Comparing mating designs to restore seed production of interspecific hybrids between Trifolium repens (white clover) and Trifolium uniflorum

Interspecific hybrids between Trifolium uniflorum and cultivated white clover (Trifolium repens) have highly useful characteristics for temperate pastoral systems derived from both parent species. However, the early hybrids (F₁ and BC₁) also have unacceptably poor seed production for commerce. This study analysed the basis for the poor seed production and investigated breeding strategies for overcoming the problem. The BC₁F₁ generation produced lower‐than‐expected numbers of heads per plant and seeds per floret. Backcrossing of selected hybrids to white clover corrected these deficiencies and created new variation. Seed numbers were also returned to near target levels by recurrent selection within the BC₁ generation. Thus, it was possible to retain a theoretical average of 25% of T. uniflorum genome and still achieve high seed production per plant. The BC₁F₂ and BC₂F₁ generations produced high seed numbers per plant, along with reasonable variation. Both of these second‐generation hybrid forms have high reproductive potential and should be the focus for the selection of the desired combinations of agronomic and seed production traits.     

CRISPR-Cas Technology in Plant Science

CRISPR-Cas technology has raised considerable interest among plant scientists, both in basic science and in plant breeding. Presently, the generation of random mutations at a predetermined site of the genome is well mastered, just like the targeted insertion of transgenes, although both remain restricted to species or genotypes amenable for plant transformation. On the other hand, true genome editing, i.e. the deliberate replacement of one or several nucleotides of the genome in a predetermined fashion, is limited to some rather particular examples that generally concern genes allowing positive selection, for example tolerance to herbicides. Therefore, further technological developments are necessary to fully exploit the potential of genome editing in enlarging the gene pool beyond the natural variability available in a given species. In principle, the technology can be applied to any quality related, agronomical or ecological trait, under the condition of upstream knowledge on the genes to be targeted and the precise modifications necessary to improve alleles. Published proof of concepts concern a wide range of agronomical traits, the most frequent being disease resistance, herbicide tolerance and the biochemical composition of harvested products. The regulatory status of the plants obtained by CRISPR-Cas technology raises numerous questions, in particular with regard to the plants that carry in their genomes the punctual modifications caused by the presence of the Cas9 nuclease but not the nuclease itself. Without clarification by the competent authorities, CRISPR-Cas technology would continue to be a powerful tool in functional genomics, but its potential in plant breeding would remain untapped.     

System for quasi‐continuous simultaneous measurement of oxygen diffusion rate and redox potential in soil

Oxygen diffusion rate (ODR) and redox potential (E_H) are quantitative indices representing oxygen availability and redox status in soils, which is valuable information for better understanding causes and effects of soil aeration. Because these indices are spatially and temporally highly variable, continuous measurements and adequate numbers of repetitions are essential for accurate in situ monitoring. Here, we present a new, fully automated recording system for in situ measurements where ODR and E_H are measured at the same platinum electrode. The conflict between electrode polarization for ODR and the resulting biased E_H readings is solved by reducing the polarization time and introducing a recovery interval between two consecutive measurement cycles. The shorter polarization time ensures accurate E_H readings. It also results in moderately overestimated ODR readings, but this can be corrected before data analysis. The recovery interval restricts temporal resolution of the E_H‐ODR data pairs to 8 h. We illustrate the use of the system with measurements in a field experiment in Zürich, Switzerland. ODR curves at different depths ran roughly parallel to the corresponding curves of O₂ concentration in soil air but ODR was much more sensitive to precipitation. Low ODR was a necessary but not a sufficient condition for declining E_H. E_H ran parallel to O₂ concentration in soil air rather than to ODR. The fully automated system allows for time series of replicate measurements in multifactorial field studies with reasonable labor requirements. It may be particularly suitable for studies examining the effects of soil tillage, compaction, and irrigation, where structure‐related soil properties such as porosity, gas permeability, and soil aeration play a dominant role.     

Microbial remediation of a pentachloronitrobenzene-contaminated soil under Panax notoginseng: A field experiment

Pentachloronitrobenzene (PCNB) is an organochlorine fungicide that is mainly used in the prevention and control of diseases in crop seedlings. Microbial removal is used as a promising method for in-situ removal of many organic pesticides and pesticide residues. A short-term field experiment (1 year) was conducted to explore the potential role of a PCNB-degrading bacterial isolate, Cupriavidus sp. YNS-85, in the remediation of a PCNB-contaminated soil on which Panax notoginseng was grown. The following three treatments were used:i) control soil amended with wheat bran but without YNS-85, ii) soil with 0.15 kg m^-2 of solid bacterial inoculum (A), and iii) soil with 0.30 kg m^-2 of solid bacterial inoculum (B). The removal of soil PCNB during the microbial remediation was monitored using gas chromatography. Soil catalase and fluorescein diacetate (FDA) esterase activities were determined using spectrophotometry. In addition, cultivable bacteria, fungi, and actinomycetes were counted by plating serial dilutions, and the microbial biodiversity of the soil was analyzed using BIOLOG. After 1 year of in-situ remediation, the soil PCNB concentrations decreased significantly by 50.3% and 74.2% in treatments A and B, respectively, when compared with the uninoculated control. The soil catalase activity decreased in the presence of the bacterial isolate, the FDA esterase activity decreased in treatment A, but increased in treatment B. No significant changes in plant biomass, diversity of the soil microbial community, or physicochemical properties of the soil were observed between the control and inoculated groups (P<0.05). The results indicate that Cupriavidus sp. YNS-85 is a potential candidate for the remediation of PCNB-contaminated soils under P. notoginseng.     

Artificial Diet Influences Population Growth of the Root Maggot Bradysia impatiens (Diptera: Sciaridae)

In order to investigate the effects of artificial diets on the population growth of root maggot Bradysia impatiens, its population growth parameters were assayed on eight artificial diets (Diet 1, D2, D3, D4, D5, D6, D7, and D8). Results showed that developmental duration from egg to pupa was successfully completed on all eight artificial diets. However, the egg to pupal duration was shortest, while the survival rate of four insect stages was lowest when B. impatiens was reared on D1. When B. impatiens was reared on D7 and D8, the survival rate, female longevity, and female oviposition were higher than those reared on other diets. When B. impatiens was reared on D7, the intrinsic rate of increase (r_m = 0.19/d), net reproductive rate (R₀ = 39.88 offspring per individual), and finite rate of increase (λ = 1.21/d) were higher for its population growth with shorter generation time (T = 19.49 d) and doubling time (Dt = 3.67 d). The findings indicate that the D7 artificial diet is more appropriate for the biological parameters of B. impatiens and can be used an indoor breeding food for population expansion as well as further research. We propose that vitamin C supplement added to the D7 is critical for the improvement of the B. impatiens growth.     

Validating an empiric sulfadiazine–trimethoprim dosage regimen for treatment of Escherichia coli and Staphylococcus delphini infections in mink (Neovison vison)

Antimicrobial agents are used extensively off‐label in mink, as almost no agents are registered for this animal species. Pharmacokinetic (PK) and pharmacodynamic (PD) data are required to determine antimicrobial dosages specifically targeting mink bacterial pathogens. The aims of this study were to assess, in a PKPD framework, the empirical dosage regimen for a combination of trimethoprim (TMP) and sulfadiazine (SDZ) in mink, and secondarily to produce data for future setting of clinical breakpoints. TMP and SDZ PK parameters were obtained experimentally in 22 minks following IV or oral administration of TMP/SDZ (30 mg/kg, i.e. 5 mg/kg TMP and 25 mg/kg SDZ). fAUC/MIC with a target value of 24 hr was selected as the PKPD index predictive of TMP/SDZ efficacy. Using a modeling approach, PKPD cutoffs for TMP and SDZ were determined as 0.062 and 16 mg/L, respectively. By incorporating an anticipated potentiation effect of SDZ on TMP against Escherichia coli and Staphylococcus delphini, the PKPD cutoff of TMP was revised to 0.312 mg/L, which is above the tentative epidemiological cutoffs (TECOFF) for these species. The current empirical TMP/SDZ dosage regimen (30 mg/kg, PO, once daily) therefore appears adequate for treatment of wild‐type E. coli and S. delphini infections in mink.