Chang-Ying Niu

Use of an innovative T-tube maze assay and the proboscis extension response assay to assess sublethal effects of GM products and pesticides on learning capacity of the honey bee Apis mellifera L.

Transgenic Cry1Ac+CpTI cotton (CCRI41) is a promising cotton cultivar throughout China but side effects and especially sublethal effects of this transgenic cultivar on beneficial insects remain poorly studied. More specifically potential sublethal effects on behavioural traits of the honey bee Apis mellifera L. have not been formally assessed despite the importance of honey bees for pollination. The goal of our study was to assess potential effects of CCRI41 cotton pollen on visual and olfactory learning by honey bees. After a 7-day oral chronic exposure to honey mixed with either CCRI41 pollen, imidacloprid-treated conventional pollen (used as positive sublethal control) or conventional pollen (control), learning performance was evaluated by the classical proboscis extension reflex (PER) procedure as well as a T-tube maze test. The latter assay was designed as a new device to assess potential side effects of pesticides on visual associative learning of honey bees. These two procedures were complementary because the former focused on olfactory learning while the latter was involved in visual learning based on visual orientation ability. Oral exposure to CCRI41 pollen did not affect learning capacities of honey bees in both the T-tube maze and PER tests. However, exposure to imidacloprid resulted in reduced visual learning capacities in T-tube maze evaluation and decreased olfactory learning performances measured with PER. The implications of these results are discussed in terms of risks of transgenic CCRI41 cotton crops for honey bees.

Sexual communication and related behaviours in Tephritidae: current knowledge and potential applications for Integrated Pest Management

Tephritidae are an enormous threat to fruit and vegetable production throughout the world, causing both quantitative and qualitative losses. Investigating mating sequences could help to unravel mate choice dynamics, adding useful information to improve behaviour-based control strategies. We review current knowledge about sexual communication and related behaviours in Tephritidae, with a focus on six key agricultural pests: Anastrepha ludens, Bactrocera cucurbitae, Bactrocera dorsalis, Bactrocera oleae, Ceratitis capitata and Rhagoletis pomonella. We examine features and the role of male–male combat in lekking sites, cues affecting mating dynamics, and some fitness-promoting female behaviours that occur at oviposition sites [the use of oviposition marking pheromones (OMPs) and female–female fights for single oviposition sites]. We outline future perspectives and potential contributions of knowledge about sexual communication to Integrated Pest Management programs for tephritid pests. Sexually selected traits are frequently good indicators of male fitness and knowledge of sexual selection processes may contribute to the improvement of the sterile insect technique (SIT), to select genotypes with high reproductive success and to promote sexually selected phenotypes through mass-rearing optimization. Furthermore, males’ exposure to parapheromones, such as phenyl propanoids (PPs), ginger root oil and trimedlure can enhance the mating success of sterile flies used in SIT programs. PPs are also a powerful tool to improve reduced-risk monitoring dispensers and the male annihilation technique, with low side effects on non-target insects. Lastly, we outline the possibility to sensitise or train mass-reared parasitoids on OMPs during the pre-release phase, in order to improve their post-release performance in the field.

RNAi Silencing of the HaHMG-CoA Reductase Gene Inhibits Oviposition in the Helicoverpa armigera Cotton Bollworm

RNA interference (RNAi) has considerable promise for developing novel pest control techniques, especially because of the threat of the development of resistance against current strategies. For this purpose, the key is to select pest control genes with the greatest potential for developing effective pest control treatments. The present study demonstrated that the 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase; HMGR) gene is a potential target for insect control using RNAi. HMGR is a key enzyme in the mevalonate pathway in insects. A complete cDNA encoding full length HMGR (encoding an 837-aa protein) was cloned from Helicoverpa armigera (Lepidoptera: Noctuidae). The HaHMGR (H. armigera HMGR) knockdown using systemic RNAi in vivo inhibited the fecundity of the females, effectively inhibited ovipostion, and significantly reduced vitellogenin (Vg) mRNA levels. Moreover, the oviposition rate of the female moths was reduced by 98% by silencing HaHMGR compared to the control groups. One-pair experiments showed that both the proportions of valid mating and fecundity were zero. Furthermore, the HaHMGR-silenced females failed to lay eggs (approximate 99% decrease in oviposition) in the semi-field cage performance. The present study demonstrated the potential implications for developing novel pest management strategies using HaHMGR RNAi in the control of H. armigera and other insect pests.

Pyrosequencing reveals a shift in symbiotic bacteria populations across life stages of Bactrocera dorsalis

Bactrocera dorsalis is one of the most economically important fruit flies around the world. In this study, 454 pyrosequencing was used to identify the bacteria associated with different developmental stages of B. dorsalis. At ≥ 97% nucleotide similarity, total reads could be assigned to 172 Operational Taxonomic Units belonging to six phyla. Proteobacteria dominated in immature stages while Firmicutes dominated in adult stages. The most abundant families were Enterococcaceae and Comamondaceae. The genus Comamonas was most abundant in pupae whereas completely absent in adults. Some identified species had low sequence similarity to reported species indicating the possibility of novel taxa. However, a majority sequence reads were similar to sequences previously identified to be associated with Bactrocera correcta, suggesting a characteristic microbial fauna for this insect genus. The type and abundance of different bacterial groups varied across the life stages of B. dorsalis. Selection pressure exerted by the host insect as a result of its habitat and diet choices could be the reason for the observed shift in the bacteria groups. These findings increase our understanding of the intricate symbiotic relationships between bacteria and B. dorsalis and provide clues to develop potential biocontrol techniques against this fruit fly.

Population dynamics, phenology, and overwintering of Bactrocera dorsalis (Diptera: Tephritidae) in Hubei Province, China

Since 2004 the oriental fruit fly, Bactrocera dorsalis (Hendel) has become widespread and economically important in Wuhan, Hubei Province, central China. In this article, the population dynamics of B. dorsalis were monitored using methyl eugenol-baited traps during 2008–2009 in Wuhan. Adults were captured from early July to the end of December in a citrus orchard and peaked in October and early November. Adult population peak coincided with the ripeness period of sweet oranges in October. Infestation with B. dorsalis was more serious in 2009 than in 2008. Field surveys of host plants for this fly species combined with rearing experiments indicated that pear was the first host plant infested by B. dorsalis. We recorded the following host shift pattern: pear (Pyrus communis L.), jujube (Zizyphus jujuba L.), persimmon (Diospyros kaki L.), and sweet orange (Citrus unshiu Marcor). Our findings suggest that B. dorsalis completes four to five generations per year in this area and most serious damage occurred in the 4th generation in the citrus orchards. The availability of preferred host fruits and the low winter temperature are key factors influencing population fluctuations. Overwintering potential experiments of adults, larvae, and pupae were also carried out in the field. Results suggest that a small proportion of pupae were able to survive winter in Wuhan and emerged successfully the following year. Practical and theoretical implications of the results regarding the invasion potential and management strategy of B. dorsalis in Hubei Province are discussed.

Identification of Top-Down Forces Regulating Cotton Aphid Population Growth in Transgenic Bt Cotton in Central China

The cotton aphid Aphis gossypii Glover is the main aphid pest in cotton fields in the Yangtze River Valley Cotton-planting Zone (YRZ) in central China. Various natural enemies may attack the cotton aphid in Bt cotton fields but no studies have identified potential specific top-down forces that could help manage this pest in the YRZ in China. In order to identify possibilities for managing the cotton aphid, we monitored cotton aphid population dynamics and identified the effect of natural enemies on cotton aphid population growth using various exclusion cages in transgenic Cry1Ac (Bt)+CpTI (Cowpea trypsin inhibitor) cotton field in 2011. The aphid population growth in the open field (control) was significantly lower than those protected or restricted from exposure to natural enemies in the various exclusion cage types tested. The ladybird predator Propylaea japonica Thunberg represented 65% of Coccinellidae predators, and other predators consisted mainly of syrphids (2.1%) and spiders (1.5%). The aphid parasitoids Aphidiines represented 76.7% of the total count of the natural enemy guild (mainly Lysiphlebia japonica Ashmead and Binodoxys indicus Subba Rao & Sharma). Our results showed that P. japonica can effectively delay the establishment and subsequent population growth of aphids during the cotton growing season. Aphidiines could also reduce aphid density although their impact may be shadowed by the presence of coccinellids in the open field (likely both owing to resource competition and intraguild predation). The implications of these results are discussed in a framework of the compatibility of transgenic crops and top-down forces exerted by natural enemy guild.

Transcriptome Characterization Analysis of Bactrocera minax and New Insights into Its Pupal Diapause Development with Gene Expression Analysis

Bactrocera minax is a major citrus pest distributed in China, Bhutan and India. The long pupal diapause duration of this fly is a major bottleneck for artificial rearing and underlying mechanisms remain unknown. Genetic information on B. minax transcriptome and gene expression profiles are needed to understand its pupal diapause. High-throughput RNA-seq technology was used to characterize the B. minax transcriptome and to identify differentially expressed genes during pupal diapause development. A total number of 52,519,948 reads were generated and assembled into 47,217 unigenes. 26,843 unigenes matched to proteins in the NCBI database using the BLAST search. Four digital gene expression (DGE) libraries were constructed for pupae at early diapause, late diapause, post-diapause and diapause terminated developmental status. 4,355 unigenes showing the differences expressed across four libraries revealed major shifts in cellular functions of cell proliferation, protein processing and export, metabolism and stress response in pupal diapause. When diapause was terminated by 20-hydroxyecdysone (20E), many genes involved in ribosome and metabolism were differentially expressed which may mediate diapause transition. The gene sets involved in protein and energy metabolisms varied throughout early-, late- and post-diapause. A total of 15 genes were selected to verify the DGE results through quantitative real-time PCR (qRT-PCR); qRT-PCR expression levels strongly correlated with the DGE data. The results provided the extensive sequence resources available for B. minax and increased our knowledge on its pupal diapause development and they shed new light on the possible mechanisms involved in pupal diapause in this species.

Pollen from Cry1Ac/CpTI-transgenic cotton does not affect the pollinating beetle Haptoncus luteolus

With the widespread distribution of transgenic cotton plants, concerns over their potential effects on beneficial insects and other non-target pests have been raised. In the present study, the field and laboratory effects of Cry1Ac/CpTI transgenic cotton pollen on the pollinating beetle Haptoncus luteolus were investigated. Abundance of adult beetles was studied in cotton plots at two sites in Wuhan in 2009, and the average number of adults per flower was recorded. Life-history parameters including developmental duration, hatch rate, pupation rate, emergence rate and egg number per female, as well as the survivorship curves of H. luteolus were studied in laboratory experiments. The results showed that the adult abundance in the field did not differ significantly between non-transgenic and transgenic cotton, the exposure to Cry1Ac/CpTI pollen in laboratory had no effect on the developmental time, hatching rate, pupation rates and emergence rates of H. luteolus, no significant differences were found in the mating rates and the fecundity between treatments, and the survival curves of both treatments also resulted in no significant differences. It indicated there were no significant direct adverse effects of transgenic cotton pollen on H. luteolus both in the field and in the laboratory. Our study bridged the gap of environmental risk assessment of transgenic cotton for pollinators other than pollinating bees.

Field Evaluation of Attractive Lures for the Fruit Fly Bactrocera minax (Diptera: Tephritidae) and Their Potential use in Spot Sprays in Hubei Province (China)

ABSTRACT The Chinese citrus fruit fly, Bactrocera minax (Enderlein) is a univoltine Tephritidae pest that infests Citrus species. Field trials were conducted in 2010 to determine the potential use of a lure based on enzymatical-hydrolyzed beer yeast as liquid bait (hereafter named H-protein bait) for B. minax in the Hubei province, China. In a citrus orchard, we compared the attractiveness among aqueous solutions of H-protein bait, GF-120 fruit fly bait, sugar-vinegar-wine mixture, torula yeast, and Jufeng attractant when used in traps and in spot sprays, that is, lures used in combination with the insecticide trichlorphon. The H-protein bait was the most attractive lure in traps, ensnaring significantly more adults than sugar-vinegar-wine mixture, torula yeast, and Jufeng attractant, in decreasing efficiency order. In spot sprays those with Ç-protein bait killed significantly more female and male flies within 40 min than those with sugar-vinegar-wine mixture, GF-120, Jufeng attractant, and the control. In addition, the total number of flies killed by H-protein bait during the spot spray duration was higher than other treatments. Our results demonstrated that the H-protein bait may be a useful tool in citrus orchards in China to monitor B. minax populations as well as to manage this pest when used in spot sprays.

The effect of silencing 20E biosynthesis relative genes by feeding bacterially expressed dsRNA on the larval development of Chilo suppressalis.

RNA interference (RNAi) is a robust tool to study gene functions as well as potential for insect pest control. Finding suitable target genes is the key step in the development of an efficient RNAi-mediated pest control technique. Based on the transcriptome of Chilo suppressalis, 24 unigenes which putatively associated with insect hormone biosynthesis were identified. Amongst these, four genes involved in ecdysteroidogenesis i.e., ptth, torso, spook and nm-g were evaluated as candidate targets for function study. The partial cDNA of these four genes were cloned and their bacterially expressed dsRNA were fed to the insects. Results revealed a significant reduction in mRNA abundance of target genes after 3 days. Furthermore, knocked down of these four genes resulted in abnormal phenotypes and high larval mortality. After 15 days, the survival rates of insects in dsspook, dsptth, dstorso, and dsnm-g groups were significantly reduced by 32%, 38%, 56%, and 67% respectively, compared with control. Moreover, about 80% of surviving larvae showed retarded development in dsRNA-treated groups. These results suggest that oral ingestion of bacterially expressed dsRNA in C. suppressalis could silence ptth, torso, spook and nm-g. Oral delivery of bacterially expressed dsRNA provides a simple and potential management scheme against C. suppressalis.