Effect of water availability on changes in root amino acids and associated rhizospere on root exudation of amino acids in Pisum sativum L

Root exudation is considered to regulate the abundance of the microbial community. It may vary both qualitatively and quantitatively in response to the environment in which the plant is growing. A part of exuded N derives from amino acids (AAs). This, in turn, may help plants to cope with abiotic stresses by favouring positive interactions with the rhizosphere environment, thus playing a potential role in maintaining healthy plants. In this respect, an under-investigated area is the effect of stress due to water deficit (WD). It is proposed that the AA profile in the rhizosphere may be altered by WD, reflecting a modulation of root AA exudation linked to a physiological response of the plant to water stress. To investigate this, Pisum sativum L. plants, grown in unsterilised Rhizobium leguminosarum-enriched soil, were stem-labelled with 15N-urea for 96 h, and then subjected/not subjected to 72 h of WD. The concentrations and abundance of 15N-labelling in individual AAs were determined in both roots and the associated rhizosphere at 24, 48 and 72 h after stress application. It was found that both AAs metabolism in the pea root and AAs exudation were strongly modified in WD conditions. After 24 h of WD, the concentrations of all measured AAs increased in the roots, accompanied by a dramatic stress-related increase in the 15N-labelling of some AAs. Furthermore, after 48–72 h of WD, the concentrations of Pro, Ala and Glu increased significantly within the rhizosphere, notably with a concomitant increase in 15N-enrichment in Pro, Ser, Asn, Asp, Thr and Ile. These results support the concept that, in response to WD, substantial amounts of recently assimilated N are rapidly translocated from the shoots to the roots, a portion of which is exuded as AAs. This leads to the rhizosphere being relatively augmented by specific AAs (notably HSer, Pro and Ala) in WD conditions, with a potential impact on soil water retention

Modulation de l'exsudation racinaire de composés azotés chez les Fabacées et réponse aux stress hydriques

Les légumineuses ont des rhizodépôts riches en azote (N). Leur emploi dans les rotations de cultures permet de réduire l\u27usage de fertilisants de synthèse. Parmi les composés azotés exsudés par les racines, les acides aminés (AA) sont reconnus pour leur rôle dans la régulation des communautés microbiennes de la rhizosphère, avec un impact sur la nutrition des plantes et leur résistance aux stress abiotiques. Cette exsudation est difficile à mesurer en raison des interactions avec les microorganismes et les particules minérales du sol. Nous avons fait l\u27hypothèse que l\u27exsudation racinaire d\u27AA varie en fonction du substrat, de la croissance et de l\u27humidité sol. Une méthode d\u27extraction des AA du sol a été mise au point pour estimer l\u27empreinte rhizosphérique en acides aminés (ERAA) dans le sol non stérilisé, comme résultat net des interactions entre l\u27exsudation d\u27AA et le milieu rhizosphérique. L\u27évolution de l\u27ERAA a ainsi été étudiée chez Medicago truncatula cultivée sur sable stérilisé ou sol non stérilisé, à 3 stades de croissance végétative, et chez Pisum sativum soumis ou non à un déficit hydrique ou un excès d\u27eau. Le marquage au 15N a été utilisé pour appréhender le mouvement des 15N-AA libérés par les racines vers la rhizosphère. L\u27ERAA augmente au cours de la croissance végétative et est dépendante en quantité et en qualité, du stade de la plante. Le déficit hydrique modifie l\u27ERAA avec la présence de plus de N récemment assimilé par la plante, plus de proline, d\u27alanine et de valine. En tenant compte des limites expérimentales, ces travaux ont permis de mieux comprendre les facteurs qui influencent l\u27ERAA et d\u27appréhender la rhizodéposition azotée des légumineuses

Amino acid fingerprint in the rhizosphere of Pisum sativum in response to water stress

In cropping systems, legumes release substantial amounts of nitrogen (N) into the soil, via rhizodeposition, and constitute a sustainable source of N, instead of synthetic N fertilisers (Fustec et al. 2010). More frequent or/and intense droughts and floodings, due to climate change and intensification of agriculture, may affect N rhizodeposition (Preece & Peñuelas 2016). However, the effects of water stress on this process are poorly documented. A part of N derived from root exudates, mainly in amino acids (AAs) form, is suspected shape and regulate rhizosphere microbial community, thus playing a potential role in maintaining plant health in case of abiotic stress (Moe 2013). We hypothesized that root AA exudation could change significantly, according to water availability, and would help to understand N metabolism changes in plant-rhizosphere interactions. Because studying exudation from plant grown in unsterilized soil is challenging (Oburger et al. 2013), we have measured the rhizosphere AA fingerprint (RAAF), as the result of interactions between AA exudation and rhizospheric environment. In addition, plants were stem-labeled (cotton-wick) with 15N-urea for 72 h to provide direct evidence of a link between root AA and exudation in the soil. The RAAF was measured in Pisum sativum rhizosphere, under either a water deficit or a water excess for 72 h. Water deficit decreases biomass accumulation in shoots but not in roots. Then, water deficit had no significant effect on total AAs released into the rhizosphere but, it significantly modified the composition of RAAF, with a preferential increase of proline, alanine and glutamate and a rise in isotopic enrichment of AAs derived from oxaloacetate in tricarboxylic acidic cycle (asparagine, aspartate, threonine and isoleucine). These results support the idea that, under the early stages of water deficit, recently assimilated N is rapidly translo-cated to the roots, and part of it is exudated in AAs. Most of the exudated AAs are known to have a specific role in increasing the water holding capacity around the root and to favour the establishment of positive interactions with plant-growth promoting bacteria (Apostel et al. 2013, Hinsinger et al. 2003). A study aimed at establishing a better understanding of the relationship between microorganisms and AA release under water deficit is now necessary

Assessing dengue transmission risk and a vector control intervention using entomological and immunological indices in Thailand: study protocol for a cluster-randomized controlled trial.

BACKGROUND: Dengue fever is the most common and widespread mosquito-borne arboviral disease in the world. There is a compelling need for cost-effective approaches and practical tools that can reliably measure real-time dengue transmission dynamics that enable more accurate and useful predictions of incidence and outbreaks. Sensitive surveillance tools do not exist today, and only a small handful of new control strategies are available. Vector control remains at the forefront for combating dengue transmission. However, the effectiveness of many current vector control interventions is fraught with inherent weaknesses. No single vector control method is effective enough to control both vector populations and disease transmission. Evaluations of novel larval and adult control interventions are needed. METHODS/DESIGN: A cluster-randomized controlled trial will be carried out between 2017 and 2019 in urban community clusters in Khon Kaen and Roi Et cities, northeastern Thailand. The effectiveness of a pyriproxyfen/spinosad combination treatment of permanent water storage containers will be evaluated on epidemiological and entomological outcomes, including dengue incidence, number of female adult dengue vectors infected or not infected with dengue virus (DENV), human exposure to Aedes mosquito bites, and several other indices. These indices will also be used to develop predictive models for dengue transmission and impending outbreaks. Epidemiological and entomological data will be collected continuously for 2 years, with the intervention implemented after 1 year. DISCUSSION: The aims of the trial are to simultaneously evaluate the efficacy of an innovative dengue vector control intervention and developing predictive dengue models. Assessment of human exposure to mosquito bites by detecting antibodies generated against Aedes saliva proteins in human blood samples has, so far, not been applied in dengue epidemiological risk assessment and disease surveillance methodologies. Likewise, DENV detection in mosquitoes (adult and immature stages) has not been used in any practical way for routine disease surveillance strategies. The integration of multiple outcome measures will assist health authorities to better predict outbreaks for planning and applying focal and timely interventions. The trial outcomes will not only be important for Thailand, but also for the entire Southeast Asian region and further afield. TRIAL REGISTRATION: ISRCTN, ISRCTN73606171 . Registered on 23 June 2017

Spatial and temporal patterns of dengue incidence in northeastern Thailand 2006-2016.

BACKGROUND: Dengue, a viral disease transmitted by Aedes mosquitoes, is an important public health concern throughout Thailand. Climate variables are potential predictors of dengue transmission. Associations between climate variables and dengue have usually been performed on large-scale first-level national administrative divisions, i.e. provinces. Here we analyze data on a finer spatial resolution in one province, which is often more relevant for effective disease control design. The objective of this study was to investigate the effect of seasonal variations, monthly climate variability, and to identify local clusters of symptomatic disease at the sub-district level based on reported dengue cases. METHODS: Data on dengue cases were retrieved from the national communicable disease surveillance system in Thailand. Between 2006 and 2016, 15,167 cases were recorded in 199 sub-districts of Khon Kaen Province, northeastern Thailand. Descriptive analyses included demographic characteristics and temporal patterns of disease and climate variables. The association between monthly disease incidence and climate variations was analyzed at the sub-district level using Bayesian Poisson spatial regression. A hotspot analysis was used to assess the spatial patterns (clustered/dispersed/random) of dengue incidence. RESULTS: Dengue was predominant in the 5-14 year-old age group (51.1%). However, over time, dengue incidence in the older age groups (> 15 years) gradually increased and was the most affected group in 2013. Dengue outbreaks coincide with the rainy season. In the spatial regression model, maximum temperature was associated with higher incidence. The hotspot analysis showed clustering of cases around the urbanized area of Khon Kaen city and in rural areas in the southwestern portion of the province. CONCLUSIONS: There was an increase in the number of reported dengue cases in older age groups over the study period. Dengue incidence was highly seasonal and positively associated with maximum ambient temperature. However, climatic variables did not explain all the spatial variation of dengue in the province. Further analyses are needed to clarify the detailed effects of urbanization and other potential environmental risk factors. These results provide useful information for ongoing prediction modeling and developing of dengue early warning systems to guide vector control operations

Underestimated role of legume roots for soil N fertility

Research ArticleNitrogen (N) is a major fertilizing element for plants. The distribution of N in legumes is influencing the efficiency of the next crop. Nitrogen storage in legumes is actually estimated by N fixation in shoots, whereas there is little knowledge on the contribution of roots and nodules to legume N and soil N. Here, we studied the contribution of roots and nodules of grain and pasture legumes to plant N and soil N in Mediterranean fields. Experiments were run under rainfed conditions for a 2-year period in three regions of Portugal. Entire plants including top plant and visible roots and nodules were sampled at the end of the growing seasons for grain legumes, sweet and yellow lupine, and over two harvests in case of pastures. N2 fixation was measured for grain legumes and pasture legumes using 15N tracing. Our results show that aboveground N concentration did not vary among legumes, but differed in the belowground tissues. Field studies show that 7–11%of total legume N was associated with roots and nodules. Data also show an allocation of 11– 14 kg N fixed t−1 belowground dry matter in indeterminate legumes, which represents half the amount of total aboveground plant. This finding demonstrates that investigation relying only on shoot Nunderestimates the role of legumes for soil N fertilityinfo:eu-repo/semantics/publishedVersio

Complex relationships between Aedes vectors, socio-economics and dengue transmission-Lessons learned from a case-control study in northeastern Thailand.

BACKGROUND/OBJECTIVES: Dengue fever is an important public health concern in most tropical and subtropical countries, and its prevention and control rest on vector surveillance and control. However, many aspects of dengue epidemiology remain unclear; in particular, the relationship between Aedes vector abundance and dengue transmission risk. This study aims to identify entomological and immunological indices capable of discriminating between dengue case and control (non-case) houses, based on the assessment of candidate indices, as well as individual and household characteristics, as potential risk factors for acquiring dengue infection. METHODS: This prospective, hospital-based, case-control study was conducted in northeastern Thailand between June 2016 and August 2019. Immature and adult stage Aedes were collected at the houses of case and control patients, recruited from district hospitals, and at patients' neighboring houses. Blood samples were tested by RDT and PCR to detect dengue cases, and were processed with the Nterm-34 kDa salivary peptide to measure the human immune response to Aedes bites. Socioeconomic status, and other individual and household characteristics were analyzed as potential risk factors for dengue. RESULTS: Study findings showed complex relationships between entomological indices and dengue risk. The presence of DENV-infected Aedes at the patient house was associated with 4.2-fold higher odds of dengue. On the other hand, Aedes presence (irrespective of infectious status) in the patient's house was negatively associated with dengue. In addition, the human immune response to Aedes bites, was higher in control than in case patients and Aedes adult abundance and immature indices were higher in control than in case houses at the household and the neighboring level. Multivariable analysis showed that children aged 10-14 years old and those aged 15-25 years old had respectively 4.5-fold and 2.9-fold higher odds of dengue infection than those older than 25 years. CONCLUSION: DENV infection in female Aedes at the house level was positively associated with dengue infection, while adult Aedes presence in the household was negatively associated. This study highlights the potential benefit of monitoring dengue viruses in Aedes vectors. Our findings suggest that monitoring the presence of DENV-infected Aedes mosquitoes could be a better indicator of dengue risk than the traditional immature entomological indices

Serological biomarker for assessing human exposure to Aedes mosquito bites during a randomized vector control intervention trial in northeastern Thailand.

BACKGROUND: Aedes mosquitoes are vectors for several major arboviruses of public health concern including dengue viruses. The relationships between Aedes infestation and disease transmission are complex wherein the epidemiological dynamics can be difficult to discern because of a lack of robust and sensitive indicators for predicting transmission risk. This study investigates the use of anti-Aedes saliva antibodies as a serological biomarker for Aedes mosquito bites to assess small scale variations in adult Aedes density and dengue virus (DENV) transmission risk in northeastern Thailand. Individual characteristics, behaviors/occupation and socio-demographics, climatic and epidemiological risk factors associated with human-mosquito exposure are also addressed. METHODS: The study was conducted within a randomized clustered control trial in Roi Et and Khon Kaen provinces over a consecutive 19 months period. Thirty-six (36) clusters were selected, each of ten houses. Serological and entomological surveys were conducted in all houses every four months and monthly in three sentinel households per cluster between September 2017 and April 2019 for blood spot collections and recording concurrent immature and adult Aedes indices. Additionally, the human exposure to Aedes mosquito bites (i.e., Mosquito Exposure Index or MEI) was estimated by ELISA measuring levels of human antibody response to the specific Nterm-34 kDa salivary antigen. The relationships between the MEI, vector infestation indices (adult and immature stages) and vector DENV infection were evaluated using a two-level (house and individual levels) mixed model analysis with one-month lag autoregressive correlation. RESULTS: There was a strong positive relationship between the MEI and adult Aedes (indoor and outdoor) density. Individuals from households with a medium mosquito density (mean difference: 0.091, p<0.001) and households with a high mosquito density (mean difference: 0.131, p<0.001) had higher MEI's compared to individuals from households without Aedes. On a similar trend, individuals from households with a low, medium or high indoor Aedes densities (mean difference: 0.021, p<0.007, 0.053, p<0.0001 and 0.037, p<0.0001 for low, medium and high levels of infestation, respectively) had higher MEI than individuals from houses without indoor Aedes. The MEI was driven by individual characteristics, such as gender, age and occupation/behaviors, and varied according to climatic, seasonal factors and vector control intervention (p<0.05). Nevertheless, the study did not demonstrate a clear correlation between MEI and the presence of DENV-infected Aedes. CONCLUSION: This study represents an important step toward the validation of the specific IgG response to the Aedes salivary peptide Nterm-34kDa as a proxy measure for Aedes infestation levels and human-mosquito exposure risk in a dengue endemic setting. The use of the IgG response to the Nterm-34 kDa peptide as a viable diagnostic tool for estimating dengue transmission requires further investigations and validation in other geographical and transmission settings