Abdelkarim Filali-Maltouf

The Genotypic Population Structure of Mycobacterium tuberculosis Complex from Moroccan Patients Reveals a Predominance of Euro-American Lineages

Background Tuberculosis (TB) remains a major health problem in Morocco. Characterization of circulating Mycobacterium tuberculosis genotypic lineages, important to understand the dynamic of the disease, was hereby addressed for the first time at a national level. Methodology/Principal Findings Spoligotyping was performed on a panel of 592 M. tuberculosis complex strains covering a 2-year period (2004–2006). It identified 129 patterns: 105 (n = 568 strains) corresponded to a SIT number in the SITVIT2 database, while 24 patterns were labeled as orphan. A total of 523 (88.3%) strains were clustered vs. 69 or 11.7% unclustered. Classification of strains within 3 large phylogenetical groups was as follows: group 1– ancestral/TbD1+/PGG1 (EAI, Bovis, Africanum), group 2– modern/TbD1−/PGG1 group (Beijing, CAS), group 3– evolutionary recent/TbD1−/PGG2/3 (Haarlem, X, S, T, LAM; alternatively designated as the Euro-American lineage). As opposed to group 3 strains (namely LAM, Haarlem, and T) that predominated (86.5% of all isolates), 6 strains belonged to group 2 (Beijing n = 5, CAS n = 1), and 3 strains (BOV_1 n = 2, BOV_4-CAPRAE) belonged to ancestral group 1 (EAI and AFRI lineage strains were absent). 12-loci MIRU-VNTR typing of the Casablanca subgroup (n = 114 strains) identified 71 patterns: 48 MITs and 23 orphan patterns; it allowed to reduce the clustering rate from 72.8% to 29.8% and the recent transmission rate from 64% to 20.2%. Conclusion The M. tuberculosis population structure in Morocco is highly homogeneous, and is characterized by the predominance of the Euro-American lineages, namely LAM, Haarlem, and T, which belong to the “evolutionary recent” TbD1−/PGG2/3 phylogenetic group. The combination of spoligotyping and MIRUs decreased the clustering rate significantly, and should now be systematically applied in larger studies. The methods used in this study appear well suited to monitor the M. tuberculosis population structure for an enhanced TB management program in Morocco.

Molecular Typing of Mycobacterium Tuberculosis Complex by 24-Locus Based MIRU-VNTR Typing in Conjunction with Spoligotyping to Assess Genetic Diversity of Strains Circulating in Morocco.

Background Standard 24-locus Mycobacterial Interspersed Repetitive Unit Variable Number Tandem Repeat (MIRU-VNTR) typing allows to get an improved resolution power for tracing TB transmission and predicting different strain (sub) lineages in a community. Methodology During 2010–2012, a total of 168 Mycobacterium tuberculosis Complex (MTBC) isolates were collected by cluster sampling from 10 different Moroccan cities, and centralized by the National Reference Laboratory of Tuberculosis over the study period. All isolates were genotyped using spoligotyping, and a subset of 75 was genotyped using 24-locus based MIRU-VNTR typing, followed by first line drug susceptibility testing. Corresponding strain lineages were predicted using MIRU-VNTRplus database. Principal Findings Spoligotyping resulted in 137 isolates in 18 clusters (2–50 isolates per cluster: clustering rate of 81.54%) corresponding to a SIT number in the SITVIT database, while 31(18.45%) patterns were unique of which 10 were labelled as “unknown” according to the same database. The most prevalent spoligotype family was LAM; (n = 81 or 48.24% of isolates, dominated by SIT42, n = 49), followed by Haarlem (23.80%), T superfamily (15.47%), >Beijing (2.97%), > U clade (2.38%) and S clade (1.19%). Subsequent 24-Locus MIRU-VNTR typing identified 64 unique types and 11 isolates in 5 clusters (2 to 3isolates per cluster), substantially reducing clusters defined by spoligotyping only. The single cluster of three isolates corresponded to two previously treated MDR-TB cases and one new MDR-TB case known to be contact a same index case and belonging to a same family, albeit residing in 3 different administrative regions. MIRU-VNTR loci 4052, 802, 2996, 2163b, 3690, 1955, 424, 2531, 2401 and 960 were highly discriminative in our setting (HGDI >0.6). Conclusions 24-locus MIRU-VNTR typing can substantially improve the resolution of large clusters initially defined by spoligotyping alone and predominating in Morocco, and could therefore be used to better study tuberculosis transmission in a population-based, multi-year sample context.

Genetic Diversity within a Global Panel of Durum Wheat (Triticum durum) Landraces and Modern Germplasm Reveals the History of Alleles Exchange

Durum wheat is the 10th most important crop in the world, and its use traces back to the origin of agriculture. Unfortunately, in the last century only part of the genetic diversity available for this species has been captured in modern varieties through breeding. Here, the population structure and genetic diversity shared among elites and landraces collected from 32 countries was investigated. A total of 370 entries were genotyped with Axiom 35K array to identify 8,173 segregating single nucleotide polymorphisms (SNPs). Of these, 500 were selected as highly informative with a PIC value above 0.32 and used to test population structure via DAPC, STRUCTURE, and neighbor joining tree. A total of 10 sub-populations could be identified, six constituted by modern germplasm and four by landraces of different geographical origin. Interestingly, genomic comparison among groups indicated that Middle East and Ethiopia had the lowest level of allelic diversity, while breeding programs and landraces collected outside these regions were the richest in rare alleles. Further, phylogenetic analysis among landraces indicated that Ethiopia might represent a second center of origin of durum wheat, rather than a second domestication site as previously believed. Together, the analyses carried here provide a global picture of the available genetic diversity for this crop and shall guide its targeted use by breeders.

Recovery of symbiotic nitrogen fixing acacia rhizobia from Merzouga Desert sand dunes in South East Morocco – Identification of a probable new species of Ensifer adapted to stressed environments ☆

Bacteria capable of nodulating Acacia tortilis and A. gummifera could be recovered from sand dunes collected in the Moroccan Merzouga desert. The trapping approach enabled the recovery of 17 desert rhizobia that all clustered within the Ensifer (Sinorhizobium) genus. Four isolates of the dominant genotype comprising 15 strains as well as 2 divergent strains were further characterized by MLSA. Phylogenetic analyzes indicated that the dominant genetic type was belonging to a new and yet undefined species within the Ensifer genus. Interestingly, housekeeping gene phylogenies showed that this possibly new species is also present in another desert but in India. Phylogenetic analyses of nifH and nodC sequences showed high sequence conservation among the Moroccan strains belonging to the dominant genotype but high divergence with sequences from Indian isolates suggesting acquisition of symbiotic genes through Horizontal Gene Transfer. These desert rhizobia were capable of growing in media containing high salt concentrations, under high pH and most of the strains showed growth at 45°C. Only recovered from desert type of Biome, yet, this new taxon appears particularly adapted to such harsh environment.

Differential physiological and antioxidative responses to drought stress and recovery among four contrasting Argania spinosa ecotypes

ABSTRACT Our study was undertaken to ascertain whether the change of the water status and the activation of superoxide dismutase and their isoenzymes in Argan tree can support edaphic drought tolerance and its recovery under rehydration. An experiment was conducted on four contrasting ecotypes of Argania spinosa plants: two contrasting coastal ecotypes (Admine (Adm) and Rabia (Rab)) and two contrasting inland ecotypes (Aoulouz (Alz) and Lakhssas (Lks)). Drought stress significantly decreased the leaf water potential and stomatal conductance in the four contrasted ecotypes. In terms of biochemical responses, significant accumulation of carbonyl groups, hydrogen peroxide and superoxide radical has been recorded in the leaves of stressed plants reflecting oxidative stress. In parallel, the activities of total superoxide dismutase (SOD) and their isoenzymes Cu/Zn-SOD, Cu/Zn-SOD and Fe-SOD were also found to have increased to scavenging ROS and protecting the cell against induced oxidative stress. The recovery kinetics of A. spinosa, as a response to rehydration, were significant and rapid. According to the traits having the most discriminating power, both inland ecotypes (Lks and Alz) showed a better upregulation of its protective mechanisms compared to coastal ecotypes (Rab and Adm). All these adaptive traits make the inland ecotypes as an elite resource of drought tolerance and might become the new focus of domestication research of argan tree in arid and semi-arid environments.

Real-Time PCR for Measles Virus Detection on Clinical Specimens with Negative IgM Result in Morocco

Since the confirmation of measles cases represents an important indicator regarding the performance of the measles-elimination program, the aim of this study was to evaluate the effectiveness of the routine procedures followed in Morocco for the laboratory confirmation of measles cases. Suspected cases reported between January 2010 and December 2012 were assessed for the timeliness of the sample collection, occurrence of measles clinical symptoms, and the results of the laboratory diagnoses. For 88% of the 2,708 suspected cases, a clinical specimen was collected within 7d of rash onset, of which 50% were IgM-positive and 2.6% were equivocal. The measles symptoms were reported in 91.4% of the cases; the occurrence of symptoms showed a positive association with the serological results (odds ratio [OR] = 2.9883, 95% confidence interval [CI] 2.2238–4.0157). Of the negative samples, 52% (n = 116) tested positive by real-time polymerase chain reaction (PCR). These results are in favor of using molecular detection to complement serological diagnosis in the context of measles surveillance approach in Morocco. In addition, the introduction of additional laboratory methods for differential diagnosis is required for the final classification of suspected cases with maculopapular rash and fever in the context of the measles elimination program.

Transcriptome Changes in Hirschfeldia incana in Response to Lead Exposure

Hirschfeldia incana, a pseudometallophyte belonging to the Brassicaceae family and widespread in the Mediterranean region, was selected for its ability to grow on soils contaminated by lead (Pb). The global comparison of gene expression using microarrays between a plant susceptible to Pb (Arabidopsis thaliana) and a Pb tolerant plant (H. incana) enabled the identification of a set of specific genes expressed in response to lead exposure. Three groups of genes were particularly over-represented by the Pb exposure in the biological processes categorized as photosynthesis, cell wall, and metal handling. Each of these gene groups was shown to be directly involved in tolerance or in protection mechanisms to the phytotoxicity associated with Pb. Among these genes, we demonstrated that MT2b, a metallothionein gene, was involved in lead accumulation, confirming the important role of metallothioneins in the accumulation and the distribution of Pb in leaves. On the other hand, several genes involved in biosynthesis of ABA were shown to be up-regulated in the roots and shoots of H. incana treated with Pb, suggesting that ABA-mediated signaling is a possible mechanism in response to Pb treatment in H. incana. This latest finding is an important research direction for future studies.

Electrolyte ions and glutathione enzymes as stress markers in Argania spinosa subjected to drought stress and recovery

Understanding the mechanisms underlying Argania spinosa responses to drought stress is essential for its regeneration and domestication. Toward that end, an integrative study of tolerance responses to drought stress in four A. spinosa ecotypes (2 contrasting coastal ecotypes (Adm and Rab) and 2 contrasting inland ecotypes (Alz and Lks)) have been conducted. Responses to soil drying and re -watering were measured at physiological and biochemical levels. Soil drying resulted in significant increase in leaf concentrations of potassium (K + ), calcium (Ca 2+ ) and magnesium (Mg 2+ ) with differential responses between ecotypes. The glutathione-related enzymes: glutathione peroxidase (GP), glutathione reductase (GR) and glutathione S-transferase (GST) showed a significant increase in their enzymatic activity in A. spinosa plants subjected to drought stress. Additionally, a significant increase in thiol protein content in the four ecotypes was recorded, during drought stress. These antioxidant traits responded differently depending on ecotype. However, rapid and significant changes in the studied physiological and biochemical traits were observed during recovery from drought, only after four days. According to the traits having the most discriminating power, the both inland ecotypes, especially Lks ecotype, seem to be potential candidates for regeneration of argan forest and their domestication in arid and semi-arid environments. Key words : Argania spinosa, drought stress, glutathione enzymes, thiol compounds, recovery.

Diversity and spatial genetic structure of natural Moroccan Quercus susber L. assessed by ISSR markers for conservation

Morocco is one of the most important regions of the world in terms of Quercus suber L. number and variation. This species is in decline due to several factors, which can lead to permanent loss of this resource. It would be essential to evaluate the genetic diversity in order to conserve maximum genetic variability of this species. The genetic diversity and differentiation of 16 sites from five regions representing the entire range of Moroccan Cork Oak were assessed. Twenty-three ISSR primers used generated 985 polymorphic fragments with an average of 42.8 bands per primer and showed 100% of polymorphism. The 173 individuals revealed a moderate level of genetic diversity at species level (I = 0.27, He = 0.161). The AMOVA showed that the highest level of diversity occurred within populations (64%), this was also confirmed by the coefficient of differentiation (Gst = 0.47). The estimated gene flow (Nm = 0.56) and the Mantel test revealed a significant correlation between geographic and genetic diversity (r = 0.266; p = 0.001). This genetic structure was further shown by the topology of the UPGMA, sPCA and STRUCTURE analysis. In addition, a core collection of 34 genotypes was established representing the essential diversity detected. This research advocates populations and individuals to preserve in order to improve and conserve this resource in the future.

Seaweed polysaccharides as bio-elicitors of natural defenses in olive trees against verticillium wilt of olive

ABSTRACT Ulvan, carrageenan, alginate and laminarin were tested in olive trees’ twigs to elicit phenolic metabolism and control verticillium wilt of olive (VWO) caused by Verticillium dahliae. The elicitation effect was determined through phenylalanine ammonia-lyase activity, total polyphenol content and lignin content. VWO was assessed in twigs previously elicited (24 h) and maintained in a solution containing bio-elicitors (2 g/L) and conidial suspension (106 conidia/mL). Our results showed stimulation of the phenolic metabolism and the decline of wilt symptoms. Ulvan reduced significantly the area under the disease progress curve for severity to 39.9% and the final incidence to 28.9%. Ulvan and alginate produced significant inhibitory rates on mycelial growth of the fungus in vitro. Seaweed polysaccharides might help to overcome VWO by strengthening the host defense metabolism and restricting the pathogen’s growth.