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Iron (Fe) toxicity is a major abiotic stress in lowland rice production. Breeding tolerant varieties has proven challenging due to the complex genetic architecture of Fe toxicity tolerance and the strong genotype-by-environment interactions. Additionally, conventional methods for phenotyping visible stress symptoms are often inaccurate, inconsistent, and lack reproducibility. In our previous work, we identified that ascorbate redox regulation, mediated by the activities of dehydroascorbate reductase (DHAR) and ascorbate oxidase (AO), contributed to high tolerance in an indica rice genotype across various environments. To explore whether this mechanism is common among other rice genotypes, we selected ten genotypes with contrasting stress symptoms under Fe-toxic conditions to examine the roles of DHAR and AO in regulating Fe toxicity tolerance. Additionally, we aimed to develop objective and accurate image-based phenotyping methods to replace the traditional leaf bronzing scoring method. Among the ten genotypes we tested, we found significant positive correlations between DHAR activity and stress symptoms in plants grown under both Fe toxicity and control conditions, suggesting a general link between ascorbate redox regulation and Fe toxicity tolerance. Using RGB signals from leaf images of plants exposed to 1000 mg/L Fe2+, we evaluated 36 different color indices to quantify stress symptoms. We identified the normalized greenâred difference index as most significant in quantifying stress symptoms under Fe toxicity conditions. Our findings suggest that DHAR activity could be potentially employed as a biomarker in the screening of rice germplasms and breeding tolerant cultivars to Fe toxicity.
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The investigation of leachate leakage at numerous landfill sites is urgently needed. This study presents an exploration of environmental tracing methods using δ2H and δ13C-difference in dissolved carbon (δ13CDIC-DOC) to localize leachate leak points at landfill sites. δ2H, δ13CDIC, δ13CDOC, δ18O, and an array of physicochemical indices (e.g., total dissolved solids, temperature, and oxidation reduction potential) were monitored in both leachate and groundwater from different zones of a landfill site in China during the year of 2021-2023. Moreover, data for these parameters (i.e., the isotopic composition and physicochemical indices) from twelve published landfill cases were also collected, and these groundwater/leachate data points were located within 1 km away from the landfill boundary. Then statistical analyses, such as Pearson correlation analysis and redundancy analysis (RDA), were performed using both the detected and collected parameters at landfill sites. Consequently, the intensity of interaction between leachate and background groundwater was found to significantly control the isotopic fractionation features of hydrogen and carbon, and both the content of major contamination indicators (total dissolved solids, chemical oxygen demand, and ammoniacal nitrogen) and the oxidation reduction potential were the key impact factors. Accordingly, the water type used to indicate leachate leakage points was determined to be leachate that significantly interacted with the background groundwater or precipitation (LBGP). δ2H showed a perfect linear correlation (0.81 ≤ r2 < 1.0) with δ13CDIC-DOC in leachate under highly anaerobic landfill conditions, and the δ2H & δ13CDIC-DOC combinations in the LBGP were significantly different from those in the other water types. For groundwater with total dissolved solids lower than 1400 mg/L at landfill sites, a strong positive linear correlation (r = 0.83) was revealed between δ13CDIC and δ13CDOC. Based on these insights, δ2H versus δ13CDIC-DOC plots and RDA using δ2H and δ13CDIC-DOC as response variables were proposed to localize leak points at both lined landfills and leachate facilities. These findings further understanding of the isotopic fractionation features of hydrogen, carbon, and oxygen and provide novel environmental tracer methods for investigating leachate leak points at MSW landfill sites.
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Groundwater pollution at landfill sites poses a significant risk to human health and ecological security. However, efficiently tracking pollution plumes in a polluted aquifer with variable pollutants remains challenging. In order to track groundwater pollution plumes at landfill sites, an in-situ borehole hydrochemical and hydrodynamic profile (BHHP) method was developed. Total dissolved solids (TDS), oxidation-reduction potential (ORP), and ammonia nitrogen were selected as the hydrochemical indicators. Meanwhile, the hydrodynamic indicators included flow direction and flow velocity of groundwater. Among the three hydrochemical indicators, TDS and ORP were analyzed to be the prior alternative ones for the BHHP application. The BHHP method was successfully applied to track groundwater pollution plumes at a typical valley-type landfill site and its neighboring downstream zone. Consequently, four groundwater pollution plumes of different types and different scales were identified in both horizontal and vertical directions within the depth of 0-50 m, and the various pollution sources for the detected pollution plumes were revealed. Furthermore, the BHHP method was validated using sampling test results of groundwater chloride and chemical oxygen demand at the surveyed landfill site.
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Contaminantes Ambientales , Agua Subterránea , Humanos , Hidrodinámica , Contaminación Ambiental , Instalaciones de Eliminación de ResiduosRESUMEN
Contamination of paddy soils with arsenic (As) can cause phytotoxicity in rice and increase the accumulation of arsenic in grains. The uptake and accumulation of As in rice depends on the different As species present in the soil. Plants detoxify As by conjugating and sequestering xenobiotic compounds into vacuoles using various enzymes. However, the severity of damage induced by arsenite (As(III)) and arsenate (As(V)), as well as the roles of glutathione S-transferase in detoxifying these As species in rice, are not fully understood. In this study, we developed plant materials overexpressing a glutathione S-transferase gene OsGSTU40 under the control of the maize UBIL promoter. Through systematic investigations of both wild-type Nipponbare (Oryza sativa L., ssp. japonica) and OsGSTU40 overexpression lines under chronic or acute stress of As, we aimed to understand the toxic effects of both As(III) and As(V) on rice plants at the vegetative growth stage. We hypothesized that (i) As(III) and As(V) have different toxic effects on rice plants and (ii) OsGSTU40 played positive roles in As toxicity tolerance. Our results showed that As(III) was more detrimental to plant growth than As(V) in terms of plant growth, biomass, and lipid peroxidation in both chronic and acute exposure. Furthermore, overexpression of OsGSTU40 led to better plant growth even though uptake of As(V), but not As(III), into shoots was enhanced in transgenic plants. In acute As(III) stress, transgenic plants exhibited a lower level of lipid peroxidation than wild-type plants. The element composition of plants was dominated by the different As stress treatments rather than by the genotype, while the As concentration was negatively correlated with phosphorus and silicon. Overall, our findings suggest that As(III) is more toxic to plants than As(V) and that glutathione S-transferase OsGSTU40 differentially affects plant reactions and tolerance to different species of arsenic.
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Arsénico , Arsenitos , Oryza , Arseniatos/toxicidad , Arseniatos/metabolismo , Arsénico/toxicidad , Arsenitos/toxicidad , Arsenitos/metabolismo , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Plantas Modificadas Genéticamente/genética , Expresión Génica , Raíces de Plantas/metabolismoRESUMEN
Ammonium nitrogen (NH4+-N) is a typical inorganic pollutant in the groundwater at landfill sites, and high-concentration NH4+-N is toxic to humans and organisms. Zeolite can effectively remove NH4+-N in water by adsorption, and it is suitable to be used as a type of reactive materials for permeable reactive barriers (PRBs). A passive sink-zeolite PRB (PS-zPRB) with higher capture efficiency than a continuous permeable reactive barrier (C-PRB) was proposed. And a passive sink configuration was integrated with PRB in the PS-zPRB, this configuration enabled the high hydraulic gradient of groundwater at the treated sites to be fully utilized. In order to explore treatment efficiency for groundwater NH4+-N using the PS-zPRB, numerical modeling on decontamination of NH4+-N plumes at a landfill site was performed. The results indicated that the NH4+-N concentrations of PRB effluent gradually decreased from 21.0 mg/L to 0.5 mg/L within 5 y, and met the drinking water standards after treatment for 900 d. The decontamination efficiency index of PS-zPRB was consistently higher than 95% within 5 y, and the service life of PS-zPRB appeared over 5 y. The capture width of PS-zPRB effectively exceeded the PRB length by around 47%. Compared with C-PRB, the capture efficiency of PS-zPRB was increased by around 28%, and the reactive material of PS-zPRB was saved by approximately 23% in volume.
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Restauración y Remediación Ambiental , Agua Subterránea , Contaminantes Químicos del Agua , Zeolitas , Humanos , Contaminantes Químicos del Agua/análisis , Contaminación AmbientalRESUMEN
BACKGROUND: Anopheles maculatus, Anopheles minimus and Anopheles dirus are the major vectors of malaria transmission in the Greater Mekong Subregion (GMS). The malaria burden in this region has decreased significantly in recent years as all GMS countries progress towards malaria elimination. It is necessary to investigate the Anopheles diversity and abundance status and assess the Plasmodium infection rates to understand the malaria transmission potential of these vector species in GMS countries to guide the development of up-to-date vector control strategies and interventions. METHODS: A survey of mosquitoes was conducted in Stung Treng, Sainyabuli and Phongsaly Provinces on the Cambodia-Laos, Thailand-Laos and China-Laos borders, respectively. Mosquito collection was done by overnight trapping at sentinel sites in each province. After morphological identification, the 18S rRNA-based nested-PCR was performed to detect malaria parasites in the captured Anopheles mosquitoes. RESULTS: A total of 18 965 mosquitoes comprising of 35 species of 2 subgenera (Subgenus Anopheles and Subgenus Cellia) and 4 tribes (Tribes Culicini, Aedini, Armigerini and Mansoniini) were captured. Tribe Culicini accounted for 85.66% of captures, followed by Subgenus Anopheles (8.15%). Anopheles sinensis dominated the Subgenus Anopheles by 99.81%. Plasmodium-infection was found in 25 out of the 1 683 individual or pooled samples of Anopheles. Among the 25 positive samples, 19, 5 and 1 were collected from Loum, Pangkhom and Siem Pang village, respectively. Eight Anopheles species were found infected with Plasmodium, i.e., An. sinensis, Anopheles kochi, Anopheles vagus, An. minimus, Anopheles annularis, Anopheles philippinensis, Anopheles tessellatus and An. dirus. The infection rates of Plasmodium falciparum, Plasmodium vivax and mixture of Plasmodium parasite species were 0.12% (2/1 683), 1.31% (22/1 683) and 0.06% (1/1 683), respectively. CONCLUSIONS: Overall, this survey re-confirmed that multiple Anopheles species carry malaria parasites in the international border areas of the GMS countries. Anopheles sinensis dominated the Anopheles collections and as carriers of malaria parasites, therefore may play a significant role in malaria transmission. More extensive investigations of malaria vectors are required to reveal the detailed vector biology, ecology, behaviour, and genetics in GMS regions in order to assist with the planning and implementation of improved malaria control strategies.
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Anopheles , Malaria , Plasmodium , Animales , Malaria/prevención & control , Anopheles/parasitología , Tailandia/epidemiología , Laos , Cambodia , Insectos Vectores/parasitología , Mosquitos Vectores , ChinaRESUMEN
BACKGROUND: Vector control is still a pivotal method for preventing malaria, and its potency is weakened by the increasing resistance of vectors to chemical insecticides. As the most abundant and vital malaria vector in Southeast Asia, the chemical insecticide resistance status in Anopheles sinensis remains elusive in Laos, which makes it imperative to evaluate the true nature of chemical insecticide resistance-associated genetic mutations in An. sinensis in Laos. METHODS: Adult An. sinensis were collected from three border regions in Laos. DNA was extracted from individual mosquitoes. PCR amplification and DNA sequencing of a fragment containing codon 1014 of the voltage-gated sodium channel (vgsc) gene were completed to study the kdr allele frequency distribution, kdr intron polymorphism, population genetic diversity, and the evolutionary status of the kdr codon. The mitochondrial cytochrome c oxidase subunit II gene (COII) was amplified and sequenced to examine population variations, genetic differentiation, spatial population structure, population expansion, and gene flow patterns. RESULTS: Nine wild kdr haplotypes of the vgsc gene were detected in this study, and eight of them, namely 1014L1, 1014L2, 1014L4, 1014L7, 1014L9, 1014L10, 1014L11, and 1014L21, were discovered in the China-Laos border (northern Laos), while 1014L3 was only detected in the Thailand-Laos border (northwestern Laos) and Cambodia-Laos border (southern Laos). The newly identified haplotype, 1014L21, was uniquely distributed in the China-Laos border and was not identified in other countries. Based on sequence analysis of the mitochondrial COII genes, significant genetic differentiation and limited gene flow were detected between the China-Laos and Cambodia-Laos An. sinensis populations, which suggested that those two regions were genetically isolated. The distinct distribution of the kdr haplotype frequencies is probably the result of geographical isolation in mosquito populations. CONCLUSIONS: Lack of kdr mutations in the vgsc gene was probably due to genetic isolation and the absence of intense selection pressure in the three border regions of Laos. This study reveals that pyrethroid-based chemical insecticides are still appropriate for battling An. sinensis in parts of Laos, and routine monitoring of chemical insecticide resistance should be continuously implemented and focused on more restricted areas as part of chemical insecticide resistance management.
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Anopheles , Insecticidas , Malaria , Piretrinas , Canales de Sodio Activados por Voltaje , Animales , Anopheles/genética , Cambodia , China , ADN Mitocondrial/genética , Genética de Población , Resistencia a los Insecticidas/genética , Laos , Mosquitos Vectores/genética , Mutación , Tailandia , Canales de Sodio Activados por Voltaje/genéticaRESUMEN
SWEETs play important roles in intercellular sugar transport. Induction of SWEET sugar transporters by Transcription Activator-Like effectors (TALe) of Xanthomonas ssp. is key for virulence in rice, cassava and cotton. We identified OsSWEET11b with roles in male fertility and potential bacterial blight (BB) susceptibility in rice. While single ossweet11a or 11b mutants were fertile, double mutants were sterile. As clade III SWEETs can transport gibberellin (GA), a key hormone for spikelet fertility, sterility and BB susceptibility might be explained by GA transport deficiencies. However, in contrast with the Arabidopsis homologues, OsSWEET11b did not mediate detectable GA transport. Fertility and susceptibility therefore are likely to depend on sucrose transport activity. Ectopic induction of OsSWEET11b by designer TALe enabled TALe-free Xanthomonas oryzae pv. oryzae (Xoo) to cause disease, identifying OsSWEET11b as a potential BB susceptibility gene and demonstrating that the induction of host sucrose uniporter activity is key to virulence of Xoo. Notably, only three of six clade III SWEETs are targeted by known Xoo strains from Asia and Africa. The identification of OsSWEET11b is relevant for fertility and for protecting rice against emerging Xoo strains that target OsSWEET11b.
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Proteínas de Transporte de Membrana/metabolismo , Oryza , Proteínas de Plantas/metabolismo , Xanthomonas , Proteínas Bacterianas/metabolismo , Resistencia a la Enfermedad/genética , Fertilidad , Regulación de la Expresión Génica de las Plantas , Proteínas de Transporte de Membrana/genética , Oryza/metabolismo , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/genética , Sacarosa , Xanthomonas/genéticaRESUMEN
Rising tropospheric ozone concentrations can cause rice yield losses and necessitate the breeding of ozone-tolerant rice varieties. However, ozone tolerance should not compromise the resistance to important biotic stresses such as the rice blast disease. Therefore, we investigated the interactive effects of ozone and rice blast disease on nine different rice varieties in an experiment testing an ozone treatment, blast inoculation, and their interaction. Plants were exposed to an ozone concentration of 100 ppb for 7 h per day or ambient air throughout the growth period. Half of the plants were simultaneously infected with rice blast inoculum. Grain yield was significantly reduced in the blast treatment (17%) and ozone treatment (37%), while the combination of both stresses did not further decrease grain yields compared to ozone alone. Similar trends occurred for physiological traits such as vegetation indices, normalized difference vegetation index (NDVI), photochemical reflectance index (PRI), Lichtenthaler index 2 (Lic2), and anthocyanin reflectance index 1 (ARI1), as well as stomatal conductance and lipid peroxidation. Ozone exposure mitigated the formation of visible blast symptoms, while blast inoculation did not significantly affect visible ozone symptoms. Although different genotypes showed contrasting responses to the two types of stresses, no systematic pattern was observed regarding synergies or trade-offs under the two types of stresses. Therefore, we conclude that despite the similarities in physiological stress responses to ozone and blast, the tolerance to these stresses does not appear to be genetically linked in rice.
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Magnaporthe , Oryza , Ozono , Ascomicetos , Grano Comestible , Genotipo , Magnaporthe/fisiología , Fitomejoramiento , Enfermedades de las Plantas/genéticaRESUMEN
INTRODUCTION: Long QT syndrome (LQTS) is a life-threatening inherited channelopathy, and prolonged QT intervals easily trigger malignant arrhythmias, especially torsades de pointes and ventricular fibrillation. MATERIALS AND METHODS: The proband with overlapped phenotypes of LQTS and sinoatrial node dysfunction underwent some necessary examinations, including echocardiography, electrocardiogram (ECG), and Holter monitoring. Next, whole-exome sequencing was performed, and candidate genes were validated by Sanger sequencing. RNA secondary structure and protein physical-chemical parameter analyses were used to predict the possible structural change of the proteins induced by the mutations. RESULTS: We identified the digenic heterozygous mutations of KCNH2 p.307_308del (NM_001204798, c.921_923del) and SCN5A p.R1865H (NM_001160160, c.G5594A) in the female and young proband (II: 1) of LQTS and ventricular fibrillation with repeat syncope at rest. Subsequently, she occurred with obvious sinus arrest with persistent ventricular pacing of implantable cardioverter-defibrillator. The heterozygous SCN5Ap.R1865H was carried by her father and sister but not carried by I:2. II:1 carried with KCNH2 p.307_308del as a de novo mutation, but not existed in other family members. RNA secondary structure of KCNH2 p.307_308del showed a false regional double helix, and its amino acids' hydrophobicity was significantly weakened. For the Nav 1.5 protein property, SCN5A p.R1865H slightly increased the molecular weight and aliphatic index but reduced the instability index. CONCLUSIONS: The digenic heterozygous KCNH2 and SCN5A mutations were associated with young early-onset long QT syndrome and sinoatrial node dysfunction.
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Síndrome de QT Prolongado , Nodo Sinoatrial , Canal de Potasio ERG1/genética , Electrocardiografía , Canales de Potasio Éter-A-Go-Go/genética , Femenino , Humanos , Canal de Potasio KCNQ1/genética , Síndrome de QT Prolongado/genética , Mutación , Canal de Sodio Activado por Voltaje NAV1.5RESUMEN
Ascorbate (Vitamin C) fulfills various functions in plant photosynthesis and abiotic stress tolerance. The four key enzymes involved in the ascorbate-turnover pathway are ascorbate peroxidase, ascorbate oxidase, monodehydroascorbate reductase, and dehydroascorbate reductase. Several reports have shown the pivotal roles of these enzymes in plant development and stress tolerance. Therefore, reliable and rapid assay protocols are required for researchers to investigate their enzymatic activities during plant development and stress responses. Previously published methods for analyzing these enzymatic activities rely on cuvette spectrophotometers, which can only handle one sample per test, leading to a prolonged investigation. In this protocol, we employed a 96-well microplate reader to analyze at least eight samples with two technical replicates simultaneously. We analyzed two rice (Oryza sativa L.) genotypes with distinct ascorbate oxidase and dehydroascorbate reductase activities to demonstrate the assay process, including plant growth, sample preparation, reaction setup, and data analysis. Our protocol provides a high throughput method for investigating ascorbate turnover-related enzymatic activities in plants.
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Rice remains a major staple food source for the rapidly growing world population. However, regular occurrences of carcinogenic arsenic (As) minerals in waterlogged paddy topsoil pose a great threat to rice production and consumers across the globe. Although As contamination in rice has been well recognized over the past two decades, no suitable rice germplasm had been identified to exploit in adaptive breeding programs. Therefore, this current study identified suitable rice germplasm for As tolerance and exclusion based on a variety of traits and investigated the interlinkages of favorable traits during different growth stages. Fifty-three different genotypes were systematically evaluated for As tolerance and accumulation. A germination screening assay was carried out to identify the ability of individual germplasm to germinate under varying As stress. Seedling-stage screening was conducted in hydroponics under varying As stress to identify tolerant and excluder genotypes, and a field experiment was carried out to identify genotypes accumulating less As in grain. Irrespective of the rice genotypes, plant health declined significantly with increasing As in the treatment. However, genotype-dependent variation in germination, tolerance, and As accumulation was observed among the genotypes. Some genotypes (WTR1-BRRI dhan69, NPT-IR68552-55-3-2, OM997, and GSR IR1-5-Y4-S1-Y1) showed high tolerance by excluding As in the shoot system. Arsenic content in grain ranged from 0.12 mg kg-1 in Huang-Hua-Zhan (indica) from China to 0.48 mg kg-1 in IRAT 109 (japonica) from Brazil. This current study provides novel insights into the performance of rice genotypes under varying As stress during different growth stages for further use in ongoing breeding programs for the development of As-excluding rice varieties for As-polluted environments.
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As a bio-based counterpart of poly(butylene adipate-co-terephthalate) (PBAT), the well-known commercially available biodegradable aliphatic-aromatic copolyester, poly(butylene succinate-co-terephthalate) (PBST) has comparable physical and mechanical properties, but its gas barrier properties, which are very important for packaging material and mulch film applications, have not yet been reported in literature. In this paper, the O2, CO2 and water vapor barrier properties of PBST vs. PBAT were comparatively studied and reported for the first time. Theoretical calculation of O2 and CO2 permeation coefficients via group contribution method was also conducted. The barrier properties of PBST show clear copolymer composition dependence due to different contribution of BS and BT repeat units and composition-dependent crystallinity. Comparing with PBAT, PBST with close copolymer and three-phase (crystalline, amorphous, rigid amorphous) compositions shows 3.5 times O2 and CO2 and 1.5 times water vapor barrier properties. The slower segment movement and less free volume of PBST, and therefore slower gas diffusion in PBST, accounts for its superior O2 and CO2 barrier, while the better hydrophilicity of PBST counteracts partial contribution of slower segment movement so that the improvement in water vapor barrier is not as high as in O2 and CO2 barrier.
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BACKGROUND: The Anopheles hyrcanus group, which includes 25 species, is widely distributed in the Oriental and Palaearctic regions. Given the difficulty in identifying cryptic or sibling species based on their morphological characteristics, molecular identification is regarded as an important complementary approach to traditional morphological taxonomy. The aim of this study was to reconstruct the phylogeny of the Hyrcanus group using DNA barcoding markers in order to determine the phylogenetic correlations of closely related taxa and to compare these markers in terms of identification efficiency and genetic divergence among species. METHODS: Based on data extracted from the GenBank database and data from the present study, we used 399 rDNA-ITS2 sequences of 19 species and 392 mtDNA-COII sequences of 14 species to reconstruct the molecular phylogeny of the Hyrcanus group across its worldwide range. We also compared the performance of rDNA-ITS2 against that of mtDNA-COII to assess the genetic divergence of closely related species within the Hyrcanus group. RESULTS: Average interspecific divergence for the rDNA-ITS2 sequence (0.376) was 125-fold higher than the average intraspecies divergence (0.003), and average interspecific divergence for the mtDNA-COII sequence (0.055) was eightfold higher than the average intraspecies divergence (0.007). The barcoding gap ranged from 0.015 to 0.073 for rDNA-ITS2, and from 0.017 to 0.025 for mtDNA-COII. Two sets of closely related species, namely, Anophels lesteri and An. paraliae, and An. sinensis, An. belenrae and An. kleini, were resolved by rDNA-ITS2. In contrast, the relationship of An. sinensis/An. belenrae/An. kleini was poorly defined in the COII tree. The neutrality test and mismatch distribution revealed that An. peditaeniatus, An. hyrcanus, An. sinensis and An. lesteri were likely to undergo hitchhiking or population expansion in accordance with both markers. In addition, the population of an important vivax malaria vector, An. sinensis, has experienced an expansion after a bottleneck in northern and southern Laos. CONCLUSIONS: The topology of the Hyrcanus group rDNA-ITS2 and mtDNA-COII trees conformed to the morphology-based taxonomy for species classification rather than for that for subgroup division. rDNA-ITS2 is considered to be a more reliable diagnostic tool than mtDNA-COII in terms of investigating the phylogenetic correlation between closely related mosquito species in the Hyrcanus group. Moreover, the population expansion of an important vivax malaria vector, An. sinensis, has underlined a potential risk of malaria transmission in northern and southern Laos. This study contributes to the molecular identification of the Anopheles hyrcanus group in vector surveillance.
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Anopheles/clasificación , Anopheles/genética , ADN Mitocondrial/genética , ADN Ribosómico/genética , Mosquitos Vectores/clasificación , Mosquitos Vectores/genética , Filogenia , Animales , Código de Barras del ADN Taxonómico/métodos , ADN Intergénico/genéticaRESUMEN
The rice black-streaked dwarf virus (RBSDV) disease causes severe rice yield losses in Asia. RNA interference (RNAi) has been widely applied to develop antiviral varieties in plants. So far, only a few studies reported the application of RNAi in rice against RBSDV and most of them are lack of enough data to support its breeding potential, which limited the progress on developing RBSDV-resistant variety. In this study, we generated three RNAi constructs to specifically target three RBSDV genes (S1, S2 and S6), respectively. We confirmed that RNAi targeting RBSDV S6 conferred rice with almost full immunity to RBSDV through phenotyping test in eight consecutive years in both artificial inoculation and field trials, while RNAi of S1 or S2 only leads to partially increased resistance. The S6RNAi was also found conferring strong resistance to southern rice black-streaked dwarf virus (SRBSDV), a novel species closely related to RBSDV that outbroke recently in Southern China. In particular, no adverse effects on agronomical and developmental traits were found in S6RNAi transgenic lines. The marker-free transgenic lines with S6RNAi, driven by either maize ubiquitin-1 promoter or rice rbcS green tissue expression promoter, in elite rice background should have great potential in breeding of resistant varieties to both RBSDV and SRBSDV and provide a basis for further safety evaluation and commercial application.
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Oryza , Virosis , China , Oryza/genética , Fitomejoramiento , Enfermedades de las Plantas/genética , Interferencia de ARNRESUMEN
Iron (Fe) toxicity is one of the most common mineral disorders affecting rice (Oryza sativa) production in flooded lowland fields. Oryza meridionalis is indigenous to northern Australia and grows in regions with Fe-rich soils, making it a candidate for use in adaptive breeding. With the aim of understanding tolerance mechanisms in rice, we screened a population of interspecific introgression lines from a cross between O. sativa and O. meridionalis for the identification of quantitative trait loci (QTLs) contributing to Fe-toxicity tolerance. Six putative QTLs were identified. A line carrying one introgression from O. meridionalis on chromosome 9 associated with one QTL was highly tolerant despite very high shoot Fe concentrations. Physiological, biochemical, ionomic, and transcriptomic analyses showed that the tolerance of the introgression lines could partly be explained by higher relative Fe retention in the leaf sheath and culm. We constructed the interspecific hybrid genome in silico for transcriptomic analysis and identified differentially regulated introgressed genes from O. meridionalis that could be involved in shoot-based Fe tolerance, such as metallothioneins, glutathione S-transferases, and transporters from the ABC and MFS families. This work demonstrates that introgressions of O. meridionalis into the O. sativa genome can confer increased tolerance to excess Fe.
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Oryza , Australia , Hierro , Oryza/genética , Fitomejoramiento , Sitios de Carácter Cuantitativo/genéticaRESUMEN
Simultaneous stiffening, strengthening, and toughening of biodegradable polymers, such as poly(butylene adipate-co-terephthalate) (PBAT) and others, is necessary for their use in packaging and agriculture applications. However, a high content of nanoinclusions is usually required, leading to a tradeoff between composite toughness and strength or stiffness in the reinforcement. Herein, we report an iterative reinforcement strategy that uses one nanocomposite to reinforce PBAT. An in-situ grafting polymerized cellulose nanocrystal (CNC)/PBAT (CNC-g-PBAT) nanocomposite consisting of ungrafted/free PBAT (PBATf) was used as an inclusion directly to reinforce a commercial PBAT. At an exceptionally low CNC usage of 0.02 wt.%, we achieved a simultaneous enhancement of the Young's modulus by 26 %, tensile strength by 27 %, elongation at break by 37 %, and toughness by 56 % over those for PBAT. To the best of our knowledge, such reinforcement efficiency is the highest among similar biodegradable polymer nanocomposites reported in the literature. The rheology, differential scanning calorimetry, and wide-angle X-ray diffraction measurements confirmed the mechanical reinforcement attributed to a synergistic contribution from PBATf and CNC-g-PBAT. In particular, the use of PBATf enhanced both stiffness and toughness of the composites, while the CNC-g-PBAT interacted within the polymer matrix and increased the crystallinity of the polymer matrix, leading to the strengthening and toughening effect. The strategy proposed here is greatly beneficial to producing high-performance biodegradable polymer nanocomposite films for packaging and agricultural applications using a very low amount of nanoinclusion.
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Plásticos Biodegradables/química , Nanocompuestos/química , Poliésteres/química , Agricultura/métodos , Plásticos Biodegradables/síntesis química , Rastreo Diferencial de Calorimetría , Celulosa/química , Elasticidad , Embalaje de Alimentos/métodos , Espectroscopía de Resonancia Magnética , Microscopía Electrónica de Transmisión , Nanocompuestos/ultraestructura , Nanopartículas/química , Reología , Espectroscopía Infrarroja por Transformada de Fourier , Resistencia a la Tracción , Termogravimetría , Difracción de Rayos XRESUMEN
Rice wild relatives (RWR) constitute an extended gene pool that can be tapped for the breeding of novel rice varieties adapted to abiotic stresses such as iron (Fe) toxicity. Therefore, we screened 75 Oryza genotypes including 16 domesticated O. sativa genotypes, one O. glaberrima, and 58 RWR representing 21 species, for tolerance to Fe toxicity. Plants were grown in a semi-artificial greenhouse setup, in which they were exposed either to control conditions, an Fe shock during the vegetative growth stage (acute treatment), or to a continuous moderately high Fe level (chronic treatment). In both stress treatments, foliar Fe concentrations were characteristic of Fe toxicity, and plants developed foliar stress symptoms, which were more pronounced in the chronic Fe stress especially toward the end of the growing season. Among the genotypes that produced seeds, only the chronic stress treatment significantly reduced yields due to increases in spikelet sterility. Moreover, a moderate but non-significant increase in grain Fe concentrations, and a significant increase in grain Zn concentrations were seen in chronic stress. Both domesticated rice and RWR exhibited substantial genotypic variation in their responses to Fe toxicity. Although no RWR strikingly outperformed domesticated rice in Fe toxic conditions, some genotypes scored highly in individual traits. Two O. meridionalis accessions were best in avoiding foliar symptom formation in acute Fe stress, while an O. rufipogon accession produced the highest grain yields in both chronic and acute Fe stress. In conclusion, this study provides the basis for using interspecific crosses for adapting rice to Fe toxicity.
Asunto(s)
Resistencia a la Enfermedad/genética , Hierro/toxicidad , Oryza/genética , Enfermedades de las Plantas/genética , Animales , Grano Comestible/efectos de los fármacos , Grano Comestible/genética , Grano Comestible/crecimiento & desarrollo , Etanol , Genotipo , Oryza/efectos de los fármacos , Oryza/crecimiento & desarrollo , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/genética , Hojas de la Planta/crecimiento & desarrollo , Semillas/genética , Semillas/crecimiento & desarrollo , Estrés Fisiológico/efectos de los fármacos , Estrés Fisiológico/genéticaRESUMEN
Genetic transformation is crucial for both investigating gene functions and for engineering of crops to introduce new traits. Rice (Oryza sativa L.) is an important model in plant research, since it is the staple food for more than half of the world's population. As a result, numerous transformation methods have been developed for both indica and japonica rice. Since breeders continuously develop new rice varieties, transformation protocols have to be adapted for each new variety. Here we provide an optimized transformation protocol with detailed tips and tricks for a new African variety Komboka using immature embryos. In Komboka, we obtained an apparent transformation rate of up to 48% for GUS/GFP reporter gene constructs using this optimized protocol. This protocol is also applicable for use with other elite indica rice varieties.