In order to develop a rapid and specific detection test for bacteria in soil, we improved a method based on the polymerase chain reaction (PCR). Each step of the protocol, including direct lysis of cells, DNA purification, and PCR amplification, was optimized. To increase the efficiency of lysis, a step particularly critical for some microorganisms which resist classical techniques, we used small soil samples (100 mg) and various lytic treatments, including sonication, microwave heating, and thermal shocks. Purification of nucleic acids was achieved by passage through up to three Elutip d columns. Finally, PCR amplifications were optimized via biphasic protocols using booster conditions, lower denaturation temperatures, and addition of formamide. Two microorganisms were used as models: Agrobacterium tumefaciens, which is naturally absent from the soil used and was inoculated to calibrate the validity of the protocol, and Frankia spp., an actinomycete indigenous to the soil used. Specific primers were characterized either in the plasmid-borne vir genes for A. tumefaciens or in the variable regions of the 16S ribosomal gene for Frankia spp. Specific detection of the inoculated A. tumefaciens strain was routinely obtained when inocula ranged from 107 to 103 cells. Moreover, the strong correlation we observed between the size of the inocula and the results of the PCR reactions permitted assessment of the validity of the protocol in enumerating the number of microbial cells present in a soil sample. This allowed us to estimate the indigenous population of Frankia spp. at 0.2 x 105 genomes (i.e., amplifiable target sequences) per g of soil.

Academics researchers and "citizen scientists" from 22 countries confirmed that yellow mealworms, the larvae of Tenebrio molitor Linnaeus, can survive by eating polystyrene (PS) foam. More detailed assessments of this capability for mealworms were carried out by12 sources: five from the USA, six from China, and one from Northern Ireland. All of these mealworms digested PS foam. PS mass decreased and depolymerization was observed, with appearance of lower molecular weight residuals and functional groups indicative of oxidative transformations in extracts from the frass (insect excrement). An addition of gentamycin (30 mg g), a bactericidal antibiotic, inhibited depolymerization, implicating the gut microbiome in the biodegradation process. Microbial community analyses demonstrated significant taxonomic shifts for mealworms fed diets of PS plus bran and PS alone. The results indicate that mealworms from diverse locations eat and metabolize PS and support the hypothesis that this capacity is independent of the geographic origin of the mealworms, and is likely ubiquitous to members of this species.

Selection of modern varieties has typically been performed in standardized, high fertility systems with a primary focus on yield. This could have contributed to the loss of plant genes associated with efficient nutrient acquisition strategies and adaptation to soil-related biotic and abiotic stresses if such adaptive strategies incurred a cost to the plant that compromised yield. Furthermore, beneficial interactions between plants and associated soil organisms may have been made obsolete by the provision of nutrients in high quantity and in readily plant available forms. A review of evidence from studies comparing older traditional varieties to modern high yielding varieties indeed showed that this has been the case. Given the necessity to use scarce and increasingly costly fertilizer inputs more efficiently while also raising productivity on poorer soils, it will be crucial to reintroduce desirable rhizosphere-related traits into elite cultivars. Traits that offer possibilities for improving nutrient acquisition capacity, plantmicrobe interactions and tolerance to abiotic and biotic soil stresses in modern varieties were reviewed. Despite the considerable effort devoted to the identification of suitable donors and of genetic factors associated with these beneficial traits, progress in developing improved varieties has been slow and has so far largely been confined to modifications of traditional breeding procedures. Modern molecular tools have only very recently started to play a rather small role. The few successful cases reviewed in this paper have shown that novel breeding approaches using molecular tools do work in principle. When successful, they involved close collaboration between breeders and scientists conducting basic research, and confirmation of phenotypes in field tests as a 'reality check'. We concluded that for novel molecular approaches to make a significant contribution to breeding for rhizosphere related traits it will be essential to narrow the gap between basic sciences and applied breeding through more interdisciplinary research that addresses rather than avoids the complexity of plant-soil interactions.

A Pseudomonas 2,4-diacetylphloroglucinol (DAPG)-producing population that occurred naturally on the roots, in rhizosphere soil of Zea mays and in the nonrhizosphere soil was investigated in order to assess the microbial diversity at five stages of plant growth. A total of 1,716 isolates were obtained, and 188 of these isolates were able to produce DAPG. DAPG producers were isolated at each stage of plant growth, indicating that the maize rhizosphere is colonized by natural DAPG producers throughout development. The frequency of DAPG producers was very low in the first stage of plant growth and increased over time. An analysis of the level of biodiversity of the DAPG producers at the species level was performed by comparing the AluI restriction patterns of the 16S ribosomal DNAs (rDNAs) amplified by PCR from 167 isolates. This comparison allowed us to cluster the isolates into four amplified rDNA restriction analysis (ARDRA) groups, and the main group (ARDRA group 1) contained 89.8% of the isolates. The diversity of the 150 isolates belonging to ARDRA group 1 was analyzed by the random amplified polymorphic DNA (RAPD) technique. An analysis of RAPD patterns by a molecular variance method revealed that there was a high level of genetic diversity in this population and that the genetic diversity was related to plant age. Finally, we found that some of the DAPG producers, which originated from all stages of plant growth, had the same genotype. These DAPG producers could be exploited in future screening programs for biocontrol agents.Some rhizobacteria, which are commonly called plant growth-promoting rhizobacteria, interact with plant roots and protect the roots against pathogenic microorganisms (26). Pseudomonads are common members of the plant growthpromoting rhizobacterial microflora in the rhizospheres of protected plants (13,25,42). The ability of pseudomonads to suppress soilborne fungal pathogens depends on their ability to produce antibiotic metabolites, such as pyoluteorin, pyrrolnitrin, phenazine-1-carboxylic acid, and 2,4-diacetylphloroglucinol (DAPG) (14,32,38,47,49,55,57). One of these metabolites, DAPG, is a major factor in the biological control of a range of plant pathogens (7,23,24,57). The antibiotic DAPG is produced by pseudomonads of worldwide origin, and its biosynthetic locus is conserved in pseudomonads obtained from diverse geographic locations (25,40). Bacteria that produce DAPG play a key role in agricultural environments, and their potential for use in sustainable agriculture is promising. However, introduction of these bacteria in the field often fails because the organisms are not able to recolonize the roots or colonize the roots but do not produce antibiotic compounds in the new environment (11,41).It is well known that root exudates are sources of nutrition for rhizosphere microorganisms (6, 43). The composition of root exudates is affected by the stage of plant development (18), which results in changes in the patterns and activities of rhizobacterial populations (9, 18, 33). Thus, t...

Background The black soldier fly (Hermetia illucens) is the most promising insect candidate for nutrient-recycling through bioconversion of organic waste into biomass, thereby improving sustainability of protein supplies for animal feed and facilitating transition to a circular economy. Contrary to conventional livestock, genetic resources of farmed insects remain poorly characterised. We present the first comprehensive population genetic characterisation of H. illucens. Based on 15 novel microsatellite markers, we genotyped and analysed 2862 individuals from 150 wild and captive populations originating from 57 countries on seven subcontinents. Results We identified 16 well-distinguished genetic clusters indicating substantial global population structure. The data revealed genetic hotspots in central South America and successive northwards range expansions within the indigenous ranges of the Americas. Colonisations and naturalisations of largely unique genetic profiles occurred on all non-native continents, either preceded by demographically independent founder events from various single sources or involving admixture scenarios. A decisive primarily admixed Polynesian bridgehead population serially colonised the entire Australasian region and its secondarily admixed descendants successively mediated invasions into Africa and Europe. Conversely, captive populations from several continents traced back to a single North American origin and exhibit considerably reduced genetic diversity, although some farmed strains carry distinct genetic signatures. We highlight genetic footprints characteristic of progressing domestication due to increasing socio-economic importance of H. illucens, and ongoing introgression between domesticated strains globally traded for large-scale farming and wild populations in some regions. Conclusions We document the dynamic population genetic history of a cosmopolitan dipteran of South American origin shaped by striking geographic patterns. These reflect both ancient dispersal routes, and stochastic and heterogeneous anthropogenic introductions during the last century leading to pronounced diversification of worldwide structure of H. illucens. Upon the recent advent of its agronomic commercialisation, however, current human-mediated translocations of the black soldier fly largely involve genetically highly uniform domesticated strains, which meanwhile threaten the genetic integrity of differentiated unique local resources through introgression. Our in-depth reconstruction of the contemporary and historical demographic trajectories of H. illucens emphasises benchmarking potential for applied future research on this emerging model of the prospering insect-livestock sector.

Understanding how ecological conditions influence physiological responses is fundamental to forensic entomology. When determining the minimum postmortem interval with blow fly evidence in forensic investigations, using a reliable and accurate model of development is integral. Many published studies vary in results, source populations, and experimental designs. Accordingly, disentangling genetic causes of developmental variation from environmental causes is difficult. This study determined the minimum time of development and pupal sizes of three populations of Lucilia sericata Meigen (Diptera: Calliphoridae; from California, Michigan, and West Virginia) at two temperatures (20 degrees C and 33.5 degrees C). Development times differed significantly between strain and temperature. In addition, California pupae were the largest and fastest developing at 20 degrees C, but at 33.5 degrees C, though they still maintained their rank in size among the three populations, they were the slowest to develop. These results indicate a need to account for genetic differences in development, and genetic variation in environmental responses, when estimating a postmortem interval with entomological data.

There is very little information concerning carrion fly population genetic structure. We generated amplified fragment length polymorphism (AFLP) profiles for the common blowfly, Phormia regina (Meigen), from sites spanning the contiguous United States. Analysis of molecular variance (AMOVA) based on 232 loci found significant variation (phi(SC) = 23%) among discrete samples (those collected at a bait in one location over a short period of time). Samples collected in the same location but at different times were also distinct. When samples were pooled into geographic regions (east, central, west), the variation was negligible (phi(CT) = 0%). A Mantel test found only a very weak correlation between individual genetic and geographic distances. Relative relatedness coefficients based on shared allele proportions indicated individual samples were likely to contain close relatives. P. regina arriving at an individual carcass typically represent a nonrandom sample of the population despite a lack of geographic structure. A female blow fly produces hundreds of offspring at one time; therefore, newly emerged siblings may respond in concert to an odor plume. These results may be of interest to forensic entomologists, many of whom use a laboratory colony founded from a small sample for the growth studies that support casework. Discrepancies between published growth curves may reflect such random differences in the founding individuals.

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