Impact of genotype, age of tree and environmental temperature on androgenesis induction of Aesculus hippocastanum L.

Influence of ten different genotypes, age of trees and environmental temperature on induction of androgenesis and apperance albino horse chestnut embryos were studied. Efficiency of in vitro androgenesis via anther and microspore culture had been investigated. Microspores and anthers were used from the same closed flower bud. Androgenic response of different genotypes was measured and compared. Anther induction rates were from 5 to 37.6%, depending on genotype. The number ofembryos per isolated anther varied between 0.5 to 5.0 embryos in anther culture, while in microspore culture varied between 3.0 to 27 embryos, depending on genotype. A microspore culture was 5 - 6 times efficient than anther culture for same genotype. Age of the trees had no influence on androgenesis induction. Temperature of about 4 - 5°C was optimal for androgenic embryo induction. Albino horse chestnut embryos phenomenon depended on genotype. The number of albino appearing in anther washigher than in microspore culture. The same correlation of appearing albino was observed between short and long day. Flow cytogenetic analysis of androgenic embryos originating from anther and microspore culture was done after a first generation of regenerants. All androgenic embryos, the first generation from microspore culture were haploid, while 50% of the regenerants originating from anther culture were haploid, and the other half diploid

High-efficiency Agrobacterium rhizogenes-mediated genetic transformation in Artemisia vulgaris: hairy root production and essential oil analysis

Agrobacterium rhizogenes-mediated genetic transformation of Artemisia vulgaris was investigated using four bacterial strains: A4GUS, R1000, R1601and ATCC15834 and three explant types: shoot tip, leaf and node. The A4GUS proved to be more competent than other strains and the highest transformation rates were observed in leaf explant (92.6%). Among 92 hairy root clones acquired, clones AV1 and AV2 exhibited the highest growth. Growth of hairy roots was assessed on the basis of total root elongation, lateral root density and biomass accumulation. Out of four media composition, the ½ MS medium was found to be most suitable for biomass production. Biomass accumulation was the highest when the medium was supplemented with 40 g L−1 sucrose. PCR and Southern hybridization analysis revealed both the TL- and TR-DNA integration in the root clones AV1 and AV2. Hairy roots showed higher growth rate and accumulated substantial amount of essential oils than non-transformed roots. Eighty-seven compounds were identified from transformed root essential oils, whereas 77 compounds were identified from nontransformed root essential oils. The main compounds identified were camphor, camphene, -thujone, germacrene D, 1,8-cineole and -caryophyllen

Fertile transgenic Lotus corniculatus resistant to the non-selective herbicide phosphinothricin

Resistance to the non-selective herbicide dl-phosphinothricin (PPT) was introduced into commercial Lotus corniculatus cv. Bokor by co-cultivation of cotyledons with Agrobacterium tumefaciensAGL1 harbouring the binary vector pDM805 which contains the bialaphos resistance gene (bar) from Streptomyces hygroscopicus encoding phosphinothricin acetyltransferase (PAT) and the uidA gene encoding -glucuronidase. The half-cotyledon explants were precultured on regeneration Murashige and Skoog's (MS) medium supplemented with 6-benzyladenine (BA) and 1-naphthaleneacetic acid (NAA) at 0.5mgL(-1) each, 3days prior to infection. Upon co-cultivation, the explants were cultured on PPT-free regeneration medium for 10days, and then subcultured on regeneration/selection media with increasing PPT concentrations (5-7mgL(-1)) for about 18weeks. Out of 480 initially co-cultivated explants, 272 regenerated shoots survived the entire PPT selection procedure. Resistant shoots were grown further, multiplied by tillering that was additionally promoted by PPT and rooted on hormone-free MS medium containing 5mgL(-1) PPT. Established shoot cultures, continuously maintained on the same medium, have preserved PPT resistance up to now (more than 2years). Transformed plants assessed in vitro and in a greenhouse were tolerant to the herbicide PPT at 300mgL(-1) equivalent to more than twofold the recommended field dosage for weed eradication. Applied PPT treatment did not affect the activities of glutamine synthetase (GS; EC 6.3.1.2) and NADH-dependent glutamate dehydrogenase (NADH-GDH; EC 1.4.1.2) in transformed plants. However, PPT did increase the mobility of glutamine synthetase isoforms GS1 and GS2 as well as the inhibition of an additional high mobility GS (hmGS) activity. In untransformed plants, PPT treatment reduced total GS activity by 4.4-fold while contrary the activity of NADH-GDH was increased by ninefold. All transformed herbicide-resistant plants were phenotypically normal and exhibited genomic stability, as were the untransformed plants analysed by flow cytometry. Under greenhouse conditions, they grew to maturity, flowered and set seeds. Stable integration and expression of the bar gene in T0 and T1 plants were confirmed by Southern and Western blot analysis, while integration of the reporter uidA gene did not occur. The bar gene was inherited in a Mendelian fashion by the progeny, as detected by PPT resistance. The production of PPT-resistant plants may have significant practical applications in weed control in fields of L. corniculatus

Perspectives: Rationale and design of the ISACS-TC (International Survey of Acute Coronary Syndromes in Transitional Countries) project

During the past 10 years, the health of people in Eastern Europe and the former Soviet Union has undergone changes very different from the health patterns seen in their Western counterparts. Mortality from cardiovascular disease has been decreasing continuously in the USA and many Western European countries, but it has increased or remained unchanged in many of the states of Eastern Europe. Analysis of this phenomenon has been hindered by insufficient information. The International Registry of Acute Coronary Syndromes registry study in Transitional Countries (ISACS-TC) is both a retrospective - over a 1-year period - and prospective study which was designed in order to obtain data of patients with acute coronary syndromes (ACSs) in countries with economy in transition in Central and Eastern Europe, and herewith control and optimize internationally guideline recommended therapies in these countries. Adhesion to the project was given by 112 Collaborating Centres in 17 countries with economy in transition (Albania, Bosnia and Herzegovina, Belarius, Bulgaria, Croatia, Hungary, Kosovo, Latvia, Lithuania, Macedonia, Moldova, Montenegro, Romania, Russian Federation, Serbia, Slovakia, Slovenia, and Ukraine). A total of 47 cluster sites in 11 countries in Central and Eastern Europe are currently collaborating in ISACS-TC. The registry encourages optimal individualization of evidence-based therapies and the international patient body ensures good representation of multiple practice patterns. It may help to make an additional improvement in clinical outcomes of countries with economy in transition

Agrobacterium rhizogenes-Mediated Transformation and Its Biotechnological Applications in Crops

The history of Agrobacterium-related plant biotechnology goes back for more than three decades with the discovery of molecular mechanisms of crown gall disease in plants. After 1980s, gene technologies began developing rapidly and today, related with the improved gene transfer methods, plant biotechnology has become one of the most important branches in science. Till now, the most important genes related with agricultural affairs have been utilized for cloning of plants with the deployment of different techniques used in genetic engineering. Especially, Agrobacterium tumefaciens was used extensively for transferring desired genetic materials to plants rapidly and effectively by the researchers to create transgenic plants. Recognition of the biology of Agrobacterium species and newly developed applications of their T-DNA systems has been a great step in plant biotechnology. This chapter provides the reader with extensive information on A. rhizogenes which is responsible for the development of hairy root disease in a wide range of dicotyledonous plants and its T-DNA system. This knowledge will be useful in improving utilization of crops and the formulation of new and up-graded transgenic based food products

Molecular aspects of somatic-to-embryogenic transition in plants

Somatic embryogenesis (SE) is a model system for understanding the physiological, biochemical, and molecular biological events occurring during plant embryo development. Plant somatic cells have the ability to undergo sustained divisions and give rise to an entire organism. This remarkable feature is called plant cell totipotency. SE is a notable illustration of plant totipotency and involves reprogramming of development in somatic cells toward the embryogenic pathway. Plant growth regularities, especially auxins, are key components as their exogenous application recapitulates the embryogenic potential of the mitotically quiescent somatic cells. It has been observed that there are genetic and also physiological factors that trigger in vitro embryogenesis in various types of plant somatic cells. Analysis of the proteome and transcriptome has led to the identification and characterization of certain genes involved in SE. Most of these genes, however, are upregulated only in the late developmental stages, suggesting that they do not play a direct role in the vegetative-to-embryogenic transition. However, the molecular bases of those triggering factors and the genetic and biochemical mechanisms leading to in vitro embryogenesis are still unknown. Here, we describe the plant factors that participate in the vegetative-to-embryogenic transition and discuss their possible roles in this process