Characterization of the symbiontRickettsiain the mirid bugNesidiocoris tenuis(Reuter) (Heteroptera: Miridae)

2014 ◽  
Vol 104 (6) ◽  
pp. 681-688 ◽  
Author(s):  
A. Caspi-Fluger ◽  
M. Inbar ◽  
S. Steinberg ◽  
Y. Friedmann ◽  
M. Freund ◽  
...  
Keyword(s):  
Biological Control ◽  
Developmental Stages ◽  
Bacterial Symbionts ◽  
Nesidiocoris Tenuis ◽  
Augmentative Release ◽  
Polymerase Chain ◽  
Bacterial Genes ◽  
Rickettsia Bellii

AbstractNesidiocoris tenuis(Reuter) (Heteroptera: Miridae) is an omnivorous insect used for biological control. Augmentative release and conservation ofN. tenuishave been used for pest control in tomato crops. Intracellular bacterial symbionts of arthropods are common in nature and have diverse effects on their hosts; in some cases they can dramatically affect biological control. Fingerprinting methods showed that the symbiotic complex associated withN. tenuisincludesWolbachiaandRickettsia. RickettsiaofN. tenuiswas further characterized by sequencing the16S rRNAandgltAbacterial genes, measuring its amount in different developmental stages of the insect by real-time polymerase chain reaction, and localizing the bacteria in the insect's body by fluorescencein situhybridization. TheRickettsiainN. tenuisexhibited 99 and 96% similarity of both sequenced genes toRickettsia belliiandRickettsiareported fromBemisia tabaci, respectively. The highest amount ofRickettsiawas measured in the 5th instar and adult, and the symbionts could be detected in the host gut and ovaries. Although the role played byRickettsiain the biology ofN. tenuisis currently unknown, their high amount in the adults and localization in the gut suggest that they may have a nutritional role in this insect.

scholarly journals Molecular Characterization of hobo-Mediated Inversions in Drosophila melanogaster

Genetics ◽  
1996 ◽  
Vol 144 (2) ◽  
pp. 647-656
Author(s):  
William B Eggleston ◽  
Nac R Rim ◽  
Johng K Lim
Keyword(s):  
X Chromosome ◽  
Polymerase Chain Reaction Amplification ◽  
Polytene Chromosomes ◽  
Chromosomal Inversions ◽  
Hobo Element ◽  
Reaction Amplification ◽  
Notch Locus ◽  
Polymerase Chain

Abstract The structure of chromosomal inversions mediated by hobo transposable elements in the Uc-1 X chromosome was investigated using cytogenetic and molecular methods. Uc-1 contains a phenotypically silent hobo element inserted in an intron of the Notch locus. Cytological screening identified six independent Notch mutations resulting from chromosomal inversions with one breakpoint at cytological position 3C7, the location of Notch. In situ hybridization to salivary gland polytene chromosomes determined that both ends of each inversion contained hobo and Notch sequences. Southern blot analyses showed that both breakpoints in each inversion had hobo-Notch junction fragments indistinguishable in structure from those present in the Uc-1 X chromosome prior to the rearrangements. Polymerase chain reaction amplification of the 12 hobo-Notch junction fragments in the six inversions, followed by DNA sequence analysis, determined that each was identical to one of the two hobo-Notch junctions present in Uc-1. These results are consistent with a model in which hobo-mediated inversions result from homologous pairing and recombination between a pair of hobo elements in reverse orientation.

Differential expression sites of TGF-β isoforms in chicken limb buds during morphogenesis

2005 ◽  
Vol 83 (4) ◽  
pp. 620-625 ◽  
Author(s):  
Shinya Aramaki ◽  
Fuminori Sato ◽  
Tomoki Soh ◽  
Nobuhiko Yamauchi ◽  
Masa-aki Hattori
Keyword(s):  
Limb Development ◽  
Developmental Stages ◽  
Polymerase Chain Reaction Analysis ◽  
Weak Signal ◽  
Temporal Expression ◽  
Chain Reaction ◽  
White Leghorn Chicken ◽  
Whole Mount ◽  
Polymerase Chain

TGF-β gene is expressed at various developmental stages and its principle role may be an involvement in organogenesis. The present study was performed to investigate the temporal expression of these TGF-β isoforms in the developing limb of White Leghorn Chicken, Gallus gallus (L., 1758). TGF-β isoforms were expressed in the developing limb as revealed by whole-mount in situ hybridization, but each showed a different pattern of expression. TGF-β2 was the dominant isoform compared with the other two isoforms. TGF-β2 first appeared along the proximodistal axis of the limb at stage 24 and condensed at the tip at stage 26. At stages 29–31, expression appeared in digits and then was extended to the interdigital spaces. A weak signal for TGF-β3 was first shown in the developing limb at stage 26, but there was no interdigital expression, unlike for TGF-β2. TGF-β4 was expressed in the developing limb at stage 26 and only in the interdigital spaces at stage 29. Reverse transcription – polymerase chain reaction analysis also showed that the transcript levels of TGF-β isoforms, especially TGF-β2, drastically increased at stage 29. These results suggest that TGF-β isoforms, with their patterns of expression, are specific regulatory factors that participate in limb development and digit morphogenesis.

scholarly journals Identification and Characterization of Four Autophagy-Related Genes That Are Expressed in Response to Hypoxia in the Brain of the Oriental River Prawn (Macrobrachium nipponense)

2019 ◽  
Vol 20 (8) ◽  
pp. 1856 ◽  
Author(s):  
Shengming Sun ◽  
Ying Wu ◽  
Hongtuo Fu ◽  
Xianping Ge ◽  
Hongzheng You ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Open Reading Frames ◽  
Dependent Manner ◽  
Macrobrachium Nipponense ◽  
Oriental River Prawn ◽  
Adverse Environmental Conditions ◽  
The Brain ◽  
Identification And Characterization

Autophagy is a cytoprotective mechanism triggered in response to adverse environmental conditions. Herein, we investigated the autophagy process in the oriental river prawn (Macrobrachium nipponense) following hypoxia. Full-length cDNAs encoding autophagy-related genes (ATGs) ATG3, ATG4B, ATG5, and ATG9A were cloned, and transcription following hypoxia was explored in different tissues and developmental stages. The ATG3, ATG4B, ATG5, and ATG9A cDNAs include open reading frames encoding proteins of 319, 264, 268, and 828 amino acids, respectively. The four M. nipponense proteins clustered separately from vertebrate homologs in phylogenetic analysis. All four mRNAs were expressed in various tissues, with highest levels in brain and hepatopancreas. Hypoxia up-regulated all four mRNAs in a time-dependent manner. Thus, these genes may contribute to autophagy-based responses against hypoxia in M. nipponense. Biochemical analysis revealed that hypoxia stimulated anaerobic metabolism in the brain tissue. Furthermore, in situ hybridization experiments revealed that ATG4B was mainly expressed in the secretory and astrocyte cells of the brain. Silencing of ATG4B down-regulated ATG8 and decreased cell viability in juvenile prawn brains following hypoxia. Thus, autophagy is an adaptive response protecting against hypoxia in M. nipponense and possibly other crustaceans. Recombinant MnATG4B could interact with recombinant MnATG8, but the GST protein could not bind to MnATG8. These findings provide us with a better understanding of the fundamental mechanisms of autophagy in prawns.

The Ikaros gene, a central regulator of lymphoid differentiation, fuses to the BCL6 gene as a result of t(3;7)(q27;p12) translocation in a patient with diffuse large B-cell lymphoma

Blood ◽  
2000 ◽  
Vol 95 (8) ◽  
pp. 2719-2721 ◽  
Author(s):  
Yoshitaka Hosokawa ◽  
Yumiko Maeda ◽  
Ryo Ichinohasama ◽  
Ikuo Miura ◽  
Masafumi Taniwaki ◽  
...  
Keyword(s):  
B Cell ◽  
Molecular Genetic ◽  
Regulatory Region ◽  
B Cell Lymphoma ◽  
Molecular Genetic Analysis ◽  
B Cell Differentiation ◽  
Polymerase Chain ◽  
Large B Cell

The BCL6 gene, isolated from the breakpoints of 3q27-associated chromosomal translocations, has been implicated in diffuse large B-cell lymphomas (DLBL). Here we describe the molecular characterization of novel t(3;7)(q27;p12) translocations in 2 patients with DLBL. Molecular genetic analysis of the breakpoint area involving BCL6 revealed the presence of the Ikaros gene, a central regulator of lymphoid differentiation that had been mapped to human chromosome 7 band p13-p11.1. As a molecular consequence of the translocation, the 5′ regulatory region of the BCL6 gene was replaced by the putative 5′ regulatory region of theIkaros gene, probably leading to deregulated expression of theBCL6 gene throughout B-cell differentiation. Reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH) analyses of a patient sample established that the t(3;7)(q27;p12) results in fusion of the Ikaros andBCL6 genes. This study provides the first evidence that the Ikaros gene is rearranged in human hematopoietic malignant disorders.

scholarly journals Candidates for Symbiotic Control of Sugarcane White Leaf Disease

2012 ◽  
Vol 78 (19) ◽  
pp. 6804-6811 ◽  
Author(s):  
Jureemart Wangkeeree ◽  
Thomas A. Miller ◽  
Yupa Hanboonsong
Keyword(s):  
Developmental Stages ◽  
Cloning And Sequencing ◽  
Content Type ◽  
Leaf Disease ◽  
White Leaf ◽  
Symbiotic Control ◽  
Sugarcane White Leaf ◽  
White Leaf Disease ◽  
Bacterial Genes

ABSTRACTThe leafhopperMatsumuratettix hiroglyphicus(Matsumura) is the most important vector of a phytoplasma pathogen causing sugarcane white leaf (SCWL) disease. The purpose of this study was to evaluate candidate bacterial symbionts for possible use as vehicles in the control of the disease. 16S rRNA bacterial genes were amplified from whole bodies ofM. hiroglyphicusleafhoppers and analyzed by cloning and sequencing. Two dominant groups were found: one belonged to theBetaproteobacteriathat did not closely match any sequences in the database and was named bacterium associated withM. hiroglyphicus(BAMH). Another one found to be abundant in this leafhopper is “CandidatusSulcia muelleri” in the orderBacteroidetes, which was previously reported in the insect members of the Auchenorrhyncha. MostM. hiroglyphicusleafhoppers carry both BAMH and “Ca. Sulcia muelleri.” Fluorescentin situhybridization showed that BAMH and “Ca. Sulcia muelleri” colocalized in the same bacteriomes. BAMH was present in the midgut and ovaries of the leafhopper and was found in all developmental stages, including eggs, nymphs, and adults. Because BAMH appears to be specific for the SCWL vector, we evaluated it as a candidate for symbiotic control of sugarcane white leaf disease.

Characterization of a microsatellite locus in the parasitoid wasp Aphelinus abdominalis (Hymenoptera: Aphelinidae)

1997 ◽  
Vol 87 (3) ◽  
pp. 313-318 ◽  
Author(s):  
F. Vanlerberghe-Masutti ◽  
P. Chavigny
Keyword(s):  
Microsatellite Locus ◽  
Genomic Library ◽  
Parasitoid Wasp ◽  
Dna Amplification ◽  
Chain Reaction ◽  
Aphid Parasitoid ◽  
Augmentative Release ◽  
Polymerase Chain ◽  
Partial Genomic Library

AbstractPrimers for DNA amplification using the polymerase chain reaction (PCR) were synthesized for a microsatellite locus isolated from a partial genomic library of the aphid parasitoid Aphelinus abdominalis (Dalman). Screening for genetic polymorphism at this locus in two laboratory strains of this wasp revealed the presence of two alleles different in the number of (GT) and (GGC) repeats. The relative frequencies of the two alleles were not significantly different between the two strains or between diploid females and haploid males. Heterozygosity at this microsatellite locus was estimated to be 0.40 which is within the range in other hymenopterous species. Given that A. abdominalis is a good candidate for augmentative release programmes in greenhouses against aphids, we suggest that microsatellite markers may have application in discriminating among aphelinid sibling species and strains. The markers provide a means for studying the performance and impact of selected parasitoid lines on pest dynamics in field release experiments.

Ultrastructural and molecular characterization of endosymbionts of the reed beetle genusMacroplea(Chrysomelidae, Donaciinae), and proposal of “CandidatusMacropleicola appendiculatae” and “CandidatusMacropleicola muticae”

2009 ◽  
Vol 55 (11) ◽  
pp. 1250-1260 ◽  
Author(s):  
Gregor Kölsch ◽  
Corinna Matz-Grund ◽  
Bo V. Pedersen
Keyword(s):  
Phylogenetic Analysis ◽  
16S Rrna ◽  
Fluorescent In Situ Hybridization ◽  
Malpighian Tubules ◽  
Rrna Gene ◽  
Beetle Species ◽  
Bacterial Symbionts ◽  
The 16S Rrna Gene

Intracellular bacterial symbionts are known from various insect groups, particularly from those feeding on unbalanced diets, where the bacteria provide essential nutrients to the host. In the case of reed beetles (Coleoptera: Chrysomelidae, Donaciinae), however, the endosymbionts appear to be associated with specialized “glands” that secrete a material used for the beetles’ unusual water-tight cocoon. These glands were discovered over a century ago, but the bacteria they contain have yet to be characterized and placed in a phylogenetic context. Here, we describe the ultrastructure of two endosymbiotic species (“ Candidatus Macropleicola appendiculatae” and “ Candidatus Macropleicola muticae”) that reside in cells of the Malpighian tubules of the reed beetle species Macroplea appendiculata and Macroplea mutica , respectively. Fluorescent in situ hybridization using oligonucleotides targeting the 16S rRNA gene specific to Macroplea symbionts verified the localization of the symbionts in these organs. Phylogenetic analysis of 16S rRNA placed “Candidatus Macropleicola” in a clade of typically endosymbiotic Enterobacteriaceae (γ-proteobacteria). Finally, we discuss the evidence available for the hypothesis that the beetle larvae use a secretion produced by the bacteria for the formation of an underwater cocoon.

scholarly journals Cloning and Characterization of Two Anthocyanin Biosynthetic Genes from Dendrobium Orchid

2005 ◽  
Vol 130 (4) ◽  
pp. 611-618 ◽  
Author(s):  
Rasika G. Mudalige-Jayawickrama ◽  
Michele M. Champagne ◽  
A. David Hieber ◽  
Adelheid R. Kuehnle
Keyword(s):  
Developmental Stages ◽  
Color Difference ◽  
Cdna Clones ◽  
Rt Pcr ◽  
Flower Bud ◽  
Floral Tissue ◽  
Polymerase Chain ◽  
Possible Cause ◽  
Anthocyanin Biosynthetic Genes

Two full-length cDNA clones, Den-CHS-4 and Den-DFR-1, encoding chalcone synthase (CHS) and dihydroflavonol 4-reductase (DFR) were obtained from flower bud RNA of a lavender cyanidin-accumulating Dendrobium Sw. hybrid using reverse transcription-polymerase chain reaction (RT-PCR). Northern analyses indicated that both genes are expressed in all developmental stages of buds, with highest expression in the medium-sized buds. RT-PCR analyses showed that DFR expression was confined to floral tissue while CHS was expressed in floral and vegetative tissues but not in pseudobulbs. The nucleotide sequence of a DFR clone isolated from a pale orange pelargonidin-accumulating Dendrobium hybrid was exactly the same as Den-DFR-1, ruling out the substrate specificity of DFR as a possible cause of the color difference.

scholarly journals Characterization of Putative Effectors from the Cereal Cyst Nematode Heterodera avenae

2018 ◽  
Vol 108 (2) ◽  
pp. 264-274 ◽  
Author(s):  
Jiang-Kuan Cui ◽  
Huan Peng ◽  
Fen Qiao ◽  
Gao-Feng Wang ◽  
Wen-Kun Huang ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Target Genes ◽  
Developmental Stages ◽  
Heterodera Avenae ◽  
Parasitic Nematodes ◽  
Sequencing Analysis ◽  
Rnai Technology

Few molecular details of effectors of Heterodera avenae parasitism are known. We performed a high-throughput sequencing analysis of the H. avenae transcriptome at five developmental stages. A total of 82,549 unigenes were ultimately obtained, and 747 transcripts showed best hits to genes putatively encoding carbohydrate-active enzymes in plant-parasitic nematodes that play an important role in the invasion process. A total of 1,480 unigenes were homologous to known phytonematode effectors, and 63 putative novel effectors were identified in the H. avenae transcriptomes. Twenty-three unigenes were analyzed by qRT-PCR and confirmed to be highly expressed during at least one developmental stage. For in situ hybridization, 17 of the 22 tested putative effectors were specifically expressed and located in the subventral gland cells, and five putative novel effectors were specifically expressed in the dorsal gland. Furthermore, 115 transcripts were found to have putative lethal RNA interference (RNAi) phenotypes. Three target genes with lethal RNAi phenotypes and two of the four tested putative effectors were associated with a decrease in the number of cysts through in vitro RNAi technology. These transcriptomic data lay a foundation for further studies of interactions of H. avenae with cereal and H. avenae parasitic control.

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