Change in Chemical Composition of Sweet Basil (Ocimum basilicum L.) Essential Oil Caused by Alfalfa mosaic virus

The effects of Alfalfa mosaic virus (AMV) infection on essential oil (EO) content and composition of a Sweet Basil cv. Gigante di Napoli were evaluated. A 10‐fold lower extraction yield from infected plants was observed, suggesting that morphological alterations induced by the disease may affect abundance and efficacy of secretive tissues. Organoleptic properties and thus quality of EO were severely affected and EO composition resulted severely altered, with a great increase in sesquiterpenes (from 72.8 to 19.8%) and a decrease in both monoterpenes (from 35 to 11%) and phenylpropanoids (from 44.5 to 15.8%, despite a slight increase in eugenol). Such report is indicative of possible direct or indirect metabolic consequences of AMV in a commercially important species like Ocimum basilicum is. The possible consequences of linalool and trans‐β‐farnesene content changes on the dispersion of viruliferous aphids are also examined and discussed.     

Effects of Temperature on Disease Severity in Plants of Subterranean Clover Infected Singly or in Mixed Infection with Bean yellow mosaic virus and Kabatiella caulivora

Many epidemics involve plants infected with more than one pathogen, but few experiments address climate change scenarios that influence mixed infections. This study addresses the interactive effects of co‐infection and temperature on disease development in plants of the annual pasture species subterranean clover (Trifolium subterraneum), which is widely sown in different world regions. Bean yellow mosaic virus (BYMV) and the fungus Kabatiella caulivora are two important pathogens causing considerable production losses in pastures containing this species. Both occur together in such pastures causing a severe necrotic disease when mixed infection occurs. Effects of temperature on symptom expression were investigated in subterranean clover plants infected singly or in mixed infection with these pathogens. Plants were maintained in controlled environment rooms at 18°C, 20°C or 22.5°C after sap inoculation with BYMV. K. caulivora conidia suspensions were inoculated to plants once systemic BYMV symptoms developed. Plants were assessed for three disease assessment parameters, dead petioles numbers, marginal leaflet necrosis and overall plant damage. In general, mixed infection caused most severe symptoms, K. caulivora least severe symptoms, and BYMV symptoms of intermediate severity. In single infections, effects of temperature on disease severity differed between pathogens: BYMV symptoms were most pronounced at 18°C, but K. caulivora induced more severe symptoms at 20°C and 22.5°C. In mixed infections, disease severity generally followed the pattern developed with BYMV alone as temperature increased. Also, synergistic increase in disease severity sometimes occurred at 18°C, but increases were only additive at 20°C and 22.5°C. These results reflected the greater BYMV multiplication detected in infected leaves at 18°C compared with 20°C or 22.5°C. Our findings indicate that in rainfed subterranean clover pastures, as global warming progresses disease severity from infection with BYMV and K. caulivora alone may decline or increase, respectively, and mixed infection with them may become less damaging.     

Begomoviruses Infecting Tomato Crops in Panama

The key regions in Panama involved in open field‐ and greenhouse‐grown commercial tomato production, including the Chiriquí, Veraguas, Herrera, Los Santos, Coclé and Panama Oeste provinces, were surveyed for the incidence and distribution of begomoviruses in the growing seasons of 2011 and 2012. The surveys took place in 14 of the 51 districts of the above‐mentioned provinces and comprised all relevant tomato production areas of the provinces. A total of 28 tomato plots were surveyed. The exact location of each plot was geo‐referenced using a hand‐held Global Positioning System unit. In total, 319 individual tomato plants (181 in 2011 and 138 in 2012) were sampled. Plants displayed diverse combinations of virus‐like symptoms of different severity, including necrosis, yellowing, mosaic, mottling, rolling, curling, distortion and puckering of leaves, reduced leaf size, and stunted growth. DNA was extracted from each plant for a subsequent polymerase chain reaction (PCR) analysis, using two sets of degenerate primers able to detect members of the genus Begomovirus. The samples displaying a positive reaction were subsequently analysed with specific primer pairs to identify the affecting begomoviruses. A total of 42.3% of all collected samples showed a positive signal to PCRs. Three begomovirus species were detected with the species‐specific set of primers; in particular, in the samples obtained in 2011, Potato yellow mosaic Panama virus (PYMPV), Tomato leaf curl Sinaloa virus (ToLCSiV) and Tomato yellow mottle virus (TYMoV) were detected, while in the 2012 samples, only PYMPV and ToLCSiV were found. To our knowledge, this is the first reported incidence of ToLCSiV and TYMoV in Panamanian tomato crops.     

Construction of an efficient genomic editing system with CRISPR/Cas9 in the vector mosquito Aedes albopictus

Aedes (Stegomyia) albopictus, also known as the Asian tiger mosquito, is a mosquito which originated in Asia. In recent years, it has become increasingly rampant throughout the world. This mosquito can transmit several arboviruses, including dengue, Zika and chikungunya viruses, and is considered a public health threat. Despite the urgent need of genome engineering to analyze specific gene functions, progress in genetical manipulation of Ae. albopictus has been slow due to a lack of efficient methods and genetic markers. In the present study, we established targeted disruptions in two genes, kynurenine hydroxylase (kh) and dopachrome conversion enzyme (yellow), to analyze the feasibility of generating visible phenotypes with genome editing by the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein 9 (Cas9) system in Ae. albopictus. Following Cas9 single guide RNA ribonucleoprotein injection into the posterior end of pre-blastoderm embryos, 30%-50% of fertile survivors produced alleles that failed to complement existing kh and yellow mutations. Complete eye and body pigmentation defects were readily observed in GI pupae and adults, indicating successful generation of highly heritable mutations. We conclude that the CRISPR/Cas9-mediated gene editing system can be used mAe. albopictus and that it can be adopted as an efficient tool for genome-scale analysis and biological study.     

Development of Beet necrotic yellow vein virus‐based vectors for multiple‐gene expression and guide RNA delivery in plant genome editing

Many plant viruses with monopartite or bipartite genomes have been developed as efficient expression vectors of foreign recombinant proteins. Nonetheless, due to lack of multiple insertion sites in these plant viruses, it is still a big challenge to simultaneously express multiple foreign proteins in single cells. The genome of Beet necrotic yellow vein virus (BNYVV) offers an attractive system for expression of multiple foreign proteins owning to a multipartite genome composed of five positive‐stranded RNAs. Here, we have established a BNYVV full‐length infectious cDNA clone under the control of the Cauliflower mosaic virus 35S promoter. We further developed a set of BNYVV‐based vectors that permit efficient expression of four recombinant proteins, including some large proteins with lengths up to 880 amino acids in the model plant Nicotiana benthamiana and native host sugar beet plants. These vectors can be used to investigate the subcellular co‐localization of multiple proteins in leaf, root and stem tissues of systemically infected plants. Moreover, the BNYVV‐based vectors were used to deliver NbPDS guide RNAs for genome editing in transgenic plants expressing Cas9, which induced a photobleached phenotype in systemically infected leaves. Collectively, the BNYVV‐based vectors will facilitate genomic research and expression of multiple proteins, in sugar beet and related crop plants.     

In vitro protein digestion kinetics of protein sources for pigs

In current feed evaluation systems, the nutritional value of protein sources in diets for pigs is based on the ileal digestibility of protein and amino acids, which does not account for the kinetics of protein digestion along the gastrointestinal tract. The objective of the present study was to determine the in vitro protein digestion kinetics of different protein sources (soya bean meal (SBM), wheat gluten (WG), rapeseed meal (RSM), whey powder (WP), dried porcine plasma protein, yellow meal worm larvae and black soldier fly larvae (BSF)). Protein sources were incubated with pepsin at pH 3.5 for 0 to 90 min and subsequently with pancreatin at pH 6.8 for 0 to 210 min at 39°C. The in vitro protein digestion kinetics were described as the kinetics of nitrogen (N) solubilisation and the release of low molecular weight peptides (LMW) (<500 Da). The N solubilisation rate ranged from 0.025 min⁻¹ for BSF to 0.685 min⁻¹ for WP during the incubation with pepsin, and from 0.027 min⁻¹ for RSM to 0.343 min⁻¹ for WP during the incubation with pancreatin. The release rate of LMW peptides ranged from 0.027 min⁻¹ for WG to 0.093 min⁻¹ for WP during the incubation with pepsin, and from 0.029 min⁻¹ for SBM to 0.385 min⁻¹ for WP. Black soldier fly larvae showed a similar release rate of LMW peptides as WP during the incubation with pancreatin. At the end of the sequential incubation with pepsin (90 min) and pancreatin (210 min), WG and WP showed the highest percentage of N present in LMW peptides relative to total N (78% and 79%, respectively), whereas SBM showed the lowest (35%). In conclusion, protein sources for pig diets show substantial differences in in vitro protein digestion kinetics as measured by the kinetics of N solubilisation and the release of LMW peptides. The rate of release of LMW peptides was not correlated to the rate of N solubilisation for each of the protein sources evaluated.     

黄壤稻田土壤微生物生物量碳磷对长期不同施肥的响应

本研究以进行了22年的黄壤稻田长期定位试验为依托,分析了不同施肥模式下土壤碳(C)、磷(P)与微生物生物量C(MBC)、P(MBP)的变化及其耦合特征,旨在探讨黄壤地区合理培肥模式以提高土壤磷素有效性.试验包括10个处理,分别为不施肥(CK)、单施氮肥(N)、磷钾配合(PK)、氮钾配合(NK)、氮磷配合(NP)、氮磷钾配合(NPK)、单施有机肥(M)、3种有机无机肥配施(1/4M+3/4NP、0.5MNP、MNPK).结果表明:与不施肥相比,N和NK处理土壤有机碳(TOC)、全磷(TP)、MBC、MBP均有不同程度的降低,施磷处理(PK、NP、NPK)则有不同程度的提升;与不施肥和施用无机肥相比,施用有机肥各处理TOC、MBC、MBP及MBP/TP均显著增加,其中M和MNPK处理增幅最大.MBC/MBP、TOC/MBP、MBC/TP以施用有机肥处理最低,N处理最高.土壤MBC、MBP及其耦合关系与土壤TOC和有效磷均呈极显著相关,TOC是影响MBC、MBP、MBP/TP的直接因素,而有效磷则是影响MBC/MBP、TOC/MBP、MBC/TP的直接因素.土壤MBP及碳、磷耦合关系各指标可以有效区分单施化肥和施用有机肥的不同施肥方式,可作为评价黄壤稻田磷素肥力的生物学指标.配施有机肥是增加黄壤稻田磷有效性、提高土壤供磷潜力和保持土壤生物健康的有效途径.     

From canopy to seed: Loss of snow drives directional changes in forest composition

Climate change is altering the conditions for tree recruitment, growth, and survival, and impacting forest community composition. Across southeast Alaska, USA, and British Columbia, Canada, Callitropsis nootkatensis (Alaska yellow‐cedar) is experiencing extensive climate change‐induced canopy mortality due to fine‐root death during soil freezing events following warmer winters and the loss of insulating snowpack. Here, we examine the effects of ongoing, climate‐driven canopy mortality on forest community composition and identify potential shifts in stand trajectories due to the loss of a single canopy species. We sampled canopy and regenerating forest communities across the extent of C. nootkatensis decline in southeast Alaska to quantify the effects of climate, community, and stand‐level drivers on C. nootkatensis canopy mortality and regeneration as well as postdecline regenerating community composition. Across the plot network, C. nootkatensis exhibited significantly higher mortality than co‐occurring conifers across all size classes and locations. Regenerating community composition was highly variable but closely related to the severity of C. nootkatensis mortality. Callitropsis nootkatensis canopy mortality was correlated with winter temperatures and precipitation as well as local soil drainage, with regenerating community composition and C. nootkatensis regeneration abundances best explained by available seed source. In areas of high C. nootkatensis mortality, C. nootkatensis regeneration was low and replaced by Tsuga. Our study suggests that climate‐induced forest mortality is driving alternate successional pathways in forests where C. nootkatensis was once a major component. These pathways are likely to lead to long‐term shifts in forest community composition and stand dynamics. Our analysis fills a critical knowledge gap on forest ecosystem response and rearrangement following the climate‐driven decline of a single species, providing new insight into stand dynamics in a changing climate. As tree species across the globe are increasingly stressed by climate change‐induced alteration of suitable habitat, identifying the autecological factors contributing to successful regeneration, or lack thereof, will provide key insight into forest resilience and persistence on the landscape.     

Distinct roles of TIR and non-TIR regions in the subcellular localization and signaling properties of MyD88

MyD88 is a cytoplasmic adaptor protein that is critical for Toll-like receptor (TLR) signaling. The subcellular localization of MyD88 is characterized as large condensed forms in the cytoplasm. The mechanism and significance of this localization with respect to the signaling function, however, are currently unknown. Here, we demonstrate that MyD88 localization depends on the entire non-TIR region and that the correct cellular targeting of MyD88 is indispensable for its signaling function. The Toll-interleukin I receptor-resistance (TIR) domain does not determine the subcellular localization, but it mediates interaction with specific TLRs. These findings reveal distinct roles for the TIR and non-TIR regions in the subcellular localization and signaling properties of MyD88.     

The Arabidopsis thaliana trehalase is a plasma membrane-bound enzyme with extracellular activity

The lack of trehalose accumulation in most plant species has been partly attributed to the presence of an active trehalase. Although trehalose synthesis enzymes are thought to be cytosolic, and previous studies have indicated that trehalase activity is extracellular, the exact location of the enzyme has not yet been established in plant cell. We present evidence that the yet uncharacterised full-length Arabidopsis trehalase is a plasma membrane-bound protein, probably anchored to the membrane through a predicted N-terminal membrane spanning domain. The full-length AtTRE1, when expressed in yeast can functionally substitute for the extracellularly active trehalase Ath1p, by sustaining the growth of an ath1 null mutant strain on trehalose and at pH 4.8. We further demonstrate that AtTRE1 expressed in yeast is plasma membrane-bound as in plant cell. In light of these findings, the regulation of plant cell endogenous trehalose by trehalase is discussed.