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Ophraella communa (Coleoptera: Chrysomelidae), the ragweed leaf beetle, is a biological control agent of the invasive common ragweed, Ambrosia artemisiifolia (Asterales: Asteraceae). Adults can survive cold conditions that occur during winter. The adults mate before entering overwintering. Understanding the connection between copulation and overwintering will be useful for determining O. communa seasonality. Determining the relationship between overwintering and copulation required comparison of mated and unmated beetles at mean lethal temperature (LTemp50) exposures for 2 h. Cold-related physiological indices, including the water ratio, super cooling point (SCP), cryoprotectant levels, and energy reserve levels, were also measured. Mating treatment decreased the LTemp50 of both sexes by reducing their mean SCP and water ratios. Although the changes of cryoprotectant levels in mated adults were not precisely consistent in between the genders, they increased greatly in both males and females. Body sugar may play a role in copulation and may also elevate cold hardiness in O. communa.

Life-history strategies of animals are governed by fitness-related trait trade-offs. In particular, the age at which either one of the two sexes copulate has been shown to affect insect reproductive output and longevity. This suggests that trade-offs between longevity and reproduction might drive the choice of a mating partner based on their age and ultimately such choices might select for different life-histories. Although several studies indicate that female age at mating dictates subsequent insect longevity, fecundity, and egg hatch rate, it is unclear how male age at copulation affects these life-history traits. In this study, we simultaneously investigated the effects of female and male age on female fecundity, eggs hatch rate, and adult longevity in Ophraella communa LeSage (Coleoptera: Chrysomellidae), the primary biological control agent of the invasive common ragweed, Ambrosia artemisiifolia L. We found that young mature females lived significantly longer when they were mated with older m...

Determining the best ratio of females to males of an insect's natural enemy is important for maximising population increase and promoting population establishment of a natural enemy. In this study, copulation behaviour, fecundity, progeny fitness and rate of population increase for the flea beetle, Agasicles hygrophila Selman & Vogt (Coleoptera: Chrysomelidae), were compared at different female percentage treatments (i.e., 80%, 66.7%, 50%, 33.3% and 20% females). The results showed that the copulation frequency and duration in males decreased, whereas those in females increased as the number of males increased. At 20%, 33.3% and 66.7% females, the rates of population increase were 3.4-, 2.17- and 0.79-fold higher than that at 50% females. Females at 20% and 33.3% were found to be optimal for mass rearing of the beetle.

Ophraella communa LeSage is native to North America and a biological control agent of the invasive weed Ambrosia artemisiifolia L. Since A. artemisiifolia plants grow old and die after September annually, O. communa suffers from food shortage. To understand the effect of food shortage or deprivation on population fitness of O. communa, the development and fecundity and hatchability of its progeny eggs were observed when larvae were offered A. artemisiifolia plants for either 3, 6, 12 or 24 hours daily. The results showed that larval food deprivation significantly influenced survival and developmental durations of larvae and pupae. Survival rates and developmental durations of larvae and pupae decreased and were prolonged significantly with decreasing time of larval daily food intake. Longevity and fecundity of adults shortened and decreased significantly with decreasing time of larval daily food intake. In addition, the hatch rates of progeny eggs decreased significantly with decreasing time of larval dai...

Ambrosia artemisiifolia L. is native to North America, and was unintentionally introduced into China in the 1930s, where it has become invasive. The two insect species Epiblema strenuana and Ophraella communa have been considered as biological control agents of A. artemisiifolia in China. The purpose of this study was to examine the control effect of O. communa + E. strenuana on A. artemisiifolia in the field. The mortality of A. artemisiifolia plants increased with time and increasing initial release densities of O. communa and/or E. strenuana in 2008 and 2009. The treatments of 0.53 O. communa + 0.53 E. strenuana per plant and 12 O. communa + 16 E. strenuana per plant at early (60–70 cm tall) and later (90–100 cm tall) growth stages could subsequently kill all plants prior to seed production in both 2008 and 2009. Thus, the two initial densities of the two insect species may be recommended when they are jointly used for biological control of A. artemisiifolia at the two growth stages. However, all or so...

Ophraella communa LeSage (Coleoptera: Chrysomelidae) is an effective biological control agent of the invasive common ragweed, Ambrosia artemisiifolia L. Body size, one of the most important life-history characters for many insects, affects O. communa mating choice. Temperature is one of the most important factors on body size, especially high temperatures. Adult body lengths were significantly inhibited after eggs, larvae and pupae of O. communa experienced high temperature stresses. With exception of subsequent female body after exposure of larvae to high temperatures, the body weights of females and males were not affected by temperatures. Since adult insect fecundities are often in proportion to their body sizes, we concluded that high temperatures may influence the population expansion of O. communa in the field.

China has the highest greenhouse-based production in the world. In 2010, the area of greenhouses devoted to vegetable production was estimated at 4.7 million ha. With the increasing costs of pest control, expanding pesticide resistance and the growing consumer concern regarding pesticide residues in fresh vegetables, a strong demand for applying non-chemical control methods is emerging in China. Biological control in the greenhouse environment is a viable alternative to pesticide use from both environmental and economic perspectives. Although we have only 17 cases of fully documented, successful biological control operations from China, involving 8 crops, 8 pest species and 14 species of natural enemies, the use of the biological control agents is much more widespread. There are 7 commercial companies and facilities producing 21 species of natural enemies, and most of them are available country-wide. Several of these employ a rearing system using artificial diets, and many now move to an integrated production system, including the mass production of the biocontrol agents, quality control, methods of long-distance transportation, release recommendations, and user feedback. While initially these systems relied on introduced natural enemies, they increasingly develop modified systems using native natural enemies. The increasing demand for pesticide-free, high quality vegetable produce year-round and the existing certification schemes make it very likely that the use of biocontrol agents will continue to increase in China.

Insects are ectotherms and their ability to resist temperature stress is limited. The immediate effects of sub-lethal heat stress on insects are well documented, but longer-term effects of such stresses are rarely reported. In this study, survival, development and reproduction of the whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) biotype B, were compared over five consecutive generations at 27, 31 and 35 °C and for one generation at 37 °C. Both temperature and generation significantly affected the fitness of the whitefly. These impacts were more dramatic with increasing generations and temperatures. Among the experimental temperatures, the most favorable for development and reproduction were 27 °C and 31 °C. At 27 °C, survival, development and fecundity were all stable over these five generations. At 31 °C, immature survival rate was the highest in the fifth generation, but female fecundities decreased in the fourth and fifth generations. At 35 °C, egg hatching rate, immature survival rate and female fecundity decreased significantly in the fourth and fifth generations. At 37 °C, survival of B. tabaci was not adversely affected, but female fecundity at 37 °C was less than 10% of that at 27 °C or 31 °C. These results demonstrate that the lethal high temperature for B. tabaci is over 37 °C, and the whitefly population continued expanding in the five generations at 35 °C. The ability of B. tabaci biotype B to survive high temperature stress will play an important role in its population extension under global warming.

Thermal adaptation plays a fundamental role in shaping the distribution and abundance of insects, and heat shock proteins (Hsps) play important roles in the temperature adaptation of various organisms. To better understand the temperature tolerance of the indigenous ZHJ2-biotype of whitefly Bemisia tabaci species complex, we obtained complete cDNA sequences for hsp90, hsp70, and hsp20 and analyzed their expression profiles under different high temperature treatments by real-time quantitative polymerase chain reaction. The high temperature tolerance of B. tabaci ZHJ2-biotype was determined by survival rate after exposure to different high temperatures for 1 h. The results showed that after 41°C heat-shock treatment for 1 h, the survival rates of ZHJ2 adults declined significantly and the estimated temperature required to cause 50% mortality (LT50) is 42.85°C for 1 h. Temperatures for onset (Ton) or maximal (Tmax) induction of hsps expression in B. tabaci ZHJ2-biotype were 35 and 39°C (or 41°C). Compared with previous studies, indigenous ZHJ2-biotype exhibits lower heat temperature stress tolerance and Ton (or Tmax) than the invasive B-biotype.

Ophraella communa is a biological control agent of invasive common ragweed, Ambrosia artemisiifolia. To understand the mate choice tactic of the beetle and improve mass-rearing of high-quality populations, the effects of morphological traits, age and copulation experience on mate choice in this beetle were studied in the laboratory. The results showed thatmate choice of male or female was related to certain adult morphological traits. Wing length, black streak width of frons and metaleg femur length of males were central to female mate choice. Females with longer wing length, black streak width of frons, pronotum width and foreleg tibia length were more attractive to males than those with the smaller ones. The chosen rate of older males was significantly higher than those of younger ones, and the chosen rate of females reached a maximum value on the 8th day after eclosion. The newly copulated females were less attractive to males than virgins, but females that have copulated in the distant past were more ...

Ophraella communa (Coleoptera: Chrysomelidae), originally from North America, has been used for biological control of common ragweed, Ambrosia artemisiifolia, in China since 2007. However, there is still a debate on whether O. communa can attack sunflowers under field conditions. To re-evaluate the biosecurity of O. communa against sunflower (Helianthus annuus), we investigated the population density of O. communa on three sunflower varieties that were intercropped with or planted in circumambience of A. artemisiifolia under field conditions. Our results showed that only very few O. communa eggs (

Ophraella communa LeSage (Coleoptera: Chrysomelidae), an effective biological control agent of A. artemisiifolia, was unintentionally introduced into China. To understand the biological control potential of O. communa on A. artemisiifolia, plant height, the number of branches and leaf control index of A. artemisiifolia injured by the beetle were determined. The results showed that plant height and number of branches decreased and leaf control index increased with the increasing initial release density of O. communa adults, when they were released at 90–100-cm plant height stage in 2008 and 60–70-cm stage in 2009. Even when O. communa was released at a low average density of 1.07 adults per plant at early growth stage of A. artemisiifolia in 2009, it could significantly suppress plant height and number of branches and led to a higher leaf control index compared with the herbicide treatment with Roundup. Prior to the fructicative period, the leaf control index of A. artemisiifolia by O. communa was...

Since insects are ectothermic, they are highly vulnerable to the sudden increase of temperature. Indeed, it has been hypothesized that the survival, development, fecundity, and even population expansion of insects are all affected significantly by extremely high temperature. We studied the effect of short-term high temperature stress on the survival and development of different stages, adult longevity and fecundity of Ophraella communa (Coleoptera: Chrysomelidae), a biological control agent of the invasive plant, the common ragweed, Ambrosia artemisiifolia (Asterales: Asteraceae) in the laboratory. The results showed that egg, larval, pupal and adult survival rates were significantly affected after 2 hour-short-stress at high temperatures (35 to 47°C) when compared to the 28°C control. With the exceptions of the control and 35°C stress, survival rate of females was significantly higher than that of males after short-stress at any high temperature. Short-term high temperature stress also significantly impa...

In China, an unintentionally introduced beetle Ophraella communa shows good control efficiency against invasive common ragweed, Ambrosia artemisiifolia. With the aim of understanding the capacity of O. communa to overcome sub-zero low temperature rapidly, the longevity, survival and fecundity of the beetle adults were studied at conditions of different rapid cold-hardening in the laboratory. The results showed that approximately 20% female or male individuals still survive at −12°C for 2 h, thus the discriminating temperature for rapid cold-hardening of both female and male beetles is −12°C. The survival, longevity and fecundity of adult beetles were significantly affected by rapid cold-hardening treatments. Compared to the control, the survival rates of adult beetles experiencing 2 h of rapid cold-hardening at 2, 5 and 8°C prior to exposure to its discriminating temperature (−12°C) were significantly increased. Adult longevity and fecundity were significantly higher after pre-treatment at 2, 5 a...

An invasive alien weed, Ageratina adenophora, has become widespread in the southwest of China due to its high environmental adaptation. In the present study, in order to understand the molecular aspects of resistance to heat and cold stress, we investigated the characterization and expression of heat shock protein genes (designated as Aahsp70 and Aahsp90) in A. adenophora. The cDNA of A. adenophora Aahsp70 and Aahsp90 was cloned by the combination of homology cloning and rapid amplification of cDNA ends (RACE). The Basic Local Alignment Search Tool analyses revealed that Aahsp70 and Aahsp90 shared high levels of identity with corresponding proteins from other species. Hybridization was employed to examine the expression patterns of Aahsp70 and Aahsp90. The relative expression levels of the Aahsp70 and Aahsp90 transcripts were both up-regulated and reached maximal levels at 2-4 h after heat shock. Then, Aahsp70 dropped progressively to the original level at ~4 h after heat shock. The transcriptional changes of Aahsp70 and Aahsp90 both were more obvious under cold stress. Recombinant Aahsp70 and Aahsp90 improved the viability, in comparison with the control cultures, of Escherichia coli under stress conditions. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the cell lysates suggested that the protective effect in vivo was related to increased thermo-stability of the soluble cytosolic proteins. This study emphasizes that the relationship between, and regulation of, heat shock proteins might incorporate variable strengths to increase the adaptation of A. adenophora.

To reveal the multi-generation effects of transgenic Bt cotton on the metabolism of secondary target herbivores, the activities of digestive and detoxification enzymes in beet armyworm, Spodoptera exigua (Hubner), fed with transgenic Bt (cv. GK-12) vs. non-Bt cotton (cv. Simian-3) for three successive generations were examined. Significantly lower activities of lipase, carboxylesterase (CarE) and acetylcholinesterase (AChE), and higher activities of trypsin and total superoxide dismutase (T-SOD) were observed in Bt-fed S. exigua than in non-Bt fed individuals during the three generations. But the activity of amylase did not differ between the Bt-fed and non-Bt fed S. exigua. Among the three generations of Bt-fed S. exigua, the activities of lipase and trypsin decreased significantly in the second or third generation than in the first generation. But those of CarE and AChE were increased significantly. Among the three generations of non-Bt fed S. exigua, the activity of lipase was significantly higher in the second and third generations than in the first generation. The increasing adaptation and fitness of S. exigua after continuous exposure to Bt cotton was supposed to be associated with the enhanced activities of detoxification enzymes. Measuring multi-generational enzymes activities of S. exigua in response to Bt cotton can provide proofs of its adaptation development, which is suggested to be a mandatory part of risk assessment of transgenic Bt plants on secondary target herbivores.

Ophraella communa, an unintentionally introduced leaf beetle in China, has good control efficiency on ragweed, Ambrosia artemisiifolia. Aspects of the climatic requirements for development, survival, longevity and fecundity of O. communa were studied under the conditions of constant temperature (25 ± 1°C), photoperiod of 14 L:10 D and three relative humidities (60%, 75% and 90% RHs). The results showed that the developmental periods of O. communa at different stages shortened along with the increasing relative humidity, except that of the pupal stage. Although no differences were observed in the pupal survival rate, ovipositional period, fecundity, longevity and adult female age-specific survivorship of O. communa under the three humidity conditions, the survival rates during the egg, larva and entire immature stage were significantly higher at 75% RH and 90% RH than at 60% RH. The innate rate of increase (rm), net reproductive rate (R0), finite rate of increase (λ) reached the maximum at 75% RH, with values of 0.181, 1116.4 and 1.198, respectively. These results indicated that the optimum relative humidity for the development of O. communa ranged from 75% RH to 90% RH. Thus O. communa prefers moist microclimate habitats. Its population may expand rapidly during mid-May to late August in south, east and central China, when the humidity is relatively high.

The effects of transgenic Bacillus thuringiensis (Bt) cotton on non-target Aphis gossypii were assessed by comparing the life-table parameters of aphids that fed on Bt vs. non-Bt cotton cultivars for the first and 37th generations. The enzyme-linked immunosorbent assay (ELISA) method was used to detect the transmission of the Bt protein from Bt cotton to A. gossypii and their honeydew. We found that the life-table parameters of A. gossypii that fed on Bt cotton for the first and 37th generations did not differ significantly from those of the non-Bt-fed individuals. However, the Bt protein was detected by ELISA in the Bt cotton leaves, and the content varied significantly at different growth stages. Furthermore, trace amounts of the Bt protein were detected in some of the Bt-fed aphids, and the honeydew of the Bt-fed aphids contained over 10 ng/g Bt protein. These results indicate that although trace amounts of the Bt protein were ingested, the Bt cotton had no significant negative impacts on A. gossypii in either the short or long term. Key words: Transgenic Bacillus thuringiensis cotton, Aphis gossypii, Bacillus thuringiensis detection, life-table parameters, multiple-generation, non-target effects.