Crosslinked Polypeptide Films via RAFT-Mediated Continuous Assembly of Polymers

Polypeptide coatings are a cornerstone in the field of surface modification due to their widespread biological potential. As their properties are dictated by their structural features, subsequent control thereof using unique fabrication strategies is important. Herein, we report a facile method of precisely creating densely crosslinked polypeptide films with unusually high random coil content through continuous assembly polymerization via reversible addition–fragmentation chain transfer (CAP‐RAFT). CAP‐RAFT was fundamentally investigated using methacrylated poly‐l‐lysine (PLLMA) and methacrylated poly‐l‐glutamic acid (PLGMA). Careful technique refinement resulted in films up to 36.1±1.1 nm thick which could be increased to 94.9±8.2 nm after using this strategy multiple times. PLLMA and PLGMA films were found to have 30–50 % random coil conformations. Degradation by enzymes present during wound healing reveals potential for applications in drug delivery and tissue engineering

Validation of Reference Genes for Robust qRT-PCR Gene Expression Analysis in the Rice Blast Fungus \u3ci\u3eMagnaporthe oryzae\u3c/i\u3e

The rice blast fungus causes significant annual harvest losses. It also serves as a genetically- tractable model to study fungal ingress. Whilst pathogenicity determinants have been unmasked and changes in global gene expression described, we know little about Magnaporthe oryzae cell wall remodelling. Our interests, in wall remodelling genes expressed during infection, vegetative growth and under exogenous wall stress, demand robust choice of reference genes for quantitative Real Time-PCR (qRT-PCR) data normalisation. We describe the expression stability of nine candidate reference genes profiled by qRT-PCR with cDNAs derived during asexual germling development, from sexual stage perithecia and from vegetative mycelium grown under various exogenous stressors. Our Minimum Information for Publication of qRT-PCR Experiments (MIQE) compliant analysis reveals a set of robust reference genes used to track changes in the expression of the cell wall remodelling gene MGG_Crh2 (MGG_00592). We ranked nine candidate reference genes by their expression stability (M) and report the best gene combination needed for reliable gene expression normalisation, when assayed in three tissue groups (Infective, Vegetative, and Global) frequently used in M. oryzae expression studies. We found that MGG_Actin (MGG_03982) and the 40S 27a ribosomal subunit MGG_40s (MGG_02872) proved to be robust reference genes for the Infection group and MGG_40s and MGG_Ef1 (Elongation Factor1-α) for both Vegetative and Global groups. Using the above validated reference genes, M. oryzae MGG_Crh2 expression was found to be significantly (p\u3c0.05) elevated three-fold during vegetative growth as compared with dormant spores and two fold higher under cell wall stress (Congo Red) compared to growth under optimal conditions. We recommend the combinatorial use of two reference genes, belonging to the cytoskeleton and ribosomal synthesis functional groups, MGG_Actin, MGG_40s, MGG_S8 (Ribosomal subunit 40S S8) or MGG_Ef1, which demonstrated low M values across heterogeneous tissues. By contrast, metabolic pathway genes MGG_Fad (FAD binding domain-containing protein) and MGG_Gapdh (Glyceraldehyde-3-phosphate dehydrogenase) performed poorly, due to their lack of expression stability across samples

Emerging health threat and cost of Fusarium mycotoxins in European wheat

Mycotoxins harm human and livestock health, while damaging economies. Here we reveal the changing threat of Fusarium head blight (FHB) mycotoxins in European wheat, using data from the European Food Safety Agency and agribusiness (BIOMIN, World Mycotoxin Survey) for ten years (2010–2019). We show persistent, high, single- and multi-mycotoxin contamination alongside changing temporal-geographical distributions, indicative of altering FHB disease pressure and pathogen populations, highlighting the potential synergistic negative health consequences and economic cost

Insect pollination is important in a smallholder bean farming system

Background: Many crops are dependent on pollination by insects. Habitat management in agricultural landscapes can support pollinator services and even augment crop production. Common bean (Phaseolus vulgaris L.) is an important legume for the livelihoods of smallholder farmers in many low-income countries, particularly so in East Africa. While this crop is autogamous, it is frequently visited by pollinating insects that could improve yields. However, the value of pollination services to common beans (Kariasii) yield is not known. Methods: We carried out pollinator-exclusion experiments to determine the contribution of insect pollinators to bean yields. We also carried out a fluorescent-dye experiment to evaluate the role of field margins as refuge for flower-visitors. Results: Significantly higher yields, based on pods per plant and seeds per pod, were recorded from open-pollinated and hand-pollinated flowers compared to plants from which pollinators had been excluded indicating that flower visitors contribute significantly to bean yields. Similarly, open and hand-pollinated plants recorded the highest mean seed weight. Extrapolation of yield data to field scale indicated a potential increase per hectare from 681 kg in self-pollinated beans to 1478 kg in open-pollinated beans indicating that flower visitors contributed significantly to crop yield of beans. Our marking study indicated that flower-visiting insects including bees, flies and lepidopterans moved from the field margin flowers into the bean crop. Overall, these results show that insect pollinators are important for optimising bean yields and an important food security consideration on smallholder farms. Field margin vegetation also provides habitat for flower-visiting insects that pollinate beans. Hence, non-crop habitats merit further research focusing on establishing which field margin species are most important and their capacity to support other ecosystem services such as natural pest regulation

A lipophilic cation protects crops against fungal pathogens by multiple modes of action

This is the final version. Available from Nature Research via the DOI in this record. The authors confirm that all relevant data are included in the paper or in the Supplementary Information file. The source data underlying Figs. 1a–k, 2a–k, 3a–d, 4a–h, 5a–h, 6a–f, 7a–l, 8a–g and Supplementary Figs. 2, 3a, b, 4a, b, 5, 7a–e, 8a, b, 9 are provided as a Source Data file. Additional information is available from the authors upon request.The emerging resistance of crop pathogens to fungicides poses a challenge to food security and compels discovery of new antifungal compounds. Here, we show that mono-alkyl lipophilic cations (MALCs) inhibit oxidative phosphorylation by affecting NADH oxidation in the plant pathogens Zymoseptoria tritici, Ustilago maydis and Magnaporthe oryzae. One of these MALCs, consisting of a dimethylsulfonium moiety and a long alkyl chain (C18-SMe2+), also induces production of reactive oxygen species at the level of respiratory complex I, thus triggering fungal apoptosis. In addition, C18-SMe2+ activates innate plant defense. This multiple activity effectively protects cereals against Septoria tritici blotch and rice blast disease. C18-SMe2+ has low toxicity in Daphnia magna, and is not mutagenic or phytotoxic. Thus, MALCs hold potential as effective and non-toxic crop fungicides.BBSR

Characterization of hymenopteran parasitoids of aphis fabae in an African smallholder bean farming system through sequencing of COI 'mini-barcodes'

Parasitoids are among the most frequently reported natural enemies of insect pests, particularly aphids. The efficacy of parasitoids as biocontrol agents is influenced by biotic and abiotic factors. For example, hyperparasitoids can reduce the abundance of the primary parasitoids as well as modify their behavior. A field study was conducted at three contrasting elevations on Mount Kilimanjaro, Tanzania, to identify the parasitoids of aphids in smallholder bean farming agroecosystems. Sentinel aphids (Aphis fabae) on potted bean plants (Phaseolus vulgaris) were exposed in 15 bean fields at three elevations for 2 days. The sentinel aphids were then kept in cages in a greenhouse until emergence of the parasitoids, which were collected and preserved in 98% ethanol for identification. Of the 214 parasitoids that emerged from sentinel aphids, the greatest abundance (44.86%) were from those placed at intermediate elevations (1000–1500 m a.s.l), compared to 42.52% from the lowest elevations and only 12.62% from the highest elevation farms. Morphological identification of the parasitoids that emerged from parasitized aphids showed that 90% were Aphidius species (Hymenoptera: Braconidae: Aphidiinae). Further characterization by sequencing DNA ‘mini-barcodes’ identified parasitoids with ≥99% sequence similarity to Aphidius colemani, 94–95% sequence similarity to Pachyneuron aphidis and 90% similarity to a Charipinae sp. in the National Center for Biotechnology Information (NCBI) database. These results confidently identified A. colemani as the dominant primary aphid parasitoid of A. fabae in the study area. A Pachyneuron sp., which was most closely related to P. aphidis, and a Charipinae sp. occurred as hyperparasitoids. Thus, interventions to improve landscapes and farming practice should monitor specifically how to augment populations of A. colemani, to ensure any changes enhance the delivery of natural pest regulation. Further studies are needed for continuous monitoring of the hyperparasitism levels and the dynamics of aphids, primary parasitoids, and secondary parasitoids in different cropping seasons and their implications in aphid control

Zymoseptoria tritici white-collar complex integrates light, temperature and plant cues to initiate dimorphism and pathogenesis

Transitioning from spores to hyphae is pivotal to host invasion by the plant pathogenic fungus Zymoseptoria tritici. This dimorphic switch can be initiated by high temperature in vitro (~27 °C); however, such a condition may induce cellular heat stress, questioning its relevance to field infections. Here, we study the regulation of the dimorphic switch by temperature and other factors. Climate data from wheat-growing areas indicate that the pathogen sporadically experiences high temperatures such as 27 °C during summer months. However, using a fluorescent dimorphic switch reporter (FDR1) in four wild-type strains, we show that dimorphic switching already initiates at 15-18 °C, and is enhanced by wheat leaf surface compounds. Transcriptomics reveals 1261 genes that are up- or down-regulated in hyphae of all strains. These pan-strain core dimorphism genes (PCDGs) encode known effectors, dimorphism and transcription factors, and light-responsive proteins (velvet factors, opsins, putative blue light receptors). An FDR1-based genetic screen reveals a crucial role for the white-collar complex (WCC) in dimorphism and virulence, mediated by control of PCDG expression. Thus, WCC integrates light with biotic and abiotic cues to orchestrate Z. tritici infection

Beneficial insects are associated with botanically rich margins with trees on small farms

Beneficial insect communities on farms are influenced by site- and landscape-level factors, with pollinator and natural enemy populations often associated with semi-natural habitat remnants. They provide ecosystem services essential for all agroecosystems. For smallholders, natural pest regulation may be the only affordable and available option to manage pests. We evaluated the beneficial insect community on smallholder bean farms (Phaseolus vulgaris L.) and its relationship with the plant communities in field margins, including margin trees that are not associated with forest fragments. Using traps, botanical surveys and transect walks, we analysed the relationship between the floral diversity/composition of naturally regenerating field margins, and the beneficial insect abundance/diversity on smallholder farms, and the relationship with crop yield. More flower visits by potential pollinators and increased natural enemy abundance measures in fields with higher plant, and particularly tree, species richness, and these fields also saw improved crop yields. Many of the flower visitors to beans and potential natural enemy guilds also made use of non-crop plants, including pesticidal and medicinal plant species. Selective encouragement of plants delivering multiple benefits to farms can contribute to an ecological intensification approach. However, caution must be employed, as many plants in these systems are introduced species

Mucedorus: the last ludic playbook, the first stage Arcadia

This article argues that two seemingly contradictory factors contributed to and sustained the success of the anonymous Elizabethan play Mucedorus (c. 1590; pub. 1598). First, that both the initial composition of Mucedorus and its Jacobean revival were driven in part by the popularity of its source, Philip Sidney's Arcadia. Second, the playbook's invitation to amateur playing allowed its romance narrative to be adopted and repurposed by diverse social groups. These two factors combined to create something of a paradox, suggesting that Mucedorus was both open to all yet iconographically connected to an elite author's popular text. This study will argue that Mucedorus pioneered the fashion for “continuations” or adaptations of the famously unfinished Arcadia, and one element of its success in print was its presentation as an affordable and performable version of Sidney's elite work. The Jacobean revival of Mucedorus by the King's Men is thus evidence of a strategy of engagement with the Arcadia designed to please the new Stuart monarchs. This association with the monarchy in part determined the cultural functions of the Arcadia and Mucedorus through the Interregnum to the close of the seventeenth century