Characterization of copy numbers of 16S rDNA and 16S rRNA of Candidatus Liberibacter asiaticus and the implication in detection in planta using quantitative PCR

<p>Abstract</p> <p>Background</p> <p>Citrus Huanglongbing (HLB) is one of the most devastating diseases on citrus and is associated with <it>Candidatus </it>Liberibacter spp.. The pathogens are phloem limited and have not been cultured <it>in vitro</it>. The current management strategy of HLB is to remove infected citrus trees and reduce psyllid populations with insecticides to prevent the spreading. This strategy requires sensitive and reliable diagnostic methods for early detection.</p> <p>Results</p> <p>We investigated the copy numbers of the 16S rDNA and 16S rRNA of the HLB pathogen and the implication of improving the diagnosis of HLB for early detection using Quantitative PCR. We compared the detection of HLB with different Quantitative PCR based methods with primers/probe targeting either 16S rDNA, beta-operon DNA, 16S rRNA, or beta-operon RNA. The 16S rDNA copy number of <it>Ca</it>. Liberibacter asiaticus was estimated to be three times of that of the beta-operon region, thus allowing detection of lower titer of <it>Ca</it>. L. asiaticus. Quantitative reverse transcriptional PCR (QRT-PCR) indicated that the 16S rRNA averaged 7.83 times more than that of 16S rDNA for the same samples. Dilution analysis also indicates that QRT-PCR targeting 16S rRNA is 10 time more sensitive than QPCR targeting 16S rDNA. Thus QRT-PCR was able to increase the sensitivity of detection by targeting 16S rRNA.</p> <p>Conclusion</p> <p>Our result indicates that <it>Candidatus </it>Liberibacter asiaticus contains three copies of 16S rDNA. The copy number of 16S rRNA of <it>Ca</it>. L. asiaticus <it>in planta </it>averaged about 7.8 times of 16S rDNA for the same set of samples tested in this study. Detection sensitivity of HLB could be improved through the following approaches: using 16S rDNA based primers/probe in the QPCR assays; and using QRT-PCR assays targeting 16S rRNA.</p

Genomic analysis of sorghum by fluorescence in situ hybridization

The reliability of genome analysis and proficiency of genetic manipulation in vivo and in vitro are increased by assignment of linkage groups to specific chromosomes, placement of centromeres, orientation with respect to telomeres, and linear alignment with respect to chromosomal features and dimensions. I undertook five studies aimed at integrating sorghum genomics and cytogenetics at several levels. The results help establish an entirely new "cyto-genomics" resource, impacts of which are likely to be broad. In the first study, I developed a FISH-based karyotyping system for Sorghum bicolor Moench. I used integrated structural genomic resources, including linkage maps and large-insert clonal libraries of sorghum genomic DNA to develop a 17-locus probe cocktail for simultaneous fluorescent in situ hybridization (FISH). This probe enabled facile identification of all chromosome pairs in mitotic chromosome spreads. Perhaps just as important, I established time-efficient means to select sorghum BAC clones for multi-probe FISH. Thus, an integrated cyto-genomics system for sorghum can be constructed without need of chromosome flow sorting or microdissection, both of which are difficult and costly. In the second study, hybridization of DNA clones from 37 different genomic regions enabled the assignment of linkage groups and orientation of linkage maps to chromosomes. Comparisons between genetic and physical distances throughout the genome enabled a new nomenclature for linkage group designation in sorghum. The results provide an integrated nomenclature system of Sorghum bicolor chromosomes and linkage groups. In the third study, I created high-resolution maps by FISH to pachytene bivalents for two linkage groups (B and H), and defined relationships between pericentromeric heterochromatin, centromeres, mapped markers and recombination rates. These relationships will help guide the development and use of sorghum genomics. In the fifth study, I used FISH in two ongoing gene-targeted efforts. For the maturity gene ma5 and fertility restoration gene rfl, I estimated physical lengths between currently available flanking molecular markers. This enables estimation of recombination densities in these regions and assessment of the applicability of map-based and -assisted cloning

Physical characterization of amorphous In-Ga-Zn-O thin-film transistors with direct-contact asymmetric graphene electrode

High performance a-IGZO thin-film transistors (TFTs) are fabricated using an asymmetric graphene drain electrode structure. A-IGZO TFTs (channel length = 3 μm) were successfully demonstrated with a saturation field-effect mobility of 6.6 cm2/Vs without additional processes between the graphene and a-IGZO layer. The graphene/a-IGZO junction exhibits Schottky characteristics and the contact property is affected not only by the Schottky barrier but also by the parasitic resistance from the depletion region under the graphene electrode. Therefore, to utilize the graphene layer as S/D electrodes for a-IGZO TFTs, an asymmetric electrode is essential, which can be easily applied to the conventional pixel electrode structure. © 2014 Author(s).1

Monitoring of multi-frequency polarization of gamma-ray bright AGNs

We started two observing programs with the Korean VLBI Network (KVN) monitoring changes in the flux density and polarization of relativistic jets in gamma-ray bright AGNs simultaneously at 22, 43, 86, 129 GHz. One is a single-dish weekly-observing program in dual polarization with KVN 21-m diameter radio telescopes beginning in 2011 May. The other is a VLBI monthly-observing program with the three-element VLBI network at an angular resolution range of 1.0--9.2 mas beginning in 2012 December. The monitoring observations aim to study correlation of variability in gamma-ray with that in radio flux density and polarization of relativistic jets when they flare up. These observations enable us to study the origin of the gamma-ray flares of AGNs.Comment: 4 pages, 4 figures, Proceedings of the conference "The innermost regions of relativistic jets and their magnetic fields", Granada, Spai

Overexpression of Arabidopsis thaliana brassinosteroid-related acyltransferase 1 gene induces brassinosteroid-deficient phenotypes in creeping bentgrass

Brassinosteroids (BRs) are naturally occurring steroidal hormones that play diverse roles in various processes during plant growth and development. Thus, genetic manipulation of endogenous BR levels might offer a way of improving the agronomic traits of crops, including plant architecture and stress tolerance. In this study, we produced transgenic creeping bentgrass (Agrostis stolonifera L.) overexpressing a BR-inactivating enzyme, Arabidopsis thaliana BR-related acyltransferase 1 (AtBAT1), which is known to catalyze the conversion of BR intermediates to inactive acylated conjugates. After putative transgenic plants were selected using herbicide resistance assay, genomic integration of the AtBAT1 gene was confirmed by genomic PCR and Southern blot analysis, and transgene expression was validated by northern blot analysis. The transgenic creeping bentgrass plants exhibited BR-deficient phenotypes, including reduced plant height with shortened internodes (i.e., semi-dwarf), reduced leaf growth rates with short, wide, and thick architecture, high chlorophyll contents, decreased numbers of vascular bundles, and large lamina joint bending angles (i.e., erect leaves). Subsequent analyses showed that the transgenic plants had significantly reduced amounts of endogenous BR intermediates, including typhasterol, 6-deoxocastasterone, and castasterone. Moreover, the AtBAT1 transgenic plants displayed drought tolerance as well as delayed senescence. Therefore, the results of the present study demonstrate that overexpression of an Arabidopsis BR-inactivating enzyme can reduce the endogenous levels of BRs in creeping bentgrass resulting in BR-deficient phenotypes, indicating that the AtBAT1 gene from a dicot plant is also functional in the monocot crop.111Ysciescopu

Force-sensitive resistors to measure the distribution of weight in the pads of sound dogs in static standing position

The purpose of this study was to measure how weight is distributed in the pads of each of the 4 limbs of dogs and evaluate the intra-investigator reproducibility and inter-investigator reliability of the measurement method. Eight dogs were examined 3 times a day by 3 investigators at 1 week intervals for 3 weeks to determine the weight distribution to each of the pads. The force-sensitive resistor was used for measurement and specific software (PetLAB2) was used to calculate the weight applied to each pad. The intra-investigator reproducibility showed moderate to good reliability (ICC range, 0.575-0.873) and the inter-investigator reliability was moderate (ICC range, 0.525-0.746). Based on this study, it can be observed whether the weight distributed to each pad approaches the normal value after treatment in patients with orthopaedic and neurologic diseases. It is expected that this experimental method will be one of the objective indicators to evaluate the degree of recovery in patients with orthopaedic and neurologic diseases

Enhanced blue photoluminescence realized by copper diffusion doping of ZnO thin films

ZnO thin films with blue photoluminescence (PL) have been fabricated through Cu diffusion doping. A CuOx-ZnO mixture, and Cu/ZnO double layer, films were prepared on amorphous SiOx/Si substrates by pulsed laser deposition (PLD), and electron beam (e-beam) deposition, respectively. After sequential oxygen annealing, CuOx-ZnO mixture films exhibited green emission centered at 523 nm. However, Cu/ZnO double layer films differed in producing a blue emission centered at 480 nm. Detailed analysis identified that this blue shift in the emission center resulted from increased blue emissions attributed to Cu dopants in the film by e-beam deposition. Luminescence intensity was increased to 6 cd/m2 for a sample annealed at 700 deg;C. Color points were close to the locus of points following the line of a black-body-radiator on the CIE 1931 XY chromaticity diagram. The present results show that Cu-doped ZnO has strong potential as a cost effective phosphor for use in down converting LEDs. © 2013 Optical Society of America.1