RAPID SCREENING OF PHYTOPATHOGENIC Erwinia sp.  OF TWO POTATO VARIETIES (SPUNTA AND DESIREE)  FROM ALGERIAN AGRICULTURAL FIELDS

Rapid screening of phytopathogenic Erwinia sp. of two potato varieties (Spunta and Desiree) from Algerian agricultural fileds. Isolation, phenotypic identification and in vitro phytopathogenicity screening of Erwinia sp. from agricultural field of two potato varieties (Spunta and Desiree) in Algeria. The current study aims to isolate, identify and screen phytopathogenic isolates of Erwinia sp. causing potato diseases. The techniques presented in this study for isolation and characterization of phytopathogenic Erwinia sp. are conventional methods that are used in this field of research. Seven phytopathogenic bacteria were recovered from potato tubers of two varieties (Spunta and Desiree). The phenotypic identification allowed characterizing typical colonies of Erwinia sp. on two semi-selective media: King’s B and TCC media. Erwinia sp. formed characteristic colonies on King’s B medium that were round, convex and representing creamy color. While, Erwinia sp. also developed specific colonies on TCC medium which were pale purple, circular, convex, even bulging; smooth and mucous. In vitro phytopathogenicity test on potato slices lead to screen the phytopathogenic isolate E5 characterized by highest rotten tissue zone of (2.33 ± 0.29 cm) and (2.33 ± 0.58 cm) toward Spunta and Desiree varieties, respectively. Followed, by isolate E4 characterized by rotten tissue zone of (1.83 ± 0.58 cm) and (2.17 ± 0.29 cm) toward Spunta and Desiree varieties, respectively; compared to their corresponding uninfected controls. The RTZ (Rotten tissue zone) evidently is proportional to the specific pathogenicity of Erwinia sp. isolates and the characteristic sensitivity of various varieties (Spunta and Desiree). Thus, make determining RTZ a rapid screening technique for the selection of the highest phytopathogenic isolates. This investigation provides valuable information for rapid screening (infected potato tuber) and characterization (isolation using semi-selective media) of pathogenic Erwinia sp. engendering potato disease compared to existing methods like infection of leaves or plants; and phytopathogenic Erwinia sp. identification through PCR amplification or in situ hybridation.

Download Full-text

A comprehensive study on the isolation and characterization of the HeLa S3 nuclear matrix

Journal of Cell Science ◽

10.1242/jcs.98.3.281 ◽

1991 ◽

Vol 98 (3) ◽

pp. 281-291

Author(s):

P. Belgrader ◽

A.J. Siegel ◽

R. Berezney

Keyword(s):

Nuclear Matrix ◽

Intermediate Filament ◽

Structural Integrity ◽

Rnase A ◽

Matrix Proteins ◽

Intermediate Filament Proteins ◽

Isolation And Characterization ◽

Hela S3

Different agents have been employed to extract the histones and other soluble components from isolated HeLa S3 nuclei during nuclear matrix isolation. We report that 0.2M (NH4)2SO4 is a milder extracting agent than NaCl and LIS (lithium 3,5-diiodosalicylate), on the basis of the apparent preservation of the elaborate fibrogranular network and the residual nucleolus that resemble the in situ structures in whole cells and nuclei, minimal aggregation, and sufficient solubilization of DNA and histones. The importance of intermolecular disulfide bonds, RNA and 37 degrees C stabilization on the structural integrity of the nuclear matrix was examined in detail using sulfydryl alkylating, reducing and oxidizing agents, and RNase A. The data suggest that any disulfides formed during the isolation are not essential for maintaining the structural integrity of the in vitro matrix. However, structural integrity of the matrix is dependent upon RNA and to some degree on disulfides that presumably existed in situ. Sodium tetrathionate and 37 degrees C stabilization of isolated nuclei resulted in nuclear matrices containing an approximately twofold greater amount of protein, RNA and DNA than control preparations. The 37 degrees C incubation, unlike the sodium tetrathionate stabilization, does not appear to induce intermolecular disulfide bond formation. Neither stabilizations resulted in significant differences of the major matrix polypeptide pattern on two-dimensional (2-D) gels stained with Coomassie Blue as compared to that of unstabilized matrix. The major nuclear matrix proteins, other than the lamins, did not react to the Pruss murine monoclonal antibody (IFA) that recognizes all known intermediate filament proteins, suggesting that the internal matrix proteins are not related to the lamins in intermediate filament-like quality.

Download Full-text

Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010005161x ◽

1974 ◽

Vol 32 ◽

pp. 320-321

Author(s):

J. P. Revel

Keyword(s):

Developmental Stages ◽

Three Dimensional ◽

Cell Movement ◽

Cellular Interactions ◽

Serial Sections ◽

Cell Sheets ◽

Freeze Etching ◽

Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

Download Full-text

In vitro and in vivo polysaccharides assembly: ultrastructural and cytochemical study of growing plant cell wall components.

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100083035 ◽

1978 ◽

Vol 36 (2) ◽

pp. 434-435

Author(s):

D. Reis ◽

B. Vian ◽

J. C. Roland

Keyword(s):

Cell Wall ◽

Self Assembly ◽

Plant Cell Wall ◽

Higher Plants ◽

Three Dimensional ◽

Cytochemical Study ◽

Wall Components

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.

Download Full-text

In Situ Localization of PPAR Gamma and Uncoupling Protein in Mouse Embryo Sections Using Digoxigenin-Labeled Riboprobes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162624 ◽

1996 ◽

Vol 54 ◽

pp. 32-33

Author(s):

C. Jennermann ◽

S. A. Kliewer ◽

D. C. Morris

Keyword(s):

Alkaline Phosphatase ◽

Room Temperature ◽

Uncoupling Protein ◽

Ppar Gamma ◽

Nuclear Hormone Receptor ◽

Full Length ◽

Northern Analysis ◽

Peroxisome Proliferator

Peroxisome proliferator-activated receptor gamma (PPARg) is a member of the nuclear hormone receptor superfamily and has been shown in vitro to regulate genes involved in lipid metabolism and adipocyte differentiation. By Northern analysis, we and other researchers have shown that expression of this receptor predominates in adipose tissue in adult mice, and appears first in whole-embryo mRNA at 13.5 days postconception. In situ hybridization was used to find out in which developing tissues PPARg is specifically expressed.Digoxigenin-labeled riboprobes were generated using the Genius™ 4 RNA Labeling Kit from Boehringer Mannheim. Full length PPAR gamma, obtained by PCR from mouse liver cDNA, was inserted into pBluescript SK and used as template for the transcription reaction. Probes of average size 200 base pairs were made by partial alkaline hydrolysis of the full length transcripts. The in situ hybridization assays were performed as described previously with some modifications. Frozen sections (10 μm thick) of day 18 mouse embryos were cut, fixed with 4% paraformaldehyde and acetylated with 0.25% acetic anhydride in 1.0M triethanolamine buffer. The sections were incubated for 2 hours at room temperature in pre-hybridization buffer, and were then hybridized with a probe concentration of 200μg per ml at 70° C, overnight in a humidified chamber. Following stringent washes in SSC buffers, the immunological detection steps were performed at room temperature. The alkaline phosphatase labeled, anti-digoxigenin antibody and detection buffers were purchased from Boehringer Mannheim. The sections were treated with a blocking buffer for one hour and incubated with antibody solution at a 1:5000 dilution for 2 hours, both at room temperature. Colored precipitate was formed by exposure to the alkaline phosphatase substrate nitrobluetetrazoliumchloride/ bromo-chloroindlylphosphate.

Download Full-text

CONSERVATION OF RARE SPECIES PLANTS AND LICHENS IN SITU WITH PLANNED ANTHROPOGENIC ACTIVITIES: BASIC APPROACHES AND IMPLEMENTATION PRINCIPLES

Engineering survey ◽

10.25296/1997-8650-2018-12-7-8-38-45 ◽

2018 ◽

Vol 12 (7-8) ◽

pp. 38-45

Author(s):

A. N. EFREMOV ◽

N. V. PLIKINA ◽

T. ABELI

Keyword(s):

Rare Species ◽

Technology Implementation ◽

Technology Development ◽

Anthropogenic Activities ◽

Natural Habitat ◽

Local Population ◽

Main Stage ◽

Social Risks

Rare species are most vulnerable to man-made impacts, due to their biological characteristics or natural resource management. As a rule, the economic impact is associated with the destruction and damage of individual organisms, the destruction or alienation of habitats. Unfortunately, the conservation of habitat integrity is an important protection strategy, which is not always achievable in the implementation of industrial and infrastructural projects. The aim of the publication is to summarize the experience in the field of protection of rare species in the natural habitat (in situ), to evaluate and analyze the possibility of using existing methods in design and survey activities. In this regard, the main methodological approaches to the protection of rare species in the natural habitat (in situ) during the proposed economic activity were reflected. The algorithm suggested by the authors for implementing the in situ project should include a preparatory stage (initial data collection, preliminary risk assessments, technology development, obtaining permitting documentation), the main stage, the content of which is determined by the selected technology and a long monitoring stage, which makes it possible to assess the effectiveness of the taken measures. Among the main risks of in situ technology implementation, the following can be noted: the limited resources of the population that do not allow for the implementation of the procedure without prior reproduction of individuals in situ (in vitro); limited knowledge of the biology of the species; the possibility of invasion; the possibility of crossing for closely related species that сo-exist in the same habitat; social risks and consequences, target species or population may be important for the local population; financial risks during the recovery of the population. The available experience makes it possible to consider the approach to the conservation of rare species in situ as the best available technology that contributes to reducing negative environmental risks.

Download Full-text