Sofia Kourmpetli

Over-expression of a gibberellin 2-oxidase gene from Phaseolus coccineus L. enhances gibberellin inactivation and induces dwarfism in Solanum species

Gibberellins (GAs) are endogenous hormones that play a predominant role in regulating plant stature by increasing cell division and elongation in stem internodes. The product of the GA 2-oxidase gene from Phaseolus coccineus (PcGA2ox1) inactivates C19-GAs, including the bioactive GAs GA1 and GA4, by 2β-hydroxylation, reducing the availability of these GAs in plants. The PcGA2ox1 gene was introduced into Solanum melanocerasum and S. nigrum (Solanaceae) by Agrobacterium-mediated transformation with the aim of decreasing the amounts of bioactive GA in these plants and thereby reducing their stature. The transgenic plants exhibited a range of dwarf phenotypes associated with a severe reduction in the concentrations of the biologically active GA1 and GA4. Flowering and fruit development were unaffected. The transgenic plants contained greater concentrations of chlorophyll b (by 88%) and total chlorophyll (11%), although chlorophyll a and carotenoid contents were reduced by 8 and 50%, respectively. This approach may provide an alternative to the application of chemical growth retardants for reducing the stature of plants, particularly ornamentals, in view of concerns over the potential environmental and health hazards of such compounds.

Artificial microRNAs reveal cell-specific differences in small RNA activity in pollen

Pollen formation, while critical for the success of plant reproduction, also represents an important paradigm for differential cellular development within small groups of cells. In Arabidopsis thaliana pollen, the male meiotic product first divides asymmetrically to form a vegetative and a generative (germ) cell, the latter then dividing to generate two sperm cells. Here we have used artificial microRNAs to study small RNA processing in the different pollen cell types. Our data suggest that translational repression by small RNAs is enhanced in the sperm. This work also provides insights into germline RNA movement and the cell-autonomous action of microRNAs.

Analysis of grain characters in temperate grasses reveals distinctive patterns of endosperm organization associated with grain shape

Members of the core pooids represent the most important crops in temperate zones including wheat, barley, and oats. Their importance as crops is largely due to the grain, particularly the storage capabilities of the endosperm. In this study, a comprehensive survey of grain morphology and endosperm organization in representatives of wild and cultivated species throughout the core pooids was performed. As sister to the core pooid tribes Poeae, Aveneae, Triticeae, and Bromeae within the Pooideae subfamily, Brachypodium provides a taxonomically relevant reference point. Using macroscopic, histological, and molecular analyses distinct patterns of grain tissue organization in these species, focusing on the peripheral and modified aleurone, are described. The results indicate that aleurone organization is correlated with conventional grain quality characters such as grain shape and starch content. In addition to morphological and organizational variation, expression patterns of candidate gene markers underpinning this variation were examined. Features commonly associated with grains are largely defined by analyses on lineages within the Triticeae and knowledge of grain structure may be skewed as a result of the focus on wheat and barley. Specifically, the data suggest that the modified aleurone is largely restricted to species in the Triticeae tribe.

Small RNA activity and function in angiosperm gametophytes

Small non-coding RNAs are key post-transcriptional and transcriptional regulators of plant gene expression in angiosperm sporophytes. In recent years, gametophytic small RNAs have also been investigated, predominantly in Arabidopsis male gametophytes, revealing features in common with the sporophyte as well as some surprising differences. Transcriptomic and deep-sequencing studies confirm that multiple small RNA pathways operate in male gametophytes, with over 100 miRNAs detected throughout development. Trans-acting siRNA pathways that are associated with novel phased transcripts in pollen, and the nat-siRNA pathway have important roles in pollen maturation and gamete function. Moreover, a role for siRNA-triggered silencing of transposable elements in male and female germ cells has been established, a feature in common with the role of piRNAs in animal germlines. Current evidence supports an integral role for small RNAs in angiosperm gametophyte development and it can be anticipated that novel small RNAs with significant roles in germline development and genome integrity await discovery.

Practical Applications of Manipulating Plant Architecture by Regulating Gibberellin Metabolism

The international trade in floriculture is estimated to be worth about US

Characterization of the Fungal Gibberellin Desaturase as a 2-Oxoglutarate-Dependent Dioxygenase and Its Utilization for Enhancing Plant Growth

150 billion, with the global demand for ornamentals steadily increasing. Consumer choice is influenced by factors such as plant architecture and flower colour. Conventional breeding has been responsible for the introduction of novel traits into ornamental plants and has played an important role in the development of new cultivars. However, a restricted gene pool and failure of distant crosses have led to the exploitation of somatic cell techniques, particularly genetic transformation, to generate plants with desirable traits. Gibberellins (GAs) are endogenous plant hormones that control key aspects of growth and development. Chemical growth regulators that modify GA biosynthesis are used extensively in horticulture to control plant stature, increasing production costs, manpower, and environmental risks. An alternative strategy involves genetic manipulation of GA metabolism to induce phenotypic changes, particularly alteration of stature. Because ornamentals are not used for human consumption, genetic manipulation approaches with these plants may be more acceptable in the immediate future to the general public, in certain parts of the world, than genetically manipulated food crops.

The fruit, the whole fruit, and everything about the fruit

The biosynthesis of gibberellic acid (GA3) by the fungus Fusarium fujikuroi is catalyzed by seven enzymes encoded in a gene cluster. While four of these enzymes are characterized as cytochrome P450 monooxygenases, the nature of a fifth oxidase, GA4 desaturase (DES), is unknown. DES converts GA4 to GA7 by the formation of a carbon-1,2 double bond in the penultimate step of the pathway. Here, we show by expression of the des complementary DNA in Escherichia coli that DES has the characteristics of a 2-oxoglutarate-dependent dioxygenase. Although it has low amino acid sequence homology with known 2-oxoglutarate-dependent dioxygenases, putative iron- and 2-oxoglutarate-binding residues, typical of such enzymes, are apparent in its primary sequence. A survey of sequence databases revealed that homologs of DES are widespread in the ascomycetes, although in most cases the homologs must participate in non-gibberellin (GA) pathways. Expression of des from the cauliflower mosaic virus 35S promoter in the plant species Solanum nigrum, Solanum dulcamara, and Nicotiana sylvestris resulted in substantial growth stimulation, with a 3-fold increase in height in S. dulcamara compared with controls. In S. nigrum, the height increase was accompanied by a 20-fold higher concentration of GA3 in the growing shoots than in controls, although GA1 content was reduced. Expression of des was also shown to partially restore growth in plants dwarfed by ectopic expression of a GA 2-oxidase (GA-deactivating) gene, consistent with GA3 being protected from 2-oxidation. Thus, des has the potential to enable substantial growth increases, with practical implications, for example, in biomass production.

Generation and Deployment of Transgenic Crop Plants: An Overview

Fruits come in an impressive array of shapes, sizes, and consistencies, and also display a huge diversity in biochemical/metabolite profiles, wherein lies their value as rich sources of food, nutrition, and pharmaceuticals. This is in addition to their fundamental function in supporting and dispersing the developing and mature seeds for the next generation. Understanding developmental processes such as fruit development and ripening, particularly at the genetic level, was once largely restricted to model and crop systems for practical and commercial reasons, but with the expansion of developmental genetic and evo-devo tools/analyses we can now investigate and compare aspects of fruit development in species spanning the angiosperms. We can superimpose recent genetic discoveries onto the detailed characterization of fruit development and ripening conducted with primary considerations such as yield and harvesting efficiency in mind, as well as on the detailed description of taxonomically relevant characters. Based on our own experience we focus on two very morphologically distinct and evolutionary distant fruits: the capsule of opium poppy, and the grain or caryopsis of cereals. Both are of massive economic value, but because of very different constituents; alkaloids of varied pharmaceutical value derived from secondary metabolism in opium poppy capsules, and calorific energy fuel derived from primary metabolism in cereal grains. Through comparative analyses in these and other fruit types, interesting patterns of regulatory gene function diversification and conservation are beginning to emerge.

A FILAMENTOUS FLOWER orthologue plays a key role in leaf patterning in opium poppy.

As biotechnology increasingly affects almost all aspects of human life, it is essential that the science behind this technology is explained in simple terms to the public to eliminate the misconceptions that may inhibit its acceptability. The basic question that is often asked is what is a gene, a promoter and a terminator? Genes are the basic units of heredity, composed of DNA sequences, which are transmitted from parents to offspring and which, independently or in combination with other genes, control specific traits in an organism. These traits may be, for example, plant height, flower color, fruit and seed size together with regulatory processes, such as assimilate partitioning and drought resistance. Genes are the basis for both the similarity and differences that exist among organisms, and are transmitted from one generation to another. Promoters are DNA sequences that are recognized by RNA polymerase in plant cells and that initiate and regulate transcription, the initial and most important step of gene expression. Terminators are those sequences that command or signal the termination of transcription.

A FILAMENTOUS FLOWER orthologue plays a key role in leaf patterning in opium poppy

The plant-specific YABBY genes were initially defined by their roles in determining abaxial/adaxial cell fate in lateral organs of eudicots, and repressing meristematic genes in differentiating tissues such as leaves. In Arabidopsis thaliana FILAMENTOUS FLOWER (FIL) is also required for inflorescence and floral meristem establishment and flower development in a pathway involving the floral transition and identity genes. Here we describe the characterization of a FIL orthologue from the basal eudicot, Papaver somniferum (the opium poppy), and demonstrate a role for the gene in patterning the highly lobed leaf of the poppy. Silencing of PapsFIL using viral-induced gene silencing resulted in leaves of reduced laminar area, more pronounced margin serration and, in some cases, leaf bifurcation. In contrast, the gene does not appear to affect the development of the flower, and these variations in function are discussed in relation to its taxonomic position as a basal eudicot and its determinate growth habit.