17 research outputs found

    Karyotype Analysis of Diploid and Spontaneously Occurring Tetraploid Blood Orange [Citrus sinensis (L.) Osbeck] Using Multicolor FISH With Repetitive DNA Sequences as Probes

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    Blood orange [Citrus sinensis (L.) Osbeck] has been increasingly appreciated by consumers worldwide owing to its brilliant red color, abundant anthocyanin and other health-promoting compounds. However, there is still relatively little known about its cytogenetic characteristics, probably because of the small size and similar morphology of metaphase chromosomes and the paucity of chromosomal landmarks. In our previous study, a naturally occurring tetraploid blood orange plant was obtained via seedling screening. Before this tetraploid germplasm can be manipulated into a citrus triploid seedless breeding program, it is of great importance to determine its chromosome characterization and composition. In the present study, an integrated karyotype of blood orange was constructed using sequential multicolor fluorescence in situ hybridization (FISH) with four satellite repeats, two ribosomal DNAs (rDNAs), a centromere-like repeat and an oligonucleotide of telomere repeat (TTTAGGG)3 as probes. Satellite repeats were preferentially located at the terminal regions of the chromosomes of blood orange. Individual somatic chromosome pairs of blood orange were unambiguously identified by repetitive DNA-based multicolor FISH. These probes proved to be effective chromosomal landmarks. The karyotype was formulated as 2n = 2x = 18 = 16m+2sm (1sat) with the karyotype asymmetry degree belonging to 2B. The chromosomal distribution pattern of these repetitive DNAs in this spontaneously occurring tetraploid was identical to that of the diploid, but the tetraploid carried twice the number of hybridization sites as the diploid, indicating a possible pathway involving the spontaneous duplication of chromosome sets in nucellar cells. Our work may facilitate the molecular cytogenetic study of blood orange and provide chromosomal characterization for the future utilization of this tetraploid germplasm in the service of seedless breeding programs

    Molecular cytogenetic analysis of genome-specific repetitive elements in Citrus clementina Hort. Ex Tan. and its taxonomic implications

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    Abstract Background Clementine mandarin (Citrus clementina Hort. ex Tan.) is one of the most famous and widely grown citrus cultivars worldwide. Variations in relation to the composition and distribution of repetitive DNA sequences that dominate greatly in eukaryote genomes are considered to be species-, genome-, or even chromosome-specific. Repetitive DNA-based fluorescence in situ hybridization (FISH) is a powerful tool for molecular cytogenetic study. However, to date few studies have involved in the repetitive elements and cytogenetic karyotype of Clementine. Results A graph-based similarity sequence read clustering methodology was performed to analyze the repetitive DNA families in the Clementine genome. The bioinformatics analysis showed that repetitive DNAs constitute 41.95% of the Clementine genome, and the majority of repetitive elements are retrotransposons and satellite DNAs. Sequential multicolor FISH using a probe mix that contained CL17, four satellite DNAs, two rDNAs and an oligonucleotide of (TTTAGGG)3 was performed with Clementine somatic metaphase chromosomes. An integrated karyotype of Clementine was established based on unequivocal and reproducible chromosome discriminations. The distribution patterns of these probes in several Citrus, Poncirus and Fortunella species were summarized through extensive FISH analyses. Polymorphism and heterozygosity were commonly observed in the three genera. Some asymmetrical FISH loci in Clementine were in agreement with its hybrid origin. Conclusions The composition and abundance of repetitive elements in the Clementine genome were reanalyzed. Multicolor FISH-based karyotyping provided direct visual proof of the heterozygous nature of Clementine chromosomes with conspicuous asymmetrical FISH hybridization signals. We detected some similar and variable distribution patterns of repetitive DNAs in Citrus, Poncirus, and Fortunella, which revealed notable conservation among these genera, as well as obvious polymorphism and heterozygosity, indicating the potential utility of these repetitive element markers for the study of taxonomic, phylogenetic and evolutionary relationships in the future

    Biobased High-Performance Rotary Micromotors for Individually Reconfigurable Micromachine Arrays and Microfluidic Applications

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    In this work, we report an innovative type of rotary biomicromachines by using diatom frustules as integrated active components, including the assembling, operation, and performance characterization. We further investigate and demonstrate unique applications of the biomicromachines in achieving individually reconfigurable micromachine arrays and microfluidic mixing. Diatom frustules are porous cell walls of diatoms made of silica. We assembled rotary micromachines consisting of diatom frustules serving as rotors and patterned magnets serving as bearings in electric fields. Ordered arrays of micromotors can be integrated and rotated with controlled orientation and a speed up to ∼3000 rpm, one of the highest rotational speeds in biomaterial-based rotary micromachines. Moreover, by exploiting the distinct electromechanical properties of diatom frustules and metallic nanowires, we realized the first reconfigurable rotary micro/nanomachine arrays with controllability in individual motors. Finally, the diatom micromachines are successfully integrated in microfluidic channels and operated as mixers. This work demonstrated the high-performance rotary micromachines by using bioinspired diatom frustules and their applications, which are essential for low-cost bio-microelectromechanical system/nanoelectromechanical system (bio-MEMS/NEMS) devices and relevant to microfluidics

    Development of SARS-CoV-2 Isothermal Amplification Detection Kits

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    The SARS-CoV-2 isothermal amplification detection kits based on loop-mediated isothermal amplification (LAMP) were developed and evaluated on three types samples of SARS-CoV-2. The kits included enzyme reaction mixtures and chromogenic agents. After the isothermal amplification reactions were completed, the reaction results were judged by using the chromogenic agents to determine whether SARS-CoV-2 exists in the samples to be tested. The detection kits have the advantages of convenient operation, fast detection speed and high sensitivity up to 1 copy of virus particles per reaction, which can speed up the detection speed of suspected cases, and avoid the missing detection problems caused by the low detection sensitivity

    Designing a Novel Nano-Vaccine against SARS-CoV-2

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    The new coronavirus SARS-CoV-2 has become a global pandemic, which has had a huge impact on the lives of people around the world and has caused huge impacts and losses on global economic development. To now, there is still no effective drug or therapy against coronavirus. A large number of studies have shown that vaccines are the ultimate weapon to eliminate major infectious diseases. The development of new vaccines against new coronaviruses is the best way to prevent new coronavirus infections. In this study, we developed a new vaccine against the new coronavirus by combining our self-developed nano adjuvant loaded with carnosine graphene oxide adjuvant with loaded with CpG molecule and RBD protein antigen. Our results showed that this vaccine can produce high titer anti-SARS-CoV-2 RBD antibody neutralizing SARS-CoV-2 in mice within 2 weeks

    Next‐Generation Energy Harvesting and Storage Technologies for Robots Across All Scales

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    Self‐powered untethered robots that can meander unrestrictedly, squeeze into small spaces, and operate in diverse harsh environments have received immense attention in recent years. As there is not a universal solution that can be applied to power robots with diverse forms, service functions, and a broad size range from nanometers to meters, the design, fabrication, and implementation of power systems with a suitable weight, desired power and operation duration, and adaptiveness to confined spaces and operation conditions represent one of the greatest challenges in robotic research. Herein, an overview of recent progress and challenges in developing the next‐generation energy harvesting and storage technologies is provided, including direct energy harvesting, energy storage and conversion, and wireless energy transmission for robots across all scales

    A Rapid and High-Sensitive Real-Time Reverse Transcription-Polymerase Chain Reaction Assay Used for the Detection of Severe Acute Respiratory Syndrome Coronavirus 2

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    The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a public health emergency of international concern. Real-time reverse transcription-polymerase chain reaction (RT-PCR) is widely used as the gold standard method for the diagnosis of SARS-CoV-2 infection. However, the reliability of current real-time RT-PCR assays is questioned due to some false-negative reports. In this study, we improved the real-time RT-PCR method based on three target regions (ORF1ab, E, and N) of SARS-CoV-2. Results showed that real-time RT-PCR assays herein could complete detection within one hour after viral RNA preparation and had high sensitivity down to 5 copies of viral RNA. In addition, six clinical specimens were detected to evaluate the availability of this method. Among them, four samples were 3-plex SARS-CoV-2 positive and two were negative by real-time RT-PCR. The sensitivity was 100% (4/4), and specificity was 100% (2/2). These results demonstrate that we develop a rapid and high-sensitive real-time RT-PCR method for SARS-CoV-2 detection, which will be a powerful tool for COVID-19 identification and for monitoring suspected patients
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