Assessing the effects of elevated ozone on physiology, growth, yield and quality of soybean in the past 40 years: A meta-analysis.

Soybean (Glycine max) production is seriously threatened by ground-level ozone (O3) pollution. The goal of our study is to summarize the impacts of O3 on physiology, growth, yield, and quality of soybean, as well as root parameters. We performed meta-analysis on the collated 48 peer-reviewed papers published between 1980 and 2019 to quantitatively summarize the response of soybean to elevated O3 concentrations ([O3]). Relative to charcoal-filtered air (CF), elevated [O3] significantly accelerated chlorophyll degradation, enhanced foliar injury, and inhibited growth of soybean, evidenced by great reductions in leaf area (-20.8%), biomass of leaves (-13.8%), shoot (-22.8%), and root (-16.9%). Shoot of soybean was more sensitive to O3 than root in case of biomass. Chronic ozone exposure of about 75.5 ppb posed pronounced decrease in seed yield of soybean (-28.3%). In addition, root environment in pot contributes to higher reduction in shoot biomass and yield of soybean. Negative linear relationships were observed between yield loss and intensity of O3 treatment, AOT40. The larger loss in seed yield was significantly associated with higher reduction in shoot biomass and other yield component. This meta-analysis demonstrates the effects of elevated O3 on soybean were pronounced, suggesting that O3 pollution is still a soaring threat to the productivity of soybean in regions with high ozone levels.

Small-Molecule Screens Reveal Novel Haustorium Inhibitors in the Root Parasitic Plant Triphysaria versicolor.

Root parasitic weeds in Orobanchaceae pose a tremendous threat to agriculture worldwide. We used an in vitro assay to screen libraries of small molecules for those capable of inhibiting or enhancing haustorium development in the parasitic plant Triphysaria versicolor. Several redox-modifying molecules and one structural analog of 2,6-dimethoxybenzoquine (DMBQ) inhibited haustorium development in the presence of the haustorium-inducing factor DMBQ, some of these without apparent growth inhibition to the root. Triphysaria seedlings were able to acclimate to some of these redox inhibitors. Transcript levels of four early-stage haustorium genes were differentially influenced by inhibitors. These novel haustorium inhibitors highlight the importance of redox cycling for haustorium development and suggest the potential of controlling parasitic weeds by interrupting early-stage redox-signaling pathways.

The influence of chosen fungicides on the activity of aminopeptidases in winter oilseed rape during pods development.

Cultivation of oilseed rape requires application of specific fungicides. Besides their protective role, they can potentially influence the expression and activity of crucial enzymes in the plant. Among the large number of enzymes expressed in plants, aminopeptidases play a key role in all crucial physiological processes during the whole life cycle (e.g. storage protein mobilization and thus supplying plant with needed amino acids, as well as plant aging, protection and defense responses). In the present paper, we evaluate for the first time, the influence of the treatment of winter oilseed rape with commercially available fungicides (Pictor 400 SC, Propulse 250 SE and Symetra 325 SC), on the activity of aminopeptidases expressed in each plant organ (flowers, leaves, stems and pods separately). Fungicides were applied once, at one of the three stages of oilseed rape development (BBCH 59-61, BBCH 63-65 and BBCH 67-69). The aminopeptidase activity was determined using six different amino acid p-nitroanilides as substrates. The results have shown, that in control plants, at the beginning of intensive pods development and seeds production, hydrophobic amino acids with bulky side chains (Phe, Leu) were preferentially hydrolysed. In control plants, the activity was ~3.5 times higher in stems and pods, compared to leaves. The treatment with all pesticides caused significant increase in aminopeptidases hydrolytic activity toward small amino acids Gly, Ala as well as proline, mostly in flowers and leaves. These amino acids are proven to be crucial in the mechanisms of delaying of plant aging, development of better resistance to stress and plant defense. It can be suggested, that studied fungicides enhance such mechanisms, by activating the expression of genes coding for aminopeptidases, which are active in hydrolysis of N-terminal amino acids such as Gly, Ala, Pro from storage peptides and proteins. Depending on fungicide, the major increase of aminopeptidase activity was observed after application at BBCH 67-69 (Pictor 400 SC and Symetra 325 SC) and BBCH 63-65 (Propulse 250 SE) stages of development. Our study revealed, that agrochemical treatment and time of application, influenced the expression and activity of aminopeptidases, even though they were not molecular targets of applied fungicides. Since aminopeptidases are widely distributed throughout all organisms and are crucial in many key physiological processes, it can be expected, that factors influencing their expression and activity in plants, can also influence these enzymes in other organisms, especially humans and other mammals.

Randomly amplified polymorphic-DNA analysis for detecting genotoxic effects of Boron on maize (Zea mays L.).

This study was carried out to investigate the genotoxic effect of boron (B) on maize using randomly amplified polymorphic DNA (RAPD) method. Experimental design was conducted under 0, 5, 10, 25, 50, 100, 125, and 150 ppm B exposures, and physiological changes have revealed a sharp decrease in root growth rates from 28% to 85%, starting from 25 ppm to 150 ppm, respectively. RAPD-polymerase chain reaction (PCR) analysis shows that DNA alterations are clearly observed from beginning to 100 ppm. B-induced inhibition in root growth had a positive correlation with DNA alterations. Total soluble protein, root and stem lengths, and B content analysis in root and leaves encourage these results as a consequence. These preliminary findings reveal that B causes chromosomal aberration and genotoxic effects on maize. Meanwhile, usage of RAPD-PCR technique is a suitable biomarker to detect genotoxic effect of B on maize and other crops for the future.

Physiological response of Cu and Cu mine tailing remediation of Paulownia fortunei (Seem) Hemsl.

The physiological responses and Cu accumulation of Paulownia fortunei (Seem) Hemsl. were studied under 15.7-157 µmol L(-1) Cu treatments in liquid culture for 14 days; the impacts of Cu concentration in the seedlings were evaluated under Cu mine tailing culture with acetic acid and EDTA treatment for 60 days. Results showed that the concentrations of Chl-a, Chl-b and Carotenoids significantly increased (p < 0.05) at 15.7-78.7 µmol L(-1)Cu treatment and significantly decreased at 157 µmol L(-1) treatment after 14 days of Cu exposure. The activities of superoxide dismutase (SOD) and catalase (CAT) significantly increased as Cu levels were enhanced and the activities of both SOD and CAT under 157 µmol L(-1) Cu stress were 2.9 and 1.9 times higher than that of control, respectively. The concentrations of proline and soluble sugars in the leaves of P. fortunei significantly increased as the Cu concentrations were elevated. Cu concentrations in roots, stems and leaves of P. fortunei increased significantly as Cu levels increased and reached 1911, 101 and 93 µg g(-1) dry weights (DW) at 157 µmol L(-1) Cu treatment, respectively. The seedlings of P. fortunei cultivated in Cu tailing experienced unsuccessful growth and loss of leaves in all treatments due to poor nutrition of the Cu tailing. The dry weight of P. fortunei increased under all the treatments of acetic acid after 60 days exposure. However, dry weight significantly decreased under both levels of EDTA. The Cu concentrations increased significantly in roots and decreased in leaves when each was treated with both concentrations of acetic acid. The Cu concentrations in the roots, stems and leaves increased significantly, and the concentrations of Cu in the stems and leaves under the treatment of 2 µmol L(-1) EDTA reached 189.5 and 763.1 µg g(-1) DW, respectively. The result indicated that SOD, CAT, proline and soluble sugars played an important role in coping with the oxidative stress of copper. Acetic acid could promote growth and EDTA at the experimental levels, which could also enhance Cu absorption and translocation into the stems and leaves of P. fortune. Furthermore, acetic acid and EDTA could be rationally utilized in Cu-contaminated soil.

Regulation of gemma formation in the copper moss Scopelophila cataractae by environmental copper concentrations.

Considerable attention has recently been focused on the use of hyperaccumulator plants for the phytoremediation of soils contaminated with heavy metals. The moss, Scopelophila cataractae (Mitt.) Broth., is a typical hyperaccumulator that is usually observed only in copper-rich environments and which accumulates high concentrations of copper in its tissues. However, many of the physiological processes and mechanisms for metal hyperaccumulation in S. cataractae remain unknown. To address this issue, we examined the mechanisms regulating gemma formation, which is considered the main strategy by which S. cataractae relocates to new copper-rich areas. From this study we found that treatment of S. cataractae with high concentrations of copper suppressed gemma formation but promoted protonemal growth. The suppressive effect was not observed by treatment with heavy metals other than copper. These results suggest the importance of copper-sensitive asexual reproduction in the unique life strategy of the copper moss, S. cataractae.

A calcium sensor and its interacting protein kinase are global regulators of abscisic acid signaling in Arabidopsis.

The phytohormone abscisic acid (ABA) triggers an oscillation in the cytosolic Ca(2+) concentration, which is then perceived by unknown Ca(2+) binding proteins to initiate a series of signaling cascades that control many physiological processes, including adaptation to environmental stress. We report here that a Ca(2+) binding protein, SCaBP5, and its interacting protein kinase, PKS3, function as global regulators of ABA responses. Arabidopsis mutants with silenced SCaBP5 or PKS3 are hypersensitive to ABA in seed germination, seedling growth, stomatal closing, and gene expression. PKS3 physically interacts with the 2C-type protein phosphatase ABI2 (ABA-insensitive 2) and to a lesser extent with the homologous ABI1 (ABA-insensitive 1) protein. Thus, SCaBP5 and PKS3 are part of a calcium-responsive negative regulatory loop controlling ABA sensitivity.

Rop GTPase-dependent dynamics of tip-localized F-actin controls tip growth in pollen tubes.

Tip-growing pollen tubes provide a useful model system to study polar growth. Although roles for tip-focused calcium gradient and tip-localized Rho-family GTPase in pollen tube growth is established, the existence and function of tip-localized F-actin have been controversial. Using the green fluorescent protein-tagged actin-binding domain of mouse talin, we found a dynamic form of tip-localized F-actin in tobacco pollen tubes, termed short actin bundles (SABs). The dynamics of SABs during polar growth in pollen tubes is regulated by Rop1At, a Rop GTPase belonging to the Rho family. When overexpressed, Rop1At transformed SAB into a network of fine filaments and induced a transverse actin band behind the tip, leading to depolarized growth. These changes were due to ectopic Rop1At localization to the apical region of the plasma membrane and were suppressed by guanine dissociation inhibitor overexpression, which removed ectopically localized Rop1At. Rop GTPase-activating protein (RopGAP1) overexpression, or Latrunculin B treatments, also recovered normal actin organization and tip growth in Rop1At-overexpressing tubes. Moreover, overexpression of RopGAP1 alone disrupted SABs and inhibited growth. Finally, SAB oscillates and appears at the tip before growth. Together, these results indicate that the dynamics of tip actin are essential for tip growth and provide the first direct evidence to link Rho GTPase to actin organization in controlling cell polarity and polar growth in plants.

Effects of zinc toxicity on sugar beet (Beta vulgaris L.) plants grown in hydroponics.

The effects of high Zn concentration were investigated in sugar beet (Beta vulgaris L.) plants grown in a controlled environment in hydroponics. High concentrations of Zn sulphate in the nutrient solution (50, 100 and 300 microm) decreased root and shoot fresh and dry mass, and increased root/shoot ratios, when compared to control conditions (1.2 microm Zn). Plants grown with excess Zn had inward-rolled leaf edges and a damaged and brownish root system, with short lateral roots. High Zn decreased N, Mg, K and Mn concentrations in all plant parts, whereas P and Ca concentrations increased, but only in shoots. Leaves of plants treated with 50 and 100 microm Zn developed symptoms of Fe deficiency, including decreases in Fe, chlorophyll and carotenoid concentrations, increases in carotenoid/chlorophyll and chlorophyll a/b ratios and de-epoxidation of violaxanthin cycle pigments. Plants grown with 300 microm Zn had decreased photosystem II efficiency and further growth decreases but did not have leaf Fe deficiency symptoms. Leaf Zn concentrations of plants grown with excess Zn were high but fairly constant (230-260 microg.g(-1) dry weight), whereas total Zn uptake per plant decreased markedly with high Zn supply. These data indicate that sugar beet could be a good model to investigate Zn homeostasis mechanisms in plants, but is not an efficient species for Zn phytoremediation.

Assessment of biomarkers of cadmium stress in lettuce.

Laboratory and field studies have provided encouraging insights into the capacity of plants to act as biomonitors of environmental quality through the use of biomarkers. However, a better understanding of the overall process of Cd-induced senescence, describing the cascade of Cd effects in plants is needed for a selection of relevant biomarkers of Cd stress. In order to approach this, 5-week old Lactuca sativa L. were exposed for 14 days to 100muM Cd(NO(3))(2) and harvested at days 0, 1, 3, 7 and 14. The parameters measured included classical endpoints (shoot and root growth) and biochemical endpoints related to photosynthesis, nutrients content, and oxidative stress. Cadmium-exposed plants displayed nutrient imbalances in leaves and roots. Photosynthetic efficiency was significantly decreased and lipid peroxidation was enhanced. Antioxidant enzymes were significantly altered during exposure-catalase was inhibited by the end of exposure and peroxidase was induced at day 1 in young leaves. These alterations culminated in a decrease in shoot growth after 14-days exposure to Cd. Biochemical alterations could be used in integrative approaches with classical endpoints in ecotoxicological tests for Cd and after further testing in real scenarios conditions, they could form the basis of a plant biomarkers battery for monitoring and predicting early effects of exposure to Cd.

[FTIR study of the endurance to Al of different soybean cultivars].

The spectra of root, stem, leaf of soybean samples with 0, 20, 40, 60 mg x L(-1) Al3+ were determined by Fourier transform infrared (FTIR) spectrometry with OMNI-sampler. Little difference was found in the spectra of leaf between two soybean cultivars, aluminum-resistant cultivar Zhechun NO. 2 and aluminum-sensitive cultivar Zhechun NO. 3, except the indices of wave number-absorbance from 928 to 1 200 cm(-1), and similar results were also observed in stem and root samples of the two soybean cultivars with 0 mg x L(-1) Al3+. However, results from the comparison of the spectra showed some distinguishable differences in the intensity and the shape of absorption peaks of their FTIR spectra from 721 to 3 366 cm(-1) of Al-stressed samples and control samples between the two soybean cultivars, and more evident differences of FTIR were exhibited in Al-stressed roots, stems and leaves with higher concentration of Al3+. The increased absorbance at 2 929 and 3 350 cm(-1) was found in root FTIR spectra with 20, 40, 60 mg x L(-10 Al3+, while roots got maximum absorbance at wave number of 1 375 cm(-1) with 20 mg x L(-1) Al3+, which decreased with higher concentration of Al3+, and the same results were showed at wave numbers of 1 410, 1 423, 1 549 and 1 645 cm(-1). Absorption peak showed maximum at wave numbers of 1 051, 2 850, 2 929 and 3 350 cm(-1) in stem FTIR spectra with 60 mg x L(-1) Al3+. There was little difference between the spectra of Al-stressed leaves and controls at wave numbers from 1 750 to 2 750 cm(-1), but visible difference in leaf spectra was exhibited at other wave number. Moreover, the results showed that the FTIR spectra of aluminum-sensitive cultivar Zhechun NO. 3 showed much more observable differences than aluminum-resistant cultivar Zhechun NO. 2 with different concentration of Al, implying that the material metabolic of aluminum-sensitive soybean was evidently affected by Al. Therefore, FTIR spectra could be used broadly for identification of the endurance of different soybean cultivars to Al.

[Effect of the complex of technogenic and recreational loads on development of trunk tissues of Scotch pine in the Krasnoyarsk forest-steppe].

The morphometric parameters of trunk tissues of Scotch pine in suburban pine forests of Krasnoyarsk are studied, which grow under the long-term impact of technogenic and recreational loads in comparison with the background. Technogenic and recreational loads decrease the intensity of work of the cambium, especially towards the xylem, its duration, the cell size, and the structure of storage tissues. In pine stands of the Krasnoyarsk forest-steppe, the effect of the complex of anthropogenic stressors is weakening and degradation of forest stands and inhibition of production process at the cell and tissue levels are taking place.

Expression of ethylene biosynthetic and receptor genes in rose floral tissues during ethylene-enhanced flower opening.

Ethylene production, as well as the expression of ethylene biosynthetic (Rh-ACS1-4 and Rh-ACO1) and receptor (Rh-ETR1-5) genes, was determined in five different floral tissues (sepals, petals, stamens, gynoecia, and receptacles) of cut rose (Rosa hybrida cv. Samantha upon treatment with ethylene or the ethylene inhibitor 1-methylcyclopropene (1-MCP). Ethylene-enhanced ethylene production occurred only in gynoecia, petals, and receptacles, with gynoecia showing the greatest enhancement in the early stage of ethylene treatment. However, 1-MCP did not suppress ethylene production in these three tissues. In sepals, ethylene production was highly decreased by ethylene treatment, and increased dramatically by 1-MCP. Ethylene production in stamens remained unchanged after ethylene or 1-MCP treatment. Induction of certain ethylene biosynthetic genes by ethylene in different floral tissues was positively correlated with the ethylene production, and this induction was also not suppressed by 1-MCP. The expression of Rh-ACS2 and Rh-ACS3 was quickly induced by ethylene in gynoecia, but neither Rh-ACS1 nor Rh-ACS4 was induced by ethylene in any of the five tissues. In addition, Rh-ACO1 was induced by ethylene in all floral tissues except sepals. The induced expression of ethylene receptor genes by ethylene was much faster in gynoecia than in petals, and the expression of Rh-ETR3 was strongly suppressed by 1-MCP in all floral tissues. These results indicate that ethylene biosynthesis in gynoecia is regulated developmentally, rather than autocatalytically. The response of rose flowers to ethylene occurs initially in gynoecia, and ethylene may regulate flower opening mainly through the Rh-ETR3 gene in gynoecia.

The Ayurvedic medicine Clitoria ternatea--from traditional use to scientific assessment.

Clitoria ternatea L. (CT) (Family: Fabaceae) commonly known as 'Butterfly pea', a traditional Ayurvedic medicine, has been used for centuries as a memory enhancer, nootropic, antistress, anxiolytic, antidepressant, anticonvulsant, tranquilizing and sedative agent. A wide range of secondary metabolites including triterpenoids, flavonol glycosides, anthocyanins and steroids has been isolated from Clitoria ternatea Linn. Its extracts possess a wide range of pharmacological activities including antimicrobial, antipyretic, anti-inflammatory, analgesic, diuretic, local anesthetic, antidiabetic, insecticidal, blood platelet aggregation-inhibiting and for use as a vascular smooth muscle relaxing properties. This plant has a long use in traditional Ayurvedic medicine for several diseases and the scientific studies has reconfirmed those with modern relevance. This review is an effort to explore the chemical constituents, pharmacological and toxicity studies of CT, which have long been in clinical use in Ayurvedic system of medicine along with a critical appraisal of its future ethnopharmacological potential in view of many recent findings of importance on this well known plant species.

Grain and starch granule morphology in superior and inferior kernels of maize in response to nitrogen.

Maize (Zea mays L.) contributes approximately 55% of China's grain production. The effects of nitrogen (N) on maize grain morphology and starch granules remain elusive. In this study, a field experiment in clay loam soil was conducted using three maize hybrids (Suyu 30, Suyu 20, and Suyu 29) and four N levels (0, 360, 450, and 540 kg ha-1) in 2010 and 2012. The results indicated that increased grain length and width, starch granule number, surface area, and volume, was associated with the application of 450 kg ha-1 of N. Differences between superior (ear base) and inferior (apical) grains decreased under highest yield treatments. The effects of N levels on inferior grains was more than that on superior grains. The starch granules of superior grains showed more polygonal, and bigger shape than inferior grains. The results revealed that N levels affected size and morphology of starch granules and grains. The application of 450 kg N ha-1 resulted in larger-sized starch granules and less difference between superior and inferior grains.

Evolutionary conservation of plant gibberellin signalling pathway components.

BACKGROUND: Gibberellins (GA) are plant hormones that can regulate germination, elongation growth, and sex determination. They ubiquitously occur in seed plants. The discovery of gibberellin receptors, together with advances in understanding the function of key components of GA signalling in Arabidopsis and rice, reveal a fairly short GA signal transduction route. The pathway essentially consists of GID1 gibberellin receptors that interact with F-box proteins, which in turn regulate degradation of downstream DELLA proteins, suppressors of GA-controlled responses. RESULTS: Arabidopsis sequences of the gibberellin signalling compounds were used to screen databases from a variety of plants, including protists, for homologues, providing indications for the degree of conservation of the pathway. The pathway as such appears completely absent in protists, the moss Physcomitrella patens shares only a limited homology with the Arabidopsis proteins, thus lacking essential characteristics of the classical GA signalling pathway, while the lycophyte Selaginella moellendorffii contains a possible ortholog for each component. The occurrence of classical GA responses can as yet not be linked with the presence of homologues of the signalling pathway. Alignments and display in neighbour joining trees of the GA signalling components confirm the close relationship of gymnosperms, monocotyledonous and dicotyledonous plants, as suggested from previous studies. CONCLUSION: Homologues of the GA-signalling pathway were mainly found in vascular plants. The GA signalling system may have its evolutionary molecular onset in Physcomitrella patens, where GAs at higher concentrations affect gravitropism and elongation growth.

Actin polymerization is essential for pollen tube growth.

Actin microfilaments, which are prominent in pollen tubes, have been implicated in the growth process; however, their mechanism of action is not well understood. In the present work we have used profilin and DNAse I injections, as well as latrunculin B and cytochalasin D treatments, under quantitatively controlled conditions, to perturb actin microfilament structure and assembly in an attempt to answer this question. We found that a approximately 50% increase in the total profilin pool was necessary to half-maximally inhibit pollen tube growth, whereas a approximately 100% increase was necessary for half-maximal inhibition of cytoplasmic streaming. DNAse I showed a similar inhibitory activity but with a threefold more pronounced effect on growth than streaming. Latrunculin B, at only 1--4 nM in the growth medium, has a similar proportion of inhibition of growth over streaming to that of profilin. The fact that tip growth is more sensitive than streaming to the inhibitory substances and that there is no correlation between streaming and growth rates suggests that tip growth requires actin assembly in a process independent of cytoplasmic streaming.

Effects of tungsten on uptake, transport and subcellular distribution of molybdenum in oilseed rape at two different molybdenum levels.

Due to the similarities of molybdenum (Mo) with tungsten (W) in the physical structure and chemical properties, studies involving the two elements have mainly examined their competitive relationships. The objectives of this study were to assess the effects of equimolar W on Mo accumulation, transport and subcellular distribution in oilseed rape at two Mo levels with four treatments: Mo1 (1µmol/L Mo, Low Mo), Mo1+W1 (1µmol/L Mo+1µmol/LW, Low Mo with Low W), Mo200 (200µmol/L Mo, High Mo) and Mo200+W200 (200µmol/L Mo+200µmol/L Mo, High Mo with high W). The fresh weight and root growth were inhibited by equimolar W at both low and high Mo levels. The Mo concentration and accumulation in root was increased by equimolar W at the low Mo level, but that in the root and shoot was decreased at the high Mo level. Additionally, equimolar W increased the Mo concentrations of xylem and phloem sap at low Mo level, but decreased that of xylem and increased that of phloem sap at the high Mo level. Furthermore, equimolar W decreased the expression of BnMOT1 in roots and leaves at the low Mo level, and only decreased its expression in leaves at the high Mo level. The expression of BnMOT2 was also decreased in root for equimolar W compared with the low Mo level, but increased compared with high Mo level. Moreover, equimolar W increased the proportion of Mo in cell wall fraction in root and that of soluble fraction in leaves when compared with the low Mo level. The results suggest that cell wall and soluble fractions might be responsible for the adaptation of oilseed rape to W stress.

beta-Carotene accumulation induced by the cauliflower Or gene is not due to an increased capacity of biosynthesis.

The cauliflower (Brassica oleracea L. var. botrytis) Or gene is a rare carotenoid gene mutation that confers a high level of beta-carotene accumulation in various tissues of the plant, turning them orange. To investigate the biochemical basis of Or-induced carotenogenesis, we examined the carotenoid biosynthesis by evaluating phytoene accumulation in the presence of norflurazon, an effective inhibitor of phytoene desaturase. Calli were generated from young seedlings of wild type and Or mutant plants. While the calli derived from wild type seedlings showed a pale green color, the calli derived from Or seedlings exhibited intense orange color, showing the Or mutant phenotype. Concomitantly, the Or calli accumulated significantly more carotenoids than the wild type controls. Upon treatment with norflurazon, both the wild type and Or calli synthesized significant amounts of phytoene. The phytoene accumulated at comparable levels and no major differences in carotenogenic gene expression were observed between the wild type and Or calli. These results suggest that Or-induced beta-carotene accumulation does not result from an increased capacity of carotenoid biosynthesis.

Expression of ZmMET1, a gene encoding a DNA methyltransferase from maize, is associated not only with DNA replication in actively proliferating cells, but also with altered DNA methylation status in cold-stressed quiescent cells.

A cDNA fragment encoding part of a DNA methyltransferase was isolated from maize. The putative amino acid sequence identically matched that deduced from a genomic sequence in the database (accession no. AF063403), and the corresponding gene was designated as ZmMET1. Bacterially expressed ZmMET1 actively methylated DNA in vitro. Transcripts of ZmMET1 could be shown to exclusively accumulate in actively proliferating cells of the meristems of mesocotyls and root apices, suggesting ZmMET1 expression to be associated with DNA replication. This was confirmed by simultaneous decrease of transcripts of ZmMET1 and histone H3, a marker for DNA replication, in seedlings exposed to wounding, desiccation and salinity, all of which suppress cell division. Cold stress also depressed both transcripts in root tissues. In contrast, however, accumulation of ZmMET1 transcripts in shoot mesocotyls was not affected by cold stress, whereas those for H3 sharply decreased. Such a differential accumulation of ZmMET1 transcripts was consistent with ZmMET1 protein levels as revealed by western blotting. Expression of ZmMET1 is thus coexistent, but not completely dependent on DNA replication. Southern hybridization analysis with a methylation-sensitive restriction enzyme revealed that cold treatment induced demethylation of DNA in the Ac/Ds transposon region, but not in other genes, and that such demethylation primarily occurred in roots. These results suggested that the methylation level was decreased selectively by cold treatment, and that ZmMET1 may, at least partly, prevent such demethylation.