The genetic diversity of SMLS (Sitobion miscanthi L type symbiont) and its effect on the fitness, mitochondrial DNA diversity and Buchnera aphidicola dynamic of wheat aphid, Sitobion miscanthi (Hemiptera: Aphididae).

SMLS (Sitobion miscanthi L type symbiont) is a recently discovered aphid secondary symbiont. Using evidence extracted from 16S rRNA sequences, previous studies indicate that SMLS is the most widely distributed and most recently transferred secondary symbiont in Chinese Sitobion miscanthi populations. Here, we further investigated genetic diversity among SMLS geographic strains with multiloci data. Furthermore, the influence of SMLS on S. miscanthi was uncovered with ecological and evolutionary evidence. The results indicated that there was limited influence of infection with SMLS on variation and evolutionary patterns of S. miscanthi mitochondrial DNA. By hemolymph injection, the SMLS-infected and SMLS-uninfected S. miscanthi clones with the identical genetic background were built in this study. Although similar Buchnera aphidicola dynamics were observed between SMLS-infected and SMLS-uninfected S. miscanthi population, B. aphidicola density of SMLS-infected S. miscanthi population was always significantly higher than SMLS-uninfected ones. The results of fitness measurements indicated that under laboratory rearing conditions, transfection of SMLS could confer modest advantages to some fitness components of S. miscanthi, that is, total number of offspring, longevity, age of first reproduction and weight of adult. However, as SMLS is not strictly associated with S. miscanthi, further investigations are needed to uncover the mechanisms responsible for this inconceivable association.

[Characteristics and Coupling Relationship of Soil Carbon and Nitrogen Transformation During In-situ Mineralization Cultivation in Forestlands in the Mountain Area of Southern Ningxia].

The study aimed to investigate the characteristics and relationship between soil carbon and nitrogen transformation of artificial forestlands, which is one type of vegetation restoration in the mountain area of Southern Ningxia. Soil samples were collected every two months in a year from three forestlands, and the characteristics of soil organic carbon, dissolved carbon, microbial biomass carbon, organic nitrogen, inorganic nitrogen, soil ammonification, nitrification and mineralization rates, microbial immobilization rates and coupling of soil carbon and nitrogen were studied by the in-situ closed-top PVC tube incubation methods. The results showed that: in the process of in-situ incubation, the most obvious changes of carbon and nitrogen were in 61-120 days which was mainly affected by soil moisture; There were significantly positive correlations between the soil organic carbon and the total nitrogen, microbial biomass carbon and microbial biomass nitrogen, dissolved carbon and dissolved nitrogen; Transformation rates of soil organic carbon had significant effects on the soil ammonification, nitrification and microbial immobilization rates. It can be well simulated by model of linear regression equation; Microbial quotient, MBN/SON were significantly increased in soil of Caragana korshinskii land. Net nitrification rates, net mineralization rates in Caragana korshinskii land were significantly higher than that in Prunus davidiana and Prunus mandshurica lands.

[Variation of soil nitrogen during in situ mineralization process under different grasslands in the mountainous area of southern Ningxia, Northwest China].

Variations in organic nitrogen, microbial biomass nitrogen, soluble organic nitrogen, NH4(+) -N, NO3(-) -N, NO2(-) -N and N mineralization were investigated under three different grasslands in the mountainous area of southern Ningxia, Northwest China (natural grassland, artificial turf and abandoned land) using the close-top tube incubation method. Microbial biomass nitrogen, soluble organic nitrogen, NH4(+)-N, NO3(-)-N, NO2(-)-N and N mineralization exhibited significant seasonal variations. The nitrogen levels remained essentially unchanged from April to June, significantly decreased in July-August, rebounded after August, and were lowest in August. The organic nitrogen content remained unchanged in the whole training process. The soil mineralization rates, nitrification and ammonification rate were lowest in June-August. The ratios of each N fraction to total N responded differently to seasonal changes. The ratios of organic N, NO2(-) -N to total N did not change, in contrast, the ratios of nitrate N, microbial biomass nitrogen and soluble organic nitrogen, to total soil N decreased from April to August, and increased from August to December. The soil organic matter, pH, and bulk density were closely related to soil N. There was a significant positive correlation among the six N fractions. The soil nitrogen content of the different grassland types followed the order of natural grassland > abandoned land > artificial turf.

Identification and expression profile analysis of putative odorant-binding proteins in Sitodiplosis mosellana (Gehin) (Diptera: Cecidomyiidae).

Odorant binding proteins (OBPs) contribute to the remarkable sensitivity of the insect's olfactory system and play important roles in the olfactory recognition. The orange blossom midge, Sitodiplosis mosellana is a cereal specialist, and utilizes pheromone and host odorant as a cue for its mating and oviposition. However, OBP genes have not been largely identified in S. mosellana. Based on the sequenced transcriptome database, twenty-six OBP genes were identified in S. mosellana for the first time. Phylogenetic analysis revealed that S. mosellana OBP genes are more closely related to Mayetiola destructor OBP genes than to Aedes aegypti OBP genes. Most OBP genes seemed to be antenna-specific, but differentially expressed in male and female antennae. Three OBP genes (OBP9, OBP19 and OBP23) are leg-specific. And also, most OBP genes have higher expression levels in adults. Only one OBP gene (OBP10) has higher expression levels in larval stages. These findings serve as an important basis for understanding the molecular mechanisms of chemosensory perception.

Sub-lethal effects of four neonicotinoid seed treatments on the demography and feeding behaviour of the wheat aphid Sitobion avenae.

BACKGROUND: Neonicotinoids are widely used as seed treatments in wheat fields against the grain aphid (Sitobion avenae F.) in China. Due to the degradation of neonicotinoids in wheat plants, wheat aphids are more likely to be exposed to low concentrations of neonicotinoids over long periods. It is therefore expected that neonicotinoids, aside from acute (lethal) effects, may also cause a range of sub-lethal effects on this pest. RESULTS: The growth and fertility of S. avenae feeding on wheat plants treated with a sub-lethal concentration (LC10 ) of imidacloprid, dinotefuran, thiacloprid and thiamethoxam were not greatly affected. However, the population growth parameters of S. avenae were significantly reduced at median lethal concentration (LC50 ). Electronic penetration graph recordings showed a higher percentage of no probing phase and shorter phloem sap ingestion phase on the wheat plants treated with LC10 and LC50 concentrations. CONCLUSION: The results indicate that even low concentrations of neonicotinoid treatments on wheat seeds have long-term, adverse effects on wheat aphid. As such, neonicotinoid seed treatments have far greater effects on wheat aphids than estimated by acute toxicity tests. These results benefit our understanding on the subtle effects of the four tested neonicotinoids when applied as seed treatments.

[Effects of different aspects on soil microbial biomass and dissolved organic carbon of the loess hilly area].

Soil samples from different aspects (southern slope and northern slope) under the same vegetation in two typical vegetation zones (forest vegetation zone and steppe vegetation zone) of Yanhe basin in the loess hilly area were chosen and analyzed, in order to investigate the content and correlation of soil microbial biomass carbon (SMBC), soil microbial biomass nitrogen (SMBN), soil microbial biomass phosphorus (SMBP) and dissolved organic carbon (DOC). The results show that, in 0-10 cm soil layer of forest vegetation zone, SMBC in southern slope and northern slope are 532.1-792.5 mg x kg(-1) and 333.6-469.8 mg x kg(-1), SMBN are 53.66-87.31 mg x kg(-1) and 47.58-61.38 mg x kg(-1) respectively, both of them are higher in southern slope than those in northern slope, but in steppe vegetation zone, SMBC and SMBN in southern slope are lower than those in northern slope, with SMBC of southern slope and northern slope are 68.90-75.34 mg x kg(-1) and 65.29-128.67 mg x kg(-1), SMBN are 13.94-18.61 mg x kg(-1) and 13.00-20.10 mg x kg(-1) respectively, SMBP in both vegetation zones have a different variation trend compared with SMBC and SMBN; the ratio of SMBC to the sum of SMBC plus DOC (SMBC + DOC) in southern slope of forest vegetation zone reaches the maximum value in two vegetation zones, which is 77.74%. In steppe vegetation zone, this ratio both decreases from northern slope to southern slope and 0-10 cm soil layer to 10-30 cm soil layer. The difference of soil moisture and temperature between different aspects under the same vegetation have a significant effect on soil microbial biomass, it also makes the ratio of SMBC to SMBC + DOC different, SMBC + DOC can reflect soil carbon availability better than SMBC, there might be an obvious change in soil microbial communities in 0-10 cm soil layer of different aspects in forest vegetation zone.

Flight performance of the orange wheat blossom midge (Diptera: Cecidomyiidae).

The orange wheat blossom midge, Sitodiplosis mosellana (Géhin) (Diptera: Cecidomyiidae), is a chronic wheat pest worldwide. Adult S. mosellana engage in short-distance flight, but also exploit weather patterns for long-distance dispersal. However, little is known about the flight performance of S. mosellana, and the effects of the biotic and abiotic factors that influence its flight activity. In this study, we explored the active flight potential of S. mosellana under various environmental factors using a 26-channel computer-monitored flight mill system. The most suitable temperature for flight and flight distance was 16-24 degrees C; flight duration peaked at 16 degrees C while speed peaked at 28 degrees C. Flight performance gradually declined between 10 and 400 lux light intensity. More than 50% individuals of 1-d-old females flew > 500 m, while only 24% of males flew > 500 m. One-day-old S. mosellana had stronger flight ability than that of 2-d-old individuals. This research showed that S. mosellana possessed strong enough flight ability that they can fly to a high altitude and then disperse via moving air currents. These results can aid in forecasting S. mosellana outbreak.

Genetic diversity and population structure of Sitodiplosis mosellana in Northern China.

The wheat midge, Sitodiplosis mosellana, is an important pest in Northern China. We tested the hypothesis that the population structure of this species arises during a range expansion over the past 30 years. This study used microsatellite and mitochondrial loci to conduct population genetic analysis of S. mosellana across its distribution range in China. We found strong genetic structure among the 16 studied populations, including two genetically distinct groups (the eastern and western groups), broadly consistent with the geography and habitat fragmentation. These results underline the importance of natural barriers in impeding dispersal and gene flow of S. mosellana populations. Low to moderate genetic diversity among the populations and moderate genetic differentiation (F ST = 0.117) between the two groups were also found. The populations in the western group had lower genetic diversity, higher genetic differentiation and lower gene flow (F ST = 0.116, Nm = 1.89) than those in the eastern group (F ST = 0.049, Nm = 4.91). Genetic distance between populations was positively and significantly correlated with geographic distance (r = 0.56, P<0.001). The population history of this species provided no evidence for population expansion or bottlenecks in any of these populations. Our data suggest that the distribution of genetic diversity, genetic differentiation and population structure of S. mosellana have resulted from a historical event, reflecting its adaptation to diverse habitats and forming two different gene pools. These results may be the outcome of a combination of restricted gene flow due to geographical and environmental factors, population history, random processes of genetic drift and individual dispersal patterns. Given the current risk status of this species in China, this study can offer useful information for forecasting outbreaks and designing effective pest management programs.

Global transcriptome analysis of orange wheat blossom midge, Sitodiplosis mosellana (Gehin) (Diptera: Cecidomyiidae) to identify candidate transcripts regulating diapause.

BACKGROUND: Many insects enter a developmental arrest (diapause) that allows them to survive harsh seasonal conditions. Despite the well-established ecological significance of diapause, the molecular basis of this crucial adaptation remains largely unresolved. Sitodiplosis mosellana (Gehin), the orange wheat blossom midge (OWBM), causes serious damage to wheat throughout the northern hemisphere, and sporadic outbreaks occur in the world. Traits related to diapause appear to be important factors contributing to their rapid spread and outbreak. To better understand the diapause mechanisms of OWBM, we sequenced the transcriptome and determined the gene expression profile of this species. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we performed de novo transcriptome analysis using short-read sequencing technology (Illumina) and gene expression analysis with a tag-based digital gene expression (DGE) system. The sequencing results generated 89,117 contigs, and 45,713 unigenes. These unigenes were annotated by Blastx alignment against the NCBI non-redundant (nr), Clusters of orthologous groups (COG), gene orthology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. 20,802 unigenes (45.5% of the total) matched with protein in the NCBI nr database. Two digital gene expression (DGE) libraries were constructed to determine differences in gene expression profiles during diapause and non-diapause developmental stages. Genes related to diapause were analyzed in detail and in addition, nine diapause-related genes were analyzed by real time PCR. CONCLUSIONS/SIGNIFICANCE: The OWBM transcriptome greatly improves our genetic understanding and provides a platform for functional genomics research of this species. The DGE profiling data provides comprehensive information at the transcriptional level that facilitates our understanding of the molecular mechanisms of various physiological aspects including development and diapause stages in OWBM. From this study it is evident that various genes coding metabolic enzymes are crucial for diapause and metamorphosis.

[Spatial pattern of Sitodiplosis mosellana and its egg parasitoids mixed population].

Geostatistis methods were adopted to analyze the spatial pattern of Sitodiplosis mosellana (Diptera: Cecidomyiidae) at its different development periods and of its egg parasitoids mixed population (Tetrastichus sp. and Platygaster error; Hymoneptera: Eulophidae and Platygastridae). The aggregated spatial arrangements for S. mosellana cocoon, adult, and larva and for egg parasitoids mixed population could be well described by spherical model, spherical-exponential model, linear sill model, and spherical-exponential model, respectively. The spatial dependence range of S. mosellana cocoon, adult at initial emergence period, adult at peak emergence period, and larva, and of egg parasitoids mixed population was 53.6, 190.6, 154.1, 4.2 and 280.3 m, and the aggregation intensity was 30.5%, 95.6%, 96.3%, 14.9% and 95.3%, respectively. The simulated maps of the spatial distribution produced by Kriging model could intuitively analyze the dynamic changes of S. mosellana at its different development periods and of egg parasitoids mixed population from the two aspects of time and space.