Detection of Yeast-like Symbionts in Brown Planthopper Reared on Different Resistant Rice Varieties Combining DGGE and Absolute Quantitative Real-Time PCR

The brown planthopper (BPH), Nilaparvata lugens, is a serious pest of rice throughout Asia. Yeast-like symbionts (YLS) are endosymbionts closely linked with the development of BPH and the adapted mechanism of BPH virulence to resistant plants. In this study, we used semi-quantitative DGGE and absolute quantitative real-time PCR (qPCR) to quantify the number of the three YLS strains (Ascomycetes symbionts, Pichia-like symbionts, and Candida-like symbionts) that typically infect BPH in the nymphal stages and in newly emerged female adults. The quantities of each of the three YLS assessed increased in tandem with the developing nymphal instar stages, peaking at the fourth instar stage, and then declined significantly at the fifth instar stage. However, the amount of YLS present recovered sharply within the emerging adult females. Additionally, we estimated the quantities of YLS for up to eight generations after their inoculation onto resistant cultivars (Mudgo, ASD7, and RH) to reassociate the dynamics of YLS with the fitness of BPH. The minimum number of each YLS was detected in the second generation and gradually increased from the third generation with regard to resistant rice varieties. In addition, the Ascomycetes symbionts of YLS were found to be the most abundant of the three YLS strains tested for all of the development stages of BPH.

Effect of elevated CO2 and temperature on thermal constants and lower threshold temperatures of maize aphid, Rhopalosiphum maidis (Fitch.) (Aphididae:Hemiptera) on maize, Zea mays (Linn.)

The objective of this study was to examine the development of corn leaf aphid,Rhopalosiphum maidis Fitch (Aphididae: Hemiptera) on maize Zea mays Linnaeus at elevated and ambient concentrations of CO2 (550 and 380ppm ± 25 ppm, respectively) at six temperatures (20, 25, 27, 30, 33 and 35°C) and to estimate thermal constants and lower temperature thresholds for the forecasting models based on heat accumulation units which could be developed for use in forecasting. The duration of different growth stages of R.maidis were reduced with an increase of temperature from 20°C to 35°C under both ambient and elevated CO2 conditions. The lower development threshold for first nymphal instar,second nymphal instar, third nymphal instar, fourth nymphal instar, adult duration and total development period required 10.1, 5.04, 13.42, 26.96, 10.9, 23.22 and 20.20°C under eCO2 whereas it was 13.32, 9.41, 19.13, 30.48, 16.38, 22.88 and 20.89°C under aCO2 conditions,respectively. The mean lower temperature threshold for nymph was slightly higher (16.38°C) at aCO2 compared to that of eCO2 (10.90°C) whereas for adult the mean lower temperature threshold was slightly higher (23.22°C) at eCO2 compared to that of aCO2 (22.88°C). The thermal requirement of R. maidis from first nymphal instar to adult (total development period) was found to be 100.00 degree days (DD) under eCO2 conditions as against 111.11degree days under aCO2 conditions. These estimated temperature thresholds and thermal constant can predict the pest scenarios and population dynamics of R. maidis.

Regulation of metamorphosis in neopteran insects is conserved in the paleopteran Cloeon dipterum (Ephemeroptera)

In the Paleozoic era, more than 400 Ma, a number of insect groups continued molting after forming functional wings. Today, however, flying insects stop molting after metamorphosis when they become fully winged. The only exception is the mayflies (Paleoptera, Ephemeroptera), which molt in the subimago, a flying stage between the nymph and the adult. However, the identity and homology of the subimago still is underexplored. Debate remains regarding whether this stage represents a modified nymph, an adult, or a pupa like that of butterflies. Another relevant question is why mayflies have the subimago stage despite the risk of molting fragile membranous wings. These questions have intrigued numerous authors, but nonetheless, clear answers have not yet been found. By combining morphological studies, hormonal treatments, and molecular analysis in the mayfly Cloeon dipterum, we found answers to these old questions. We observed that treatment with a juvenile hormone analog in the last nymphal instar stimulated the expression of the Kr-h1 gene and reduced that of E93, which suppress and trigger metamorphosis, respectively. The regulation of metamorphosis thus follows the MEKRE93 pathway, as in neopteran insects. Moreover, the treatment prevented the formation of the subimago. These findings suggest that the subimago must be considered an instar of the adult mayfly. We also observed that the forelegs dramatically grow between the last nymphal instar, the subimago, and the adult. This necessary growth spread over the last two stages could explain, at least in part, the adaptive sense of the subimago.

The mayfly subimago explained. The regulation of metamorphosis in Ephemeroptera

ABSTRACTIn the Paleozoic era, more than 400 million years ago, insects continued molting after forming functional wings. Today, however, all flying insects stop molting after metamorphosis when they become fully winged. The only exception is the mayflies (Ephemeroptera), which molt in the subimago, a flying intermediate stage between the nymph and the adult. However, the identity and homology of the subimago remains underexplored. Debate remains regarding whether this stage represents a modified nymph, an adult, or a pupa like that of butterflies. Another relevant question is why do mayflies maintain the subimago stage despite the risk of molting fragile membranous wings. These questions have intrigued numerous authors but nonetheless, clear answers have not yet been found. However, by combining morphological studies, hormonal treatments, and molecular analysis in the mayfly species Cloeon dipterum, we found new answers to these old questions. We observed that treatment with a juvenile hormone analog in the last nymphal instar stimulated the expression of Kr-h1 gene and reduced that of E93, which suppress and trigger metamorphosis, respectively. Consequently, the subimago is not formed in these treated mayflies. This indicates that metamorphosis is determined prior to the formation of the subimago, which must therefore be considered an instar of the adult stage. We also observed that the forelegs dramatically grow between the last nymphal instar, the subimago, and the adult. This necessary growth is spread over the last two stages, which could explain, at least in part, the adaptive sense of the subimago.

E93-depleted adult insects preserve the prothoracic gland and molt again

ABSTRACTInsect metamorphosis originated around the middle Devonian, associated with the innovation of the final molt; this occurs after histolysis of the prothoracic gland (PG; which produces the molting hormone) in the first days of adulthood. We previously hypothesized that transcription factor E93 is crucial in the emergence of metamorphosis, because it triggers metamorphosis in extant insects. This work on the cockroach Blattella germanica reveals that E93 also plays a crucial role in the histolysis of PG, which fits the above hypothesis. Previous studies have shown that the transcription factor FTZ-F1 is essential for PG histolysis. We have found that FTZ-F1 depletion towards the end of the final nymphal instar downregulates the expression of E93, whereas E93-depleted nymphs molt to adults that retain a functional PG. Interestingly, these adults are able to molt again, which is exceptional in insects. The study of insects able to molt again in the adult stage may reveal clues about how nymphal epidermal cells definitively become adult cells, and whether it is possible to reverse this process.

Energy reserves in Brevicoryne brassicae (Lin.) (Homoptera: Aphididae) instars and their effect on predation and longevity of three coccinellid species under laboratory conditions

Background Contents of carbohydrates, protein, and lipids in tissues of insect prey constitute the required energy reserve utilized by their predators. Their amounts define the preference of the predator to certain prey on which its biological performance is the optimum. Feeding preferences and longevity of the 3 coccinellid beetles; Coccinella septempunctata L., C. undecimpunctata L., and Scymnus interruptus L. were studied in relation to energy reserve in tissues of the cabbage aphid, Brevicoryne brassicae (Lin.). Main body The level of energy reserve increased with increasing of aphid nymphal instars showing the highest in 4th instar (0.10844 cal/mg). Lipids and carbohydrates represented the highest energy reserve average rate in B. brassicae adults (0.29139 and 0.07404 cal/mg), respectively. C. septempunctata larval stage recorded the highest efficiency in daily predation (89.42 prey), when fed on 1st instar nymphs of the aphid. It required 21 days when fed on the 3rd nymphal instar of B. brassicae. S. interruptus larval stage reflected the lowest consumption and longevity in predation where it consumed less than 69.25 prey/day and lasted a period less than 17.5 days. Generation time for C. undecimpunctata lasted the longest (20.89 days) when fed on B. brassicae 1st nymphal instar. It was decreased in S. interruptus recording (12.22 days) when fed on B. brassicae 3rd nymphal instar. Conclusion Only C. septempunctata was efficient in predation and its biological performance was the best compared to the other tested coccinellid species. Use of C. septempunctata can be recommended when applying biological control programs in the field against the cabbage aphid.

E93-depleted adult insects preserve the prothoracic gland and molt again

ABSTRACTInsect metamorphosis originated around the middle Devonian, associated with the innovation of the final molt; this occurs after the histolysis of the prothoracic gland (PG; which produces the molting hormone) in the first days of adulthood. We previously hypothesized that transcription factor E93 was crucial in the emergence of metamorphosis, since it triggers metamorphosis in extant insects. This work on the cockroach Blattella germanica reveals that E93 also plays a crucial role in the histolysis of PG, which fits the above hypothesis. Previous studies have shown that the transcription factor FTZ-F1 is essential for PG histolysis. We have found that FTZ-F1 depletion, towards the end of the final nymphal instar, downregulates the expression of E93, while E93-depleted nymphs molt to adults that retain a functional PG. Interestingly, these adults are able to molt again, which is exceptional in insects. The study of insects able to molt again in the adult stage may reveal clues as to how nymphal epidermal cells definitively become adult cells, and if it is possible to revert this process.Summary statementThe prothoracic gland disintegrates after insect metamorphosis. It was believed that the factor FTZ-F1 determines this disintegration. This work reveals that FTZ-F1 action is mediated by the factor E93.

The Plecoptera of Panama. IV. Description of the last nymphal instar for two species of Anacroneuria Klapálek (Plecoptera: Perlidae) from the Caldera River, Chiriquí, Panamá

The last nymphal instars of Anacroneuria planicollis Klapálek, 1923 and A. quetzali Gutiérrez-Fonseca and Springer, 2015 are described from the Caldera River, Chiriquí Province, Panamá. The adult female of A. quetzali is associated for the first time and described also from the Caldera River, Chiriquí Province, Panamá.

Regulatory mechanisms underlying the specification of the pupal-homologous stage in a hemimetabolous insect

Juvenile hormones and the genetic interaction between the transcription factors Krüppel homologue 1 ( Kr-h1 ) and Broad ( Br ) regulate the transformation of insects from immature to adult forms in both types of metamorphosis (holometaboly with a pupal stage versus hemimetaboly with no pupal stage); however, knowledge about the exact instar in which this occurs is limited. Using the hemimetabolous cricket Gryllus bimaculatus ( Gb ), we demonstrate that a genetic interaction occurs among Gb ′ Kr-h1 , Gb ′ Br and the adult-specifier transcription factor Gb ′ E93 from the sixth to final (eighth) nymphal instar. Gb ′ Kr-h1 and Gb ′ Br mRNAs were strongly expressed in the abdominal tissues of sixth instar nymphs, with precocious adult moults being induced by Gb ′ Kr-h1 or Gb ′ Br knockdown in the sixth instar. The depletion of Gb ′ Kr-h1 or Gb ′ Br upregulates Gb ′ E93 in the sixth instar. By contrast, Gb ′ E93 knockdown at the sixth instar prevents nymphs transitioning to adults, instead producing supernumerary nymphs. Gb ′ E93 also represses Gb ′ Kr-h1 and Gb ′ Br expression in the penultimate nymphal instar, demonstrating its important role in adult differentiation. Our results suggest that the regulatory mechanisms underlying the pupal transition in holometabolous insects are evolutionarily conserved in hemimetabolous G . bimaculatus , with the penultimate and final nymphal periods being equivalent to the pupal stage. This article is part of the theme issue ‘The evolution of complete metamorphosis’.

The Effects of Different Diets and Transgenerational Stress on Acyrthosiphon pisum Development

Despite the fact that sap-feeding hemipterans are major agricultural pests, little is known about the pea aphid’s (Acyrthosiphon pisum) nymphal development, compared to other insect models. Given our limited understanding of A. pisum nymphal development and variability in the naming/timing of its developmental events between different environmental conditions and studies, here, we address developmental knowledge gaps by elucidating how diet impacts A. pisum nymphal development for the LSR1 strain when it develops on its universal host plant (Vicia faba), isolated leaves, and artificial diet. Moreover, we test how plant age and transgenerational stressors, such as overcrowding and low plant vigor, can affect nymphal development. We also validate a morphological method to quickly confirm the life stage of each nymphal instar within a mixed population. Overall, we found extremely high variation in the timing of developmental events and a significant delay in nymphal (~5–25-h/instar) and pre-reproductive adult (~40-h) development when reared on isolated leaves and artificial diets, compared to intact host plants. Also, delays in development were observed when reared on older host plants (~9–17-h/event, post 2nd instar) or when previous generations were exposed to overcrowding on host plants (~20-h delay in nymph laying) compared to controls.