Numerical Study on Characteristics of Convection and Temperature Evolution in Microchannel of Thermal Flowmeter.

During practical usage, thermal flowmeters have a limited range of applications. The present work investigates the factors influencing thermal flowmeter measurements and observes the effects of buoyancy convection and forced convection on the flow rate measurement sensitivity. The results show that the gravity level, inclination angle, channel height, mass flow rate, and heating power affect the flow rate measurements by influencing the flow pattern and the temperature distribution. Gravity determines the generation of convective cells, while the inclination angle affects the location of the convective cells. Channel height affects the flow pattern and temperature distribution. Higher sensitivity can be achieved with smaller mass flow rates or higher heating power. According to the combined influence of the aforementioned parameters, the present work investigates the flow transition based on the Reynolds number and the Grashof number. When the Reynolds number is below the critical value corresponding to the Grashof number, convective cells emerge and affect the accuracy of flowmeter measurements. The research on influencing factors and flow transition presented in this paper has potential implications for the design and manufacture of thermal flowmeters under different working conditions.

Melanin Synthesis Pathway Interruption: CRISPR/Cas9-mediated Knockout of dopa decarboxylase (DDC) in Harmonia axyridis (Coleoptera: Coccinellidae).

CRISPR/Cas9 technology is a very powerful genome editing tool and has been used in many insect species for functional genomics studies through targeted gene mutagenesis. Here, we successfully established CRISPR/Cas9 research platform in Asian multi-colored ladybird beetle, Harmonia axyridis, an important natural enemy in biological control. In this study, one pivotal gene dopa decarboxylase (DDC) in melanin synthesis was targeted by CRISPR/Cas9 to generate mutants in H. axyridis by CRISPR/Cas9 technology. Our results showed that injection of single guide RNA of the DDC and Cas9 protein into preblastoderm eggs induced one insertion and four deletion (indels) mutant H. axyridis. Mutations of HaDDC gene generated 25% mutant rate with melanin missing phenotype in larva, pupa,l and adult stage. The predation ability of the fourth instar larvae has no significant difference between wild (control) and mutant H. axyridis (G0), while these mutant fourth instar larvae had longer developmental period than that of the wild type. Consequently, the total predation of the fourth instar larvae was significantly increased in H. axyridis mutants comparing with the wild type. These results indicated that the success of CRISPR/Cas9 gene editing in H. axyridis. The gene editing platform in H. axyridis would facilitate the gene function research and promote special strain of predatory ladybird beetle generation.

Optimizing the Use of Basil as a Functional Plant for the Biological Control of Aphids by Chrysopa pallens (Neuroptera: Chrysopidae) in Greenhouses.

Effective biological control agents that can provide sustainable pest control need to be researched in further detail; functional plants (or non-crop insectary plants), in particular, are garnering increased research interest. Much remains to be learned as to how non-crop plants can augment biological control in greenhouse systems. In this study, we combined laboratory and greenhouse assays to assess the extent to which basil (Ocimum basilicum L.) (Lamiales: Lamiaceae) affected the biological control of aphids by the predatory lacewing Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae). In the presence of the target prey (peach aphid; Myzus persicae (Sulzer)), both the vegetative and flowering stages of basil enhanced C. pallens longevity and (early-age) fecundity as compared to a control treatment. When basil plants were established near aphid infested eggplants (Solanum melongena L.), the C. pallens colonization rate improved by 72-92% in the short-term. Lacewing colonization patterns were modulated by the basil planting density and spatial arrangement (i.e., perimeter planting vs. intercropping). Under high density intercrop arrangements, C. pallens colonization rates were highest, its populations persisted longer in the crop, and the aphid numbers declined more rapidly. Our work shows how basil enhanced the key fitness attributes of a generalist predatory lacewing and benefitted aphid biological control in a greenhouse setting.

The Molecular Properties and Roles of Pannier in Harmonia axyridis's Metamorphosis and Melanin Synthesis.

The GATA transcription factor Pannier is identified as the major regulatory gene in color pattern formation in the Asian multi-colored ladybird beetle (Harmonia axyridis). however, the mechanisms of Pannier in regulating melanin synthesis and development in H. axyridis remain elusive. In this study, we identified and characterized Pannier in H. axyridis (HaPnr) and showed it to have two alternative spliced variants named HaPnr-α and HaPnr-ß. Analyses of developmental stage expression patterns revealed that HaPnr, HaPnr-α and HaPnr-ß were constitutively expressed throughout all developmental stages. To examine the role of HaPnr in H. axyridis development, RNA interference was performed in late larvae (the fourth instar) and early pupae (the first day of pupa stage). The transcript levels of HaPnr were effectively suppressed after the injection of double-stranded RNA of HaPnr (dsHaPnr). The fourth instar larvae injected with dsHaPnr reduced the pupation rates to only 61.50%, compared with 88.5% in the dsGFP-injected group. The un-pupated larvae gradually died after 1 week, and visually unaffected pupae emerged into abnormal adults with malformed hind wings and melanin absent from the cuticle. These abnormal adults gradually died 10 days after eclosion. However, when early pupae were injected with dsHaPnr, the normal eclosion rate was achieved at 88.41% on day 6 after the injection. In addition, these successful eclosion adults also showed an absence of melanin in the cuticle, but they could mate normally and have normal fecundity as compared with the control. We further demonstrated that the suppression of HaPnr-α or HaPnr-ß individually did not affect the pupation and eclosion process. The suppression of HaPnr-α expression resulted in elytra melanin decreasing in both the conspicua and the succinea subgroup in H. axyridis. Even though the suppression of HaPnr-ß expression only affected the melanin synthesis in the succinea subgroup, it significantly prolonged the time taken for melanin synthesis to occur in the conspicua subgroup in H. axyridis. These results indicate that HaPnr plays an essential role in insect development, especially during their metamorphosis, and also support our hypothesis that HaPnr could regulate melanin synthesis in H. axyridis under the combined action with its two splicing variants, HaPnr-α and HaPnr-ß.

Ecological function of key volatiles in Vitex negundo infested by Aphis gossypii.

Herbivore induced plant volatiles (HIPVs) are key components of plant-herbivorous-natural enemies communications. Indeed, plants respond to herbivores feeding by releasing HIPVs to attract natural enemies. The present study analyses the effect of HIPVs of Vitex negundo (Lamiaceae), an indigenous plant species in northern China, on the predatory ladybug species Harmonia axyridis. Y-tube olfactometer bioassay showed that H. axyridis adults were significantly attracted by V. negundo infested by the aphid Aphis gossypii. We analyzed and compared volatile profiles between healthy and A. gossypii infested V. negundo, screened out the candidate active HIPVs mediated by A. gossypii which could attract H. axyridis, and tested the olfactory behavior of the candidate active compounds on H. axyridis. The gas chromatography-mass spectrometry analysis showed that five volatile compounds were significantly up-regulated after V. negundo infestation by A. gossypii, and five substances were significantly down-regulated in the terpenoid biosynthesis pathway. The olfactory behavior response showed that H. axyridis has significant preference for sclareol, eucalyptol, nonanal and α-terpineol, indicating that this chemical compounds are the important volatiles released by V. negundo to attract H. axyridis. This study preliminarily clarified that V. negundo release HIPVs to attract natural enemies when infected by herbivorous insects. The description of the volatile emission profile enriches the theoretical system of insect-induced volatile-mediated plant defense function of woody plants. Applications in crop protection would lie in designing original strategies to naturally control aphids in orchards.

Predatory Ability of Harmonia axyridis (Coleoptera: Coccinellidae) and Orius sauteri (Hemiptera: Anthocoridae) for Suppression of Fall Armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae).

The lepidopteran pest, Spodoptera frugiperda (JE Smith), spread rapidly after its first detection in China and has caused significant yield loss to maize production in the southwestern part of the country. Although natural enemies of S. frugiperda are present in the field, biological control using naturally distributed predators is ineffective because their underlying populations are too low. To enhance our understanding of the potential role of natural enemies in regulating this invasive pest, functional response experiments were conducted to quantify the response of two predators, Orius sauteri (Poppius) (Hemiptera: Anthocoridae) and Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), in terms of consumption of S. frugiperda. Experimental results revealed that the predatory effects of nymphs of O. sauteri and H. axyridis on the eggs and larvae of S. frugiperda fitted Holling's Type II functional response model. Importantly, the theoretical maximum number of prey consumed per day (Na-max), the instantaneous attack rate (a') and the handling time (Th) of O. sauteri nymphs on S. frugiperda eggs were 15.19, 0.7444 and 0.049 d, respectively; and the parameters on first instar larvae of S. frugiperda were 700.24, 0.5602 and 0.0008 d, respectively. These data contrast to those of H. axyridis, where the Na-max, a' and Th of adults on eggs of S. frugiperda were 130.73, 1.1112 and 0.085 d, respectively, and on the first instar larvae of S. frugiperda were 1401.1, 0.8407 and 0.0006 d, respectively. These results revealed that H. axyridis is a highly voracious predator of the eggs and young larvae of S. frugiperda and O. sauteri could also be used as biocontrol agent of this pest. Our work provides a theoretical framework for the application of natural enemies to control S. frugiperda in the field. Further research is required to strategize conservation biological control approaches in the field to increase populations of these predators and enhance the suppression of S. frugiperda.

Study on properties of wooden capillary electrospray ionization mass spectrometry.

RATIONALE: In view of the unique properties of wooden materials as electrospray emitters, a novel wooden capillary electrospray ionization (WC-ESI) device was fabricated. The performance of a wooden capillary as an electrospray emitter was investigated by using a wooden capillary instead of the metal emitter of commercial ESI sources. METHODS: The mass spectrometric measurement of baicalein, emodin and myoglobin was carried out by using wooden capillary (WC) and metal capillary (MC) ESI sources. Contrasting analysis of signal intensity between WC and MC electrospray ionization mass spectrometry (ESI-MS) was implemented at different sample flow rates. The effect of WC-ESI-MS and MC-ESI-MS was evaluated experimentally with electrospray solutions in different solvent ratios. RESULTS: Generally, the signal generated by WC-ESI-MS was much stronger than that obtained by MC-ESI-MS. In particular, the MS signal in negative ion mode was very strong, which may solve the long-standing problem of low MS signals in negative ion mode, and fully improve the detection efficiency of ESI-MS. CONCLUSIONS: The signal intensity produced by WC-ESI-MS is significantly higher than that from MC-ESI-MS, and polymerization and electrolysis are reduced; therefore, the spectra become simpler. In addition, it is also tolerant to high flow rates and high aqueous phase samples.

Effects of simulated climate warming on the population dynamics of Sitobion avenae (Fabricius) and its parasitoids in wheat fields.

BACKGROUND: Climate warming has considerable effects on crop development and pest population dynamics. Crucially, the tri-trophic responses of plants, herbivores and their natural enemies to warming are poorly understood. To delineate these interactive properties, a three-system approach and integrating life table methodology were used to examine the responses of wheat plants, English grain aphid and parasitoids under open-field infrared heating to simulate warming. RESULTS: Warming significantly increased wheat biomass and grain weight, causing a phenological shift in plant growth. Importantly, warming significantly increased the number of aphids and the reproductive period, coupled with a higher net reproductive rate and intrinsic growth rate. Otherwise, duration of development, generation span, and population doubling time all decreased significantly. Warming had no effect on parasitoid abundance but resulted in a significant decrease in the rate of parasitism. CONCLUSION: Warming may strengthen bottom-up effects on aphids by increasing wheat biomass, resulting in reduced regulation of aphid populations. Warming had a different effect on parasitoids between 2015 and 2016. These findings provide an important characterization of ecological mechanisms in plant-herbivore-parasitoid systems and give a theoretical foundation for improved forecasting of aphid population dynamics under climate change. © 2019 Society of Chemical Industry.

Differential Expression Analysis of Olfactory Genes Based on a Combination of Sequencing Platforms and Behavioral Investigations in Aphidius gifuensis.

Aphidius gifuensis Ashmead is a dominant endoparasitoid of aphids, such as Myzus persicae and Sitobion avenae, and plays an important role in controlling aphids in various habitats, including tobacco plants and wheat in China. A. gifuensis has been successfully applied for the biological control of aphids, especially M. persicae, in green houses and fields in China. The corresponding parasites, as well as its mate-searching behaviors, are subjects of considerable interest. Previous A. gifuensis transcriptome studies have relied on short-read next-generation sequencing (NGS), and the vast majority of the resulting isotigs do not represent full-length cDNA. Here, we employed a combination of NGS and single-molecule real-time (SMRT) sequencing of virgin females (VFs), mated females (MFs), virgin males (VMs), and mated males (MMs) to comprehensively study the A. gifuensis transcriptome. Behavioral responses to the aphid alarm pheromone (E-ß-farnesene, EBF) as well as to A. gifuensis of the opposite sex were also studied. VMs were found to be attracted by female wasps and MFs were repelled by male wasps, whereas MMs and VFs did not respond to the opposite sex. In addition, VFs, MFs, and MMs were attracted by EBF, while VMs did not respond. According to these results, we performed a personalized differential gene expression analysis of olfactory gene sets (66 odorant receptors, 25 inotropic receptors, 16 odorant-binding proteins, and 12 chemosensory proteins) in virgin and mated A. gifuensis of both sexes, and identified 13 candidate genes whose expression levels were highly consistent with behavioral test results, suggesting potential functions for these genes in pheromone perception.

UPLC-MS/MS Method for the Determination of 14 Compounds in Rat Plasma and Its Application in a Pharmacokinetic Study of Orally Administered Xiaoyao Powder.

Xiaoyao Powder (XYP), a common Chinese medicine, comprises eight traditional Chinese herbs and has been widely used clinically to treat liver damage and mental disorders. An ultra-performance liquid chromatography⁻tandem mass spectrometry method was developed to investigate the pharmacokinetics of 14 compounds (albiflorin, paeoniflorin, ferulic acid, senkyunolide I, quercetin, isoliquiritigenin, atractylenolide III, ligustilide, atractylenolide II, liquiritin, liquiritigenin, saikosaponin c, glycyrrhizic acid, and saikosaponin a) in XYP. Naringenin was used as the internal standard. The compounds were separated using an ACQUITY UPLCTM BEH C18 column (1.7 µm, 50 × 2.1 mm) with a mobile phase consisting of acetonitrile and 0.1% formic acid in water at a flow rate of 0.3 mL/min. Detection was performed on a triple-quadrupole tandem mass spectrometer using multiple reaction monitoring and an electrospray ionization source in both positive and negative ionization modes. All calibration curves exhibited good linearity (r² > 0.9974) over the measured ranges. The intra- and inter-day precisions were within 12%, and the accuracy ranged from 89.93% to 106.64%. Extraction recovery and matrix effect results were satisfactory. The method was successfully applied in a pharmacokinetic study of the 14 compounds in rat plasma after the oral administration of XYP.

Pharmacokinetics of Eight Flavonoids in Rats Assayed by UPLC-MS/MS after Oral Administration of Drynariae rhizoma Extract.

As a traditional Chinese medicine, Drynariae rhizoma (Kunze ex Mett.) J. Sm. has been used to treat osteoporosis and bone resorption for 2500 years. Based on the previous study and literature references, flavonoids were proved to be the most abundant and main active compounds of Drynariae rhizoma for osteoporosis treatment. In order to make good and rational use of Drynariae rhizoma in future, a rapid, sensitive, and selective ultraperformance liquid chromatography-mass spectrometry (UPLC-MS/MS) method was developed to investigate the pharmacokinetics of eight main flavonoids in rat plasma after oral administration of the Drynariae rhizoma extract, including neoeriocitrin, luteolin-7-O-ß-D-glucoside, astragalin, naringin, eriodictyol, luteolin, naringenin, and kaempferol. Plasma samples' pretreatment involved a solid-phase extraction column. The separation was performed on an ACQUITY UPLCTM BEH C18 column with a gradient mobile-phase system of acetonitrile and 1% acetic acid in water. The detection was performed using a triple quadrupole tandem mass spectrometer equipped with an electrospray ionization interface (ESI) by multiple reaction monitoring (MRM) in the positive ion mode. All calibration curves exhibited good linearity (r 2 > 0.9990) over the measured ranges. The intraday and interday precisions (RSD) were within 13.87%, and the accuracy (RE) ranged from -14.57% to -0.25% at three quality control levels. Extraction recovery, matrix effect, and stability were satisfactory. The pharmacokinetic characteristics of the eight flavonoids of interest were clearly elucidated.

Identification and Expression Analysis of Candidate Odorant-Binding Protein and Chemosensory Protein Genes by Antennal Transcriptome of Sitobion avenae.

Odorant-binding proteins (OBPs) and chemosensory proteins (CSPs) of aphids are thought to be responsible for the initial molecular interactions during olfaction that mediate detection of chemical signals. Analysis of the diversity of proteins involved comprises critical basic research work that will facilitate the development of sustainable pest control strategies. To help us better understand differences in the olfactory system between winged and wingless grain aphids, we constructed an antennal transcriptome from winged and wingless Sitobion avenae (Fabricius), one of the most serious pests of cereal fields worldwide. Among the 133,331 unigenes in the antennal assembly, 13 OBP and 5 CSP putative transcripts were identified with 6 OBP and 3 CSP sequences representing new S. avenae annotations. We used qPCR to examine the expression profile of these genes sets across S. avenae development and in various tissues. We found 7 SaveOBPs and 1 SaveCSP were specifically or significantly elevated in antennae compared with other tissues, and that some transcripts (SaveOBP8, SaveCSP2 and SaveCSP5) were abundantly expressed in the legs of winged or wingless aphids. The expression levels of the SaveOBPs and SaveCSPs varied depending on the developmental stage. Possible physiological functions of these genes are discussed. Further molecular and functional studies of these olfactory related genes will explore their potential as novel targets for controlling S. avenae.

Structure identification of five unknown dilipo-alkaloids extracted from processed tuber of Aconitum Carmiechaeli by electrospray ionization tandem mass spectrometry.

The alkaloids in processed aconite tuber of Aconitum Carmiechaeli were studied, and five novel alkaloids in extract from processed aconite tuber were found. The first step involved the use of electrospray ionization mass spectrometry (ESI-MS), and then multi-stage tandem mass spectrometry (MSn) was used to provide structural information. Based on their MSn spectra, the structures of the five novel compounds were elucidated to be C3,C8-difatty acid esters of mesaconitine, aconitine and 10-hydroxyaconitine.

Characterization of aconitine-type alkaloids in the flowers of Aconitum kusnezoffii by electrospray ionization tandem mass spectrometry.

The fragmentation mechanism of aconitine-type alkaloids in the flowers of Aconitum kusnezoffii (FAK) was investigated using electrospray ionization tandem mass spectrometry (ESI-MS(n)) firstly. The analysis of the collision-induced dissociation (CID) spectra of three purified aconitine standards and six previously reported aconitines indicated that the fragmentation of the protonated aconitines at low-energy CID follows a similar pathway. The elimination of a C(8)-substituent such as an acetic acid or a fatty acid is the dominant fragmentation mode in MS2. Successive losses of CH(3)COOH, CH(3)OH, H(2)O, BzOH, and CO are the main fragmentation pathways of aconitine-type alkaloids in MS(3) spectra. Based on these features, a rapid method for the direct detection and characterization of alkaloids from an ethanolic extract of FAK is described. All the known aconitum alkaloids are detected and a series of lipo-aconitines has been found for the first time in this plant.