The genetic basis of parallel and divergent phenotypic responses in evolving populations of Escherichia coli

Pleiotropy plays a central role in theories of adaptation, but little is known about the distribution of pleiotropic effects associated with different adaptive mutations. Previously, we described the phenotypic effects of a collection of independently arising beneficial mutations in Escherichia coli. We quantified their fitness effects in the glucose environment in which they evolved and their pleiotropic effects in five novel resource environments. Here we use a candidate gene approach to associate the phenotypic effects of the mutations with the underlying genetic changes. Among our collection of 27 adaptive mutants, we identified a total of 21 mutations (18 of which were unique) encompassing five different loci or gene regions. There was limited resolution to distinguish among loci based on their fitness effects in the glucose environment, demonstrating widespread parallelism in the direct response to selection. However, substantial heterogeneity in mutant effects was revealed when we examined their pleiotropic effects on fitness in the five novel environments. Substitutions in the same locus clustered together phenotypically, indicating concordance between molecular and phenotypic measures of divergence.     

Compatibility of Steinernema feltiae (Nematoda: Steinernematidae) with Pesticides and Plant Growth Regulators Used in Glasshouse Plant Production

Compatibility of the entomopathogenic nematode, Steinernema feltiae Filipjev with 17 different pesticides and three plant growth regulators, recommended for drench application in glasshouses, was evaluated. The products tested were two biofungicides, Streptomyces griseoviridis (Mycostop) and Trichoderma harzianum (Rootshield), nine chemical fungicides, iprodione (Chipco 26GT), thiophanate-methyl (Fungo Flo 4.5F), azoxystrobin (Heritage), fludioxonil (Medallion), flutolanil (Prostar), mefenoxam (SudDue Maxx 21.3ME), PCNB (Terraclor 400F), triflumizole (Terraguard), and etridiazole (Terrazole), two bioinsecticides, spinosad (Conserve SC) and Bacillus thuringiensis subsp. israelensis (Gnatrol), three chemical insecticides diflubenzuron (Adept IGR), acephate (Orthene), and fenoxycarb (Precision 25WP), one herbicide, clethodim (Envoy) and three plant growth regulators, ancymidol (A-Rest), paclobutrazol (Bonzi), and uniconazole-P (Sumagic). Infective juvenile nematodes were exposed to each product at the highest recommended concentration in 24-well plates at 22°C. Observations on the viability and infectivity of the nematodes were made at 4, 24, and 72 h after exposure. We found that S. feltiae is compatible with the majority of the tested formulations with no loss in viability and infectivity up to 24 h of exposure. The viability of S. feltiae was more than 80% in all the products even after 72 h of exposure. Three pesticide formulations, Prostar (1%), Gnatrol (9%), and Terrazole (10%) decreased the viability of S. feltiae significantly within 24 h compared to the controls. However, during the 24 to 72 hr incubation period, eight pesticides affected the viability of S. feltiae with Gnatrol and Terrazole causing the highest decrease (17% and 15%, respectively). Only Terrazole decreased the infectivity of S. feltiae to Galleria mellonella larvae compared to the control when tested after 24 h exposure. At 72 hr, Orthene and Terrazole caused significant decrease in the infectivity of S. feltiae (10% and 15%, respectively) and Gnatrol caused a significant increase in the infectivity (11%) compared to the control. Our results suggest that S. feltiae can be tank-mixed and applied in combination with all the tested formulations, except Terrazole.     

Impact of oilseed rape expressing the insecticidal cysteine protease inhibitor oryzacystatin on the beneficial predator Harmonia axyridis (multicoloured Asian ladybeetle)

Insect-resistant transgenic plants have been suggested to have deleterious effects on beneficial predators through transmission of the transgene product by the pest to the predator. To test this hypothesis, effects of oilseed rape expressing the cysteine protease inhibitor oryzacystatin-1 (OC-1) on the predatory ladybird Harmonia axyridis were investigated using diamondback moth Plutella xylostella as the pest species. As expected, oilseed rape expressing OC-1 had no effects on either development or survival of the pest, which utilizes serine digestive proteases. Immunoassays confirmed accumulation of the transgene product in pest larval tissues at levels of up to 3 ng per gut. Characterization of proteolytic digestive enzymes of H. axyridis demonstrated that larvae and adults utilize cysteine and aspartic proteases; the former activity was completely inhibited by oryzacystatin in vitro. However, when H. axyridis larvae consumed prey reared on OC-1 expressing plants over their entire life cycle, no significant effects upon survival or overall development were observed. The inhibitor initially stimulated development, with a shortening of the developmental period of the second instar by 27% (P < 0.0001) accompanied by a 36% increase in weight of second instar larvae (P = 0.007). OC-1 had no detrimental effects on reproductive fitness of adult H. axyridis. Interestingly there was a significant increase in consumption of OC-1 dosed prey. The results show that prey reared on transgenic plants expressing a protein which inhibited ladybird digestive enzymes in vitro had no effects in vivo; the ladybird was able to up-regulate digestive proteases in response to the inhibitor.     

Effects of potato plants expressing a barley cystatin on the predatory bug Podisus maculiventris via herbivorous prey feeding on the plant

The aim of this study was to assess the effects of potato plants expressing a barley cystatin on a potentially cystatin-susceptible natural enemy by predation on susceptible and non-susceptible preys feeding on the plant. We have focussed on the impact of the variant HvCPI-1 C⁶⁸ → G, in which the only cysteine residue was changed by a glycine, on the growth and digestive physiology of the Colorado potato beetle (CPB), Leptinotarsa decemlineata, and the Egyptian cotton leafworm (ECW), Spodoptera littoralis. Moreover, we have studied the prey-mediated effects of the barley cystatin at the third trophic level, using the predatory spined soldier bug (SSB), Podisus maculiventris, as a model. Feeding trials conducted with CPB larvae reared on transgenic potato plants expressing the C⁶⁸ → G variant resulted in significantly lower weight gains compared to those fed on non-transformed (NT) plants. On the contrary, larger weight gains were obtained when ECW larvae, that lack digestive cysteine proteases, were reared on transgenic potato expressing the cystatin, as compared to larvae fed on NT plants. No negative effects on survival and growth were observed when SSB nymphs were exposed to HvCPI-1 C⁶⁸ → G by predation on either CPB or ECW larvae reared on transgenic potato plants expressing the barley cystatin, despite the fact that the inhibitor suppressed in vitro gut proteolysis of the predatory bug. To investigate the physiological background, biochemical analysis were carried out on guts of insects dissected at the end of the feeding assays. Fernando álvarez-Alfageme and Manuel Martínez are contributed equally to this work.     

草原蘑菇圈的土壤-植物系统研究

在内蒙古锡林郭勒草原,比较研究了蒙古口蘑形成的蘑菇圈各环带的植物生长状况和土壤生态因子的变化。结果表明,绿草环上的植物体内营养元素和叶绿素的含量明显增加,其地上部分植物生物量显著高于正常草区,产生增产效应;枯草环上的植物生长则受到抑制。蘑菇圈土植物的区系组成有所变化。蘑菇圈上土壤养分特别是速效态的N和P含量有显著增加。蒙古口蘑菌丝体生长对土壤理化性质产生影响并产生明显的土壤生物学效应。     

Management practices influence biocontrol potential of generalist predators in maize cropping systems

Intensification of conventional agriculture is a leading cause of negative environmental impact, loss of biodiversity, and reduced delivery of ecosystem services in agroecosystems. Maize, due to its growth habits and cultivation management, provides a poor habitat for beneficial arthropods. Several strategies are available to make maize cropping systems more sustainable and to promote biodiversity at field level. The present study evaluates the effects of various maize cropping systems – precisely, maize continuous monoculture, maize multiple cropping, and three mixed cropping systems (maize-runner bean, maize-sorghum, and maize-flower strips) – on generalist predators and their biological control potential. Overall, we found that the reference system with maize continuous monoculture and conventional crop management had the lowest activity of generalist predators compared in particular to the low-input mixed cropping systems. Higher activity density and biocontrol potential were found in the systems that provided a dense and permanent vegetation cover of the ground (i.e., maize/sorghum, maize/flower strip). Although these effects were not consistent for all the parameters investigated and for every sampling date, we conclude that generalist predators can benefit from more conservative management practices in maize systems, thereby enhancing their biological pest control potential. Furthermore, spatial intercropping may represent a valid alternative to the conventional monocultural crop system to support the delivery of biodiversity-mediated ecosystem services towards a more sustainable system.     

Monitoring Intraspecies Competition in a Bacterial Cell Population by Cocultivation of Fluorescently Labelled Strains

Many microorganisms such as bacteria proliferate extremely fast and the populations may reach high cell densities. Small fractions of cells in a population always have accumulated mutations that are either detrimental or beneficial for the cell. If the fitness effect of a mutation provides the subpopulation with a strong selective growth advantage, the individuals of this subpopulation may rapidly outcompete and even completely eliminate their immediate fellows. Thus, small genetic changes and selection-driven accumulation of cells that have acquired beneficial mutations may lead to a complete shift of the genotype of a cell population. Here we present a procedure to monitor the rapid clonal expansion and elimination of beneficial and detrimental mutations, respectively, in a bacterial cell population over time by cocultivation of fluorescently labeled individuals of the Gram-positive model bacterium Bacillus subtilis. The method is easy to perform and very illustrative to display intraspecies competition among the individuals in a bacterial cell population.