scholarly journals A Survey of Potential Insect Vectors of Mountain Pine Proliferation Decline Phytoplasma in Curonian Spit, Lithuania

2020 ◽  
Vol 3 (1) ◽  
pp. 81
Author(s):  
Algirdas Ivanauskas ◽  
Jolanta Rimsaite ◽  
Jurij Danilov ◽  
Guy Soderman ◽  
Donatas Sneideris ◽  
...  
Keyword(s):  
Rflp Analysis ◽  
Insect Vector ◽  
Insect Vectors ◽  
Curonian Spit ◽  
Nested Polymerase Chain Reaction ◽  
Mountain Pine ◽  
Rflp Profile ◽  
Pcr Rflp ◽  
Pine Trees ◽  
Polymerase Chain

Mountain pine (Pinus mugo Turra) is a coniferous native to the highlands of central Europe. Our previous study revealed that mountain pine proliferation decline (MPPD) disease in the Curonian Spit of Lithuania is caused by a ‘Candidatus Phytoplasma pini’-related strain (16SrXXI-A). However, the insect vector of MPPD has not been identified. In this study, we conducted a survey to determine potential insect vectors of MPPD phytoplasma for three consecutive years (2016–2019). More than 1000 insect samples were collected from four locations in the Curonian Spit. These insects were identified as belonging to six families and ten genera. The presence of phytoplasma in insect samples was examined by nested polymerase chain reaction (PCR) using phytoplasma-specific primers (P1A/16S-SR and R16F2n/R16R2n). Phytoplasmas were detected in Cinara (Cinara) pini (Scots pine aphid), Cinara (Cinara) piniphila and Cinara (Schizolachnus) pineti (waxy grey pine needle aphid) insect samples. Subsequent restriction fragment length polymorphism (RFLP) analysis showed that the PCR-RFLP profile of these positive insect samples was consistent with that of the MPPD of diseased pine trees. These results suggest that C. (C.) pini, C. (C.) piniphila and C. (S.) pineti may be potential insect vectors of MPPD phytoplasma. The findings from this survey will provide useful information for the management of MPPD disease.

scholarly journals Genetic Diversity Among Viruses Associated with Sugarcane Mosaic Disease in Tucumán, Argentina

2009 ◽  
Vol 99 (1) ◽  
pp. 38-49 ◽  
Author(s):  
M. F. Perera ◽  
M. P. Filippone ◽  
C. J. Ramallo ◽  
M. I. Cuenya ◽  
M. L. García ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Mosaic Virus ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Mosaic Disease ◽  
Rt Pcr ◽  
Rflp Profile ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Sugarcane Mosaic Disease

Sugarcane leaves with mosaic symptoms were collected in 2006–07 in Tucumán (Argentina) and analyzed by reverse-transcriptase polymerase chain reaction (RT-PCR) restriction fragment length polymorphism (RFLP) and sequencing of a fragment of the Sugarcane mosaic virus (SCMV) and Sorghum mosaic virus (SrMV) coat protein (CP) genes. SCMV was detected in 96.6% of samples, with 41% showing the RFLP profile consistent with strain E. The remaining samples produced eight different profiles that did not match other known strains. SCMV distribution seemed to be more related to sugarcane genotype than to geographical origin, and sequence analyses of CP genes showed a greater genetic diversity compared with other studies. SrMV was detected in 63.2% of samples and most of these were also infected by SCMV, indicating that, unlike other countries and other Argentinean provinces, where high levels of co-infection are infrequent, co-existence is common in Tucumán. RFLP analysis showed the presence of SrMV strains M (68%) and I (14%), while co-infection between M and H strains was present in 18% of samples. Other SCMV subgroup members and the Sugarcane streak mosaic virus (SCSMV) were not detected. Our results also showed that sequencing is currently the only reliable method to assess SCMV and SrMV genetic diversity, because RT-PCR-RFLP may not be sufficiently discriminating.

scholarly journals New genetically distinct phytoplasmas and insect carriers associated with pine tree disease revealed by a survey in Curonian Spit, Lithuania

2021 ◽  
Author(s):  
Algirdas Ivanauskas ◽  
Deividas Valiunas ◽  
Jolanta Rimsaite ◽  
Jurij Danilov ◽  
Donatas Sneideris ◽  
...  
Keyword(s):  
Rflp Analysis ◽  
Universal Primers ◽  
Insect Vectors ◽  
Pine Needle ◽  
Curonian Spit ◽  
Tree Disease ◽  
Pine Tree ◽  
Pine Trees ◽  
Virtual Rflp ◽  
Phloem Feeding

Our previous studies reported that phytoplasma was the causative agent of the pine disease in Curonian spit, Lithuania. In this study, insects from diseased pine trees and their adjacent areas were collected from 2016 to 2019 to further identify potential insect vectors that spread phytoplasmas. A total of 1018 phloem-feeding insects (order Hemiptera) were identified, 98.62% of which were aphids (Aphididae), and no known phytoplasma vectors were found. Results from semi-nested PCR using phytoplasma-universal primers revealed that phytoplasmas were detected in scots pine aphids (Cinara pini), waxy grey pine needle aphids (Cinara pineti), and species-unknown aphids. Further sequence analysis and virtual RFLP analysis of aphid-harbored phytoplasma strains indicated that they were closely related to ‘Candidatus Phytoplasma pini’ (16SrXXI-A), but mainly 16SrXXI-A variants, which were also main strains identified in diseased pine trees. In addition, three new phytoplasma subgroups were delineated in the present study. Subgroups 16SrXXI-C and 16SrXXI-D were unveiled from previously identified (but classification was overlooked) Lithuanian pine phytoplasma strains. Subgroup 16SrXXI-E was discovered from the newly identified aphid-harbored phytoplasmas. Further transmission trial study on these aphids will provide insights into the epidemiology, and pathosystem of pine phytoplasma diseases, as well as the disease management.

scholarly journals Association of growth hormone (GH) gene polymorphism with growth and carcass in Sumba Ongole (SO) cattle

2017 ◽  
Vol 42 (3) ◽  
pp. 153 ◽  
Author(s):  
P. P. Agung ◽  
S. Anwar ◽  
W. P. B. Putra ◽  
M. S. A. Zein ◽  
A. S. Wulandari ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Gene Polymorphism ◽  
Fragment Length Polymorphism ◽  
Growth Parameters ◽  
Rflp Analysis ◽  
Cattle Breeding ◽  
Pcr Rflp ◽  
Gh Gene ◽  
Polymerase Chain ◽  
Dressing Percentage

A study was conducted to identify the polymorphism in the intron 3 of the Growth Hormone (GH) gene and also to evaluate the association of the GH gene polymorphism with growth parameters and dressing percentage in the Sumba Ongole (SO) cattle. A total of 267 individual DNA samples were used in the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis. The SO cattle growth parameters data (n=44) including birth weight (BW), weaning weight at 205 days of age (WW205), yearling weight at 365 days of age (YW365) and also dressing percentage (DP) (n=122) were investigated in this study. There were three genotypes (AA, AB, and BB) of the GH gene based on the PCR-RFLP analysis with allele frequency was 0.87 and 0.13 for A allele and B allele respectively. The highest genotype frequency in the SO cattle is AA (0.76) and the lowest is BB (0.02). The Heterozygosity Observed (Ho) value in the SO cattle population is 0.23 and Polymorphism Information Content (PIC) value is 0.20. Therefore, the genetic diversity in the SO cattle based on the GH gene polymorphism is quite low. There is no association (P>0.05) in BW, WW205, YW365, and DP with genotypes of the GH gene. As the result, the GH gene in this study cannot be used as a genetic marker in the SO cattle breeding program.

scholarly journals MMP-7 A-181G Polymorphism in Breast Cancer Patients from Western Iran

Breast Care ◽  
2015 ◽  
Vol 10 (6) ◽  
pp. 398-402 ◽  
Author(s):  
Kheirollah Yari ◽  
Ziba Rahimi ◽  
Mehrdad Payandeh ◽  
Zohreh Rahimi
Keyword(s):  
Breast Cancer ◽  
Cancer Patients ◽  
Fragment Length Polymorphism ◽  
Ductal Carcinoma ◽  
Lobular Carcinoma ◽  
Rflp Analysis ◽  
Breast Cancer Patients ◽  
Western Iran ◽  
Pcr Rflp ◽  
Polymerase Chain

Background: Matrix metalloproteinases (MMPs) are upregulated in tumors. The MMP-7 A-181G polymorphism is associated with increased expression of the MMP-7 gene. Aim of the present study was to investigate the association between the MMP-7 A-181G polymorphism and susceptibility to breast cancer. Patients and Methods: The MMP-7 A-181G variants were studied in a cohort of 251 subjects consisting of 100 breast cancer patients and 151 healthy controls; all were from Western Iran. The MMP-7 A-181G genotypes were identified using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Results: The frequencies of the MMP-7 AA, AG, and GG genotypes in healthy individuals were 34.4, 50.4, and 15.2%, respectively. In breast cancer patients, the frequencies of AA (34%), AG (52%), and GG (14%) genotypes (p = 0.95) were similar to those in the controls. There was a trend toward an increased frequency of the combined genotype of MMP-7 AG+GG in patients with lymph node metastasis (70.4%) compared to those without metastasis (66.7%). Also, in patients with invasive lobular carcinoma, the frequency of the MMP-7 AG+GG genotype tended to be higher (71.4%) compared to that in patients with invasive ductal carcinoma (66.2%) (p = 0.78). Conclusion: Our findings indicate that the MMP-7 A-181G polymorphism may not be correlated with susceptibility to breast cancer in our population.

scholarly journals Molecular Detection of Toxoplasma gondii and Neospora caninum in Domestic Ducks in Hunan Province, China

2021 ◽  
Vol 8 ◽  
Author(s):  
Qiu-Yan Lv ◽  
He-Liang Zheng ◽  
Wen-He Yang ◽  
Guo-Hua Liu
Keyword(s):  
Toxoplasma Gondii ◽  
Neospora Caninum ◽  
Rflp Analysis ◽  
Hunan Province ◽  
Economic Losses ◽  
Domestic Ducks ◽  
New Information ◽  
B1 Gene ◽  
Pcr Rflp ◽  
Polymerase Chain

Toxoplasma gondii and Neospora caninum are protozoan parasites that infect warm-blooded animals, and cause major economic losses in livestock industries worldwide. However, little is known about the genotypes of T. gondii and N. caninum in domestic ducks in China. Herein, brain samples from 588 domestic ducks from Hunan province in China were examined for the presence of T. gondii and N. caninum. Polymerase chain reaction (PCR) was used to detect T. gondii B1 gene and N. caninum NC-5 gene. Forty-five DNA samples (7.7%; 95% CI: 5.5–9.9) were positive for B1 gene, and two (0.3%; 95% CI: 0–0.7) were positive for NC-5 gene. The risk factors significantly associated with T. gondii infection were age and sex. The 45 samples positive for T. gondii were genotyped using multi-locus PCR-RFLP analysis and only one sample was fully genotyped as ToxoDB#9 (Chinese I). These results provide new information about the epidemiology of T. gondii and N. caninum in ducks in Hunan province in China. The data also highlight the importance of a “One Health” approach to dealing with toxoplasmosis.

scholarly journals Molecular Detection and PCR-RFLP Analysis of Mucoviscosity-Associated Gene A (magA) in Clinical Isolates of Multidrug-Resistant Klebsiella pneumoniae in Bangladesh

2020 ◽  
Vol 14 (1) ◽  
pp. 196-204
Author(s):  
Md. Hazrat Ali ◽  
Saeed Anwar ◽  
Nusrat Jahan Toma ◽  
Ikram Rafid ◽  
Md. Kamrul Hasan ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Clinical Specimen ◽  
Rflp Analysis ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Specific Gene ◽  
Multicenter Cohort ◽  
Pcr Rflp ◽  
Invasive Infections ◽  
Polymerase Chain

Background and Objective: The mucoviscosity associated gene A (magA) in the hypermucoviscous variants of K. pneumoniae is reported to be associated with invasive infections and considered a virulence factor. We sought to analyze the magA genes in K. pneumoniae isolates in the clinical specimen collected from Bangladesh. Methods: We established a multicenter cohort of patients with Klebsiella infection hospitalized at 05 different hospitals between September 2016 and April 2017. We collected 313 K. pneumoniae isolates from patients who consented to participate in the study. The isolates were evaluated for harboring the magA genes using a single-tube multiplexed polymerase chain reaction. The magA genes were analyzed by PCR-RFLP technique using two enzymes, namely PciI and SmaI. Antibiogram assay using 12 commercially available antibiotic discs was performed on all the isolates. Results: The presence of K. pneumoniae specific gene (ureD) was confirmed in all the isolates. The percentage of isolates harboring the magA gene was 7.34%(23 isolates), the majority of which was collected from the patients admitted in intensive care units (16 isolates, 69.6%), and infectious diseases wards (5 isolates, 21.7%). PCR-RFLP analysis revealed that for 7 out of 23 isolates, where Sma1 could not cleave the magA gene. All the isolates showed resistance to ampicillin, carbenicillin cefradine, chloramphenicol, erythromycin, kanamycin, and sulphamethoxazole, though the extent was varying. However, imipenem showed 100% sensitivity to all the tested isolates. Conclusion: This study demonstrates the presence of the magA gene in multidrug-resistant clinical isolates of K. pneumoniae collected from Bangladesh.

Genetic determination of Saccharomyces cerevisiae et Saccharomyces bayanus species by PCR/RFLP analysis of the MET2 gene

OENO One ◽  
1996 ◽  
Vol 30 (1) ◽  
pp. 15
Author(s):  
Isabelle Masneuf-Pomarède ◽  
Michel Aigle ◽  
Denis Dubourdieu
Keyword(s):  
Dna Hybridization ◽  
Rflp Analysis ◽  
Distinct Species ◽  
Genetic Determination ◽  
Yeast Strains ◽  
Saccharomyces Bayanus ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Em Gene

<p style="text-align: justify;">Several yeast strains of the species <em>S. cerevisiae</em>, <em>S. bayanus</em> and <em>S. paradoxus</em>, first identified by hybridization experiments and DNA/DNA hybridization were characterized by using Polymerase Chain Reaction/Restriction Fragment Length Polymorphism (PCR/RFLP) of the <em>MET2 gene</em>. The concordance between this tool and classical genetic analyses did not reveal any exception for all the strains analysed, so PCR/RFLP of the MET2 gene proves to be a reliable and fast tool for delimiting <em>S. cerevisiae</em> and <em>S. bayanus</em>. OEnological strains race <em>bayanus</em>, <em>chevalieri</em>, <em>capensis</em> gave <em>S. cerevisiae</em> restriction patterns, whereas most of strains race <em>S. uvarum</em> belong to <em>S. bayanus</em> and displayed a specific chromosomal band patterns different from band patterns of <em>S. cerevisiae</em> strains. To avoid confusion in oenological terminology, oenologists should no longer use the name of <em>bayanus</em> to designate industrial or wild <em>S. cerevisiae</em> Gal- strains, and should consider <em>S. bayanus</em> as a distinct species.</p>

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