Development of specific primers for the detection of HVA1 from barley in transgenic durum wheat by polymerase chain reaction (PCR) technology

Genetic transformation is a widely employed tool in both basic research and commercial plant breeding programs. Its application requires that transgenes be stably integrated and expressed in the plant genome. When transgenic plants are developed, it is essential to determine which plants contain the transgene. Detection methods are usually based on amplification of the target transgene. This paper describes a development of detection method based on conventional and real time polymerase chain reaction (PCR) for simultaneous detection of barley HVA1 transgene and its transcript in transformed durum wheat. Since there exist a high homology between the barley HVA1 gene and the wheat gene, development of a specific sets of primers is needed for PCR-based characterizations, and the study of the transgene. Based on the alignment of the two genes sequences obtained from public databases, several primers were designed to detect and distinguish between the transformed and non-transformed plants. Real time PCR has been employed because of its inherent sensitivity and quantitative nature. It has been possible to design the following primers pairs F2/MMR, F2/R10 and F14/R10 as highly specific and suitable for the detection of HVA1 DNA by conventional and real-time PCR. Nonetheless, the primers used were allowed to reach high efficiencies and did not show any cross-reactivity with DNAs extracted from various plants. The sensitivity achieved was 6.4 pg. The primer pair F2/R10 was considered as highly specific for the detection of both DNA and mRNA of the HVA1 by real-time PCR. The assays proved to be accurate, specific, sensitive and sufficiently reproducible for further application in high-throughput molecular characterization of transgenic lines.Keywords: HVA1, durum wheat, transgenic plant, real time polymerase chain reaction (PCR), droughtAfrican Journal of Biotechnology, Vol. 13(4), pp. 581-592, 22 January, 201

In Silico Investigation of Aristolochia longa Anticancer Potential against the Epidermal Growth Factor Receptor (EGFR) in the Tyrosine Kinase Domain

This study employed in silico methods to identify potential therapeutic targets for the inactive EGFR tyrosine kinase domain in complex with Erlotinib (PDB: 4HJO) which is known to cause cancer, using natural extracts from Aristolochia longa root. A library comprising five natural compounds (Luteolin, 4-hydroxycinnamic acid, Kaempferol, ferulic acid, citric acid, and quinic acid) and the standard Erlotinib (control) were subjected to Lipinski's rule of five, ADMET parameter analyses, molecular docking and molecular dynamics simulation. Results revealed comparable pharmacological responses between the five compounds and the standard drug, demonstrating promising outcomes without limitations. Notably, Luteolin, Kaempferol, and quinic acid exhibited higher binding energies than the reference molecule, with binding affinities of -9.083 kcal/mol, -8.260 kcal/mol, and -5.857 kcal/mol, respectively. Molecular dynamics simulations confirmed the stability of the most effective EGFR protein-ligand, displaying consistent interaction profiles, favorable molecular properties, and a stable trajectory (RMSD, RMSF). Overall, these in silico analyses highlight the potential of aromatic and medicinal plant-derived compounds to inhibit EGFR protein associated with cancer development, emphasizing the need for further in vitro and in vivo investigations to explore their therapeutic applications in cancer patients

Utilisation Des Caractères Biochimiques, Agronomiques Et Morphologiques Pour L’évaluation De La Diversité Génétique Des Variants D’orangers Dans La Région Du Gharb, Maroc

20 clones of orange trees, identified at the level of the collection of nucellar citrus varieties, have been the subject of several tests of behavior at the level of different citrus fruit sites in Morocco. Indeed, the present work aims to evaluate these variants on a clay soil at the Gharb region (INRA/Kenitra). The variants grafted on Citrange troyer were divided into three groups. Each group contains variants, genetically similar, from seedlings of the same parent variety. Clonal selection is based on the synthesis of the results of the study of the variability of the criteria of fruit quality and production. The average yield varies from 44.25 kg / tree to 217.30 kg / tree. While the average weight of a fruit varies from 95 g to 360 g. The results showed a predominance of juicy and medium-sized fruits with an equatorial diameter greater than 56 mm, acceptable for the fresh fruit market or the juice industry. The maturity coefficient study of the variants within each group, or between the three groups, showed that the harvest period extends over four months, from December to March. The final selection was focused on six productive and good quality variants. These promising variants have been described according to UPOV, and then presented for registration in the official catalog

Effects of Temperature and Relative Humidity on the In Vitro and In Vivo Radial Growth of Penicillium italicum and on the Biocontrol Activity of Pichia guilliermondii, Strain Z1

peer reviewedThe objective of this study was to assess the effect of temperature (5-25°C) on the ‘in vitro’ and ‘in vivo’ growth rates of Penicillium italicum and to determine the combined effect of temperature and relative humidity (45 to 100%) on lesion size of this pathogenic fungus on Valencia late oranges, either alone or in combination with the antagonistic yeast strain Z1 of Pichia guilliermondii Wickerham. Statistical analysis showed a significant effect of temperature on the ‘in vitro’ and ‘in vivo’ radial growth of P. italicum with the maximum growth observed at temperature of 25°C. In both cases, no growth was observed at a temperature of 35°C. These factors had a significant effect on P. italicum lesion size when it was applied alone on Valencia late oranges and insignificant when yeast strain Z1 was applied 24 h before P. italicum inoculation. Our results confirm previous ‘in vitro’ findings that aw has a greater influence than temperature on P. italicum growth and highlight that the strain Z1 showed high antagonistic potential against this pathogen over a range of temperature-relative humidity regimes favouring P. italicum development

Effet de la salinité sur les paramètres morphophysiologiques de deux variétés de bananier (Musa acuminata L)

Objectifs : Le présent travail a pour objectif de déterminer l’effet du stress salin sur le taux de régénération des explants, et sur les caractères morphophysiologiques de deux variétés de bananier (Musa acuminata) grande et petite naine afin de sélectionner des vitro plants tolérants à la salinité.Méthodologie et résultats : Des rejets de deux variétés de bananier (Musa acuminata) « la grande naine et la petite naine issus de la région du Gharb ont été cultivés sur le milieu de Murashige et Skoog, additionné ou non de NaCl (0, 2, 4 et 6 g/l). Les résultats obtenus montrent que les deux variétés ont une régénération des pousses malgré le traitement salin. Cependant, le sel affecte, la longueur des feuilles des explants et le taux de régénération. L’augmentation de la concentration saline entraine une diminution du nombre de pousse par bourgeon. De même, le comportement physiologique de ces deux variétés de bananier sous l’effet de doses croissantes de NaCl se traduit par des teneurs faibles en chlorophylle a, b et totale, et par des teneurs élevées en proline racinaire et en sucres solubles totaux, au niveau des racines et des feuilles, Par ailleurs la teneur en sucres solubles totaux dans les feuilles est plus élevée chez la grande que chez la petite naine. Par contre, dans les racines, c’est l’inverse de ce constat qui se produit.Conclusion et application de la recherche : La salinité a entrainé des modifications morpho physiologique qui n’affectent pas les variétés de bananier de la même manière. Le degré de sensibilité au sel dépend de la variété, de l’intensité du stress et de type d’organe (racine, feuille). La sélection du matériel végétal tolérant au sel est, par conséquent, tributaire d’une connaissance approfondie des mécanismes physiologiques et biochimiques.Mots clé : bananier, salinité, chlorophylle, proline, sucres solubles totaux, nombre de pousses par bourgeon

Effet de la salinité sur la croissance des deux variétés de bananier « grande naine » et « petite naine » et leur nutrition minérale au Maroc

Objectifs : Le travail présenté ici a pour objectif de décrire l’effet de la salinité sur la croissance et la distribution des principaux éléments minéraux chez le bananier. Méthodologie et résultats : L’étude de l’effet de la salinité a été réalisée sur deux variétés du bananier (Musa acuminata L), la grande naine et la petite naine issues de culture in vitro. Les variétés testées ont été combinées à quatre niveaux de chlorure de sodium (NaCl) à savoir : 0, 2, 4 et 6 grammes par litre (g/l). Les résultats ont montré la réduction de la biomasse aérienne et racinaire, une accumulation de potassium dans la partie aérienne et de sodium dans la partie racinaire, ainsi qu’une réduction d’absorption de calcium et de magnésium et une légère variation d’azote dès la concentration 4g/l. Une certaine tolérance aux sels a été notée chez la variété grande naine comparativement à la petite naine. Cette tolérance pourrait être liée à une bonne sélectivité vis-à vis du potassium. Les variétés étudiées se comportent comme des plantes exclusives. Conclusion et application de la recherche : Les caractéristiques ioniques de la variété de bananier, la grande naine, diffèrent de celles de la petite naine, Les résultats obtenus confirment que l’amélioration de la productivité des bananiers au Maroc ne peut être réalisée que par des travaux destinés à élaborer un matériel génétique adapté à partir de la variété grande naine. Le choix doit s’effectuer sur la base d’un grand nombre de caractères (morphologiques, biochimiques physiologiques) contrôlant les déterminismes génétiques complexes de la tolérance à la salinité. Mots clés : Bananier, chlorure de sodium, éléments minéraux

A Layered Cooperative Architecture and its Application in TeleTeaching Framework

Various cooperative applications have been developed like shared editors, teleconferences, and more generally multimedia applications. These applications are based on similar mechanisms. This paper presents a model of cooperation based on two layers. The application layer uses the services provided by the cooperative layer regardless what type of application is used. Some common points and their functionnalities are respectively explicated. In this project, Teleteaching will be used as an example to illustrate and validate the proposed layer. Automata describe the objects of layers and the behaviors. Keywords: Cooperation, Layered Model, Services, Specification, TeleTeaching, Finite State Machines. 1. Introduction The Computer Supported Cooperative Work (CSCW) [2,6,7] concerns systems that allow a group of users, geographically distributed, to work together in the most natural manner. This needs the management of the interface between a user and a machine and also between two users ..

Effects of Supplemental Irrigation and Nitrogen Applied on Yield and Yield Components of Bread Wheat at the Sais Region of Morocco

The objective of this study is to determinate the optimal growth stage for applying supplemental irrigation and nitrogen to enhance bread wheat (Triticum aestivum L.) productivity and water use efficiency under rainfed condition in Morocco. Field trial was conducted during two years (2007-2008 and 2008-2009). Three genotypes (G) of Moroccan bread wheat; Achtar, Arrehane and an Advanced Line II were combined with five nitrogen doses (N); 0, 40, 80, 120 and 160 kg N ha–1 and three water treatments (I); rainfed (I0), irrigated (60 mm) at 21 according to the Zadoks scale (I1) and irrigated (60 mm) at 59 according to the Zadoks scale (I2). Results in the drought year (2007-2008) show that I2 improves water use efficiency, grain yield and its components by 91 and 60% respectively, compared to I0 and I1 treatments. I2 has limited the effects of the Chergui –hot and dry wind coming from the Sahara- which has a negative impact on growth mainly at the end of the cycle of the crop. Grain yield and other yield components increased simultaneously with the increase in nitrogen inputs during the second 2008-2009 crop year which was rainy. The 120 kg N ha–1 dose allowed the highest results, with no significant difference with the yield obtained after a 160 kg N ha–1 input. Genotype appeared to have no significant effect neither on grain yield nor on water use efficiency in both the years

Genetic diversity and structure of a barley collection predominatly from North African region

The cultivated barley ranks fourth cereal crop in the world. The demand for higher-yielding, nutritious, and better-adapted crop varieties has increased the need to exploit genebanks diversity. Thus, assessing the genetic diversity of barley is essential to determine the genetic relationship between lines and deploy novel alleles in breeding programs. Here, we used 14 polymorphic Simple Sequence Repeat (SSR) markers to assess genetic diversity, genetic relationship, and population structure of 113 barley lines originated from 14 countries, and the majority from Africa. The average alleles per locus of 5.36, Polymorphism Information Content (PIC) of 0.57, and genetic diversity of 0.61 indicate a high degree of genetic variation in this collection. The Analysis of Molecular Variance AMOVA showed genetic variation within countries to be higher (74%) than among countries (26%). The STRUCTURE analysis was consistent with neighbor-joining clustering and principal components analysis, which identified three main subpopulations clustered primarily according to their geographic origin. Growing environments, migration between and within countries might have caused a considerable genetic diversity observed in the North African barley germplasm. The results of this study contribute to the conservation and utilization of these barley germplasm