Genome-wide identification of the BmAKR gene family in the silkworm (Bombyx mori) and their expression analysis in diapause eggs and nondiapause eggs / 生物工程学报

The aldo-keto reductase super family (AKRs) has a wide range of substrate specificity. However, the systematic identification of insect AKR gene family members has not been reported. In this study, bioinformatics methods were used to predict the phylogenetic evolution, physical and chemical properties, chromosome location, conserved motifs, and gene structure of AKR family members in Bombyx mori (BmAKR). Transcriptome data or quantitative real time polymerase chain reaction (qRT-PCR) were used to analyze the expression level of BmAKR genes during different organizational periods and silkworm eggs in different developmental states. Moreover, Western blotting was used to detect the expression level of the BmAKR in silkworm eggs. The results showed that 11 BmAKR genes were identified. These genes were distributed on 4 chromosomes of the silkworm genome, all of which had the (α/β) 8-barrel motif, and their physical and chemical characteristics were relatively similar. Phylogenetic analysis showed that the BmAKR genes could be divided into 2 subgroups (AKR1 and AKR2). Transcriptome data analysis showed that the expression of BmAKR genes were quite different in different tissues and periods. Moreover, the expression analysis of BmAKR genes in silkworm eggs showed that some genes were expressed significantly higher in nondiapause eggs than in diapause eggs; but another gene, BmAKR1-1, was expressed significantly higher in diapause eggs than in nondiapause eggs. The detection of protein level found that the difference trend of BmAKR1-1 in diapause eggs and non-diapause eggs was consistent with the results of qRT-PCR. In conclusion, BmAKR1-1 was screened out as candidates through the identification and analysis of the BmAKR genes in silkworm, which may regulate silkworm egg development is worthy of further investigation.

Biological aspects of Tiracola grandirena (Herrich-Schäffer, 1868) (Lepidoptera: Noctuidae): a polyphagous armyworm / Aspectos biológicos de Tiracola grandirena (Herrich-Schäffer, 1868) (Lepidoptera: Noctuidae): uma lagarta polífaga

We studied the biology of Tiracola grandirena (Herrich-Schäffer, 1868) (Lepidoptera: Noctuidae: Hadeninae) at 25 ± 1 °C, 70 ± 10% RH and 14 hours of photo phase. Three experiments, using 150 larvae each, were conducted for the larval stage. In the first, used to assess the duration and survival of all stages, insects were reared individually and fed an artificial diet (Grenee). In the second, individuals were also reared separately, but were fed leaves of 10 plants from different families. In the third, the larvae were not individualised, the food plants were rotated such as to provide three plant species every 48 hours. In the first experiment, the viability of the eggs, larvae, pupae and prepupae was 91.9, 94.7, 32.49 and 43.5%, respectively. The average duration of the egg, larvae, prepupae, pupae and adult were 6.0, 25.3, 25.7, 21.4 and 12.7 days, respectively. The prolonged prepupal period indicates that T. grandirena can develop larval (prepupal) diapause. The results of the experiments with different host plants showed that T. grandirena is polyphagous at species, population and individual level.

Estudou-se a biologia de Tiracola grandirena (Herrich-Schäffer, 1868) (Lepidoptera: Noctuidae: Hadeninae) em condições controladas de 25 ± 1 °C, 70 ± 10% UR e fotofase de 14 horas. Na fase larval foram estabelecidos três experimentos empregando 150 larvas cada. No primeiro, os insetos foram criados individualmente, com dieta artificial de Grenee, cujos dados foram utilizados para avaliar a duração e a sobrevivência das larvas, pupas e adultos. No segundo, as larvas foram criadas individualmente, alimentadas com folhas de 10 plantas de famílias diferentes avaliando-se a sobrevivência e duração larval (fase ativa). No terceiro, as larvas foram mantidas em um mesmo recipiente, variando-se a alimentação vegetal, três espécies a cada 48 horas, avaliando-se apenas a sobrevivência larval. No primeiro experimento a viabilidade de ovo, larva, pré-pupa e pupa foi de 91,93; 94,67; 32,39 e 43,48%, respectivamente. A duração média de ovo, larva, pré-pupa, pupa e adulto foi de 6,00; 25,26; 25,76; 21,35 e 12,67 dias, respectivamente. O prolongado período de pré-pupa indicou que, nas condições em que foi criada T. grandirena pode desenvolver diapausa larval (pré-pupal). Os resultados dos experimentos com diferentes plantas hospedeiras demonstraram que T. grandirena é polífaga ao nível específico, populacional e individual.

Differential gene expression during early embryonic development in diapause and non-diapause eggs of multivoltine silkworm Bombyx mori.

Quantification of the differential expression of metabolic enzyme and heat-shock protein genes (Hsp) during early embryogenesis in diapause and non-diapause eggs of the silkworm B. mori was carried out by semi-quantitative RT-PCR. Data analysis revealed that, the phosphofructokinase (PFK) expression started at a higher level in the early stage (6 h after oviposition) in non-diapause eggs, while in diapause induced eggs, it started at a lower level. However, the PFK gene expression in diapause eggs was comparatively higher than in non-diapause eggs. PFK facilitates use of carbohydrate reserves. The lower level of PFK gene expression in the early stage of diapause induced eggs but comparatively higher level of expression than in non-diapause eggs is due to enzyme inactivation via protein phosphorylation during early embryogenesis followed by de-phosphorylation in later stage. The sorbitol dehydrogenase-2 (SDH-2) gene was down regulated in diapause induced eggs up to 24 h and its expression levels in diapause induced eggs coincided with that of PFK gene at 48h in non-diapause eggs. During carbohydrate metabolism, there is an initial temporary accumulation of sorbitol which acts as protectant. The down regulation of SDH-2 gene during the first 24 hours in diapause induced eggs was due to the requirement of sorbitol as protectant. However, since the diapause process culminates by 48 h, the SDH-2 gene expression increased and coincided with that of PFK gene expression. The trehalase (Tre) gene expression was at a lower level in diapause induced eggs compared to non-diapausing eggs. The induction of Tre activity is to regulate uptake and use of sugar by the tissues. The non-diapause eggs revealed maximum expression of GPase gene with major fluctuations as well as an overall higher expression compared to diapause induced eggs. The diapause process requires less energy source which reflects lower activity of the gene. Heat shock protein (Hsp)genes (Hsp20.4, 40, 70, and 90) revealed differential levels of expression in both the eggs at all stages of embryonic development. The present study thus provides an overview of the differential expression levels of metabolic enzyme and Hsp genes in non-diapause and diapause induced eggs of multivoltine silkworm B. mori within 48 h after oviposition, confirming the major role of in early embryogenesis.

Photoperiodism and seasonality in neotropical population of Plutella xylostella L. (Lepidoptera: Yponomeutidae) / Fotoperiodismo e sazonalidade em população neotropical de Plutella xylostella L. (Lepidoptera: Yponomeutidae)

Neotropical populations of the diamondback moth Plutella xylostella L. have seasonal cycles of growth and decrease, and moth migration plays a fundamental role in generating such population dynamics. Since the oscillation of these populations is predictable, photoperiod might operate as a signal that triggers the migratory behaviour of the insect. Migration in insects is usually preceded by reproductive diapause, a photoperiodic response that can be characterised by morphological, physiological and behavioural alterations that permit to discriminate between migratory and non-migratory forms. In this study, I tested whether the pre-imaginal and reproductive development of P. xylostella from Minas Gerais (Brazil) is affected by artificial day-lengths that are equivalent to the periods of natural population growth or decrease. No evidence of photoperiodic response was found for the insect reared in laboratory on five different constant photoperiods, from 8h to 16h of light per day. There was no significant variation in survival and duration of egg, larva, and pupa stages or in pupal weight, adult size (forewing length), fecundity, and longevity. Although some species have geographically distinct photoperiodic responses, previous assumptions that cosmopolitan P. xylostella responds to photoperiod in temperate regions was questioned. Migratory and population seasonality among neotropical populations of P. xylostella certainly occurs independently of the photoperiodic announcement of seasonal changes in habitat quality.

Populações neotropicais de Plutella xylostella L. possuem ciclos sazonais de crescimento e declíneo e a migração é um fator-chave na geração da dinâmica populacional. Devido à previsibilidade sazonal das oscilações populacionais, o fotoperíodo poderia agir como o sinal que desencadea o comportamento migratório do inseto. Migrações de insetos são geralmente precedidas por diapausa reprodutiva, uma típica resposta fotoperiódica caracterizada por alterações morfológicas, fisiológicas e comportamentais que possibilitam discriminar as formas migratórias das não-migratórias. Neste estudo, foi testado se o desenvolvimento pré-imaginal e reprodutivo de P. xylostella, proveniente de Minas Gerais, é inibido por fotoperíodos equivalentes aos comprimentos de dias em que ocorrem as fases de crescimento ou de declínio das populações naturais. Nenhuma evidencia de resposta fotoperiódica foi encontrada para os insetos criados em laboratório sob cinco regimes constantes de fotoperíodos, variando de oito a dezesseis horas-luz-dia. Não houve variação significativa na sobrevivência e na duração dos estágios de ovo, larva e pupa. O peso das pupas também não foi afetado, assim como o tamanho, a fecundidade e a longevidade dos adultos. Embora algumas espécies de insetos possuam respostas fotoperiódicas geograficamente diferenciadas, a sugestão de alguns autores de que a cosmopolita P. xylostella responde ao fotoperíodo em regiões de clima temperado foi questionada. Os dados experimentais não suportaram a hipótese de que a sazonalidade migratória e populacional de P. xylostella dependem do fotoperíodo como sinal de anúncio de mudanças temporais na qualidade do habitat.

Long-term storage and viability of Ochlerotatus albifasciatus (Diptera: Culicidae)

The viability of Ochlerotatus albifasciatus (Macquart) eggs stored at room temperature and at 5ºC was studied over 31 months. After 12, 18 and 31 months of storage, eggs were acclimatized at 22ºC for ten days, and then inundated twice every seven days. The effect of the storage period on the percentage of hatching was analyzed by one way ANOVA. Differences on the hatching response between the first and second flooding were analyzed by paired t-test. Differences on the hatching response between the two storage conditions were analyzed by Mann-Whitney rank test. Results showed that (1) Oc. albifasciatus eggs were able to survive and hatch over 31 months; (2) the percent hatching of eggs stored at 5ºC was higher than that of eggs stored at room temperature; and (3) low temperatures and long periods without water favor installment hatching.

Influência do fotoperíodo na reprodução e longevidade de Orius insidiosus (Say) (Heteroptera: Anthocoridae) / Effect of photoperiod on reproduction and longevity of Orius insidiosus (Say) (Heteroptera: Anthocoridae)

Orius insidiosus (Say) is an important predator of thrips and photoperiod can affect the reproduction of this insect. The objective of this work was to verify the occurrence of reproductive diapause in O. insidiosus in laboratory conditions. The experiments were carried out in environmental chambers, at photoperiods of 9L:15D, 10L:14D, 11L:13D, 12L:12D, 13L:11D and 14L:10D, temperature of 25±1°C and RH of 70±10 percent. Fecundity and sex ratio were not affected by the different photoperiods; an average number of 162.0, 189.8, 175.8, 195.2, 132.2 and 125.1 eggs/female, respectively, were found on these photoperiods. However, the photoperiods affected longevity of the predator. Longevity varied from 77.1 days, at photoperiod 10L:14D, to 40.1 days at photoperiod 9L:15D. O. insidiosus did not enter reproductive diapause in the conditions carried out in this experiment. This information is important to understand the factors that are involved on performance and use of O. insidiosus on biological control of thrips on protected cultivation.

Orius insidiosus (Say) é um importante predador de tripes, e sua reprodução pode ser afetada pelo fotoperíodo. O objetivo deste trabalho foi verificar a ocorrência de diapausa reprodutiva em O. insidiosus submetido a diferentes fotoperíodos em laboratório. O experimento foi conduzido em câmaras climáticas, nos fotoperíodos 9L:15E, 10L:14E, 11L:13E, 12L:12E, 13L:11E e 14L:10E, temperatura de 25±1°C e UR de 70±10 por cento. A fecundidade e a razão sexual do predador não foram afetadas pelos diferentes fotoperíodos; o número médio de ovos/fêmea foi de 162,0, 189,8, 175,8, 195,2, 132,2 e 125,1 nos fotoperíodos 9L:15E, 10L:14E, 11L:13E, 12L:12E, 13L:11E e 14L:10E respectivamente. O fotoperíodo, no entanto, afetou a longevidade de O. insidiosus independentemente do sexo, sendo que a maior longevidade (77,1 dias) ocorreu no fotoperíodo 14L:10E e a menor (40,1 dias) no fotoperíodo 9L:15E. O. insidiosus não apresentou predisposição a entrar em diapausa reprodutiva nos fotoperíodos analisados, nas condições deste experimento. Essa informação é importante para a compreensão dos fenômenos que regem o desempenho e a utilização dessa espécie de predador para o controle biológico de tripes em sistemas de cultivos protegidos.

Photophase influence on the reproductive diapause, seasonal morphs, and feeding activity of Euschistus heros (Fabr., 1798) (Hemiptera: Pentatomidae)

Laboratory studies were conducted to verify the influence of photophase on diapause incidence in the Neotropical brown stink bug, Euschistus heros (Fabr., 1798), fed with soybean [Glycine max (L.)] Merrill pods. Nymphs were maintained at three different photophases: 10 h, 12 h, and 14 h, with constant temperature of 25 ± 1ºC and relative humidity of 65 ± 5%. With 14 h, aproximately 100% of the adults showed mature reproductive organs; the shoulder (spine) length was significantly greater (2.96 and 2.79 mm for females and males, respectively) than those of bugs maintained at the photophase of 12 h (2.60 mm for females and males) and 10 h (2.59 and 2.53 mm for females and males). At the longer photophase (14 h), E. heros showed better reproductive performance and greater feeding activity than insects reared at 10 h and 12 h; in all photophases bugs tended to reduce feeding from the 1st to the 6th week of life. Body color was considered an unreliable parameter to indicate diapause incidence. However, at 14 h, 60% of the insects were dark brown and 40% were reddish brown. These results indicate that E. heros enters reproductive diapause with photophase of 12 hours or less, showing immature reproductive organs or with intermediate development, with shoulder (spine) less developed and reduced feeding activity.

A influência da fotofase na diapausa do percevejo-marrom Euschistus heros (Fabr.) alimentado com soja [Glycine max (L.)] Merrill foi estudada em laboratório. Os experimentos foram conduzidos sob três fotofases, 10 h, 12 h e 14 h, mantendo-se constantes a temperatura (25 ± 1ºC) e a umidade relativa (65 ± 5%). Sob 14 h, cerca de 100% dos insetos apresentaram órgãos reprodutivos maduros; os espinhos pronotais, medidos aos 10 dias de vida adulta, foram significativamente maiores (2,96 e 2,79 mm para fêmeas e machos, respectivamente) que os espinhos dos insetos mantidos sob 12 h (2,60 mm para fêmeas e machos) ou 10 h (2,59 e 2,53 mm para fêmeas e machos, respectivamente). Com a fotofase mais longa, ocorreu desempenho reprodutivo e atividade alimentar maiores do que o observado para adultos criados sob fotofase de 10 h ou 12 h. Em todas as fotofases observou-se tendência de diminuição da atividade alimentar dos percevejos da 1ª à 6ª semana. A coloração do corpo mostrou-se como um parâmetro não confiável para indicar a diapausa, mas, sob 14 h, cerca de 60% dos insetos apresentaram corpo marrom-escuro e os demais marrom-avermelhado. Esses resultados demonstram que esse percevejo entra em diapausa reprodutiva sob fotofase de 12 horas ou menos, apresentando órgãos reprodutivos imaturos ou com desenvolvimento intermediário, espinhos pronotais menos desenvolvidos e redução na atividade alimentar.

Photoperiodic sensitivity and diapause induction during ovarian, embryonic and larval development of the flesh fly, Sarcophaga argyrostoma.

Sensitivity to the daily photoperiod, particularly with respect to pupal diapause induction, was studied during ovarian, embryonic, and larval development of the flesh fly Sarcophaga argyrostoma. Large flies were shown to have a greater number of primary follicles in their ovaries and to be capable of limited ovarian maturation in the absence of exogenous protein (autogeny). Such ovarian development occurred independently of photoperiod. However, long days experienced during embryogenesis caused more rapid development, and earlier larviposition, than short days. Short days during embryonic and subsequent larval development also induced pupal diapause, whereas long days led to continuous or non-diapause development of the pupae. Pupal diapause could not be induced by photoperiods during the vitellogenic phase of ovarian development. In Sarcophaga argyrostoma, a maternal effect preventing pupal diapause among the progeny of files with a diapause history was not observed.

Growth difference of Plasmodium vivax in Anopheles sinensis and An.anthropophagus during the periods of clinic attack and diapause of vivax malaria patients / 中国血吸虫病防治杂志

Objective To compare the growth of Plasmodium vivax in Anopheles sinensis(An.s) and Anopheles anthropophagus(An.a) during the periods of malaria clinic attack and diapause. Methods The blood samples of patients during the clinic attack and diapause of vivax malaria patients in the vivax epidemic area in China were collected, feeding the mosquitoes of An.s and An.a by using the artificial membrane feeding system in vivo in the lab, and the mosquitoes were dissected during the day 7-9th and 14th after the infection and the oocysts and sporozoites in the stomach and gland of the mosquitoes were counted, respectively. Results The oocyst positive rate in An.s fed by Plasmodium vivax during the fever stage was lower than that in non-fever stage, the sporozoite positive rate in An.a fed by Plasmodium vivax during the fever stage was lower than that in non-fever stage. The positive mosquito rate with oocyst and sporozoite infected by Plasmodium vivax in the fever stage to An.s and An.a were lower than those in non-fever stage. The infective sporozoite intensity of An.s fed by Plasmodium vivax in the fever stage was lower than that in non-fever stage, but the reverse result was found to An.a. Conclusion There is a significant difference between the periods of malaria clinic attack and diapause of the oocyst and sporozoite infection to An.s and An.a.