scholarly journals Association of “Candidatus Phytoplasma australiense” with Sudden Decline of Cabbage Tree in New Zealand

Plant Disease ◽  
2001 ◽  
Vol 85 (5) ◽  
pp. 462-469 ◽  
Author(s):  
Mark T. Andersen ◽  
Ross E. Beever ◽  
Paul W. Sutherland ◽  
Richard L. S. Forster
Keyword(s):  
New Zealand ◽  
Rrna Gene ◽  
Mature Trees ◽  
Nested Polymerase Chain Reaction ◽  
Transmission Electron ◽  
Adult Trees ◽  
Leaf Yellowing ◽  
Pcr Products ◽  
One Step ◽  
Cabbage Tree

Sudden decline of the New Zealand cabbage tree (Cordyline australis) results in the rapid death of affected plants within months of first external symptoms becoming apparent. Symptoms, which have been observed in saplings and mature trees, include vascular discoloration and leaf yellowing followed by leaf desiccation and eventual plant collapse. Previous work failed to link the disease with any causal agent. A phytoplasma has now been detected in all symptomatic saplings and some symptomatic trees tested, using one-step and nested polymerase chain reaction (PCR) to amplify portions of the 16S rRNA gene. This phytoplasma was not detected in nonsymptomatic plants. Phytoplasma DNA was found in shoot and rhizome apices, leaves and wood tissue of saplings, and in the rhizome apex and trunk tissues of adult trees. Sequencing of the PCR products from selected samples indicated that the phytoplasma is “Candidatus Phytoplasma australiense.” Phytoplasma cells were detected by transmission electron microscopy in phloem sieve tubes of the rhizomes of affected saplings. One sapling with early symptoms recovered after injection with tetracycline antibiotic, but two saplings with advanced symptoms did not recover. It is concluded that “Candidatus Phytoplasma australiense” is present in symptomatic plants and is the cause of sudden decline.

scholarly journals An Epidemic of Almond Witches'-broom in Lebanon: Classification and Phylogenetic Relationships of the Associated Phytoplasma

Plant Disease ◽  
2002 ◽  
Vol 86 (5) ◽  
pp. 477-484 ◽  
Author(s):  
Yusuf Abou-Jawdah ◽  
Armig Karakashian ◽  
Hana Sobh ◽  
Marta Martini ◽  
Ing-Ming Lee
Keyword(s):  
Rflp Analysis ◽  
Pigeon Pea ◽  
Universal Primers ◽  
Rrna Gene ◽  
Nested Polymerase Chain Reaction ◽  
Main Trunk ◽  
Stunted Growth ◽  
Rdna Sequences ◽  
Pcr Products ◽  
Almond Trees

An epidemic of almond witches'-broom has devastated almond production in Lebanon. Thousands of almond trees have died over the past 10 years due to the rapid spread of the disease. The symptoms, which include early flowering, stunted growth, leaf rosetting, dieback, off-season growth, proliferation of slender shoots, and witches'-brooms arising mainly from the main trunk and roots, resemble those caused by phytoplasmal infections. For the detection of the putative causal agent, nested polymerase chain reaction (PCR) was performed using universal primers (P1/P7, R16mF2/R16mR1, and R16F2n/R16R2) commonly used for the specific diagnosis of plant pathogenic phytoplasmas. Phytoplasmas were readily detected from infected trees with witches'-broom symptoms collected from three major almond growing regions in Lebanon. Restriction fragment length polymorphism (RFLP) analysis of PCR products amplified by the primer pair R16F2n/R16R2 revealed that the phytoplasma associated with infected almonds is similar to, but distinct from, members of the pigeon pea witches'-broom phytoplasma group (16SrIX). A new subgroup, 16SrIX-B, was designated. Sequencing of the amplified products of the phytoplasma 16S rRNA gene indicated that almond witches'-broom (AlmWB) phytoplasma is most closely related to members of the pigeon pea witches'-broom phytoplasma group (with sequence homology ranging from 98.4 to 99.0%). Phylogenetic analysis of 16S rDNA sequences from AlmWB phytoplasma and from representative phytoplasmas from GenBank showed that the AlmWB phytoplasma represents a distinct lineage within the pigeon pea witches'-broom subclade. The same phytoplasma appears also to infect peach and nectarine seedlings.

scholarly journals Co-cultivation of ‘Candidatus Liberibacter asiaticus’ with Actinobacteria from Citrus with Huanglongbing

Plant Disease ◽  
2008 ◽  
Vol 92 (11) ◽  
pp. 1547-1550 ◽  
Author(s):  
Michael J. Davis ◽  
Sachindra N. Mondal ◽  
Huiqin Chen ◽  
Michael E. Rogers ◽  
Ronald H. Brlansky
Keyword(s):  
16S Rrna ◽  
Rrna Gene ◽  
Insect Vector ◽  
Candidatus Liberibacter Asiaticus ◽  
Citrus Greening Disease ◽  
Transmission Electron ◽  
Pcr Products ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Polymerase Chain

Huanglongbing (HLB), also known as citrus greening disease, is a devastating disease of citrus caused by phloem-limited bacteria that have not been grown in culture. Three species, ‘Candidatus Liberibacter asiaticus’, ‘Ca. L. africanus’, and ‘Ca. L. americanus’, are known. ‘Ca. L. asiaticus’ and its insect vector, the psyllid Diaphorina citri, have been recently introduced into Florida. We attempted to isolate ‘Ca. L. asiaticus’ using media formulations developed in response to the growth of another bacterium that appears to be related to the liberibacters based on 16S rRNA gene identities. Cultures were obtained that were polymerase chain reaction (PCR) positive for ‘Ca. L. asiaticus’. However, transmission electron microscope examination of the culture, PCR using generic primers, and sequencing of the PCR products revealed the presence of other bacteria in the cultures. These were actinobacteria related to Propionibacterium acnes based on 16S rRNA identities. The co-cultures remained after attempts to purify the cultures by single-colony isolation, suggesting that the bacteria might be mutually beneficial to each other in culture. The co-cultures have survived more than 10 weekly passages to fresh medium. PCR using P. acnes-specific primers indicated that actinobacteria are common inhabitants of citrus and psyllids, whether or not ‘Ca. L. asiaticus’ is present.

scholarly journals First Report of “Candidatus Phytoplasma asteris”-Related Strains in Brassica rapa in Saskatchewan, Canada

Plant Disease ◽  
2006 ◽  
Vol 90 (6) ◽  
pp. 832-832 ◽  
Author(s):  
C. Y. Olivier ◽  
G. Séguin-Swartz ◽  
D. Hegedus ◽  
T. Barasubiye
Keyword(s):  
Sequence Data ◽  
Direct Sequencing ◽  
Nested Polymerase Chain Reaction ◽  
Aster Yellows ◽  
First Report ◽  
Leaf Yellowing ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Subgroup Member ◽  
Root Tissues

“Candidatus phytoplasma asteris” and related strains (i.e., aster yellows group 16SrI) have been associated with diseases of numerous plant species worldwide. Symptoms of aster yellows (AY) have been reported on rapeseed/canola (Brassica napus and B. rapa) crops in Saskatchewan (SK) and Manitoba, Canada since 1953 (2). Symptoms generally include stunting, virescence, leaf yellowing or purpling, phyllody, and formation of bladder-like siliques. A total of 120 mature B. rapa cv. AC Sunbeam plants exhibiting AY symptoms were collected in commercial fields near Medstead, SK during 2003 and 2004 (one field per year). As described previously (4), total genomic DNA was extracted from leaf, stem, roots, and seeds collected from the 120 plants, from seeds from the seed lots sown in 2003 and 2004, and from leaf and stem tissue of 20 greenhouse-grown plants from each seed lot. The latter DNA samples were assayed for phytoplasma DNA by a nested polymerase chain reaction (PCR) assay incorporating phytoplasma universal 16S rRNA primer pairs P1/P6 (1) followed by R16R2/R16F2 (4). Seed samples analyzed from the 2003 and 2004 seed lots and tissues of the 40 greenhouse-grown plants all tested negative for phytoplasma DNA using this assay. Leaf, stem, and/or root tissues of all plants collected in the field in 2003 (60 plants) and 2004 (60 plants) and 71.1% (315 of 443) of seed samples (five seeds per sample) tested positive for the presence of phytoplasma DNA, as evidenced by the presence of an expected band of 1.2 kb on the gels after the second amplification with primers R16R2/R16F2. Nested PCR products from plant samples collected in 2003 were cloned, sequenced, and compared with phytoplasma sequences archived in the GenBank nucleotide database. On this basis, phytoplasmas detected in plants or their seeds collected in 2003 were found to be most similar (98.8%) to CHRY (Accession No. AY180956), a 16SrI-A subgroup strain, or were most similar (98.9%) to isolate 99UW89 (Accession no. AF268407), a known 16SrI-B subgroup strain. Sequences of phytoplasmas detected in plants or their seeds in 2004 were obtained by direct sequencing of rRNA products amplified from samples using PCR incorporating primer pairs P1/P6 and P4/P7 (3). Analysis of sequence data revealed that phytoplasmas in these plants were all most similar (99.5%) to AY-WB (Accession no. AY389828), a 16SrI-A subgroup member. The nucleotide sequences have been deposited with GenBank under Accession nos. DQ404346, DQ404347, and DQ411470. To our knowledge, this is the first report of 16SrI-A and 16SrI-B subgroup phytoplasmas infecting plants and seed of B. rapa in Saskatchewan. References: (1) I.-M. Lee et al. Phytopathology, 83:834, 1993. (2) W. E. Sackston. Can. Plant Dis. Surv. 33:41, 1953. (3) L. B. Sharmila et al. J. Plant Biochem. Biotech. 13:1, 2004. (4) E. Tanne et al. Phytopathology, 91:741, 2001.

scholarly journals Molecular characterisation of phytoplasmas infecting roses in Poland

2013 ◽  
Vol 55 (1) ◽  
pp. 325-334
Author(s):  
Hanna Śliwa ◽  
Tadeusz Malinowski ◽  
Maria Kamińska
Keyword(s):  
Reference Strain ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Nested Polymerase Chain Reaction ◽  
Aster Yellows ◽  
Leaf Chlorosis ◽  
Leaf Yellowing ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Aster Yellows Phytoplasma

Symptoms of shoot dieback and leaf yellowing followed by leaf chlorosis were observed in naturally infected roses 'Frisco' and 'Suela', cultivated in a commercial greenhouse in Poland. The presence of phytoplasma was demonstrated in affected plants by nested polymerase chain reaction (PCR) with R16Fl/RO and Pl/P7 primer pairs in the first round followed by a second one with R16F2n/R2, fA/rA, Pc399/P1694, R16(I)Fl/Rl and Pl/fArev primer pairs. Restriction fragment length polymorphism (RFLP) analysis of PCR products (primed with primers R16F2n/R2) was done using enzymes AluI, MseI, RsaI and HpaII. Restriction profiles obtained with these enzymes were identical to those of reference strain AY1 belonging to aster yellows phytoplasma group, subgroup I-B (16SrI-B). Nested PCR products from roses 'Frisco' and 'Suela' were sequenced. Analysis of sequences confirmed that the phytoplasma infecting those roses could be classified to aster yellows phytoplasma group, subgroup B.

An Interactive Minicomputer Program for the Indexing and Simulation of Electron Diffraction Patterns

1976 ◽  
Vol 34 ◽  
pp. 542-543
Author(s):  
R.P. Goehner ◽  
W.T. Hatfield ◽  
Prakash Rao
Keyword(s):  
Electron Diffraction ◽  
Real Time ◽  
Pattern Analysis ◽  
Flow Diagram ◽  
Hard Copy ◽  
Time Analysis ◽  
Real Time Analysis ◽  
Transmission Electron ◽  
One Step ◽  
Diffraction Patterns

Computer programs are now available in various laboratories for the indexing and simulation of transmission electron diffraction patterns. Although these programs address themselves to the solution of various aspects of the indexing and simulation process, the ultimate goal is to perform real time diffraction pattern analysis directly off of the imaging screen of the transmission electron microscope. The program to be described in this paper represents one step prior to real time analysis. It involves the combination of two programs, described in an earlier paper(l), into a single program for use on an interactive basis with a minicomputer. In our case, the minicomputer is an INTERDATA 70 equipped with a Tektronix 4010-1 graphical display terminal and hard copy unit.A simplified flow diagram of the combined program, written in Fortran IV, is shown in Figure 1. It consists of two programs INDEX and TEDP which index and simulate electron diffraction patterns respectively. The user has the option of choosing either the indexing or simulating aspects of the combined program.

scholarly journals Hydrothermal Synthesis of Iridium-Substituted NaTaO3 Perovskites

Nanomaterials ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1537
Author(s):  
David L. Burnett ◽  
Christopher D. Vincent ◽  
Jasmine A. Clayton ◽  
Reza J. Kashtiban ◽  
Richard I. Walton
Keyword(s):  
Significant Proportion ◽  
Edge Length ◽  
Perovskite Oxide ◽  
Hydrogen Yield ◽  
Orthorhombic Space Group ◽  
X Ray ◽  
Profile Fitting ◽  
Transmission Electron ◽  
Scanning Transmission ◽  
One Step

Iridium-containing NaTaO3 is produced using a one-step hydrothermal crystallisation from Ta2O5 and IrCl3 in an aqueous solution of 10 M NaOH in 40 vol% H2O2 heated at 240 °C. Although a nominal replacement of 50% of Ta by Ir was attempted, the amount of Ir included in the perovskite oxide was only up to 15 mol%. The materials are formed as crystalline powders comprising cube-shaped crystallites around 100 nm in edge length, as seen by scanning transmission electron microscopy. Energy dispersive X-ray mapping shows an even dispersion of Ir through the crystallites. Profile fitting of powder X-ray diffraction (XRD) shows expanded unit cell volumes (orthorhombic space group Pbnm) compared to the parent NaTaO3, while XANES spectroscopy at the Ir LIII-edge reveals that the highest Ir-content materials contain Ir4+. The inclusion of Ir4+ into the perovskite by replacement of Ta5+ implies the presence of charge-balancing defects and upon heat treatment the iridium is extruded from the perovskite at around 600 C in air, with the presence of metallic iridium seen by in situ powder XRD. The highest Ir-content material was loaded with Pt and examined for photocatalytic evolution of H2 from aqueous methanol. Compared to the parent NaTaO3, the Ir-substituted material shows a more than ten-fold enhancement of hydrogen yield with a significant proportion ascribed to visible light absorption.

Structural, Morphological, Magnetic and Self-Heating Studies of One-Step Polyol Synthesized Manganese Ferrite (MnFe2O4) Nanoparticles

2019 ◽  
Vol 19 (01) ◽  
pp. 1950003
Author(s):  
P. R. Ghutepatil ◽  
S. H. Pawar
Keyword(s):  
Room Temperature ◽  
Synthesis Method ◽  
Average Crystallite Size ◽  
Superparamagnetic Nanoparticles ◽  
Polyol Synthesis ◽  
X Ray Diffraction ◽  
Self Heating ◽  
Transmission Electron ◽  
One Step ◽  
Mnfe2o4 Nanoparticles

In this paper, uniform and superparamagnetic nanoparticles have been prepared using one-step polyol synthesis method. Structural, morphological and magnetic properties of obtained MnFe2O4 nanoparticles have been investigated by using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), field emission scanning electron microscope (FE-SEM), transmission electron microscopy (TEM), vibrating sample magnetometry (VSM) and thermogravimetric analysis (TGA) techniques. Structural investigation showed that the average crystallite size of obtained nanoparticles was about 10[Formula: see text]nm. Magnetic study revealed that the nanoparticles were superparamagnetic at room temperature with magnetization 67[Formula: see text]emu/g at room temperature. The self-heating characteristics of synthesized MnFe2O4 nanoparticles were studied by applying external AC magnetic field of 167.6 to 335.2[Formula: see text]Oe at a fixed frequency of 265[Formula: see text]kHz. The SAR values of MnFe2O4 nanoparticles were calculated for 2, 5, 10[Formula: see text]mg[Formula: see text]mL[Formula: see text] concentrations and it is observed that the threshold hyperthermia temperature is achieved for all concentrations.

One Step Synthesis and Characterization of Zirconia-Graphene Composites

2012 ◽  
Vol 600 ◽  
pp. 174-177 ◽  
Author(s):  
Jian Fei Xia ◽  
Zong Hua Wang ◽  
Yan Zhi Xia ◽  
Fei Fei Zhang ◽  
Fu Qiang Zhu ◽  
...  
Keyword(s):  
Synthesis And Characterization ◽  
Two Dimensional ◽  
X Ray Diffraction ◽  
X Ray ◽  
Graphene Composite ◽  
Transmission Electron ◽  
One Step ◽  
Ft Ir ◽  
Xrd Techniques

Zirconia-graphene composite (ZrO2-G) has been successfully synthesized via decomposition of ZrOCl2•6H2O in a water-isopropanol system with dispersed graphene oxide (GO) utilizing Na2S as a precursor could enable the occurrence of the deposition of Zr4+ and the deoxygenation of GO at the same time. Transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) techniques were used to characterize the samples. It was found that graphene were fully coated with ZrO2, and the ZrO2 existing in tetragonal phase, which resulted in the formation of two-dimensional composite.

Species-specific duplex PCR for the detection of Pratylenchus penetrans

Nematology ◽  
2009 ◽  
Vol 11 (6) ◽  
pp. 847-857 ◽  
Author(s):  
Lieven Waeyenberge ◽  
Nicole Viaene ◽  
Maurice Moens
Keyword(s):  
Internal Control ◽  
Dna Sequences ◽  
Rrna Gene ◽  
Duplex Pcr ◽  
Specific Primers ◽  
5.8S Rrna Gene ◽  
5.8S Rrna ◽  
Wide Range ◽  
Pcr Products ◽  
Species Specific

Abstract ITS1, the 5.8S rRNA gene and ITS2 of the rDNA region were sequenced from 20 different Pratylenchus species. Additionally, the same region was sequenced from seven populations of P. penetrans. After purifying, cloning and sequencing the PCR products, all sequences were aligned in order to find unique sites suitable for the design of species-specific primers for P. penetrans. Since ITS regions showed variability between and even within populations of P. penetrans, only three small DNA sequences were suitable for the construction of three potentially useful species-specific primers. New species-specific primers were paired with existing universal ITS primers and tested in all possible primer combinations. The best performing primer set, supplemented with a universal 28S rDNA primer set that served as an internal control, was tested in duplex PCR. The ideal annealing temperature, Mg2+ concentration and primer ratios were then determined for the most promising primer set. The optimised duplex PCR was subsequently tested on a wide range of different Pratylenchus spp. and 25 P. penetrans populations originating from all over the world. To test the sensitivity, the duplex PCR was conducted on DNA extracted from a single P. penetrans nematode mixed with varying amounts of nematodes belonging to another Pratylenchus species. Results showed that a reliable and sensitive P. penetrans species-specific duplex PCR was constructed.

Two main clusters within Trypanosoma cruzi zymodeme 3 are defined by distinct regions of the ribosomal RNA cistron

Parasitology ◽  
2002 ◽  
Vol 124 (2) ◽  
pp. 177-184 ◽  
Author(s):  
M. B. A. MENDONÇA ◽  
N. S. NEHME ◽  
S. S. SANTOS ◽  
E. CUPOLILLO ◽  
N. VARGAS ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Ribosomal Rna ◽  
Southern Blotting ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
Pcr Products ◽  
Phylogenetic Lineages ◽  
Definition Of ◽  
Ribosomal Cistron ◽  
Panstrongylus Geniculatus

Trypanosoma cruzi is currently classified into 2 major phylogenetic lineages, T. cruzi I and II, that correlate with the formerly described zymodeme 1 and 2, respectively. Another isoenzymic group (zymodeme 3–Z3) was also described. In this study, we analysed the genetic diversity among Z3 isolates of the Brazilian Amazon by restriction fragment length polymorphism of the intergenic transcribed spacers (ITSs) of the ribosomal RNA cistron and the size of the divergent domain D7 of the 24Sα rRNA gene. DNAs from 12 T. cruzi Z3 isolates obtained from humans (2), Panstrongylus geniculatus (1), and Rhodnius brethesi (9) were submitted to PCR amplification of the ITSs plus the 5·8S rDNA. The PCR products were digested with 4 distinct endonucleases and the profiles analysed by a numerical methodology. The phenetic dendrogram revealed a clear dichotomy in the Z3 group, defining 2 groups that were named Z3-A and Z3-B. Dimorphism was also found in the band sizes of the amplified D7 divergent domain of the 24Sα rDNA, which showed a perfect correlation with the ITSs clustering. The organization of the ribosomal cistron was investigated by Southern blotting and shown to be conserved in the genome of the 2 Z3 groups. This study shows that the rDNA cistron allows the definition of 2 distinct subclusters in Z3 isolates.

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