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PubMed2003

The release of genetically modified crops into the environment. Part I. Overview of current status and regulations.

In the past 6 years, the global area of commercially grown, genetically modified (GM) crops has increased more than 30-fold to over 52 million hectares. The number of countries involved has more than doubled. Especially in developing countries, the GM crop area is anticipated to increase rapidly in the coming years. Despite this high adoption rate and future promises, there is a multitude of concerns about the impact of GM crops on the environment. Regulatory approaches in Europe and North America are essentially different. In the EU, it is based on the process of making GM crops; in the US, on the characteristics of the GM product. Many other countries are in the process of establishing regulation based on either system or a mixture. Despite these differences, the information required for risk assessment tends to be similar. Each risk assessment considers the possibility, probability and consequence of harm on a case-by-case basis. For GM crops, the impact of non-use should be added to this evaluation. It is important that the regulation of risk should not turn into the risk of regulation. The best and most appropriate baseline for comparison when performing risk assessment on GM crops is the impact of plants developed by traditional breeding. The latter is an integral and accepted part of agriculture.

Biotechnology↗

PubMedDate not supplied

[Interaction of Pantoea agglomerans with the agent of basal bacteriosis of wheat].

Interactions between the agent of basal bacteriosis Pseudomonas syringae pv. atrofaciens and epiphyte Pantoea agglomerans have been studied in the laboratory and field conditions. It is shown the P. agglomerans affects the infectious process caused by P. s. pv. atrofaciens that leads to intensification of symptoms of the disease development. The epiphyte cells accumulation predominates under mutual artificial infection in the infected tissue. The higher competitive ability of the epiphyte P. agglomerans as compared with phytopathogen has been found out. The epiphyte propagates more quickly and inhibits growth and development of phytopathogen under joint cultivation of the studied strains of bacteria. Competition is shown between the studied strains of bacteria on the wheat leaf surface which depended on the phase of plant development.

Pantoea↗

PubMed2006

SECRET AGENT and SPINDLY have overlapping roles in the development of Arabidopsis thaliana L. Heyn.

O-GlcNAc transferase (OGT) catalyses transfer of GlcNAc (N-acetylglucosamine) to serine or threonine of proteins. The Arabidopsis OGTs, SECRET AGENT (SEC) and SPINDLY (SPY) have overlapping functions during gametogenesis and embryogenesis. SPY functions in a number of processes including circadian, light, and gibberellin (GA) responses. The role of SEC in plant development and GA signalling was investigated by determining the phenotypes of sec-1 and sec-2 plants and the expression pattern of SEC. Similar to SPY, SEC transcripts were ubiquitous. Although there is no evidence of transcript-level regulation by other factors, SEC mRNA levels are elevated in spy plants and SPY mRNA levels are elevated in sec plants. sec-1 and sec-2 plants exhibited few of the defects observed in spy plants and had wild-type GA responses. Compared with wild type, sec plants produced leaves at a reduced rate. Haplo-insufficiency at SEC in a spy ga1 double mutant background suppressed spy during germination and enhanced the production of ovaries with four carpels by spy. By contrast, SPY haplo-insufficiency in a sec ga1 double mutant background caused a novel phenotype, production of a proliferation of pin-like structures instead of a floral shoot. These results are consistent with SEC function overlapping with SPY for leaf production and reproductive development.

Arabidopsis↗

PubMed1995

Nod factors produced by Rhizobium leguminosarum biovar viciae induce ethylene-related changes in root cortical cells of Vicia sativa ssp. nigra.

Vicia sativa ssp. nigra plants develop the "Thick short root" (Tsr) phenotype when both (i) the roots are inoculated with the root nodule inducing bacterium Rhizobium leguminosarum biovar viciae, and (ii) the plants, including the roots, are grown in the light. Tsr roots have a reduced length, are locally twice as thick as normal roots and have an increased number of root hairs. Development of the Tsr phenotype is correlated with the presence of nod (nodulation) genes in the rhizobia. Nod factors (lipochitin oligosaccharides), products of these nod genes, can induce the Tsr phenotype in the absence of rhizobia. The Tsr phenotype can be mimicked by addition of the ethylene-releasing compound ethephon. Using several microscopical techniques, we compared roots showing the Tsr phenotype (Tsr roots) with normal roots and roots grown in the presence of the ethylene inhibitor aminoethoxyvinylglycine (AVG). The thickening of Tsr roots appeared to be caused by a swelling of the cortical cells, which corresponded with (i) a reorientation of the interphase cortical microtubules from a transverse to a longitudinal direction, (ii) general cell wall modifications, (iii) frequent absence of middle lamellae, and (iv) local maceration. The same changes could be induced by ethephon and were inhibited by AVG. This strongly suggests that the Tsr phenotype is caused by excessive ethylene production. The ethylene-related changes mentioned above are also seen during infection thread formation, but only very locally. Apparently, Vicia roots when grown in the light overrespond to Nod factors leading to overproduction of ethylene and to a non-local "ripening" process. These phenomena inhibit nodulation of the main root by preventing formation of pre-infected threads and by reducing formation of root nodule primordia. Local controlled production of ethylene, as induced by Nod factors, may, however, be an essential element of the nodulation process.

Cell Wall↗

PubMed1971

Calcium Deficiency of Dark-grown Seedlings of Phaseolus vulgaris L.

Hypocotyl collapse in dark-grown seedlings of Phaseolus vulgaris cv. Pinto was due to calcium deficiency. There was no evidence of an associated pathogen. The number of seedlings with hypocotyl collapse decreased and the mean hypocotyl length increased when increasing levels of calcium (0-100 micrograms per gram) were supplied in an external nutrient solution to seedlings grown under sterile conditions.When seedlings were supplied with a complete nutrient solution, containing calcium at 100 micrograms per gram, but minus potassium, magnesium, sulfur, nitrogen, or phosphorus, occasional plants developed hypocotyl collapse symptoms; however, the lengths of hypocotyls varied little from those of controls grown in complete nutrient. When the calcium level in the deficient nutrient solutions was raised to 200 micrograms per gram, the number of plants with hypocotyl collapse was reduced markedly.With complete nutrient solution minus calcium, seedlings developed symptoms of calcium deficiency irrespective of seed size, i.e., irrespective of whether or not the seed contained a total calcium content that was low or relatively high.An increase in hypocotyl length in response to an external supply of calcium was obtained with five cultivars of Phaseolus vulgaris L. and with one of Soja max Piper. A similar response to calcium was obtained for epicotyl growth of a cultivar of Vicia faba L., but not for a cultivar of Pisum sativum L.

Journal Article↗

PubMed1991

Cytokinin content and tissue distribution in plants transformed by a reconstructed isopentenyl transferase gene.

The cytokinin gene, isopentenyl transferase (ipt), was placed under the control of a heat-inducible promoter from the Drosophila melanogaster hsp70 gene and introduced into Nicotiana plumbaginifolia by cocultivation with Agrobacterium tumefaciens. Transformants were analyzed for organ-specific expression, cytokinin levels and effects on plant development before and after the heat induction. The ipt gene transcripts were detected in leaves and stems but not roots of transgenic plants following a 2 hour, 45 degrees C treatment. Maximum mRNA levels observed occurred 2 hours after heat treatment and 46 hours later were detected only in leaves. Zeatin and zeatinriboside concentrations 2 hours after heat shock ranged from over 900 to 2000 pmol/g, representing a greater than 140- to 200-fold increase over uninduced levels. After 46 hours, approximately 50% of the cytokinins are still present in the leaves as opposed to much reduced levels in the stems. Transgenic plants were greener, shorter, had an underdeveloped root system, reduced leaf width, and increased growth of axillary buds. After a single heat treatment, plants exhibited a darker green pigment and continued growth of lateral buds. Transient accumulations of endogenous cytokinins following thermal induction did not appear to alter the plant's preprogrammed pattern of differentiation.

Adenosine↗

PubMed2005

Yield enhancement genes: seeds for growth.

Yield is a multifactorial trait, integrating various developmental and physiological processes. Despite this complexity, evidence is mounting that yield can be increased by the genetic modification of single genes. Positive results have been obtained by targeting different yield constituents, indicating that there is ample room for further yield improvement by genetic means. Successful targets include photosynthesis, starch biosynthesis, plant architecture and transcriptional networks controlling plant development. Most of the current data have been obtained in a (semi-)controlled environment and relate to yield calculated on a per plant basis. Demonstrating the ability to transfer these effects to field-grown plants and with reference to yield on a per area unit basis will be a crucial step in establishing the agronomic importance of these findings.

Arabidopsis↗

PubMed2001

Lipoxygenase H1 gene silencing reveals a specific role in supplying fatty acid hydroperoxides for aliphatic aldehyde production.

Lipoxygenases catalyze the formation of fatty acid hydroperoxide precursors of an array of compounds involved in the regulation of plant development and responses to stress. To elucidate the function of the potato 13-lipoxygenase H1 (LOX H1), we have generated transgenic potato plants with reduced expression of the LOX H1 gene as a consequence of co-suppression-mediated gene silencing. Three independent LOX H1-silenced transgenic lines were obtained, having less than 1% of the LOX H1 protein present in wild-type plants. This depletion of LOX H1 has no effect on the basal or wound-induced levels of jasmonates derived from 13-hydroperoxylinolenic acid. However, LOX H1 depletion results in a marked reduction in the production of volatile aliphatic C6 aldehydes. These compounds are involved in plant defense responses, acting as either signaling molecules for wound-induced gene expression or as antimicrobial substances. LOX H1 protein was localized to the chloroplast and the protein, expressed in Escherichia coli, showed activity toward unesterified linoleic and linolenic acids and plastidic phosphatidylglycerol. The results demonstrate that LOX H1 is a specific isoform involved in the generation of volatile defense and signaling compounds through the HPL branch of the octadecanoid pathway.

Aldehydes↗

PubMed2004

Identification of Thlaspi caerulescens genes that may be involved in heavy metal hyperaccumulation and tolerance. Characterization of a novel heavy metal transporting ATPase.

Thlaspi caerulescens is a heavy metal hyperaccumulator plant species that is able to accumulate extremely high levels of zinc (Zn) and cadmium (Cd) in its shoots (30,000 microg g(-1) Zn and 10,000 microg g(-1) Cd), and has been the subject of intense research as a model plant to gain a better understanding of the mechanisms of heavy metal hyperaccumulation and tolerance and as a source of genes for developing plant species better suited for the phytoremediation of metal-contaminated soils. In this study, we report on the results of a yeast (Saccharomyces cerevisae) complementation screen aimed at identifying candidate heavy metal tolerance genes in T. caerulescens. A number of Thlaspi genes that conferred Cd tolerance to yeast were identified, including possible metal-binding ligands from the metallothionein gene family, and a P-type ATPase that is a member of the P1B subfamily of purported heavy metal-translocating ATPases. A detailed characterization of the Thlaspi heavy metal ATPase, TcHMA4, demonstrated that it mediates yeast metal tolerance via active efflux of a number of different heavy metals (Cd, Zn, lead [Pb], and copper [Cu]) out of the cell. However, in T. caerulescens, based on differences in tissue-specific and metal-responsive expression of this transporter compared with its homolog in Arabidopsis (Arabidopsis thaliana), we suggest that it may not be involved in metal tolerance. Instead, we hypothesize that it may play a role in xylem loading of metals and thus could be a key player in the hyperaccumulation phenotype expressed in T. caerulescens. Additionally, evidence is presented showing that the C terminus of the TcHMA4 protein, which contains numerous possible heavy metal-binding His and Cys repeats residues, participates in heavy metal binding. When partial peptides from this C-terminal domain were expressed in yeast, they conferred an extremely high level of Cd tolerance and Cd hyperaccumulation. The possibilities for enhancing the metal tolerance and phytoremediation potential of higher plants via expression of these metal-binding peptides are also discussed.

Adenosine Triphosphatases↗

PubMed2002

Nucleocytoplasmic partitioning of the plant photoreceptors phytochrome A, B, C, D, and E is regulated differentially by light and exhibits a diurnal rhythm.

The phytochrome family of plant photoreceptors has a central role in the adaptation of plant development to changes in ambient light conditions. The individual phytochrome species regulate different or partly overlapping physiological responses. We generated transgenic Arabidopsis plants expressing phytochrome A to E:green fluorescent protein (GFP) fusion proteins to assess the biological role of intracellular compartmentation of these photoreceptors in light-regulated signaling. We show that all phytochrome:GFP fusion proteins were imported into the nuclei. Translocation of these photoreceptors into the nuclei was regulated differentially by light. Light-induced accumulation of phytochrome species in the nuclei resulted in the formation of speckles. The appearance of these nuclear structures exhibited distinctly different kinetics, wavelengths, and fluence dependence and was regulated by a diurnal rhythm. Furthermore, we demonstrate that the import of mutant phytochrome B:GFP and phytochrome A:GFP fusion proteins, shown to be defective in signaling in vivo, is regulated by light but is not accompanied by the formation of speckles. These results suggest that (1) the differential regulation of the translocation of phytochrome A to E into nuclei plays a role in the specification of functions, and (2) the appearance of speckles is a functional feature of phytochrome-regulated signaling.

Active Transport, Cell Nucleus↗

PubMed2006

Melatonin in Glycyrrhiza uralensis: response of plant roots to spectral quality of light and UV-B radiation.

Melatonin (N-acetyl-5-methoxytryptamine) is known to be synthesized and secreted by the pineal gland in vertebrates. Evidence for the occurrence of melatonin in the roots of Glycyrrhiza uralensis plants and the response of this plant to the spectral quality of light including red, blue and white light (control) and UV-B radiation (280-315 nm) for the synthesis of melatonin were investigated. Melatonin was extracted and quantified in seed, root, leaf and stem tissues and results revealed that the root tissues contained the highest concentration of melatonin; melatonin concentrations also increased with plant development. After 3 months of growth under red, blue and white fluorescent lamps, the melatonin concentrations were highest in red light exposed plants and varied depending on the wavelength of light spectrum in the following order red >> blue > or = white light. Interestingly, in a more mature plant (6 months) melatonin concentration was increased considerably; the increments in concentration were X4, X5 and X3 in 6-month-old red, blue and white light exposed (control) plants, respectively. The difference in melatonin concentrations between blue and white light exposed (control) plants was not significant. The concentration of melatonin quantified in the root tissues was highest in the plants exposed to high intensity UV-B radiation for 3 days followed by low intensity UV-B radiation for 15 days. The reduction of melatonin under longer periods of UV-B exposure indicates that melatonin synthesis may be related to the integrated (intensity and duration) value of UV-B irradiation. Melatonin in G. uralensis plant is presumably for protection against oxidative damage caused as a response to UV irradiation.

Glycyrrhiza uralensis↗

PubMed2003

The plant cell cycle.

Cell division in plants is controlled by the activity of cyclin-dependent kinase (CDK) complexes. Although this basic mechanism is conserved with all other eukaryotes, plants show novel features of cell-cycle control in the molecules involved and their regulation, including novel CDKs showing strong transcriptional regulation in mitosis. Plant development is characterized by indeterminate growth and reiteration of organogenesis and is therefore intimately associated with cell division. This may explain why plants have a large number of cell-cycle regulators that appear to have overlapping and distinct functions. Here we review the recent considerable progress in understanding how core cell-cycle regulators are involved in integrating and coordinating cell division at the molecular level.

Cell Cycle↗

PubMed2005

Two-component phosphorelay signal transduction systems in plants: from hormone responses to circadian rhythms.

One of the most fundamental and widespread mechanisms of signal perception/transduction in prokaryotes is generally referred to as the "two-component regulatory system (TCS)." The concept of TCS has already been introduced a decade ago from extensive studies on the model prokaryotic bacterium Escherichia coli. Results of recent studies on the model higher plant Arabidopsis thaliana have led us to learn a new scenario as to the versatility of TCS in eukaryotic species. In the plant, on the one hand, TCS are crucially involved in the signal transduction mechanism underlying the regulation of sophisticated plant development in response to hormones (e.g., cytokinin and ethylene). On the other hand, a unique TCS variant is essentially integrated into the plant clock function that generates circadian rhythms, and also tells us the time and season on this regularly spinning and revolving world. In this review, recent progress with regard to studies on TCS in higher plants will be discussed, focusing particularly on the model higher plant Arabidopsis thaliana.

Arabidopsis↗

PubMed2005

The Arabidopsis PEX12 gene is required for peroxisome biogenesis and is essential for development.

Peroxisomes perform diverse and vital functions in eukaryotes, and abnormalities in peroxisomal function lead to severe developmental disorders in humans. Peroxisomes are also involved in a wide array of physiological and metabolic functions unique to plants, yet many aspects of this important organelle are poorly understood. In yeast and mammals, various steps in peroxisome biogenesis require the function of peroxin (PEX) proteins, among which PEX12 is a RING finger peroxisomal membrane protein involved in the import of matrix proteins. To investigate the role of PEX12 in plants, we identified a T-DNA knockout allele of PEX12 and generated partial loss-of-function pex12 mutants using RNA interference. We show that pex12 null mutants are developmentally arrested during early embryogenesis, and that the embryo-lethal phenotype can be rescued by overexpression of the PEX12-cyan fluorescent protein fusion protein, which targets to the peroxisome. Using virus-induced gene-silencing techniques, we demonstrate that peroxisomal number and fluorescence of the yellow fluorescent protein-peroxisome targeting signal type 1 protein are greatly reduced when PEX12 is silenced. RNA interference plants with partial reduction of the PEX12 transcript exhibit impaired peroxisome biogenesis and function, inhibition of plant growth, and reduced fertility. Our work provides evidence that the Arabidopsis (Arabidopsis thaliana) PEX12 protein is required for peroxisome biogenesis and plays an essential role throughout plant development.

Amino Acid Sequence↗

PubMed2005

Cloning and functional characterization of NtCPK4, a new tobacco calcium-dependent protein kinase.

A cDNA clone, encoding calcium (Ca2+)-dependent protein kinase (CDPK or CPK), was isolated from tobacco (Nicotiana tabacum). The full-length cDNA of 2360 bp contains an open reading frame for NtCPK4 consisting of 572 amino acid residues. Sequence alignment indicated that NtCPK4 shared high similarities with other CPKs and some CPK-related protein kinases (CRKs). Biochemical analyses showed that NtCPK4 phosphorylated itself and calf thymus histones fraction III-S (histone III-S) in a calcium-dependent manner, and the K0.5 of calcium activation was 0.29 microM or 0.25 microM with histone III-S or syntide-2 as substrates, respectively. The Vmax and Km were 588 nmol min-1 mg-1 and 176 microg ml-1, respectively, when histone III-S was used as substrate, while they were 2415 nmol min-1 mg-1 and 58 microM, respectively, with syntide-2 as substrate. In addition, the phosphorylation of NtCPK4 occurred on threonine residue, as shown by capillary electrophoresis analyses. All of these data demonstrated that NtCPK4 was a serine/threonine protein kinase. NtCPK4 as a low copy gene was expressed in all tested organs including the root, leaf, stem, and flower of tobacco, while its expression was temporally and spatially modulated in both productive and vegetative tissues during tobacco growth and development. NtCPK4 expression was also increased in response to the treatment of gibberellin or NaCl. Our study suggested that NtCPK4 might play vital roles in plant development and responses to environmental stimuli.

Amino Acid Sequence↗

PubMed2002

OsPNH1 regulates leaf development and maintenance of the shoot apical meristem in rice.

The Arabidopsis PINHEAD/ZWILLE (PNH/ZLL) gene is thought to play an important role in the formation of the shoot apical meristem (SAM) and in leaf adaxial cell specification. To investigate the molecular mechanisms of rice development, we have isolated a rice homologue of PNH/ZLL, called OsPNH1. Around the SAM, OsPNH1 was strongly expressed in developing leaf primordia, specifically in the presumptive vascular domains, developing vascular tissues, a few cell-layers of the adaxial region, and future bundle sheath extension cells. In the SAM, only weak expression was observed in the central region, whereas strong expression was detected in the mid-vein region of leaf founder cells in the peripheral SAM domain. We produced transgenic rice plants containing the antisense OsPNH1 strand. The antisense OsPNH1 plants developed malformed leaves with an altered vascular arrangement and abnormal internal structure. These plants also formed an aberrant SAM with reduced KNOX gene expression. We examined the subcellular localization of the OsPNH1-GFP fusion protein and found that it was localized in the cytoplasm. On the basis of these observations, we propose that OsPNH1 functions not only in SAM maintenance as previously thought, but also in leaf formation through vascular development.

Amino Acid Sequence↗

PubMed2006

Analysis of the Arabidopsis rsr4-1/pdx1-3 mutant reveals the critical function of the PDX1 protein family in metabolism, development, and vitamin B6 biosynthesis.

Vitamin B6 represents a highly important group of compounds ubiquitous in all living organisms. It has been demonstrated to alleviate oxidative stress and in its phosphorylated form participates as a cofactor in >100 biochemical reactions. By means of a genetic approach, we have identified a novel mutant, rsr4-1 (for reduced sugar response), with aberrant root and leaf growth that requires supplementation of vitamin B6 for normal development. Cloning of the mutated gene revealed that rsr4-1 carries a point mutation in a member of the PDX1/SOR1/SNZ (for Pyridoxine biosynthesis protein 1/Singlet oxygen resistant 1/Snooze) family that leads to reduced vitamin B6 content. Consequently, metabolism is broadly altered, mainly affecting amino acid, raffinose, and shikimate contents and trichloroacetic acid cycle constituents. Yeast two-hybrid and pull-down analyses showed that Arabidopsis thaliana PDX1 proteins can form oligomers. Interestingly, the mutant form of PDX1 has severely reduced capability to oligomerize, potentially suggesting that oligomerization is important for function. In summary, our results demonstrate the critical function of the PDX1 protein family for metabolism, whole-plant development, and vitamin B6 biosynthesis in higher plants.

Amino Acid Sequence↗

PubMed2002

Overexpression of Pto induces a salicylate-independent cell death but inhibits necrotic lesions caused by salicylate-deficiency in tomato plants.

Tomato plants overexpressing the disease resistance gene Pto (35S::Pto) exhibit spontaneous cell death, accumulation of salicylic acid (SA), elevated expression of pathogenesis-related genes, and enhanced resistance to a broad range of pathogens. Because salicylate plays an important role in the cell death and defense activation in many lesion mimic mutants, we investigated the interaction of SA-mediated processes and the 35S::Pto-mediated defense pathway by introducing the nahG transgene that encodes salicylate hydroxylase. Here, we show that SA is not required for the 35S::Pto-activated microscopic cell death and plays a minor role in defense gene activation and general disease resistance in 35S::Pto plants. In contrast, temperature greatly affects the spontaneous cell death and general resistance in 35S::Pto plants, and high temperature inhibits the cell death. The NahG tomato plants develop spontaneous, unconstrained necrotic lesions on leaves. These lesions also are initiated by the inoculation of a virulent strain of Pseudomonas syringae pv. tomato. However, the NahG-dependent necrotic lesions are inhibited in the NahG/35S::Pto plants. This inhibition is most pronounced under conditions favoring the 35S::Pto-mediated spontaneous cell death development. These results indicate that the signaling pathways activated by Pto overexpression suppress the cellular damage that is caused by SA depletion. We also found that ethylene is dispensable for the 35S::Pto-mediated general defense.

Cell Death↗