Does geographic origin dictate ecological strategies in Acacia senegal (L.) Willd.? Evidence from carbon and nitrogen stable isotopes

Background and aims

Acacia senegal, a leguminous dryland tree, is economically and ecologically important to sub-Saharan Africa. Water-use efficiency (WUE) and biological nitrogen fixation (BNF) are fundamental to plant productivity and survival. We quantify provenance differences in WUE, BNF, photosynthesis, biomass and gum arabic production from A. senegal assessing genetic improvement potential.

Methods

Using stable isotope ratios, we determined WUE (δ¹³C) and BNF (δ¹⁵N) from provenances of mature A. senegal in field-trials (Senegal), sampling leaves at the beginning (wet) and end (dry) of the rainy season. Seedling provenance trials (UK) determined photosynthesis, and biomass and δ¹³C in relation to water table. Environmental data were characterised for all provenances at their sites of origin.

Results

Provenances differed in both δ¹³C and δ¹⁵N. Gum yield declined with increasing WUE. Virtually no BNF was detected during the dry season and seedlings and mature trees may have different WUE strategies. Wind speed and soil characteristics at provenance origin were correlated with isotope composition and gum production.

Conclusion

Provenance differences suggest that selection for desirable traits, e.g., increased gum production, may be possible. As ecological strategies relate to native locality, the environmental conditions at plantation site and provenance origin are important in assessing selection criteria.     

The genetic basis of drought tolerance in the high oil crop Sesamum indicum

Unlike most of the important food crops, sesame can survive drought but severe and repeated drought episodes, especially occurring during the reproductive stage, significantly curtail the productivity of this high oil crop. Genome‐wide association study was conducted for traits related to drought tolerance using 400 diverse sesame accessions, including landraces and modern cultivars. Ten stable QTLs explaining more than 40% of the phenotypic variation and located on four linkage groups were significantly associated with drought tolerance related traits. Accessions from the tropical area harboured higher numbers of drought tolerance alleles at the peak loci and were found to be more tolerant than those from the northern area, indicating a long‐term genetic adaptation to drought‐prone environments. We found that sesame has already fixed important alleles conferring survival to drought which may explain its relative high drought tolerance. However, most of the alleles crucial for productivity and yield maintenance under drought conditions are far from been fixed. This study also revealed that pyramiding the favourable alleles observed at the peak loci is of high potential for enhancing drought tolerance in sesame. In addition, our results highlighted two important pleiotropic QTLs harbouring known and unreported drought tolerance genes such as SiABI4, SiTTM3, SiGOLS1, SiNIMIN1 and SiSAM. By integrating candidate gene association study, gene expression and transgenic experiments, we demonstrated that SiSAM confers drought tolerance by modulating polyamine levels and ROS homeostasis, and a missense mutation in the coding region partly contributes to the natural variation of drought tolerance in sesame.     

Trait discovery and editing in tomato

Tomato (Solanum lycopersicum), which is used for both processing and fresh markets, is a major crop species that is the top ranked vegetable produced over the world. Tomato is also a model species for research in genetics, fruit development and disease resistance. Genetic resources available in public repositories comprise the 12 wild related species and thousands of landraces, modern cultivars and mutants. In addition, high quality genome sequences are available for cultivated tomato and for several wild relatives, hundreds of accessions have been sequenced, and databases gathering sequence data together with genetic and phenotypic data are accessible to the tomato community. Major breeding goals are productivity, resistance to biotic and abiotic stresses, and fruit sensorial and nutritional quality. New traits, including resistance to various biotic and abiotic stresses and root architecture, are increasingly being studied. Several major mutations and quantitative trait loci (QTLs) underlying traits of interest in tomato have been uncovered to date and, thanks to new populations and advances in sequencing technologies, the pace of trait discovery has considerably accelerated. In recent years, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing (GE) already proved its remarkable efficiency in tomato for engineering favorable alleles and for creating new genetic diversity by gene disruption, gene replacement, and precise base editing. Here, we provide insight into the major tomato traits and underlying causal genetic variations discovered so far and review the existing genetic resources and most recent strategies for trait discovery in tomato. Furthermore, we explore the opportunities offered by CRISPR/Cas9 and their exploitation for trait editing in tomato.     

Quality of Sterile Male Tsetse after Long Distance Transport as Chilled,Irradiated Pupae

Background

Tsetse flies transmit trypanosomes that cause human and African animal trypanosomosis, a debilitating disease of humans (sleeping sickness) and livestock (nagana). An area-wide integrated pest management campaign against Glossina palpalis gambiensis has been implemented in Senegal since 2010 that includes a sterile insect technique (SIT) component. The SIT can only be successful when the sterile males that are destined for release have a flight ability, survival and competitiveness that are as close as possible to that of their wild male counterparts.

Methodology/Principal Findings

Tests were developed to assess the quality of G. p. gambiensis males that emerged from pupae that were produced and irradiated in Burkina Faso and Slovakia (irradiation done in Seibersdorf, Austria) and transported weekly under chilled conditions to Dakar, Senegal. For each consignment a sample of 50 pupae was used for a quality control test (QC group). To assess flight ability, the pupae were put in a cylinder filtering emerged flies that were able to escape the cylinder. The survival of these flyers was thereafter monitored under stress conditions (without feeding). Remaining pupae were emerged and released in the target area of the eradication programme (RF group). The following parameter values were obtained for the QC flies: average emergence rate more than 69%, median survival of 6 days, and average flight ability of more than 35%. The quality protocol was a good proxy of fly quality, explaining a large part of the variances of the examined parameters.

Conclusions/Significance

The quality protocol described here will allow the accurate monitoring of the quality of shipped sterile male tsetse used in operational eradication programmes in the framework of the Pan-African Tsetse and Trypanosomosis Eradication Campaign.     

Profiling the Succession of Bacterial Communities throughout the Life Stages of a Higher Termite Nasutitermes arborum (Termitidae,Nasutitermitinae) Using 16S rRNA Gene Pyrosequencing

Previous surveys of the gut microbiota of termites have been limited to the worker caste. Termite gut microbiota has been well documented over the last decades and consists mainly of lineages specific to the gut microbiome which are maintained across generations. Despite this intimate relationship, little is known of how symbionts are transmitted to each generation of the host, especially in higher termites where proctodeal feeding has never been reported. The bacterial succession across life stages of the wood-feeding higher termite Nasutitermes arborum was characterized by 16S rRNA gene deep sequencing. The microbial community in the eggs, mainly affiliated to Proteobacteria and Actinobacteria, was markedly different from the communities in the following developmental stages. In the first instar and last instar larvae and worker caste termites, Proteobacteria and Actinobacteria were less abundant than Firmicutes, Bacteroidetes, Spirochaetes, Fibrobacteres and the candidate phylum TG3 from the last instar larvae. Most of the representatives of these phyla (except Firmicutes) were identified as termite-gut specific lineages, although their relative abundances differed. The most salient difference between last instar larvae and worker caste termites was the very high proportion of Spirochaetes, most of which were affiliated to the Treponema Ic, Ia and If subclusters, in workers. The results suggest that termite symbionts are not transmitted from mother to offspring but become established by a gradual process allowing the offspring to have access to the bulk of the microbiota prior to the emergence of workers, and, therefore, presumably through social exchanges with nursing workers.     

A 20-Amino Acid Module of Protein Kinase C? Involved in Translocation and Selective Targeting at Cell-Cell Contacts

In the pituitary gland, activated protein kinase C (PKC) isoforms accumulate either selectively at the cell-cell contact (α and ϵ) or at the entire plasma membrane (β1 and δ). The molecular mechanisms underlying these various subcellular locations are not known. Here, we demonstrate the existence within PKCϵ of a cell-cell contact targeting sequence (3CTS) that, upon stimulation, is capable of targeting PKCδ, chimerin-α1, and the PKCϵ C1 domain to the cell-cell contact. We show that this selective targeting of PKCϵ is lost upon overexpression of 3CTS fused to a (R-Ahx-R)₄ (where Ahx is 6-aminohexanoic acid) vectorization peptide, reflecting a dominant-negative effect of the overexpressed 3CTS on targeting selectivity. 3CTS contains a putative amphipathic α-helix, a 14-3-3-binding site, and the Glu-374 amino acid, involved in targeting selectivity. We show that the integrity of the α-helix is important for translocation but that 14-3-3 is not involved in targeting selectivity. However, PKCϵ translocation is increased when PKCϵ/14-3-3 interaction is abolished, suggesting that phorbol 12-myristate 13-acetate activation may initiate two sets of PKCϵ functions, those depending on 14-3-3 and those depending on translocation to cell-cell contacts. Thus, 3CTS is involved in the modulation of translocation via its 14-3-3-binding site, in cytoplasmic desequestration via the α-helix, and in selective PKCϵ targeting at the cell-cell contact via Glu-374.Activation of cytoplasmic kinases often induces their targeting to various subcellular locations where they phosphorylate their substrates and exert their biological functions. Representative examples of proteins for which targeting involves complex and various molecular mechanisms are provided by the protein kinase C (PKC)⁶ family, which comprises 10 known isoforms, displaying ubiquitous, tissue- or cell type-specific expression and playing crucial roles in signal transduction (1, 2). Depending on the cell type and the stimulus, various inactive cytoplasmic PKC isoforms may, upon activation, associate with the plasma, Golgi, or nuclear membranes (3–5). Even within a given cell type, a particular isoform can be targeted and accumulated at various subcellular locations (6, 7), and these processes involve direct interaction with phospholipids or other proteins (8, 9).In pituitary GH3B6 cells, PKC isoforms accumulate at different subcellular locations upon phorbol 12-myristate 13-acetate (PMA) stimulation or thyrotropin-releasing hormone (TRH) receptor activation (10, 11). Activated PKCα and -ϵ accumulate selectively at cell-cell contacts, whereas PKCβ1 and -δ are detected along the entire plasma membrane. The selective partitioning of specific PKC isoforms at cell-cell contacts is not restricted to the GH3B6 cell line. It was also observed in blastocysts (12), in the pituitary gland (11), at heterotypic contacts between fibroblasts and epithelial cells (13), at the interface between macrophages and IgG-coated beads (14), and at the immunological synapse (15–17). Although the molecular mechanism underlying this partitioning remains largely unknown, an interesting clue was provided by the discovery in human pituitary and thyroid tumors of a natural PKCα D294G mutant (18, 19), which is devoid of cell-cell contact targeting selectivity (20). A similar loss of selectivity is found when an E374G mutation is introduced in PKCϵ (11), indicating that the Asp-294 and Glu-374 amino acids located within the V3 region of PKCα and ϵ, respectively, are essential for proper targeting after activation. Interestingly, the PKCα D294G mutant was also shown to be a loss-of-function mutant (21). However, because replacing Phe by Glu in the corresponding position does not induce the targeting of PKCδ to the cell-cell contact, it is likely that other amino acids are required for cell-cell contact targeting selectivity.In the present work, we sought to deepen our understanding of the requirements for efficient targeting to the cell-cell contact by focusing our analysis on the sequence surrounding position Asp-294 of PKCα and Glu-374 of PKCϵ. On the basis of isoform sequence comparison, we identified a 20-aa stretch in the V3 region of PKCϵ that includes Glu-374 and contains one of the two 14-3-3-binding sites of PKCϵ and a putative amphipathic α-helix. This 20-aa module fulfills the criteria of a cell-cell contact targeting sequence, and we therefore propose to name this sequence 3CTS.     

Study of NAT2 genetic polymorphism in West African subjects: example of an healthy non-smoker Senegalese population

The NAT2 genetic polymorphism determines the individual acetylator status and, consequently, the capacity to metabolize, or not, drugs and xenobiotics which are substrates of NAT2. As the nature and frequency of the NAT2 polymorphisms vary remarkably between populations of different ethnic origins, genotyping strategies used to predict the acetylation phenotype need to be adapted for each particular population regarding their genetic backgrounds at this locus. As few data on the genetic polymorphism of NAT2 are available in the Senegalese population, we performed an extensive identification of NAT2 variants in 105 healthy non-smoker Senegalese subjects by direct PCR sequencing of the coding region. Eleven previously described SNPs were identified in this Senegalese population. Upon allele analysis, the four most frequent alleles were of the NAT2*5- (35.7?%), NAT2*6- (21.0?%), NAT2*12- (16.7?%) and NAT2*14- (10.0?%) type, the remaining alleles, including the wild-type NAT2*4, having each a frequency lower than 10?%. According to the observed genotypes, 51 and 50 subjects were predicted to be of the rapid (48.6?%) and slow (47.6?%) acetylator phenotype, respectively, while four individuals (3.8?%) were considered of unknown phenotype as they carry at least one allele with a yet unknown functional effect. These baseline data would be of particular interest to set up an efficient genotyping strategy to predict the acetylation status of Senegalese patients with tuberculosis and, thus, to optimize their isoniazid treatment.     

Genetic relationship of cowpea (Vigna unguiculata) varieties from Senegal based on SSR markers

Genetic diversity and phylogenetic relationships among 22 local cowpea (Vigna unguiculata) varieties and inbred lines collected throughout Senegal were evaluated using simple sequence repeat molecular markers. A set of 49 primer combinations were developed from cowpea genomic/expressed sequence tags and evaluated for their ability to detect polymorphisms among the various cowpea genotypes. Forty-four primer combinations detected polymorphisms, with the remaining five primer sets failing to yield PCR amplification products. From one to 16 alleles were found among the informative primer combinations; their frequencies ranged from 0.60 to 0.95 (mean = 0.79). The genetic diversity of the sample varied from 0.08 to 0.42 (mean = 0.28). The polymorphic information content ranged from 0.08 to 0.33 (mean = 0.23). The local varieties clustered in the same group, except 53-3, 58-53, and 58-57; while Ndoute yellow pods, Ndoute violet pods and Baye Ngagne were in the second group. The photosensitive varieties (Ndoute yellow pods and Ndoute violet pods) were closely clustered in the second group and so were inbred line Mouride and local cultivar 58-57, which is also one of the parents for inbred line Mouride. These molecular markers could be used for selection and identification of elite varieties for cowpea improvement and germplasm management in Senegal.