scholarly journals Proof of principle for piggyBac-mediated transgenesis in the flatworm Macrostomum lignano

Genetics ◽  
2021 ◽  
Author(s):  
Kirill Ustyantsev ◽  
Jakub Wudarski ◽  
Igor Sukhikh ◽  
Filipa Reinoite ◽  
Stijn Mouton ◽  
...  
Keyword(s):  
Insertional Mutagenesis ◽  
Single Copy ◽  
Free Living ◽  
Experimental Platform ◽  
Stem Cell Regulation ◽  
Alternative Approach ◽  
Macrostomum Lignano ◽  
Integration Sites ◽  
Transgenic Lines ◽  
First Time

Abstract Regeneration-capable flatworms are informative research models to study the mechanisms of stem cell regulation, regeneration, and tissue patterning. The free-living flatworm Macrostomum lignano is currently the only flatworm where stable transgenesis is available, and as such it offers a powerful experimental platform to address questions that were previously difficult to answer. The published transgenesis approach relies on random integration of DNA constructs into the genome. Despite its efficiency, there is room and need for further improvement and diversification of transgenesis methods in M. lignano. Transposon-mediated transgenesis is an alternative approach, enabling easy mapping of the integration sites and the possibility of insertional mutagenesis studies. Here, we report for the first time that transposon-mediated transgenesis using piggyBac can be performed in M. lignano to create stable transgenic lines with single-copy transgene insertions.

scholarly journals A platform for efficient transgenesis in Macrostomum lignano, a flatworm model organism for stem cell research

2017 ◽  
Author(s):  
Jakub Wudarski ◽  
Daniil Simanov ◽  
Kirill Ustyantsev ◽  
Katrien de Mulder ◽  
Margriet Grelling ◽  
...  
Keyword(s):  
Stem Cell ◽  
Fluorescent Proteins ◽  
Model Organism ◽  
Promoter Regions ◽  
Cell Stage ◽  
Genetic Studies ◽  
Stem Cell Regulation ◽  
Genome Wide ◽  
Macrostomum Lignano ◽  
Transgenic Lines

ABSTRACTRegeneration-capable flatworms are informative research models to study the mechanisms of stem cell regulation, regeneration and tissue patterning. However, the lack of transgenesis methods significantly hampers their wider use. Here we report development of a transgenesis method for Macrostomum lignano, a basal flatworm with excellent regeneration capacity. We demonstrate that microinjection of DNA constructs into fertilized one-cell stage eggs, followed by a low dose of irradiation, frequently results in random integration of the transgene in the genome and its stable transmission through the germline. To facilitate selection of promoter regions for transgenic reporters, we assembled and annotated the M. lignano genome, including genome-wide mapping of transcription start regions, and showed its utility by generating multiple stable transgenic lines expressing fluorescent proteins under several tissue-specific promoters. The reported transgenesis method and annotated genome sequence will permit sophisticated genetic studies on stem cells and regeneration using M. lignano as a model organism.

scholarly journals Genome Assembly of the Canadian Two-row Malting Barley Cultivar AAC Synergy

2021 ◽  
Author(s):  
Wayne Xu ◽  
James R Tucker ◽  
Wubishet A Bekele ◽  
Frank M You ◽  
Yong-Bi Fu ◽  
...  
Keyword(s):  
Reference Genome ◽  
Single Copy ◽  
Barley Cultivar ◽  
Malting Barley ◽  
Orthologous Genes ◽  
Hordeum Vulgare L ◽  
Chromosome Conformation ◽  
Mate Pair ◽  
Genome Assemblies ◽  
First Time

Abstract Barley (Hordeum vulgare L.) is one of the most important global crops. The six-row barley cultivar Morex reference genome has been used by the barley research community worldwide. However, this reference genome can have limitations when used for genomic and genetic diversity analysis studies, gene discovery, and marker development when working in two-row germplasm that is more common to Canadian barley. Here we assembled, for the first time, the genome sequence of a Canadian two-row malting barley, cultivar AAC Synergy. We applied deep Illumina paired-end reads, long mate-pair reads, PacBio sequences, 10X chromium linked read libraries, and chromosome conformation capture sequencing (Hi-C) to generate a contiguous assembly. The genome assembled from super-scaffolds had a size of 4.85 Gb, N50 of 2.32 Mb and an estimated 93.9% of complete genes from a plant database (BUSCO, benchmarking universal single-copy orthologous genes). After removal of small scaffolds (< 300 Kb), the assembly was arranged into pseudomolecules of 4.14 Gb in size with seven chromosomes plus unanchored scaffolds. The completeness and annotation of the assembly were assessed by comparing it with the updated version of six-row Morex and recently released two-row Golden Promise genome assemblies.

scholarly journals First Report of Echinococcus ortleppi in Free-Living Wild Boar (Sus scrofa) from Portugal

2021 ◽  
Vol 9 (6) ◽  
pp. 1256
Author(s):  
Teresa Letra Mateus ◽  
Maria João Gargaté ◽  
Anabela Vilares ◽  
Idalina Ferreira ◽  
Manuela Rodrigues ◽  
...  
Keyword(s):  
Wild Boar ◽  
Internal Transcribed Spacer ◽  
Sus Scrofa ◽  
One Health ◽  
Control Strategies ◽  
Free Living ◽  
First Report ◽  
Wildlife Reservoirs ◽  
First Time ◽  
Echinococcus Granulosus Sensu Lato

Cystic echinococcosis (CE) is a zoonosis that is prevalent worldwide. It is considered endemic in Portugal but few studies have been performed on Echinococcus granulosus sensu lato and their hosts. In this study, CE cysts are reported for the first time in a free-living wild boar (Sus scrofa) in Portugal. The presence of the metacestodes in the liver of the wild boar was identified by morphological features, microscopic examination and molecular analysis. The sequencing of part of the DNA nuclear ribosomal internal transcribed spacer-1 (ITS-1) region revealed a G5 genotype that presently corresponds to Echinococcus ortleppi. This is the first report of E. ortleppi in Portugal and to the best of the authors’ knowledge, in Europe. These results suggest that wild boar may be a host of CE, namely, crossing the livestock–wildlife interface, which has important public health implications. Wildlife reservoirs must be taken into account as CE hosts and surveillance of game as well as health education for hunters should be implemented using a One Health approach, with implementation of feasible and tailor-made control strategies, namely, proper elimination of byproducts in the field.

Arthropods in the nests of lesser spotted eagle (Aquila pomarina)

Biologia ◽  
2009 ◽  
Vol 64 (5) ◽  
Author(s):  
Ján Krištofík ◽  
Peter Mašán ◽  
Zbyšek Šustek ◽  
Dušan Karaska
Keyword(s):  
Trophic Group ◽  
Trophic Groups ◽  
Significant Component ◽  
Free Living ◽  
Parasitic Species ◽  
Number Of Species ◽  
Aquila Pomarina ◽  
Investigation Period ◽  
First Time

AbstractIn 2001–2007, altogether 57 nests of lesser spotted eagle were collected in the Orava region in northwestern Slovakia and four groups of arthropods were extracted from them. Richest in number of species and individuals were mites (23 species, 17,500 ind.), followed by beetles (12 species, 725 ind.), whereas pseudoscorpions were represented only by Pselaphochernes scorpioides (39 ind.) and fleas by Ceratophyllus garei (3 ind.). Unlike nests of other birds, free-living mites predominated in the nests fauna (83% of individuals), followed by nidicolous species with more or less free relationship to the nests, while parasitic species represented only a negligible part of the fauna. For the first time we observed phoresy of Nenteria pandioni, a specific and abundant mite in the eagles’ nests, on the nidicolous staphylinid Haploglossa puncticollis. The beetle fauna in the nests was much poorer than in nests of other birds. The predatory H. puncticollis was dominant in the nests (83%) and occurred continuously during the whole investigation period. Other beetles, even the widely distributed nidicols such as the histerid Gnathoncus buyssoni, were found rarely in nests. Predators were also the only abundant trophic group of beetles in the nests, while other trophic groups of beetles abundantly co-occur in nests of majority of other birds. The occurrence of all beetles was very unevenly distributed during the investigation period, but was positively correlated with occurrence of mites. The relatively low number of species and individuals of mites and beetles in the lesser spotted eagle nests resulted from their position on tree tops, at a height of 20–30 m and their quick drying out by sun and wind. It was also indicated by an enormously low number of species and individuals of mycetophagous beetles, which represent a significant component of the fauna in nests of other birds.

Ten New Records of Free-living Nematodes from Uttarakhand, India

2018 ◽  
Vol 41 (1) ◽  
pp. 39-42
Author(s):  
Vinita Sharma ◽  
Keyword(s):  
Medicinal Plants ◽  
New Records ◽  
Nematode Species ◽  
Forest Trees ◽  
Wildlife Sanctuary ◽  
Free Living ◽  
First Time

Ten nematode species of order Dorylaimida were recorded for the first time from Uttarakhand during survey of terrestrial nematodes associated with forest trees and medicinal plants in Govind Wildlife Sanctuary, Uttarkashi, Uttarakhand, India.

A new and two known species of Hatschekia Poche, 1902 (Copepoda, Siphonostomatoida, Hatschekiidae) parasitic on Japanese fishes

Crustaceana ◽  
2018 ◽  
Vol 91 (12) ◽  
pp. 1483-1493
Author(s):  
Kunihiko Izawa
Keyword(s):  
Free Living ◽  
First Time ◽  
Branchiostegus Japonicus

Abstract Hatschekia laeopsi sp. nov., H. branchiostegi Yamaguti, 1939 and H. pagrosomi Yamaguti, 1939 are (re-)described based on specimens collected from the branchial lamellae of three Japanese fishes, Laeops kitaharae (Smith & Pope, 1906) (Pleuronectidae), Branchiostegus japonicus (Houttuyn, 1782) (Malacanthidae), and Dentex tumifrons (Temminck & Schlegel, 1843) (Sparidae), respectively. The free-living stages of H. branchiostegi are described for the species for the first time.

Agrobacterium-mediated insertional mutagenesis of the ochratoxigenic fungus Aspergillus westerdijkiae

2007 ◽  
Vol 53 (1) ◽  
pp. 148-151 ◽  
Author(s):  
Marcia M Mata ◽  
Marta H Taniwaki ◽  
Beatriz T Iamanaka ◽  
Daniele Sartori ◽  
André L.M Oliveira ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Insertional Mutagenesis ◽  
Aspergillus Ochraceus ◽  
Agrobacterium Mediated Transformation ◽  
Morphological Variations ◽  
Carcinogenic Potential ◽  
Coffee Beans ◽  
Aspergillus Westerdijkiae ◽  
Polymerase Chain ◽  
First Time

Aspergillus westerdijkiae is a potent ochratoxin A (OTA) producer that has been found in coffee beans. OTA is known to have nephrotoxic effects and carcinogenic potential in animal species. Here we report for the first time the Agrobacterium-mediated transformation for Aspergillus westerdijkiae and the generation of ochratoxin-defective mutants. Conidia were transformed to hygromycin B resistance using strain AGL-1 of Agrobacterium tumefaciens. The obtained transformation frequency was up to 47 transformants per 106 target conidia. Among 600 transformants, approximately 5% showed morphological variations. Eight transformants with consistently reduced OTA production were obtained. Two of these transformants did not produce OTA (detection limit: 0.1 µg/kg); the other six mutants produced lower amounts of OTA (1%–32%) compared with the wild-type strain. By using thermal asymmetric interlaced polymerase chain reaction, we successfully identified a putative flavin adenine dinucleotide monooxygenase gene.Key words: Aspergillus ochraceus, Aspergillus westerdijkiae, Agrobacterium-mediated transformation, Agrobacterium-mediated insertional mutagenesis, ochratoxin A.

A new pathogen spillover from domestic to wild animals: Plasmodium juxtanucleare infects free-living passerines in Brazil

Parasitology ◽  
2018 ◽  
Vol 145 (14) ◽  
pp. 1949-1958 ◽  
Author(s):  
Francisco C. Ferreira-Junior ◽  
Daniela de Angeli Dutra ◽  
Patrícia Silveira ◽  
Richard Campos Pacheco ◽  
Rute Witter ◽  
...  
Keyword(s):  
Complete Mitochondrial Genome ◽  
Wild Birds ◽  
Habitat Modification ◽  
Wild Animals ◽  
Conservation Units ◽  
Free Living ◽  
Central Brazil ◽  
Blood Smears ◽  
Backyard Chickens ◽  
First Time

AbstractHabitat modification may facilitate the emergence of novel pathogens, and the expansion of agricultural frontiers make domestic animals important sources of pathogen spillover to wild animals. We demonstrate for the first time that Plasmodium juxtanucleare, a widespread parasite from domestic chickens, naturally infects free-living passerines. We sampled 68 wild birds within and at the border of conservation units in central Brazil composed by Cerrado, a highly threatened biome. Seven out of 10 passerines captured in the limits of a protected area with a small farm were infected by P. juxtanucleare as was confirmed by sequencing a fragment of the parasite's cytochrome b. Blood smears from these positive passerines presented trophozoites, meronts and gametocytes compatible with P. juxtanucleare, meaning these birds are competent hosts for this parasite. After these intriguing results, we sampled 30 backyard chickens managed at the area where P. juxtanucleare-infected passerines were captured, revealing one chicken infected by the same parasite lineage. We sequenced the almost complete mitochondrial genome from all positive passerines, revealing that Brazilian and Asian parasites are closely related. P. juxtanucleare can be lethal to non-domestic hosts under captive and rehabilitation conditions, suggesting that this novel spillover may pose a real threat to wild birds.

Transgenic Rice Expressing Isoflavone Synthase Gene from Soybean Shows Resistance against Blast Fungus (Magnaporthe oryzae)

Plant Disease ◽  
2021 ◽  
Author(s):  
Suresh Pokhrel ◽  
Sathish K Ponniah ◽  
Yulin Jia ◽  
Oliver Yu ◽  
Muthusamy Manoharan
Keyword(s):  
Disease Resistance ◽  
Transgenic Rice ◽  
Magnaporthe Oryzae ◽  
Plant Disease Resistance ◽  
Rice Blast ◽  
Plant Disease ◽  
Plant Secondary Metabolites ◽  
Single Copy ◽  
Transgenic Lines ◽  
Isoflavone Synthase

The isoflavones are a group of plant secondary metabolites primarily synthesized in legumes and are known for their role in improving human health and plant disease resistance. The isoflavones, especially genistein, act as precursors for the production of phytoalexins, which may induce broad-spectrum disease resistance in plants. In the present study, we screened transgenic rice lines expressing the isoflavone synthase (GmIFS1) gene from soybean for rice blast (Magnaporthe oryzae) resistance. Two homozygous transgenic lines (I2 and I10), based on single copy gene integration, were identified. The expression of GmIFS1 in transgenic lines was confirmed by qRT-PCR. Genistein was detected in the transgenic lines using LC-MS/MS. Subsequently, the transgenic lines were evaluated against the rice blast pathogen, isolate YJ54 (race IB-54). The results indicated that more than 60% of the plants in both the lines (I2 and I10) showed resistance against the blast pathogen. The progenies of one of the resistant transgenic lines (I10) also showed more than 65% resistance against rice blast. The resistance of these transgenic lines against rice blast may be attributed to the synthesis of isoflavone (genistein) in rice.

scholarly journals Expression of thaumatin, a new type of alternative sweetener in rice

2020 ◽  
Vol 48 (3) ◽  
pp. 1276-1291
Author(s):  
Shahina AKTER ◽  
Md. Amdadul HUQ ◽  
Yu-Jin JUNG ◽  
Kwon-Kyoo KANG
Keyword(s):  
Transgenic Rice ◽  
Oryza Sativa L ◽  
Plasmid Vector ◽  
Single Copy ◽  
Cauliflower Mosaic Virus ◽  
Pcr Analysis ◽  
35S Promoter ◽  
Alternative Sweetener ◽  
Sweet Proteins ◽  
Transgenic Lines

  Sweet proteins are the natural alternative to the artificial sweeteners as well as flavor enhancers. Among other sweet protein, thaumatin protein was isolated from Thaumatococcus daniellii Benth plant fruit. In this study, pinII Ti plasmid vector was constructed with thaumatin gene, where thaumatin was placed under the control of the duel cauliflower mosaic virus 35S promoter into rice (Oryza sativa L. var. japonica cv. ‘Dongjinbyeo’) by Agrobacterium-mediated transformation to generate transgenic plants. Thirteen plant lines were regenerated and the transgenic rice lines were confirmed by different molecular analysis. The genomic PCR result revealed that all of the plant lines were transgenic. The single copy and intergenic plant lines were selected by Taqman PCR analysis and FST analysis, respectively. Expression of thaumatin gene in transgenic rice resulted in the accumulation of thaumatin protein in the leave. Thaumatin protein was also accumulated in leave of T1 generation. Sensory analysis result suggested that the thaumatin protein expressing transgenic lines exerted sweet tasting activity. These results demonstrated that thaumatin was expressed in transgenic rice plants.

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