Uvitex2B: a rapid and efficient stain for detection of arbuscular mycorrhizal fungi within plant roots

The study of arbuscular mycorrhiza often requires the staining of fungal structures using specific dyes. Fluorescent dyes such as acid fuchsin and wheat germ agglutinin conjugates give excellent results, but these compounds are either hazardous or very expensive. Here, we show that a safer and inexpensive dye, Uvitex2B, can be efficiently used to stain intraradical fungal structures formed by the arbuscular mycorrhizal fungus Glomus intraradices in three plant species: carrot, Casuarina equisetifolia, and Medicago truncatula. The intensity and stability of Uvitex2B allow the acquisition of high-quality images using not only confocal laser scanning microscopy but also epifluorescence microscopy coupled with image deconvolution. Furthermore, we demonstrate that Uvitex2B and β-glucuronidase staining are compatible and can thus be used to reveal arbuscular mycorrhizal structures in the context of promoter activation analysis.     

Somatic deletions of genes regulating MSH2 protein stability cause DNA mismatch repair deficiency and drug resistance in human leukemia cells

DNA mismatch repair enzymes (for example, MSH2) maintain genomic integrity, and their deficiency predisposes to several human cancers and to drug resistance. We found that leukemia cells from a substantial proportion of children (～11%) with newly diagnosed acute lymphoblastic leukemia have low or undetectable MSH2 protein levels, despite abundant wild-type MSH2 mRNA. Leukemia cells with low levels of MSH2 contained partial or complete somatic deletions of one to four genes that regulate MSH2 degradation (FRAP1 (also known as MTOR), HERC1, PRKCZ and PIK3C2B); we also found these deletions in individuals with adult acute lymphoblastic leukemia (16%) and sporadic colorectal cancer (13.5%). Knockdown of these genes in human leukemia cells recapitulated the MSH2 protein deficiency by enhancing MSH2 degradation, leading to substantial reduction in DNA mismatch repair and increased resistance to thiopurines. These findings reveal a previously unrecognized mechanism whereby somatic deletions of genes regulating MSH2 degradation result in undetectable levels of MSH2 protein in leukemia cells, DNA mismatch repair deficiency and drug resistance.     

Genetic variation of Casuarina equisetifolia subsp equisetifolia and C. equisetifolia subsp incana populations on the northern coast of Senegal

The genetic variation of 70 individual samples of Casuarina equisetifolia (L. Johnson) subsp equisetifolia and C. equisetifolia subsp incana growing along the northern coast of Senegal was analyzed with RAPD markers. Of the 160 primers tested, five were chosen; they generated 1396 reproducible bands and 61 polymorphic bands that were scored. This result showed a narrow genetic variation among (4.36%) and within (5.90%) C. equisetifolia subsp equisetifolia and C. equisetifolia subsp incana plantation sites. The genetic variation at each site revealed a high degree of polymorphism in Potou (5.90%) and low diversity in Retba (3.06%). In the dendrogram analyses, each sampling site was formed by two main groups. Similar results were found for the dendrograms based on the RAPD data gathered from the five different sites. These dendrograms revealed several polytomies in one of the subgroups, suggesting replication of the same specimens in different sites along the Senegalese coast. The RAPD data support the hypothesis that these populations are of the same provenance, subject to hybridization and inbreeding depression.     

Ground‐ and satellite‐based evidence of the biophysical mechanisms behind the greening Sahel

After a dry period with prolonged droughts in the 1970s and 1980s, recent scientific outcome suggests that the decades of abnormally dry conditions in the Sahel have been reversed by positive anomalies in rainfall. Various remote sensing studies observed a positive trend in vegetation greenness over the last decades which is known as the re‐greening of the Sahel. However, little investment has been made in including long‐term ground‐based data collections to evaluate and better understand the biophysical mechanisms behind these findings. Thus, deductions on a possible increment in biomass remain speculative. Our aim is to bridge these gaps and give specifics on the biophysical background factors of the re‐greening Sahel. Therefore, a trend analysis was applied on long time series (1987–2013) of satellite‐based vegetation and rainfall data, as well as on ground‐observations of leaf biomass of woody species, herb biomass, and woody species abundance in different ecosystems located in the Sahel zone of Senegal. We found that the positive trend observed in satellite vegetation time series (+36%) is caused by an increment of in situ measured biomass (+34%), which is highly controlled by precipitation (+40%). Whereas herb biomass shows large inter‐annual fluctuations rather than a clear trend, leaf biomass of woody species has doubled within 27 years (+103%). This increase in woody biomass did not reflect on biodiversity with 11 of 16 woody species declining in abundance over the period. We conclude that the observed greening in the Senegalese Sahel is primarily related to an increasing tree cover that caused satellite‐driven vegetation indices to increase with rainfall reversal.     

Variability of key biological parameters of round sardinella Sardinella aurita and the effects of environmental changes

We examined growth rates and reproductive characteristics of Sardinella aurita off Senegal and other coastal areas over a 20 year period (1995–2014) to determine how they relate to variations in environmental characteristics of coastal waters. Based on fish length-frequency data and a coastal upwelling index, we found that S. aurita recruitment tends to occur during the periods of most intensive upwelling (March–April off Senegal). Peak reproduction corresponds to periods of low sea-surface temperature (in February or March). The sex ratio was remarkably consistent during the 30 year study period and so was not affected by environmental changes. We hypothesise that S. aurita takes advantage of the higher zooplankton productivity that occurs in coastal waters when upwelling brings nutrient-rich water to the surface (i.e., it increases its growth rate and accumulates energy reserves for spawning). Growth performance appears to be strongly dependent on environmental conditions. The timing of spawning seems to occur when food (zooplankton) is most available for supplying the energy requirements needed by adults for spawning and early development of larvae. Environmental changes seem to have a significant effect on S. aurita growth and reproduction, which endorses their high phenotypic plasticity.     

Phylogenetic signal in matK vs. trnK: a case study in early diverging eudicots (angiosperms)

The matK gene has been among the most useful loci for resolving plant phylogenetic relationships at different evolutionary time-scales, but much less is known about the phylogenetic utility of the flanking trnK intron, especially for deep level phylogenetics. We compared the relative performance of matK and trnK intron regions for resolving the relationships of the early diverging eudicots (angiosperms). The two regions display similar nucleotide compositions and distributions of rate variation among sites. The trnK intron sequences also provide similar levels of phylogenetic information per-site as matK. Combining the trnK intron sequences with matK increases overall bootstrap support for the early diverging eudicots compared to analyses of matK alone. MP, ML and Bayesian analyses provide strong support for eudicots, the sister group relationship of Ranunculales to remaining eudicots, and a Buxales+Trochodendraceae+core eudicots clade. matK and the trnK intron support conflicting positions for Buxales and Trochodendrales in relation to the core eudicots.     

Description of two new species of Nippostrongylinae (Nematoda, Trichostrongyuna, Heligmosomoidea) coparasites in Arvicanthis ansorgei (Muridae) from Cameroon

In the small intestine of a single Arvicanthis ansorgei from Cameroon, two new species of Nippostrongylinae were found: Neoheligmonella zero n. sp. and Heligmonina comerounensis n. sp. N. zero belongs to the Neoheligmonella species in which the right dorsal ridge is poorly developed. Among these species, N. bainae (Durette-Desset, 1970), a parasite of Steatomys opinus from Burkina Fasso, is a closely related species. It is differentiated by the presence of vulvar alae, the ratio uterus length/body length and the separation of rays 2 and 3 at two thirds of their length. N. zero is also closely related to N. kenyae (Yeh, 1958) a parasite of Rattus rattus kijabius from Kenya by the pattern of the caudal bursa and the ratio spicules length/body length. The synlophe of N. kenyae was not described in detail but it is differentiated from N. zero by the position of the excretory pore, situated just posteriorly to the nerve ring and in the female by the ratio ovejector length/body length which is smaller. H. camerounensis n. sp. is characterised by the ratio dorsal ridges/ventral ridges which is 4/7. It is differentiated from the species of which the synlophe has not been described by the pattern of the caudal bursa (type 1-4 with tendancy 1-3-1). It is the first report of Nippostrongylinae species in Cameroon and the first record of a species of the genus Heligmonina in an Arvicanthis.     

Phylogenetic relationships in Termitomyces (Family Agaricaceae) based on the nucleotide sequence of ITS: a first approach to elucidate the evolutionary history of the symbiosis between fungus-growing termites and their fungi

Termitomyces constitutes a very poorly known genus of fungi whose essential characteristic is that all representatives of the genus are cultivated by termites (Macrotermitinae) in their nest and that all the fungi cultivated by termites belong to this genus. For the first time, the phylogenetic relationships of several African Termitomyces species was studied by the sequencing of their internal transcriber spacer region (ITS1--5.8S--ITS2). It appeared that this group is clearly monophyletic and belongs to the Tricholomataceae family. The total homology of the ITS zone of several Termitomyces symbionts of different termite genera indicated that the specific diversity of this group is in fact less important than previously supposed. Finally, the comparison between the Termitomyces phylogenetic tree and the taxonomic tree of Macrotermitinae showed that if for certain genera the hypothesis of termite/fungus coevolution is acceptable, it should not be applied for all symbiosis.     

Impact of rhizobial inoculation on Acacia senegal (L.) Willd. growth in greenhouse and soil functioning in relation to seed provenance and soil origin

Rhizobial inoculation has a positive impact on plants growth; however, there is little information about its effect on soil microbial communities and their activity in the rhizosphere. It was therefore necessary to test the effect of inoculation of Acacia senegal (L.) Willd. seedlings with selected rhizobia on plant growth, structure and diversity of soil bacterial communities and soil functioning in relation to plant provenance and soil origin. In order to carry out this experiment, three A. senegal seeds provenance from Kenya, Niger, and Senegal were inoculated with selected rhizobial strains. They have been further grown during 4 months in greenhouse conditions in two non-disinfected soils, Dahra and Goudiry coming respectively from arid and semi-arid areas. The principal component analysis (ACP) showed an inoculation effect on plant growth, rhizospheric bacterial diversity and soil functioning. However, the performances of the rhizobial strains varied in relation to the seed provenance and the soil origin. The selected rhizobial strains, the A. senegal provenance and the soil origin have modified the structure and the diversity of soil bacterial communities as measured by principal component analysis/denaturing gradient gel electrophoresis analyses. It is interesting to note that bacterial communities of Dahra soil were highly structured according to A. senegal provenance, whereas they were structured in relation to rhizobial inoculation in Goudiry soil. Besides, the impact of inoculation on soil microbial activities measured by fluorescein diacetate analyses varied in relation to plant provenance and soil origin. Nevertheless, total microbial activity was about two times higher in Goudiry, arid soil than in Dahra, semi-arid soil. Our results suggest that the rhizobial inoculation is a suitable tool for improving plants growth and soil fertility. Yet, the impact is dependent on inoculants, plant provenance and soil origin. It will, therefore, be crucial to identify the appropriate rhizobial strains and plant provenance or species in relation to the soil type.     

Methylation analysis revealed salicylic acid affects pearl millet defense through external cytosine DNA demethylation

The cytosine DNA methylation and demethylation have a role in regulating plant responses to the environment by affecting the promoter regions of most plant defense-related genes through the CpG islands or the CCGG motifs. Salicylic acid, a defense and signaling plant hormone, is seen playing crucial role in the variation of the methylome. In this study, the effects of salicylic acid and feeding of the millet headminer (Heliocheilus albipunctella de Joannis) on pearl millet DNA methylome changes were evaluated through MSAP epigenotyping during panicle development. The results showed that millet headminer feeding increased the level of genomic methylation while application of salicylic acid caused DNA demethylation occurring mostly at external cytosine and accompanied by a decrease of the number of larvae per panicle. This suggests that hemimethylation (external cytosine methylation) has key role in regulating defense responses and conferring tolerance to pearl millet through salicylic acid application.