Evidence for a conserved microbiota across the different developmental stages of Plodia interpunctella

Diversity and composition of lepidopteran microbiotas are poorly investigated, especially across the different developmental stages. To improve this knowledge, we characterize the microbiota among different developmental stages of the Indian meal moth, Plodia interpunctella, which is considered one of the major pest of commodities world-wide. Using culture-independent approach based on Illumina 16S rRNA gene sequencing we characterized the microbiota of four developmental stages: eggs, first-, and last-instar larvae, and adult. A total of 1022 bacterial OTUs were obtained, showing a quite diversified microbiota associated to all the analyzed stages. The microbiotas associated with P. interpunctella resulted almost constant throughout the developmental stages, with approximately 77% of bacterial OTUs belonging to the phylum of Proteobacteria. The dominant bacterial genus is represented by Burkholderia (?64%), followed by Propionibacterium, Delftia, Pseudomonas, and Stenotrophomonas. A core bacterial community, composed of 139 OTUs, was detected in all the developmental stages, among which 112 OTUs were assigned to the genus Burkholderia. A phylogenetic reconstruction, based on the 16S rRNA, revealed that our Burkholderia OTUs clustered with Burkholderia cepacia complex, in the same group of those isolated from the hemipterans Gossyparia spuria and Acanthococcus aceris. The functional profiling, predicted on the base of the bacterial 16S rRNA, indicates differences in the metabolic pathways related to metabolism of amino acids between preimaginal and adult stages. We can hypothesize that bacteria may support the insect host during preimaginal stages.     

不同生长季节黑果枸杞的根际细菌群落结构

【目的】黑果枸杞是一种耐盐植物,是我国西北干旱区盐渍土改良的优良植物物种,其根际土壤细菌群落结构在不同生长时期的变化特征尚不清楚。【方法】本研究采用Illumina MiSeq高通量测序研究了黑果枸杞3个生长阶段的根际土壤细菌群落结构的动态变化。【结果】所有样品中共获得317467条序列,对应于7028个细菌/古细菌OTUs。根际土壤细菌群落的α多样性显著高于非根际土壤。衰老期根际细菌的多样性和丰富度明显低于营养生长期和花/果期。变形菌门和酸杆菌门的相对丰度随生长时期的演变而逐渐降低,而蓝细菌门则相反。厚壁菌门的丰度在衰老期明显高于营养生长期和花/果期。优势属的组成也随生长期的演变而改变,营养生长期、花/果期、衰老期的优势属数量分别为17、16、4,且组成也具有差异。相似性分析表明营养生长期和花/果期的根际细菌群落具有很高的相似性,衰老期根际细菌群落组成与生长期和花/果期具有很高差异,然而与非根际土壤的群落结构具有较高的相似性。【结论】根际土壤细菌群落多样性和组成随生长期的改变而表现出明显的动态变异性,表明黑果枸杞生长时期对根际土壤细菌群落结构具有重要的影响。     

Chromosome constitution and nucleolar behaviour in the endosperm of Cucurbita pepo L

At early stages of development the endosperm of Cucurbita pepo L. was found to be triploid with 3n=60. Higher levels of ploidy (6n, 12n, 24n, 48n) were encountered at later stages. The euploid increase in chromosome number is the result of endopolyploidy. The volume of the nucleolus increased with the increase in nuclear volume. Cells with varying numbers of nucleoli of different sizes and shapes were observed in the endosperm at all stages. Fragmentation of the nucleolus is believed to be responsible for the increase in nucleolar number. Signs of nuclear disorganisation in the endosperm were visible from the 17th day onwards. As the endosperm tissue gets absorbed by the developing embryo, the mature seed is non-endospermous.     

Stage correlation of symbiotic bacterial community and function in the development of litchi bugs (Hemiptera: Tessaratomidae)

Bacterial symbionts of insects have been shown to play important roles in host fitness. However, little is known about the bacterial community of Tessaratoma papillosa which is one of the most destructive pests of the well-known fruits Litchi chinensis Sonn and Dimocarpus longan Lour in Oriental Region, especially in South-east Asia and adjacent areas. In this study, we surveyed the bacterial community diversity and dynamics of T. papillosa in all developmental stages with both culture-dependent and culture-independent methods by the third-generation sequencing technology. Five bacterial phyla were identified in seven developmental stages of T. papillosa. Proteobacteria was the dominant phylum and Pantoea was the dominant genus of T. papillosa. The results of alpha and beta diversity analyses showed that egg stage had the most complex bacterial community. Some of different developmental stages showed similarities, which were clustered into three phases: (1) egg stage, (2) early nymph stages (instars 1–3), and (3) late nymph stages (instars 4–5) and adult stage. Functional prediction indicated that the bacterial community played different roles in these three phases. Furthermore, 109 different bacterial strains were isolated and identified from various developmental stages. This study revealed the relationship between the symbiotic bacteria and the development of T. papillosa, and may thus contribute to the biological control techniques of T. papillosa in the future.

     

Long-term stability in the interaction betweenTetranychus urticae andPhytoseiulus persimilis producing successful integrated control on roses in southeast Queensland

Phytoseiulus persimilis Athias-Henriot successfully controlledTetranychus urticae Koch on rose hedges in an integrated mite control programme in two unheated plastic-covered tunnels at Cleveland in southern Queensland. Mite populations were monitored fortnightly for nearly three years in one tunnel and for two years in a second. Once established,P. persimilis quickly suppressedT. urticae and then persisted. A relatively stable, long-term, low-level interaction developed between the two species under normal commercial conditions. In one tunnel, the production (upper) level of the four rose hedges contained fewer mites of both species per compound leaf (1.71 of all stages ofT. urticae, 0.38 motiles; 0.25 of all stages ofP. persimilis, 0.12 motiles) than the maintenance (lower) layer (2.02 of all stages ofT. urticae, 0.75 motiles; 0.35 of all stages ofP. persimilis, 0.21 motiles). On 68% of sampling occasions, 10% or less of compound leaves in the production layer were infested withT. urticae (all stages including eggs) and control was excellent throughout the 30-month period afterP. persimilis was established, there being no economic losses. In the second tunnel, there was no significant difference between mite numbers in the production and maintenance layers of the two hedges examined. The overall mean number of all stages ofT. urticae per compound leaf was 3.2 (1.45 motiles), and ofP. persimilis 0.46 (0.25 motiles). Control here was slightly less effective than in the first tunnel, but was still satisfactory, with 10% or less of leaves being infested on 45.5% of sampling occasions (20% or less on 66% of sampling occasions) withT. urticae (all stages including eggs).Phytoseiulus persimilis was not reintroduced or redistributed during the course of the observations. An auxiliary miticide (clofentezine) was applied on only one occasion to selected areas in the second tunnel. The influence of high temperatures on control byP. persimilis is discussed.     

Post-fire succession of small mammals in the Cerrado of central Brazil

In the Cerrado savannas from Brazil fire events are common and strongly influence the vegetation structure and, consequently, the associated small mammals. In this paper, we investigate changes in the structure of small mammal communities related to sites of different post-fire ages. Mammals were captured in similar Cerrado sites that differed in time since the last burn (1 to 26 yr). We sampled six sites in the wet season of 1997 (phase 1) and, three years later, six sites in the wet and dry seasons (phase 2). Six rodent species and four marsupials were captured. Community composition changed drastically as a function of time since fire. The diversity and abundance of small mammals reached maximum values in the early successional stages. The rodent Calomys tener was present only in early seral stages. The rodent Bolomys lasiurus was more frequent in mid-successional stages and decreased in later seral stages, and the rodent Oryzomys subflavus occupied all successional stages. The marsupial Gracilinanus agilis was dominant in the area that did not burn for at least 23 yr. Changes in composition of the community of small mammals were more accelerated in early successional stages, when there are more drastic vegetational changes. The ability of small mammals to cope with Cerrado fires and the great dissimilarity among post-burning seral stages suggest that a mosaic of areas representing different post-fire seral stages could increase the regional diversity of this group.     

Comparison of quantitative gonad maturation scales in a temperate oyster (Crassostrea gigas) and a sub-tropical oyster (Crassostrea corteziensis)

Quantitative scales to evaluate maturity of female gonads were compared for a temperate oyster, Crassostrea gigas and a tropical oyster, Crassostrea corteziensis grown in the same locality. C. gigas had well-defined maturation and spawning periods and a resting phase in winter; in C. corteziensis mature individuals occurred most of the year and there were several spawning peaks. Of the quantitative scales used here, average oocyte diameter and gonad coverage area, much used for C. gigas, and ovary maturity index, less used, were inadequate to distinguish the reproductive pattern of C. corteziensis, since they both skewed the degree of maturation in vitellogenic stages in favor of C. gigas. Maximum oocyte diameter and maximum cytoplasm area were different among species at vitellogenic stages and also in previtellogenic stages. Nucleus:cytoplasm ratio was significantly different in previtellogenic and spawned stages between C. gigas and C. corteziensis. The Index of gonadal development was skewed in favor of C. gigas in postvitellogenic stage. The only scale that was comparable between species was reproductive potential, but it also was one of the most laborious. Other ordinal scales commonly used, such as a visual external evaluation of the gonad, only classified correctly a quarter of the stages.     

Edge Effects Act Differentially on Multiple Early Regeneration Stages of a Shade‐tolerant Tree Tapirira mexicana

Forest fragmentation is pervasive in tropical landscapes, and one pathway by which fragmentation may negatively impact populations is via edge effects. Early life‐stages are particularly important for species regeneration as they act as bottlenecks, but how edge effects may act differentially on different life‐stages is unknown. This study evaluated edge effects on multiple early life‐stages of a currently common animal‐dispersed, shade‐tolerant tree Tapirira mexicana (Anacardiaceae). The study was conducted in tropical premontane wet forest fragments in a highly deforested region of Costa Rica. The stages assessed were pre‐dispersal predation, primary dispersal, post‐dispersal predation, secondary dispersal, ex situ germination, in situ seed longevity, first and second year seedling abundance, second year seedling survivorship, and basal diameter growth. Results showed that impacts of edge effects were not equal across stages, but were limited to specific stages and times. One stage which may act as a bottleneck for species regeneration was pre‐dispersal predation. Over 60 percent of the seeds were predated by larvae, and predation was higher near the edge than interior habitat. Seeds lost viability within 10 d in the forest. Germination to first year seedling stage was also lower near edges, but such effect was eliminated within a year after that. Primary dispersal, seedling survivorship, and growth were not affected by proximity to edges, and both secondary dispersal and post‐dispersal predation were rare. This study demonstrates that current population abundance may not guarantee future species persistence and the importance of considering multiple life‐stages for a comprehensive assessment of forest fragmentation effects on species regeneration.     

Root-hair infection and nodulation of four grain legumes as affected by the form and the application time of nitrogen fertilizer

The effects of application of combined nitrogen fertilizer (ammonium nitrate or urea) on root-hair infection and nodulation of four grain legumes were studied. Young roots of each legume were inoculated with their compatible rhizobia. The application of the two forms of combined N either at the early stages of plant growth and/or at the time of nodule formation depressed root-hair curling, infection and nodulation. Infection of hairs on the primary roots was more sensitive to the N fertilizer than hair infection of secondary roots in bothVicia faba andPisum sativum. The nodule number and total fresh mass of the four legumes were drastically affected by fertilizer application. The combined N added both at early and at later stages significantly reduced the nodulation ofV. faba, Phaseolus vulgaris andVigna sinensis. The inhibitory effect of urea on nodulation ofP. sativum was only observed when the fertilizer was applied at the late stages of plant growth. It is concluded that, although the nodulation of the four legumes was suppressed by combined N, the initial events ofRhizobium-legume symbiosis (infection of roots and nodule initiation) are more sensitive to combined N than the stages after nodule formation.     

Dynamic changes of gut bacterial communities present in larvae of Anoplophora glabripennies collected at different developmental stages

The Asian long-horned beetle, Anoplophora glabripennies (Motschulsky), is a destructive wood-boring pest that is capable of killing healthy trees. Gut bacteria in the larvae of the wood-boring pest is essential for the fitness of hosts. However, little is known about the structure of the intestinal microbiome of A. glabripennies during larval development. Here, we used Illumina MiSeq high-throughput sequencing technology to analyze the larval intestinal bacterial communities of A. glabripennies at the stages of newly hatched larvae, 1st instar larvae and 4th instar larvae. Significant differences were found in larval gut microbial community structure at different larvae developmental stages. Different dominant genus was detected during larval development. Acinetobacter were dominant in the newly hatched larvae, Enterobacter and Raoultella in the 1st instar larvae, and Enterococcus and Gibbsiella in the 4th instar larvae. The microbial richness in the newly hatched larvae was higher than those in the 1st and 4th instar larvae. Many important functions of the intestinal microbiome were predicted, for example, fermentation and chemoheterotrophy functions that may play an important role in insect growth and development was detected in the bacteria at all tested stages. However, some specific functions are found to be associated with different development stages. Our study provides a theoretical basis for investigating the function of the intestinal symbiosis bacteria of A. glabripennies.     

Molecular evolution and functional specialization of chalcone synthase superfamily from Phalaenopsis Orchid

Plant genomes appear to exploit the process of gene duplication as a primary means of acquiring biochemical and developmental flexibility. The best example is the gene encoding chalcone synthase (CHS, EC2.3.1.74), the first committed step in flavonoid biosynthesis. In this study, we examined the molecular evolution of three CHS family members of Phalaenopsis including a novel chs gene (phchs5), which is slowly evolved. The inferred phylogeny of the chs genes of Phalaenopsis with other two orchid plants, Bromoheadia finlaysoniana and Dendrobium hybrid, suggested that gene duplication and divergence have occurred before divergence of these three genera. Relatively quantitative RT-PCR analysis identified expression patterns of these three chs genes in different floral tissues at different developmental stages. Phchs5 was the most abundantly expressed chs gene in floral organs and it was specifically transcribed in petal and lip at the stages when anthocyanin accumulated (stage1–4). Phchs3 and phchs4 were expressed at much lower levels than phchs5. Phchs3 was expressed in pigmented tissue (including lip, petal and sepal) at middle stages (stages 2–4) and in colorless reproductive tissue at late stage (stage 5). Phchs4 was only expressed in petal at earlier stages (stage 1–3) and in lip at middle stage (stage 4). These results present new data on differentiation of gene expression among duplicate copies of chs genes in Phalaenopsis.     

Methylation patterns revealed by MSAP profiling in genetically stable somatic embryogenic cultures of <Emphasis Type="Italic">Ocotea catharinensis</Emphasis> (Lauraceae)

Ocotea catharinensis is a rare tree species indigenous to the Atlantic rainforest of South America. In spite of its value as a hardwood species, it is in danger of extinction. The species erratically produces seeds showing irregular flowering and slow growth. Therefore, plants are not easily replaced. Tissue culture-based techniques are commonly used for obtaining living material for tree propagation and in vitro preservation. Therefore, a high-frequency somatic embryogenic system was developed for the species. In the present work, the genetic fidelity of cell aggregates and somatic embryos at various stages of in vitro development of O. catharinensis was investigated using RAPD and AFLP markers. Both analyses confirmed the absence of genetic variation in all developmental stages of O. catharinensis embryogenic cultures, verifying that the in vitro system is genetically stable. The cultures were also analyzed for their methylation profiles at 5′-CCGG-3′ sites by identifying methylation-sensitive amplification polymorphisms. Some of these markers differentiated cell aggregates from embryo bodies. The sequencing of ten MSAP markers revealed that four sequences showed significant similarity to genes encoding plant proteins. Particularly, the predicted amino acid sequence of the fragment designated as OcEaggHMttc155 was similar to the enzyme 1–aminocyclopropane-1-carboxylate oxidase (ACO), which is involved in the biosynthesis of ethylene, and its expression was reported to occur from the beginning to the intermediate stages of plant embryo development. Here, we suggest that this enzyme is possibly involved in the control of the earliest stages of somatic embryogenesis of O. catharinensis, and an approach to study ACO expression during somatic embryogenesis is proposed.     

Midgut bacterial communities in the giant Asian honeybee (Apis dorsata) across 4 developmental stages: A comparative study

Bacterial communities are known to play important roles during the developmental stages of insects, but current knowledge of bacteria associated with the midgut of Apis dorsata, the giant Asian honeybee, is limited. Using polymerase chain reaction‐denaturing gradient gel electrophoresis analysis (PCR‐DGGE) and 16S rRNA sequencing, the aim of this study was to determine the dynamics of bacterial community structure across four A. dorsata life stages in different geographical locations. The results reveal that bacterial diversity increased as the bee progressed through larval stage to newly emerged worker and old worker. However, in the pupal stage, no bands identified as bacteria could be observed. Overall, 2 bacterial phyla (Proteobacteria and Firmicutes) and 4 classes (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, and Bacilli) were identified, but the frequency varied among the different stages and locations. The classes of Gammaproteobacteria and Bacilli dominated among larval, newly emerged worker and old worker developmental stages.     

Humerus development in moles (Talpidae,Mammalia)

The humerus of fossorial moles has a highly derived anatomy, reflecting the ecological specialization of these animals for digging. It is short and broad, with enlarged muscle attachment sites and pronounced articulations compared to non‐fossorial sister taxa and other mammals. Both condyles are rotated in opposite directions, resulting in a torsion which is unique among eutherian mammals. The development of this exceptional bone was studied in embryonic stages of the fossorial Iberian mole (Talpa occidentalis) from mesenchymal condensation to incipient ossification based on histological serial sections using 3D reconstruction methods. For comparison, embryonic stages of the semi‐fossorial Japanese shrew mole (Urotrichus talpoides) as well as a sister taxon of moles, the terrestrial North American least shrew (Cryptotis parva), were studied. Results show that the humerus of Talpa already shows its derived anatomy with broadened muscle attachment sites and distinct articulations at early cartilaginous stages, when ossification has just started in the mid‐diaphyseal region. The torsion takes place simultaneously with the medial rotation of the forelimbs. The supracondylar foramen is closed in all studied Talpa embryos, but patent in Cryptotis and Urotrichus. This is an example of developmental penetrance, suggesting that variation of adult elements can be found at early stages as well.     

Early Developmental Stages of the Green Tiger Prawn, Penaeus semisulcatus de Haan (Crustacea, Decapoda, Penaeidae)

Gravid females of Penaeus semisulcatus were spawned in the laboratory by natural means. The embryos were documented and the larvae were reared from hatching to postlarval stage at 28.2–30.0 °C and 33.5–34.5 g kg⁻¹ salinity for about 10 days (223 h 55 min). Six naupliar stages, three protozoea stages, three mysis stages and the first postlarval stage were described and illustrated. The larvae were fed only with microalgae Tetraselmis tetrathele and Chaetoceros gracilis from first protozoea until the second mysis, with about 90% survival rate; from the third mysis until the first postlarva they were fed with similar microalgae coupled with rotifer Brachionus plicatilis and Artemia nauplii. The embryonic and larval stages of P. semisulcatus are generally similar to those of other closely related species in the family Penaeidae, such as Melicertus canaliculatus, Fenneropenaeus merguiensis, and Marsupenaeus japonicus, except for the size and structure of diagnostic characters, setation of appendages and duration of metamorphoses. The change in the feeding habit during ontogeny was related to morphological transformation of the feeding apparatus of larvae and postlarvae. This paper is the first comprehensive and complete account of the early developmental stages of P. semisulcatus.     

The shape and distribution of lysosomes and endocytosis in the ciliary epithelial cells of rats

Stages of the spermatogenic cycle in human seminiferous tubules were evaluated in men with varied efficiencies of spermatogenesis to determine if the architectural arrangement of stages or the atypical cell types contributed to variation in sperm production rates. Testes were selected from men with low, intermediate, and high daily sperm production per g parenchyma (DSP/g). Round tubular cross sections were photographed by bright-field microscopy. Stages were identified for each cross section by two observers and the number of stages represented in each cross section was recorded. Number of stages per cross section in men with low efficiency of spermatogenesis were significantly (P<0.05) fewer than men with intermediate and high efficiency of spermatogenesis. Further, the percentage of stages with atypical cell types in men with high DSP/g was significantly (P<0.05) higher than men with low DSP/g. There was a significant relationship (P<0.01) between the percentages of stages with atypical cell types per stage and number of stages per cross section. The atypical cell types appear to result from high density of stages per cross section in men with high DSP/g. There was no significant difference observed between groups for tubular volume, diameter, length, volume density, and volume density of seminiferous epithelium. However, a significant (P<0.05) positive correlation between percent seminiferous epithelium per testis with DSP/g or with the number of stages per cross section was found. These findings reveal that the architectural makeup of stages within seminiferous tubules and atypical cell types within stages varies with the level of efficiency of spermatogenesis, and this variation may reflect differences in yield of early spermatogonial divisions that are responsible for generating the different stages.