In etiolated tissues, protochlorophyllide content was unaffected by FMC 57020; however, the herbicide eliminated carotenoid accumulation. There was no effect of FMC 57020 on phytoene or phytofluene content, although norflurazon [4-chloro-5-(methylamino)-2-(3-trifluoromethyl) phenyl)-3(2H)-pyridazinone] increased phytoene content in these tissues. The Shibata shift was greatly retarded in FMC 57020-treated cotyledons, suggesting that phytol levels are also reduced by the herbicide. There were no ultrastructural effects on etioplasts; however, under low white light (150 μE·m-2·s-1PAR), plastids of FMC 57020-treated seedlings did not develop into chloroplasts but rapidly developed ultrastructural symptoms of photobleaching. Starch was not mobilized in herbicide-treated plastids and sugar levels were higher in these plastids than in control plastids. Etiolated hypocotyl growth was inhibited by FMC 57020, whereas norflurazon had no effect upon it. Our results suggest that FMC 57020 blocks both diterpene and tetraterpene synthesis.

Light control of extractable nitrate reductase activity in higher plants

Light regulation of extractable nitrate reductase (NR) activity of higher plants is complicated by: 1) involvement of several photoreceptors, 2) differences in the relative importance of the several photoreceptors among species and among developmental stages of the same species, 3) two types of effects – alteration of activity of existing NR and influences on de novo synthesis of NR, and 4) differing forms of NR within the same species. The interrelationships of all of these factors are not clear. It may be that each system will have to be understood separately before a general model can be developed. Immunochemical quantification of NR from systems exposed to varied light regimes may enhance our understanding of this area. Currently few general conclusions can be made; however, we think that the following statements are true or are usually true: (1) Phytochrome influences extractable NR activity by the low irradiance response and high irradiance response in etiolated tissues. (2) In de‐etiolated tissues phytochrome can influence NR activity decay at the end of a light period by the low irradiance response. (3) The phytochrome equilibrium or the absolute level of Pfr influences extractable NR activity in green tissues under white light. (4) Blue light influences extractable NR activity through phytochrome and another, unknown, blue light‐absorbing pigment. Flavins may be involved in vitro in reactivation of inactivated NR. (5) Photosynthesis does not directly influence the induction of the forms of NR that require substrate and light for induction. (6) In some tissues there appears to be a close link between nitrite‐reducing and nitrate‐reducing capabilities. (7) Much circumstantial evidence from kinetic and protein‐synthesis‐inhibitor studies and the only available immunochemical data indicate that light induces de novo synthesis of NR, resulting in increased extractable activity.