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Characterisation of a new allele of pale cress and its role in greening in Arabidopsis thaliana.

A recessive mutant with white leaves was identified in a screen of a population of T-DNA-tagged Arabidopsis thaliana plants. The mutation is lethal, but plants develop almost to maturity under sterile conditions. The white areas in leaves are devoid of developed chloroplasts, but the plants frequently develop green sectors which contain green chloroplasts. Molecular characterisation of the affected gene revealed that the mutant is allelic to pale cress (pac), a recently described mutation, and was therefore named pac-2. Sequencing of cDNAs and the genomic region revealed several noteworthy features of this genetic locus. In pac-2 the T-DNA had inserted in the region of the promoter and abolished transcription of the PAC gene completely. Cytokinin induced greening in mature, white homozygous pac-2 plants, and therefore is likely to be responsible for the greening observed in callus and shoots induced on roots from such plants. However, the PAC transcript was found to be absent in both white leaves and green callus. Thus, since cytokinin induced greening in the absence of PAC RNA this plant hormone appears to be able to bypass PAC function.

Alleles↗

Cadmium resistance in transgenic tobacco plants expressing the Nicotiana glutinosa L. metallothionein-like gene.

To understand the function of metallothioneins (MTs) in plants, we introduced the Nicotiana glutinosa MT gene into tobacco (N. tabacum) plants via an Agrobacterium mediated transformation. Full-length MT cDNA was fused between the cauliflower mosaic virus 35S (CaMV 35S) promoter and the nopaline synthase (nos) terminator of the pMBP1 binary vector in sense orientation. Tobacco leaf discs which were cocultivated with Agrobacterium carrying the chimeric MT gene, formed kanamycin-resistant shoots on medium containing kanamycin. The kanamycin-resistant shoots were subsequently rooted on medium containing 200 microM CdSO4. Approximately 30% of individual transgenic plants developed normally. Nontransgenic plants promptly underwent leaf chlorosis, and their growth and development were inhibited on MS medium containing 50 microM CdSO4. Genomic Southern blot analysis showed that the MT gene was stably integrated into the nuclear genome of transgenic tobacco plants. The expression level of MT transcripts was analyzed by RNA gel blot analysis. Self-pollinated seeds obtained from transgenic tobacco plants showing cadmium tolerance were germinated on a medium containing 100 microM CdSO4. PCR analysis from sensitive and stably resistant T2 seedlings for cadmium sulfate confirmed a high correlation between the phenotypic expression of the MT gene and the transgenic genotype, indicating that the MT gene is inherited in the next generation.

Cadmium↗

Role of nicotianamine in the intracellular delivery of metals and plant reproductive development.

Nicotianamine (NA), a chelator of metals, is ubiquitously present in higher plants. Nicotianamine aminotransferase (NAAT) catalyzes the amino group transfer of NA in the biosynthetic pathway of phytosiderophores and is essential for iron acquisition in graminaceous plants. The gene that encodes NAAT from barley was introduced into the nongraminaceous plant tobacco, which produces NA but not phytosiderophores. Transgenic tobacco plants (naat tobacco) that constitutively expressed the NAAT gene had young leaves with interveinal chlorosis and flowers that were abnormally shaped and sterile. Endogenous NA was consumed as a result of NAAT overproduction in naat tobacco. The resulting NA shortage caused disorders in internal metal transport, leading to these abnormal phenotypes. In addition to its role in long-distance metal transport, NA may be involved in the regulation of metal transfer within the cells. These results suggest that a shortage of NA impaired the functions of metal-requiring proteins, including transcription factors.

Alkyl and Aryl Transferases↗

The Decreased apical dominance1/Petunia hybrida CAROTENOID CLEAVAGE DIOXYGENASE8 gene affects branch production and plays a role in leaf senescence, root growth, and flower development.

Carotenoids and carotenoid cleavage products play an important and integral role in plant development. The Decreased apical dominance1 (Dad1)/PhCCD8 gene of petunia (Petunia hybrida) encodes a hypothetical carotenoid cleavage dioxygenase (CCD) and ortholog of the MORE AXILLARY GROWTH4 (MAX4)/AtCCD8 gene. The dad1-1 mutant allele was inactivated by insertion of an unusual transposon (Dad-one transposon), and the dad1-3 allele is a revertant allele of dad1-1. Consistent with its role in producing a graft-transmissible compound that can alter branching, the Dad1/PhCCD8 gene is expressed in root and shoot tissue. This expression is upregulated in the stems of the dad1-1, dad2, and dad3 increased branching mutants, indicating feedback regulation of the gene in this tissue. However, this feedback regulation does not affect the root expression of Dad1/PhCCD8. Overexpression of Dad1/PhCCD8 in the dad1-1 mutant complemented the mutant phenotype, and RNA interference in the wild type resulted in an increased branching phenotype. Other differences in phenotype associated with the loss of Dad1/PhCCD8 function included altered timing of axillary meristem development, delayed leaf senescence, smaller flowers, reduced internode length, and reduced root growth. These data indicate that the substrate(s) and/or product(s) of the Dad1/PhCCD8 enzyme are mobile signal molecules with diverse roles in plant development.

Alleles↗

Expression of a chimeric CaMV 35S Bacillus thuringiensis insecticidal protein gene in transgenic tobacco.

Insecticidal transgenic tobacco plants containing a truncated Bacillus thuringiensis cryIA(b) crystal protein (ICP) gene expressed from the CaMV 35S promoter were analyzed for ICP gene expression under field and greenhouse conditions over the course of a growing season. We present new information on temporal and tissue-specific expression of a CaMV 35S/cryIA(b) gene. Levels of cryIA(b) protein and mRNA were compared in both homozygous and hemizygous lines throughout plant development. Levels of ICP mRNA and protein increased during plant development with a pronounced rise in expression at the time of flowering. Homozygous ICP lines produced higher levels of ICP than the corresponding hemizygous lines. ELISA analysis of different tissues in the tobacco plant showed ICP gene expression in most tissues with a predominance of ICP in older tissue. All transgenic ICP tobacco lines which were studied in the field and greenhouse contained 400 ng to 1 microgram ICP per gram fresh weight in leaves from the mid-section of the plant at flowering. The amounts of ICP produced by field lines were directly comparable to levels observed in greenhouse-grown plants.

Bacillus thuringiensis↗

Polymers from plants to develop biodegradable plastics.

Katrin Neumann et al. have recently shown that transgenic tobacco and potato plants can accumulate high levels of cyanophycin, a possible source for poly-aspartate. This work opens the way to the future production of biodegradable plastics using a plant-based production system. Several problems need to be overcome first, such as growth retardation as a result of cyanophycin accumulating in the cytosol, and a co-production system needs to be developed for economical reasons.

Bacterial Proteins↗

A proteoglycan mediates inductive interaction during plant vascular development.

Inductive cell-cell interactions are essential for controlling cell fate determination in both plants and animals; however, the chemical basis of inductive signals in plants remains little understood. A proteoglycan-like factor named xylogen mediates local and inductive cell-cell interactions required for xylem differentiation in Zinnia cells cultured in vitro. Here we describe the purification of xylogen and cloning of its complementary DNA, and present evidence for its role in planta. The polypeptide backbone of xylogen is a hybrid-type molecule with properties of both arabinogalactan proteins and nonspecific lipid-transfer proteins. Xylogen predominantly accumulates in the meristem, procambium and xylem. In the xylem, xylogen has a polar localization in the cell walls of differentiating tracheary elements. Double knockouts of Arabidopsis lacking both genes that encode xylogen proteins show defects in vascular development: discontinuous veins, improperly interconnected vessel elements and simplified venation. Our results suggest that the polar secretion of xylogen draws neighbouring cells into the pathway of vascular differentiation to direct continuous vascular development, thereby identifying a molecule that mediates an inductive cell-cell interaction involved in plant tissue differentiation.

Amino Acid Sequence↗

The Arabidopsis ERECTA gene is expressed in the shoot apical meristem and organ primordia.

In Arabidopsis thaliana (L.) Heynh, the mutation in ERECTA is known to confer a compact inflorescence by a reduction in the lengths of internodes and pedicels. We analyzed the expression pattern of this gene during plant development. In situ hybridization and histochemical analysis using transgenic plants carrying chimeric gene fusions, with the ERECTA promoter fused to the beta-glucuronidase (GUS) gene, showed that ERECTA was predominantly expressed in the shoot apical meristems and organ primordia. ERECTA expression in the shoot apical meristem was weak early in plant development but increased with the transition from the vegetative to the reproductive growth phase. ERECTA was also strongly expressed in organ primordia and immature organs but weakly in mature organs. Thus, ERECTA was expressed in a cell-specific and developmentally regulated manner. In order to identify the regulatory mechanism responsible for the expression pattern of ERECTA, the cis-acting regions in the ERECTA promoter were defined by study of the expression of the chimeric genes that consist of the 5'- or internal deleted promoter and a GUS reporter gene in transgenic plants. The results showed that the essential cis-regulatory elements governing the spatially and temporally specific expression of ERECTA are located between positions -462 and -228 bp and between positions -228 and -153 bp with respect to the transcriptional initiation site.

Arabidopsis↗

Interaction of the tobacco mosaic virus replicase protein with the Aux/IAA protein PAP1/IAA26 is associated with disease development.

Virus-infected plants often display developmental abnormalities that include stunting, leaf curling, and the loss of apical dominance. In this study, the helicase domain of the Tobacco mosaic virus (TMV) 126- and/or 183-kDa replicase protein(s) was found to interact with the Arabidopsis Aux/IAA protein PAP1 (also named IAA26), a putative regulator of auxin response genes involved in plant development. To investigate the role of this interaction in the display of symptoms, a TMV mutant defective in the PAP1 interaction was identified. This mutant replicated and moved normally in Arabidopsis but induced attenuated developmental symptoms. Additionally, transgenic plants in which the accumulation of PAP1 mRNA was silenced exhibit symptoms like those of virus-infected plants. In uninfected tissues, ectopically expressed PAP1 accumulated and localized to the nucleus. However, in TMV-infected tissues, PAP1 failed to accumulate to significant levels and did not localize to the nucleus, suggesting that interaction with the TMV replicase protein disrupts PAP1 localization. The consequences of this interaction would affect PAP1's putative function as a transcriptional regulator of auxin response genes. This is supported by gene expression data indicating that approximately 30% of the Arabidopsis genes displaying transcriptional alterations in response to TMV contain multiple auxin response promoter elements. Combined, these data indicate that the TMV replicase protein interferes with the plant's auxin response system to induce specific disease symptoms.

Arabidopsis Proteins↗

Genetic and epigenetic processes in seed development.

Seed development has emerged as an important area of research in plant development. Recent research has highlighted the divergent reproductive strategies of the male and female genomes and interaction between genetic and epigenetic control mechanisms. Isolation of genes involved in embryo and endosperm development is leading to an understanding of the regulation of these processes at the molecular level. A thorough grasp of these processes will not only illuminate an important area of plant development but will also have an impact on agronomy by helping to facilitate food production. An understanding of seed development is also likely to clarify the molecular mechanisms of apomixis, a fascinating process of asexual seed production present in many plants.

Chromatin↗

An antigen expressed during plant vascular development crossreacts with antibodies towards KLH (keyhole limpet hemocyanin).

An antigen present in plant vascular tissue crossreacts with antibodies towards keyhole limpet hemocyanin (KLH). The antigen is present in xylem and vascular cambium, as evidenced by immunocytochemical staining of plant sections. This cell type assignment was confirmed by staining of mesophyll cell cultures from Zinnia elegans L. undergoing tracheary cell differentiation. The strongest staining both in sections and cell cultures occurred in cells and tissues during early stages of differentiation. Although the anti-KLH antibodies can easily be removed by affinity purification, our findings suggest that a certain caution is needed when KLH is used as an immunological carrier for studies in plants.

Amino Acid Sequence↗

Turnover of Galactans and Other Cell Wall Polysaccharides during Development of Flax Plants.

We investigated the synthesis and turnover of cell wall polysaccharides of the flax (Linum usitatissimum L.) plant during development of the phloem fibers. One-month-old flax plants were exposed to a 40-min pulse with 14CO2 followed by 8-h, 24-h, and 1-month periods of chase with ambient CO2, and radioactivity in cell wall sugars was determined in various plant parts. The relative radioactivity of glucose in noncellulosic polysaccharides was the highest compared with all other cell wall sugars immediately after the pulse and decreased substantially during the subsequent chase. The relative radioactivities of the other cell wall sugars changed with differing rates, indicating turnover of specific polysaccharides. Notably, after 1 month of chase there was a marked decrease in the proportional mass and total radioactivity in cell wall galactose, indicating a long-term turnover of the galactans enriched in the fiber-containing tissues. The ratio of radiolabeled xylose to arabinose also increased during the chase, indicating a turnover of arabinose-containing polymers and interconversion to xylose. The pattern of label redistribution differed between organs, indicating that the cell wall turnover processes are tissue- and cell-specific.

Journal Article↗

LIGHT CONTROL OF SEEDLING DEVELOPMENT.

Light control of plant development is most dramatically illustrated by seedling development. Seedling development patterns under light (photomorphogenesis) are distinct from those in darkness (skotomorphogenesis or etiolation) with respect to gene expression, cellular and subcellular differentiation, and organ morphology. A complex network of molecular interactions couples the regulatory photoreceptors to developmental decisions. Rapid progress in defining the roles of individual photoreceptors and the downstream regulators mediating light control of seedling development has been achieved in recent years, predominantly because of molecular genetic studies in Arabidopsis thaliana and other species. This review summarizes those important recent advances and highlights the working models underlying the light control of cellular development. We focus mainly on seedling morphogenesis in Arabidopsis but include complementary findings from other species.

Journal Article↗

Sequence relationships, conserved domains, and expression patterns for maize homologs of the polycomb group genes E(z), esc, and E(Pc).

Polycomb group (PcG) proteins play an important role in developmental and epigenetic regulation of gene expression in fruit fly (Drosophila melanogaster) and mammals. Recent evidence has shown that Arabidopsis homologs of PcG proteins are also important for the regulation of plant development. The objective of this study was to characterize the PcG homologs in maize (Zea mays). The 11 cloned PcG proteins from fruit fly and the Enhancer of zeste [E(z)], extra sex combs (esc), and Enhancer of Polycomb [E(Pc)] homologs from Arabidopsis were used as queries to perform TBLASTN searches against the public maize expressed sequence tag database and the Pioneer Hi-Bred database. Maize homologs were found for E(z), esc, and E(Pc), but not for Polycomb, pleiohomeotic, Posterior sex combs, Polycomblike, Additional sex combs, Sex combs on midleg, polyhometoic, or multi sex combs. Transcripts of the three maize Enhancer of zeste-like genes, Mez1, Mez2, and Mez3, were detected in all tissues tested, and the Mez2 transcript is alternatively spliced in a tissue-dependent pattern. Zea mays fertilization independent endosperm1 (ZmFie1) expression was limited to developing embryos and endosperms, whereas ZmFie2 expression was found throughout plant development. The conservation of E(z) and esc homologs across kingdoms indicates that these genes likely play a conserved role in repressing gene expression.

Amino Acid Sequence↗

Phenological modifications in plants by various edaphic factors.

Various mechanical, chemical and physical soil analyses were carried out, in addition to weather observations, for 3 years at several sites along an oceanic-continental gradient in a fjord district in western Norway. All the environmental factors observed were correlated with the spring and a few late-season phenophases of many native and cultivated woody plants and some herbs by simple, linear correlations and by stepwise multiple and partial analyses. Different techniques were used to try and eliminate many intercorrelations between various environmental factors. As expected, air temperature measurements in nearly all analyses from these temperate region districts gave the most significant correlations with the phenology of the plants, the temperature during the night generally being the most important in mainly vegetative periods, e.g. to leaf bud break in spring, and the temperature during the day affecting the more generative phases, such as the period between leaf bud break and flowering. The other environmental factors, however, showed strong variation in correlation significance among the various species studied and also with different phenophases of the same species. Various hypotheses are put forward to explain such variation. Air humidity (including precipitation) and/or soil moisture (including intercorrelated parameters, e.g. soil grain size and bulk density) were relatively often found to be of importance. In the stepwise multiple analyses for leaf bud break of the birch (Betula pubescens), for instance, the amount of precipitation was the second factor to enter the analyses by a positive correlation with the developmental rate, after the most important factor, the night temperature. Positive correlations with a high clay content and bulk density in the soil indicated that high soil moisture is also favourable for early bud break in the birch. Other phenophases that seemed to be favoured by a good water supply were leaf bud break of the bird cherry (Prunus padus) and rowan (Sorbus aucuparia), and flowering of the hazel (Corylus avellana), common lilac (Syringa vulgaris), plum ('Victoria') and currant ('Red Dutch') and also, to some degree, the goat willow (Salix caprea). The amount of ions (P, K, Mg and Ca) often showed negative correlations with the developmental rate, particularly of earlier phenophases of both native and cultivated plants (except for the apple 'Gravenstein' and pear 'Moltke'), possibly, indicating that a high nutrient level delayed plant development. A similar explanation might be given for the observation that high pH in the soil often seemed to delay plant development (leaf bud break of Betula, Sorbus, Syringa and plum, and flowering of Corylus, bluebell (Campanula rotundifolia) and red currant). According to the analyses there seemed to be a tendency for plants that are particularly dependent on warm weather for leaf bud break, e.g. the ash (Fraxinus excelsior), and flowering, e.g. Prunus, pear, apple and, to some degree, the raspberry ('Preussen'), to be less dependent on other environmental factors for their development. For instance, if there were any effects of water for these plants, they were negative for moisture and soil factors intercorrelated with water.

Acclimatization↗

Signaling among neighboring plants and the development of size inequalities in plant populations.

Transgenic tobacco plants that express an oat phytochrome gene (phyA) under control of the cauliflower mosaic virus (CaMV) 35S promoter and display altered photophysiology were used to test the role of light as a vehicle of information in dominance relationships between neighboring plants. Compared with the isogenic wild type, phyA-overexpressing plants showed dramatically reduced morphological responsivity to changes in the red/far red ratio of the incident light and to the proximity of neighboring plants in spacing experiments. In transgenic canopies an increase in stand density caused the small plants of the population to be rapidly suppressed by their neighbors. In wild-type canopies, plants responded to increased density with large morphological changes, and there appeared to be an inverse relationship between the magnitude of this morphological response and the ranking of the individual plant in the population size hierarchy. In these wild-type populations, size inequality increased only moderately with density within the time frame of the experiments. Our results suggest that, in crowded stands, the ability of individual plants to acquire information about their light environment via phytochrome plays a central role in driving architectural changes that, at the population level, delay the development of size differences between neighbors.

Avena↗

The future of transgenic plants in developing countries.

Whatever their own policies may be, developing countries will inevitably be affected by the development of genetically-modified organisms in industrialized countries. While maintaining a cautious attitude, most of these countries wish to keep their options open, thus protecting themselves from the risk of being deprived of future technologies that might allow them to achieve self-sufficiency in food production, to resolve certain problems confronting their most vulnerable populations and to preserve the international competitiveness of their products. Companies should see that it is in their interest to help these countries implement their own policies, notably through an open attitude to industrial property. If the value of genetic engineering is thus confirmed, then it perhaps in this manner that GMOs will earn the legitimacy required to make them acceptable to the people of Northern countries where the majority of solvent markets are located.

Agriculture↗