Differences in morphology, mitochondrial genomes, and reproductive compatibility between two clades of parasitic wasps Aphelinus mali (Hymenoptera: Aphelindae) in China.

Aphelinus mali (Haldeman) (Hymenoptera: Aphelinidae) in China is comprised of two clades (termed, the Shandong and Liaoning clades). In order to clarify the genetic relationship between these two clades, we compared and analyzed the morphological characteristics and the mitochondrial genome of each, and performed a hybridization experiment. Morphological results showed that both males and females of the Liaoning clade were larger than Shandong clade, in terms of whole body, abdominal, wing and antennal lengths, however, there were no significant differences between clades for total length of the middle or hind leg of females. The length of the mitochondrial genome of the Shandong clade was 14415 bp and, for the Liaoning clade, it was 14804 bp. Each contained 31 genes, including 13 protein-encoded genes, 16 tRNA genes, and 2 rRNA genes. The highest AT level among the 13 protein-coding genes for the two clades were the same gene (ATP8) (Shandong clade, 91.52%; Liaoning clade, 90.91%). By hybridization and backcrossing, we found that there was no cross incompatibility between these two clades of A. mali. Our results indicate that the historic geographical isolation between these clades has not yet caused reproductive isolation of these populations, and they belong to the same species.

Efficacy of RNA interference using nanocarrier-based transdermal dsRNA delivery system in the woolly apple aphid, Eriosoma lanigerum.

RNA interference (RNAi) is an essential approach for studying gene function and has been considered as a promising strategy for pest control. However, RNAi method has not been conducted in Woolly apple aphid (Eriosoma lanigerum Hausmann), one of the most damaging apple pests in the world. In the study, we investigated the efficacy of RNAi of V-ATPase subunit D (ATPD), an efficacious target for RNAi in other insects, in E. lanigerum by a transdermal double-stranded RNA (dsRNA) delivery system with nanocarriers. Our results showed although topical application of dsATPD in E. lanigerum for 24 h produced 40.5% gene silencing, the additional help of nanocarriers extremely improved the interference efficiency with 98.5% gene silencing. Moreover, a 55.75% mortality was observed 5 days after topical application of nanocarriers and dsATPD, relative to the control (topical application of nanocarriers and double-stranded green fluorescent protein [dsGFP]). The nanocarrier-based transdermal dsRNA delivery system will promote the development of functional analysis of vital genes and also provide a potential target for RNAi-based management of E. lanigerum.

Use of Age-Stage, Two-Sex Life Table to Compare the Fitness of Bactrocera dorsalis (Diptera: Tephritidae) on Northern and Southern Host Fruits in China.

Bactrocera dorsalis (Hendel), as a quarantine pest in many countries and regions, has shown a trend of northward diffusion in the past century in China. In order to determine whether B. dorsalis will cause great harm to the dominant northern fruits, the age-stage two-sex life tables of peaches and apples were constructed, with oranges as the control. The results showed that the developmental rate, intrinsic rate of increase (r), and finite rate of increase (λ) on oranges and peaches were significantly greater than on apples. Additionally, the prediction of population growth 90 days after oviposition revealed that the whole population on oranges and peaches increased by 13,667.3 and 12,112.1 times, respectively, indicating that B. dorsalis is very likely to endanger peach orchards. The population increased on apples by 4311 times, though this is lower than that on oranges and peaches. Overall, peaches with high fitness similar to oranges are very suitable as a host for B. dorsalis and are likely to become a new favorable host, while apples may also become a potentially new host, though with lower fitness. Therefore, the most pressing solutions to take are population monitoring, comprehensive prevention, and control in the case of any potential large-scale outbreak of B. dorsalis in northern China.

EPG analysis of stylet penetration preference of woolly apple aphid on different parts of apple trees.

Woolly apple aphid (WAA), Eriosoma lanigerum (Hausmann), is an important global pest that feeds on Malus species. We studied the feeding preference of WAA on apple trees in the field for two consecutive years and in the laboratory we used electronic penetration graphs (EPG) to record the stylet penetration behavior of WAA on different parts of apple trees. We found that in the field WAA fed primarily on twigs and branches, not on leaves and fruits. Six EPG waveforms were produced during WAA probing on shoots, trunks and leaves of apple trees, including the non-penetration wave (np), the stylet pathway phase wave (C), the intracellular feeding wave (pd), the xylem feeding wave (G), waves indicative of release of saliva into the phloem (E1), and a wave indicative of ingestion from phloem (E2). In the laboratory, aphids only successfully fed on shoots, trunks and leaves, not on fruits. The EPG parameters on the phloem of shoots were significantly higher than those on trunks, indicating WAA prefer to feed on shoots. These laboratory findings explain the relative field feeding preference of WAA on different parts of apple trees, which occurs primarily on branches, barks, and young twigs in orchards, especially on young twigs.

Loss of FZO1 gene results in changes of cell dynamics in fission yeast.

Mitochondrial fission and fusion dynamics are critical cellular processes, and abnormalities in these processes are associated with severe human disorders, such as Beckwith­Wiedemann syndrome, neurodegenerative diseases, Charcot­Marie­Tooth disease type 6, multiple symmetric lipomatosis and microcephaly. Fuzzy onions protein 1 (Fzo1p) regulates mitochondrial outer membrane fusion. In the present study, Schizosaccharomyces pombe (S. pombe) was used to explore the effect of FZO1 gene deletion on cell dynamics in mitosis. The mitochondrial morphology results showed that the mitochondria appeared to be fragmented and tubular in wild­type cells; however, they were observed to accumulate in fzo1Δ cells. The FZO1 gene deletion was demonstrated to result in slow proliferation, sporogenesis defects, increased microtubule (MT) number and actin contraction defects in S. pombe. The FZO1 gene deletion also affected the rate of spindle elongation and phase time at the metaphase and anaphase, as well as spindle MT organization. Live­cell imaging was performed on mutant strains to observe three distinct kinetochore behaviors (normal, lagging and mis­segregation), as well as abnormal spindle breakage. The FZO1 gene deletion resulted in coenzyme and intermediate metabolite abnormalities as determined via metabolomics analysis. It was concluded that the loss of FZO1 gene resulted in deficiencies in mitochondrial dynamics, which may result in deficiencies in spindle maintenance, chromosome segregation, spindle breakage, actin contraction, and coenzyme and intermediate metabolite levels.

Relative Efficacy of Two Clades of Aphelinus mali (Hymenoptera: Aphelinidae) for Control of Woolly Apple Aphid (Hemiptera: Aphididae) in China.

Aphelinus mali (Haldeman) is an effective natural enemy used in China to control woolly apple aphid. Previous studies have found that populations of A. mali in China fall into two distinct genetic clades. We found that one of these, the Shandong clade, is more effective as a biological control agent than the Liaoning clade. The control ability of the Shandong clade of A. mali (a/Th = 2.447) was found to be stronger than that of the Liaoning clade (a/Th = 1.278); the search parameter (Q) and the mutual interference parameter (m) of the Shandong clade (0.1166, 0.2567) were both significantly higher than that of the Liaoning clade (0.076, 0.185). The egg production and life (64.4 ± 10.44 eggs and 16.26 ± 1.04 (female), 13.54 ± 0.75 (male) days, respectively) of the Shandong clade were both significantly higher than they were for the Liaoning clade (30.67 ± 4.02 eggs and 9.34 ± 0.67 (female), 7.64 ± 0.48 (male) days, respectively), and the Shandong clade had a longer oviposition period for one oviposition (Shandong 86.6 s, Liaoning 61.4 s). We expect, in Shandong Province, the pest control ability of the Shandong clade of A. mali to be better than the Liaoning clade.

Levels of Salivary Enzymes of Apolygus Lucorum (Hemiptera: Miridae), From 1st Instar Nymph to Adult, and Their Potential Relation to Bug Feeding.

In recent years, Apolygus lucorum has caused increasing damage to cotton and fruit trees in China. The salivary enzymes secreted by A. lucorum when sucking on host plants induce a series of biochemical reactions in plants, and the pre-oral digestion benefits the bug feeding. In this study, the food intake of A. lucorum from 1st instar nymphs to adults was measured, and the corresponding salivary activity of pectinase, amylase, cellulase, protease, polyphenol oxidase and peroxidase was determined. Daily food intake varied with developmental stage, peaking in 3rd and 4th instar nymphs. Pectinase, amylase, cellulase and protease were detected in both nymphal and adult saliva of A. lucorum, while neither polyphenol oxidase nor peroxidase was detected. Protease activity varied with food intake peaking at the 3rd-4th instar, and then slightly decreasing at the 5th instar. Levels of pectinase, amylase and cellulase increased significantly with the daily feeding level until the 3rd instar, corresponding with increasing damage to host plants. The activity of both cellulase and protease had a significant linear relationship with the average daily food intake. The increasing activity of enzymes in saliva explain stage-specific impacts of A. lucorum on the host plants, and suggest that optimal management of A. lucorum would be confined to its control threshold prior to the peak of daily feeding in the 3rd instar.

Leucobacter populi sp. nov. isolated from a symptomatic bark of Populus × euramericana canker.

A Gram-stain positive, aerobic, non-motile, rod-shaped, oxidase-negative and catalase-positive bacterial strain, designated 06C10-3-11T, was isolated from the symptomatic bark of a Populus × euramericana canker. Growth occurred at 10-45 °C (optimum, 30 °C), pH 6-11 (optimum, pH 7.0-8.0), 0-7 % (w/v) NaCl (optimum, 0-1 %) and in the presence of 20 mM Cr (VI). The major fatty acids (≥10 %) of the novel strain were identified as anteiso-C15:0, anteiso-C17:0 and iso-C16:0. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phospholipid, glycolipid and two unknown lipids. The strain contained the respiratory quinone MK-10 (71 %) as a major component and MK-11 (29 %) in lesser amounts. The cell wall amino acids were 2,4-diaminobutyric acid, alanine, glutamic acid and glycine. The genomic DNA G+C content of the type strain was 69.8 mol%. Phylogenetic analysis, based on 16S rRNA gene sequences, indicated that strain 06C10-3-11T belongs to the genus Leucobacter, showing the highest 16S rRNA gene sequence similarities with Leucobacter celer NAL101T (96.19 %), 'Leucobacter kyeonggiensis' F3-P9T (96.18 %), Leucobacter denitrificans M1T8B10T (96.10 %) and Leucobacter aridicollis CIP 108388T (96.06 %). The DNA G+C content of strain 06C10-3-11T was 69.8 mol%. Based on the molecular data and physiological and biochemical characteristics, strain 06C10-3-11T is considered to represent a novel species of the genus Leucobacter, for which the name Leucobacterpopuli sp. nov. is proposed. The type strain is 06C10-3-11T (= CFCC 12199T = KCTC 39685T).

Invasion Genetics of Woolly Apple Aphid (Hemiptera: Aphididae) in China.

In China, the woolly apple aphid, Eriosoma lanigerum (Hausmann), was first detected as an invasive species during the 1910s to 1930s, restricted to Shandong, Liaoning, and Yunnan Provinces. However, since the 1990s, the pest has spread into many other areas of China. To determine the possible spread routes of the recently established populations, the genetic diversity and genetic structure of 24 populations in 10 provinces were analyzed using eight microsatellite loci. Analyses using STRUCTURE software identified two genetic clusters overall. Three populations from Yunnan and Xinjiang consisted of individuals originating from a single cluster. Nineteen populations from eight northern provinces consisted only of individuals from another cluster, which formed a single large and panmictic population, resembling a distinct "supercolony" in Northern China. The other two populations from Yunnan consisted of individuals from both clusters. The possible routes of spread of the recently established populations of E. lanigerum in China were revealed as follows: 1) the populations in Northern China (including these from Henan, Hebei, Shanxi, Shannxi, Jiangsu, and Gansu) may have been introduced from Shandong or Liaoning Provinces; 2) the populations in Yunnan consisted of an early-established population and a population introduced secondarily from Shandong or neighboring areas, indicating that the population in Yunnan has at least two sources; and 3) the recently established populations of E. lanigerum in Xinjiang might not have been introduced from the "supercolony" in Northern China. Knowledge of these routes of spread is useful for avoiding further dissemination and/or additional introductions.

Chemical characteristics of normal, woolly apple aphid-damaged, and mechanically damaged twigs of six apple cultivars, measured in autumn wood.

Chemical characteristics of normal, woolly apple aphid-damaged, and mechanically damaged twigs of six apple cultivars: Red Fuji, Golden Delicious, Qinguan, Zhaojin 108, Starkrimson, and Red General, were examined in autumn wood to provide abetter understanding of factors related to cultivar resistance to the woolly apple aphid, Eriosoma lanigerum (Hausmann). Chemical measures examined included soluble sugars, soluble proteins and amino acids, total phenolics, and polyphenol oxidase (that enhances the resistance of plants to insects) and superoxide dismutase, peroxidase, and catalase (that degrade waste products in plants). Soluble sugar, protein, and amino acid contents in normal (undamaged) twigs of Red Fuji, aphid-susceptible cultivar, were higher than in mechanically damaged and aphid-damaged twigs. Total phenolic compounds, an important group of defensive compounds against aphids, increased by 30.5 and 6.0% in mechanically damaged twigs of Qinguan and Zhaojin 108, respectively, and decreased by 21.7 and 16.1% in aphid-damaged twigs of Red Fuji and Red General, respectively. Compared with normal twigs, in aphid-damaged twigs, superoxide dismutase, peroxidase, and polyphenol activity all decreased in Red Fuji. The resistance of some apple cultivars to woolly apple aphid during the growth of autumn shoots was related to several of the physiological indices we monitored. The thin epidermis of callus tissue over healed wounds showed increased susceptibility to the attack by woolly apple aphid. Apple cultivar Qinguan with the highest level of resistance to woolly apple aphid in autumn had increased in amino acid, total phenolic compound levels, and enzyme activity after aphid feeding.

A luminescent Eu(III) complex based on 2-(4', 4', 4'-trifluoro-1', 3'-dioxobutyl)-dibenzothiophene for light-emitting diodes.

Complex Eu(dbt)(3)(phen) (Hdbt=2-(4', 4', 4'-trifluoro-1', 3'-dioxobutyl)-dibenzothiophene, phen=1,10-phenanthroline) was synthesized. The complex emits red luminescence, characteristic of the (5)D(0)-->(7)F(J) (J=0-4) emission bands of Eu(3+) under near ultraviolet. A red conversion light-emitting diode (LED) device was fabricated by coating complex onto InGaN-based-LED chip that emits 395nm ultraviolet light. When the mass ratio of the red phosphor to the silicone is 1:25, the LED device's CIE chromaticity coordinates are x=0.5835, y=0.2857, and the luminescence efficiency is 1.29 lm/w. All the results show that this europium complex may act as a red component in fabrication of white LEDs with high color-rendering index.