Open-Source Hardware Is a Low-Cost Alternative for Scientific Instrumentation and Research

Scientific research requires the collection of data in order to study, monitor, analyze, describe, or understand a particular process or event. Data collection efforts are often a compromise: manual measurements can be time-consuming and labor-intensive, resulting in data being collected at a low frequency, while automating the data-collection process can reduce labor requirements and increase the frequency of measurements, but at the cost of added expense of electronic data-collecting instrumentation. Rapid advances in electronic technologies have resulted in a variety of new and inexpensive sensing, monitoring, and control capabilities which offer opportunities for implementation in agricultural and natural-resource research applications. An Open Source Hardware project called Arduino consists of a programmable microcontroller development platform, expansion capability through add-on boards, and a programming development environment for creating custom microcontroller software. All circuit-board and electronic component specifications, as well as the programming software, are open-source and freely available for anyone to use or modify. Inexpensive sensors and the Arduino development platform were used to develop several inexpensive, automated sensing and datalogging systems for use in agricultural and natural-resources related research projects. Systems were developed and implemented to monitor soil-moisture status of field crops for irrigation scheduling and crop-water use studies, to measure daily evaporation-pan water levels for quantifying evaporative demand, and to monitor environmental parameters under forested conditions. These studies demonstrate the usefulness of automated measurements, and offer guidance for other researchers in developing inexpensive sensing and monitoring systems to further their research.

Novel Gluten-Free Amaranth and Oat Flour Cookies Fortified with Soybean Hulls

Soybean hulls, an abundant byproduct of soybean processing, contain rich phytochemicals, fibers, proteins, and minerals. Currently soybean hulls are primarily used as animal feeds. For value-added soybean hull utilization, 25% soybean hulls were substituted for amaranth or whole oat flour (WOF) in novel gluten-free cookies. Composition, nutritional values, water-holding capacities, correlation between properties, and pasting and rheological properties of soybean hulls, amaranth, and WOF were appraised in comparison to wheat flour. Water loss, cookie texture, and geometrical properties of the cookies were examined. The results disclosed that soybean hulls, amaranth and WOF contain higher protein content, minerals, fiber, special amino acids, and critical vitamins (C and K) than wheat flour. Considerably higher total amino acid content was found in soybean hulls (18.33%) than wheat flour (12.77%). Water-holding capacities increased by replacing amaranth and WOF with soybean hulls. Soybean hulls exhibited higher rheological elastic properties than amaranth, WOF and wheat flours. The soybean hulls utilized in amaranth or WOF cookies greatly improved their nutritional value, the water retention and moisture content along with acceptable physical properties when compared to wheat flour cookies. This study explored the feasibility and potential of utilizing soybean hulls with amaranth and WOF in gluten-free bakery products and other food applications.

UAV and Satellite-Based Sensing to Map Ecological States at the Landscape Scale

Mapping ecological states in semi-arid rangelands is crucial for effective land management and conservation efforts because it identifies difference in the ecological conditions across a landscape. This study presents an innovative approach for mapping two ecological states, Large Shrub Grass (LSG) and Large Shrub Eroded (LSE), within the Sandy Loam Upland and Deep (SLUD) ecological sites using a combination of drone and satellite data. The methodology leverages the Largest Patch Index (LPI) as a proxy metric to estimate eroded areas and classify ecological states. The integration of unmanned aerial vehicle (UAV) data with satellite-based remote sensing provides a scalable approach that can benefit various stakeholders involved in rangeland management. The study demonstrates the potential of this methodology by generating spatial layers at the landscape scale to inform on the state of rangeland ecosystems. The workflow showcases the power of remote sensing technology to map ecological states and addresses limitations in spatial coverage by integrating UAV and satellite data. By utilizing the bare ground LPI metric, which indicates the connectedness of bare ground, the methodology enables the classification of ecological states at a regional scale. This cost-effective approach potentially offers a standardized and reproducible method applicable across different sites and regions. The accuracy of the classification process is evaluated by comparing the results to ground-based polygons, dirt roads, and water locations. While the model performs well in identifying eroded areas, misclassifications occur in regions with mixed vegetation cover or low biomass. Future research should focus on incorporating temporal information from historical remote sensing archives to improve understanding of ecological state dynamics. Additionally, validation efforts can be enhanced by incorporating more ground-truth data and testing the methodology in diverse rangeland areas. The workflow serves as a blueprint for scaling up ecological states mapping in similar semi-arid rangelands. Further work should involve refining the approach through additional validation and exploring new remote sensing datasets. The methodology can be replicated in other regions to inform land management decisions, promote sustainable resource use, and advance the field of ecological states mapping.

Nitrogen mobility,ammonia volatilization,and estimated leaching loss from long-term manure incorporation in red soil

Nitrogen(N) loss from fertilization in agricultural fields has an unavoidable negative impact on the environment and a better understanding of the major pathways can assist in developing the best management practices. The aim of this study was to evaluate the fate of N fertilizers applied to acidic red soil(Ferralic Cambisol) after 19 years of mineral(synthetic) and manure fertilizer treatments under a cropping system with wheat-maize rotations. Five field treatments were examined: control(CK), chemical nitrogen and potash fertilizer(NK), chemical nitrogen and phosphorus fertilizer(NP), chemical nitrogen, phosphorus and potash fertilizer(NPK) and the NPK with manure(NPKM, 70% N from manure). Based on the soil total N storage change in 0–100 cm depth, ammonia(NH_3) volatilization, nitrous oxide(N_2O) emission, N plant uptake, and the potential N leaching loss were estimated using a mass balance approach. In contrast to the NPKM, all mineral fertilizer treatments(NK, NP and NPK) showed increased nitrate(NO_3~–) concentration with increasing soil depth, indicating higher leaching potential. However, total NH_3 volatilization loss was much higher in the NPKM(19.7%) than other mineral fertilizer treatments(≤4.2%). The N_2O emissions were generally low(0.2–0.9%, the highest from the NPKM). Total gaseous loss accounted for 1.7, 3.3, 5.1, and 21.9% for NK, NP, NPK, and NPKM treatments, respectively. Estimated N leaching loss from the NPKM was only about 5% of the losses from mineral fertilizer treatments. All data demonstrated that manure incorporation improved soil productivity, increased yield, and reduced potential leaching, but with significantly higher NH_3 volatilization, which could be reduced by improving the application method. This study confirms that manure incorporationis an essential strategy in N fertilization management in upland red soil cropping system.

Induction of avirulence by AVR-Pita1 in virulent U.S. field isolates of Magnaporthe oryzae’

The AVR-Pita1 gene,from the Chinese isolate O-137 of Magnaporthe oryzae,is an effector that determines the efficacy of the Pi-ta rice blast resistance gene.In the present study,the avirulence function of AVR-Pita1 was induced by transformation of field isolates(TM2,ZN19,B2 and B8)that originally were collected from the U.S.and are virulent on Pi-ta-carrying rice cultivars.The presence of AVR-Pita1 from O-137 in independent transformants was detected by PCR using AVR-Pita1 specific primers and verified by DNA sequencing and Southern blot analysis using the AVR-Pita1 coding region as a probe.The results of pathogenicity assays showed that the AVR-Pita1-transformed isolates were not able to infect rice cultivars Katy and Drew carrying Pi-ta.Control isolates that were transformed with inserts lacking the AVR-Pita1gene remained virulent.Our findings demonstrate that AVR-Pita1 can be used to induce novel gene-specific blast resistance in nature.

Change of soil productivity in three different soils after long-term field fertilization treatments

Soil productivity(SP) without external fertilization influence is an important indicator for the capacity of a soil to support crop yield.However,there have been difficulties in estimating values of SPs for soils after various long-term field treatments because the treatment without external fertilization is used but is depleted in soil nutrients,leading to erroneous estimation.The objectives of this study were to estimate the change of SP across different cropping seasons using pot experiments,and to evaluate the steady SP value(which is defined by the basal contribution of soil itself to crop yield) after various longterm fertilization treatments in soils at different geographical locations.The pot experiments were conducted in Jinxian of Jiangxi Province with paddy soil,Zhengzhou of Henan Province with fluvo-aquic soil,and Gongzhuling of Jilin Province with black soils,China.Soils were collected after long-term field fertilization treatments of no fertilizer(control;CK-F),chemical fertilizer(NPK-F),and combined chemical fertilizer with manure(NPKM-F).The soils received either no fertilizer(F0) or chemical fertilizer(F1) for 3-6 cropping seasons in pots,which include CK-P(control;no fertilizer from long-term field experiments for pot experiments),NPK-P(chemical fertilizer from long-term field experiments for pot experiments),and NPKM-P(combined chemical and organic fertilizers from long-term field experiments for pot experiments).The yield data were used to calculate SP values.The initial SP values were high,but decreased rapidly until a relatively steady SP was achieved at or after about three cropping seasons for paddy and fluvo-aquic soils.The steady SP values in the third cropping season from CK-P,NPK-P,and NPKM-P treatments were 37.7,44.1,and 50.0% in the paddy soil,34.2,38.1,and 50.0% in the fluvo-aquic soil,with the highest value observed in the NPKM-P treatment for all soils.However,further research is required in the black soils to incorporate more than three cropping seasons.The partial least squares path mode(PLS-PM) showed that total N(nitrogen) and C/N ratio(the ratio of soil organic carbon and total N) had positive effects on the steady SP for all three soils.These findings confirm the significance of the incorporation of manure for attaining high soil productivity.Regulation of the soil C/N ratio was the other main factor for steady SP through fertilization management.

在美国使用无纺布菌条防治光肩星天牛的前景(英文)

An invasive long-horned beetle,Anoplophora glabripennis,was first reported in the northeastern and midwestern United States and eastern Canada between 1996 and 2004 and has been given the common name Asian longhorned beetle(ALB).This beetle has also been found in several countries in Europe.ALB is difficult to control because larvae are found within the wood of living trees and the long-lived adults often occur high in tree canopies.This species is native to China and Korea and,because it has been a major tree killer in China,government agencies in the U.S.and Canada are working to eradicate ALB from North America.Our laboratory has been developing a microbial control approach targeting ALB adults,based on the Japanese product Biolisa Kamikiri which is used to control cerambycids in orchards.Entomopathogenic fungi are grown within non-woven fiber bands(= fungal bands)and placed around tree trunks and branches where ALB adults become inoculated when walking across bands.We have conducted bioassays with Beauveria brongniartii,Beauveria bassiana and Metarhizium anisopliae against ALB larvae and adults to identify effective isolates and now focus our efforts on M.anisopliae F52(ARSEF 7711).Caged field trials conducted in China to compare fungal sprays with fungal bands(2000,2001)demonstrated decreased ALB longevity and fitness for both application methods but longer activity of fungi in cages treated with fungal bands compared with sprays.Uncaged field trials(2001,2002)yielded faster ALB adult mortality in fungal-treated plots and decreased fitness.Studies in New York City testing the longevity of activity of fungal bands in the field have documented that bands retain>1×107 conidia·cm-2(the threshold for activity of Biolisa Kamikiri)for over 3 months.In contrast,studies with unformulated conidia sprayed onto tree trunks in New York documented conidial survival of only a few days.Sublethal effects of exposure of adult female ALB to fungal bands have been investigated further in the laboratory.After either newly eclosed or reproductively active females are exposed to fungal bands,few viable larvae are produced before death of the females.When females are exposed to fungal bands and then caged with males,males become infected.

Weedy red rice has novel sources of resistance to biotic stress

Weedy red rice(Oryza sativa;WRR),a close relative of cultivated rice,is a highly competitive weed that commonly infests rice fields and can also naturally interbreed with rice.Useful genes for biotic stress have been maintained in WRR and can be explored for breeding.Here we describe genetic and physiological traits of WRR that can be beneficial in preventing major rice diseases.Rice blast,caused by the hemibiotrophic fungal pathogen Magnaporthe oryzae,and sheath blight disease,caused by the necrotrophic pathogen Rhizoctonia solani,are the two most damaging biotic stresses of rice.Many major and minor resistance genes and QTL have been identified in cultivated and wild rice relatives.However,novel QTL were recently found in the two major U.S.biotypes of WRR,blackhull-awned(BH)and strawhullawnless(SH),suggesting that WRR has evolved novel genetic mechanisms to cope with these biotic stresses.Twenty-eight accessions of WRR(PI 653412–PI 653439)from the southern USA were characterized and placed in the National Small Grains Collection,and are available for identification of novel genetic factors to prevent biotic stress.

Evaluation of alternative methods for estimating reference evapotranspiration

Evapotranspiration is an important component in water-balance and irrigation scheduling models. While the FAO-56 Penman-Monteith method has become the de facto standard for estimating reference evapotranspiration (ETo), it is a complex method requiring several weather parameters. Required weather data are oftentimes unavailable, and alternative methods must be used. Three alternative ETo methods, the FAO-56 Reduced Set, Hargreaves, and Turc methods, were evaluated for use in Mississippi, a humid region of the USA, using only measurements of air temperature. The Turc equation, developed for use with measured temperature and solar radiation, was tested with estimated radiation and found to provide better estimates of FAO-56 ETo than the other methods. Mean bias errors of 0.75, 0.28, and -0.19 mm, mean absolute errors of 0.92, 0.68, and 0.62 mm, and percent errors of 22.5%, 8.5%, and -5.7% were found for daily estimates for the FAO-56 Reduced Set, Hargreaves, and Turc methods, respectively.

瓦螨微生物防治的不良效果(英文)

We report six different field trials testing the efficacy of Metarhizium anisopliae,an entomopathogenic fungus,against varroa mites in honey bee hives.Varroa mites are parasitic on honey bees and cause serious damage to Apis mellifera colonies.Several control methods are available for varroa mites,none are very effective,so new,more effective methods are being sought.Varroa has previously been shown to be highly susceptible to M.anisopliae infections,and in our first two field trials,we found some efficacy from spore applications.However,in subsequent field trials,we were not able to obtain any varroa control,despite attempting several different application methods,two different strains of the fungus,and testing in different climates and during different phenological states of the honey bee colony.We conclude that microbial control of varroa using fungi is not likely to be effective unless some way is found to prolong the survival of the spores(or other infective units)in the hive environment.

Large-scale evaluation of pea(Pisum sativum L.)germplasm for cold tolerance in the field during winter in Qingdao

As a cool-season crop, pea(Pisum sativum L.) can tolerate frost at the vegetative stage but experiences yield loss when freezing stress occurs at the reproductive stage. Cold-tolerance improvement of pea varieties is important for stable yield and expansion of the winter pea planting area. Under natural low-temperature conditions during winter in Qingdao,Shandong, China, we evaluated the cold tolerance of 3672 pea germplasm accessions in the field and categorized them as displaying high resistance(214), moderate resistance(835), or susceptibility(2623). The highly and moderately resistant genotypes were validated in the following year. We found that genotypes from the winter production region showed higher cold tolerance than genotypes from the spring production region. The accessions identified as having high levels of cold tolerance are recommended as potential genetic resources in cold-tolerance breeding of pea.

Breeding system and its consequence on fruit set of a rare sand dune shrub Eremosparton songoricum(Fabaceae:Papilionoideae):implications for conservation

The breeding system and its consequence on fruit set of Eremosparton songoricum(Litv.) Vass.,a rare shrubby legume occurring in moving or semi-fixed sand dunes of Central Asian deserts,were examined by manipulative experiments and observational studies in natural populations during the period of 2007-2009.The results showed that E.songoricum exhibits a mixed mating system.It is self-compatible,but depends strictly on pollinators to set fruits.Only two effective pollinators were detected and they triggered the specialized pollination mechanism(a 'brush type' and 'tripping mechanism').Geitonogamy becomes predominant in natural populations,because(74.5±1.3)% of visiting activity happened within or between inflorescences and(24.3±1.4)% occurred between ramets.As a result,inbreeding depression caused by geitonogamous selfing inevitably happened under natural conditions,showing 2.36 times less fruit set than was achieved by hand cross-pollination.The results explained the rarity of the species due to its breeding system,and will assist to develop suitable conservation strategies in severe desert environments.

Genetics of a sex-linked recessive red eye color mutant of the tarnished plant bug, Lygus lineolaris

An inbred colony of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Miridae: Hemiptera), was observed to contain specimens with abnormal traits including red eyes, deformed antennae, and deformed legs. These specimens were isolated and back crossed to create stable phenotypic strains. The only successful strain established was a red eyed strain named Cardinal. The trait was more prevalent and stable in males, suggesting that it could be sex linked. To test the hypothesis that the trait was based on a recessive sex linked allele, classical genetic crosses were performed. The hypothesis was confirmed, and the eye color phenotype was measured and characterized using color analysis software. The trait is similar to other red eyed phenotypes described in this species, but is clearly based on a different mutation since it is sex linked rather than autosomal. The results of crossing experiments also suggest that inbreeding in this species results in substantial fitness cost to laboratory insects.

Evaluation of Reproductive Characteristics of 21 Highly Inbred Lines of White Leghorns Divergently Selected for or Segregating in Tumor Resistance

Reproduction performance of 21 inbred experimental lines of White Leghorns was evaluated based on samples of reproduction records over a period of eight consecutive years. Two lines (63 and 72) have been extensively used in studies, especially in research seeking for genetic and epigenetic factors underlying resistance to avian tumor virus-induced diseases in chickens. The other 19 lines are recombinant congenic strains (RCS), which were generated by crossing lines 63 and 72 followed by two consecutive backcrosses to the line 63 and then full-sib mating. In theory, each RCS processes 7/8 of progenitor background line 63 genome and a random sample (1/8) of the progenitor donor line 72 genome. All 21 inbred lines share a common major histocompatibility complex haplotype, B*2. The estimated average fertility of the 21 inbred lines ranged from 72.9% (RCS-J) up to 96.8% (RCS-P). Both progenitor lines 63 and 72 were observed with lower average fertility (82.4% and 81.6%, respectively) in comparison with the RCS except the RCS-J, suggesting a substantial polygenic component underlying the fertility phenotype. The average embryo mortality rate ranged from 14.5% (RCS-P) up to 47.0% (RCS-M). The background line 63 fell at about the middle of the range (28.3%) significantly higher than the donor line 72 (15.7%), which was among the group with the lowest embryo mortality. By definition, hatchability of fertile eggs is reversely correlated with embryo mortality. The average hatchability ranged from 26.5% (RCS-M) up to 66.8% (line 72) while the background line 63 remained (46.6%) at about the middle of the range. The variability of the average embryo mortality and hatchability observed among the 21 inbred lines indicated the two correlated traits also follow polygenic models of inheritance. Findings from this study paves the way for further investigation on genetic and environmental influence over reproductive performance of inbred lines of chickens, and particularly in understanding and improving the reproduction fitness of invaluable genetic resources like these inbred lines.

Detection of Adaptive Genetic Diversity in Wild Potato Populations and Its Implications in Conservation of Potato Germplasm

A better understanding on how genetic diversity is structured at natural habitats can be helpful for exploration and acquisition of plant germplasm. Historically, studies have relied on DNA markers to elucidate potato genetic diversity. Current advances in genomics are broadening applications allowing the identification of markers linked to genomic regions under selection. Those markers, known as adaptive markers, unlock additional ways to value and organize germplasm diversity. For example, conservation priorities could be given to germplasm units containing markers associated to unique geographic identity, and/or linked to traits of tolerance to abiotic stresses. This study investigated if adaptive marker loci were possible to be identified in a large AFLP marker dataset of ninety-four populations of the wild potato species S. fendleri. These populations originated from six different mountain ranges in southern Arizona, USA. A total of 2094 polymorphic AFLP markers were used to conduct genetic diversity analyses of populations and mountain ranges. Adaptive markers were detected using Bayesian methods which distinguished marker loci departing significantly from frequencies expected under neutral models of genetic differentiation. This identified 16 AFLP loci that were considered to be adaptive. To contrast diversity parameters generated with each set of markers, analyses that included all the 2094 AFLP markers, and only the 16 adaptive markers were conducted. The results showed that both were efficient for establishing genetic associations among populations and mountain ranges. However, adaptive markers were better on revealing geographic patterns and identity which would suggest these markers were linked to selection at the natural sites. An additional test to determine if adaptive markers associated to climate variables found two loci associated to specific climate variables in populations from different regions but sharing similar environmental structure. The distribution of adaptive markers among populations revealed that only two were needed to build a core subset able to keep all the markers. This preliminary assessment shows that adaptive genetic diversity could offer an additional way to measure diversity in potato germplasm and to set up options for conservation and research.

Butyrate in combination with forskolin alleviates necrotic enteritis,increases feed efficiency,and improves carcass composition of broilers

Background:The emergence of antimicrobial resistance has necessitated the development of effective alternatives to antibiotics for livestock and poultry production.This study investigated a possible synergy between butyrate and forskolin(a natural labdane diterpene)in enhancing innate host defense,barrier function,disease resistance,growth performance,and meat quality of broilers.Methods:The expressions of representative genes involved in host defense(AvBD9 and AvBD10),barrier function(MUC2,CLDN1,and TJP1),and inflammation(IL-1β)were measured in chicken HD11 macrophages in response to butyrate and forskolin in the presence or absence of bacterial lipopolysaccharides(LPS).Intestinal lesions and the Clostridium perfringens titers were also assessed in C.perfringens-challenged chickens fed butyrate and forskolincontaining Coleus forskohlii(CF)extract individually or in combination.Furthermore,growth performance and carcass characteristics were evaluated in broilers supplemented with butyrate and the CF extract for 42 d.Results:Butyrate and forskolin synergistically induced the expressions of AvBD9,AvBD10,and MUC2 in chicken HD11 cells(P<0.05)and the synergy was maintained in the presence of LPS.Butyrate and forskolin also suppressed LPS-induced IL-1βgene expression in HD11 cells in a synergistic manner(P<0.05).The two compounds significantly reduced the intestinal lesions of C.perfringens-challenged chickens when combined(P<0.05),but not individually.Furthermore,butyrate in combination with forskolin-containing CF extract had no influence on weight gain,but significantly reduced feed intake(P<0.05)with a strong tendency to improve feed efficiency(P=0.07)in a 42-d feeding trial.Desirably,the butyrate/forskolin combination significantly decreased abdominal fat deposition(P=0.01)with no impact on the carcass yield,breast meat color,drip loss,or pH of d-42 broilers.Conclusions:Butyrate and forskolin has potential to be developed as novel antibiotic alternatives to improve disease resistance,feed efficiency,and carcass composition of broilers.

Rapid Deployment of Internet-Connected Environmental Monitoring Devices

Advances in electronic sensing and monitoring systems and the growth of the communications infrastructure have enabled users to gain immediate access to information and interaction with physical devices. To facilitate the uploading, viewing, and sharing of data via the internet, while avoiding the complexities and expense of creating personal web servers, a number of service providers have created websites offering free data hosting and viewing capabilities. Simple setup and configuration routines and available application programming interfaces allow users to quickly and easily interface sensing and monitoring devices to the internet. To demonstrate the ease and feasibility of deploying internet-connected devices, an urban landscape environmental monitoring system, consisting of two wireless field sensing systems and a wireless central receiver, was developed. The field-data sensing units consisted of Arduino microcontroller platforms, sensors, and Xbee radio modems, while the receiver consisted of an Arduino microcontroller, Xbee radio, and Ethernet module connected to an internet modem. The receiver collected and stored data from the wireless transmitters, and uploaded the data to the Xively Cloud Services data hosting and sharing website. Real-time and archived sensor data were then made available for public viewing via the internet and a web browser. Data-sharing services such as Xively provide rapid and convenient means of developing internet-accessible data-collection and viewing applications, enabling enhanced access to the Internet of Things.

Determination of Particulate Matter Emissions from Cattle Feedlots Using Wind-Trax and the Flux-Gradient Technique

Large commercial cattle feedlots are significant sources of particulate matter (PM) emissions. This research compared WindTrax and the flux-gradient technique in estimating emissions of PM with aerodynamic diameter < 10 μm (PM10) from cattle feedlots. Meteorological conditions were measured and PM10 concentrations were profiled vertically (i.e., 2.0 to 7.62 m) at a large commercial beef cattle feedlot in Kansas from May through September 2011. Results show that between the two methods evaluated, WindTrax was least sensitive to changes in heights and number of heights used in the emission estimation, with calculated PM10 emission rates varying by up to 18% only. On the other hand, PM10 emission rates produced by the flux-gradient technique varied by almost 56% when changing either heights and/or number of heights in emission calculation. Both methods were sensitive to height settings, with their respective PM10 emission rates higher when the lowest height setting (2.0 m) was included. Calculating PM10 emission rates with the 7.62-m height led to lower estimates for the flux-gradient technique but no significant change in estimates was observed for WindTrax. As demonstrated in this study, for the flux-gradient technique, settings for the lowest and highest heights were the most critical in emission estimation;exclusion of other heights in between showed only to 2% to 6% change in calculated PM10 emission rates. In general, the higher PM10 emission rates were obtained with the flux-gradient technique. However, eliminating the lowest height (2.0 m) in the calculation and, at the same time, using a specific set of formulations for the flux-gradient technique made its calculated PM10 emission rates slightly lower (but not significantly different) than those from WindTrax.

An expedited method for isolation of DNA for PCR from Magnaporthe oryzae stored on filter paper

The fungus Magnaporthe oryzae is the causal agent of a wide range of cereal diseases. For long-term preservation, the fungus is grown and stored desiccated on filter papers at –20 °C.Inoculated filter papers are cut into pieces of 0.5–1.0 cm diameter prior to storage. In the present study, a fast(11 min) and simple method of preparing DNA suitable for amplifying avirulence genes of M. oryzae by polymerase chain reaction(PCR) was developed. A piece of filter paper containing the fungus was removed from a glass bottle and placed in a 0.2 mL Eppendorf tube containing 100 μL 10 × TE. The suspension was heated for 10 min at 95 °C in a PCR machine. The tube was then centrifuged for 1 min at 3000 r min-1. One μL of 10 × TE solution containing DNA was used for PCR. A total of 28 samples were PCR tested. As a positive control, fungal DNA was extracted using a conventional DNA preparation method. DNA samples obtained from both methods were stored at 4 °C. PCR was performed with DNA on the preparation day and after 4, 8,10, and 18 days of refrigerated storage. In four samples, samples 12, 13, 14, and 28, AVR-Pi9 failed to be amplified. These four samples were tested with a different set of primers for AVR-Pi9, and for AVR-Pita1, confirming that the quality of the samples was insufficient for PCR. Overall, for nearly 90%(24/28) of the samples, the quality of the DNA prepared directly from the fungus on filter paper appeared suitable for a rapid survey of genetic identity of the rice blast fungus by PCR.This method will be useful and effective for reducing cost and time and could readily be adopted worldwide for analysis of M. oryzae and possibly other fungi.

Genome-wide characterization of miRNA and siRNA pathways in the parasitoid wasp Pteromalus puparum

microRNAs(miRNAs) and small interfering RNAs(siRNAs) are small non-coding RNAs(ncRNAs) that trigger RNA interference(RNAi) in eukaryotic organisms. The biogenesis pathways for these ncRNAs are well established in Drosophila melanogaster, Aedes aegypti, Bombyx mori and other insects, but lacking in hymenopteran species, particularly in parasitoid wasps. Pteromalus puparum is a parasitoid of pupal butterflies. This study identified and analyzed two pathways by interrogating the P. puparum genome. All core genes of the two pathways are present in the genome as a single copy, except for two genes in the siRNA pathway, R2D2(two copies) and Argonaute-2(three). Conserved domain analyses showed the protein structures in P. puparum were similar to cognate proteins in other insect species. Phylogenetic analyses of hymenopteran Dicer and Argonaute genes suggested that the siRNA pathway-related genes evolved faster than those in the miRNA pathway. The study found a decelerated evolution rate of P. puparum Dicer-2 with respect to Dicer-1, which was contrary to other hymenopterans. Expression analyses revealed high mRNA levels for all miRNA pathway genes in P. puparum adults and the siRNA related genes were expressed in different patterns. The findings add valuable new knowledge of the miRNA and siRNA pathways and their regulatory actions in parasitoid wasps.