scholarly journals DETEKSI MENGGUNAKAN PCR (POLYMERASE CHAIN REACTION) CANDIDATUS LIBERIBACTER ASIATICUS, PENYEBAB HUANGLONGBING PADA JERUK SIEM DENGAN BEBERAPA TIPE GEJALA PADA DAUN

2010 ◽  
Vol 10 (2) ◽  
pp. 178-183
Author(s):  
Achmad Himawan ◽  
Y.B Sumardiyono ◽  
Susamto Somowiyarjo ◽  
Y. Andi Trisyono ◽  
Andrew Beattie
Keyword(s):  
Polymerase Chain Reaction ◽  
Candidatus Liberibacter Asiaticus ◽  
Chain Reaction ◽  
Causal Organism ◽  
Pcr Polymerase Chain Reaction ◽  
Different Types ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Polymerase Chain ◽  
Leaf Symptoms

Detection using PCR (Polymerase Chain Reaction) Candidatus Liberibacter asiaticus, Huanglongbing causal Organism on Siem Mandarin with different types of symptoms.  Huanglongbing (HLB) or Citrus Vein Phloem Degeration (CVPD) is one of major diseases on Siem mandarin in Indonesia. HLB is caused by bacteria Candidatus liberibacter asiatus (LAS). The bacteria only live in the phloem cells of host tree and only recently it was reported to be successfully cultured on agar medium. Early detection method of LAS is needed to support healthy Siem mandarin cultivation program. This research was conducted to detect LAS in different types of HLB leaf symptoms based on Polymerase Chain Reaction (PCR) method with specific primer forward MHO 353 and reverse MHO 354.  The results suggested that 8 types of HLB leaf symptoms were found on the samples used in this experiment. LAS was detected at 60% on the leaves without any symptom followed by the leaves with completely chlorosis symptom at 66%. The leaves with unevenly yellow showing higher percentage of LAS detection ranged from 80-86%. PCR technique successfully amplified DNA of LAS with the size target of 600 bp.

scholarly journals In Planta Distribution of ‘Candidatus Liberibacter asiaticus’ as Revealed by Polymerase Chain Reaction (PCR) and Real-Time PCR

2008 ◽  
Vol 98 (5) ◽  
pp. 592-599 ◽  
Author(s):  
Satyanarayana Tatineni ◽  
Uma Shankar Sagaram ◽  
Siddarame Gowda ◽  
Cecile J. Robertson ◽  
William O. Dawson ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Citrus Tree ◽  
Candidatus Liberibacter Asiaticus ◽  
In Planta ◽  
Chain Reaction ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Polymerase Chain

Huanglongbing (HLB) is one of the most devastating diseases of citrus worldwide, and is caused by a phloem-limited fastidious prokaryotic α-proteobacterium that is yet to be cultured. In this study, a combination of traditional polymerase chain reaction (PCR) and real-time PCR targeting the putative DNA polymerase and 16S rDNA sequence of ‘Candidatus Liberibacter asiaticus,’ respectively, were used to examine the distribution and movement of the HLB pathogen in the infected citrus tree. We found that ‘Ca. Liberibacter asiaticus’ was distributed in bark tissue, leaf midrib, roots, and different floral and fruit parts, but not in endosperm and embryo, of infected citrus trees. Quantification analysis of the HLB bacterium indicated that it was distributed unevenly in planta and ranged from 14 to 137,031 cells/μg of total DNA in different tissues. A relatively high concentration of ‘Ca. Liberibacter asiaticus’ was observed in fruit peduncles. Our data from greenhouse-infected plants also indicated that ‘Ca. Liberibacter asiaticus’ was transmitted systemically from infection site to different parts of the plant. Understanding the distribution and movement of the HLB bacterium inside an individual citrus tree is critical for discerning its virulence mechanism and to develop management strategies for HLB.

scholarly journals Reaction of sweet orange cultivars expressing the attacin A gene to 'Candidatus Liberibacter asiaticus' infection

2013 ◽  
Vol 48 (11) ◽  
pp. 1440-1448 ◽  
Author(s):  
Rafaella Teles Arantes Felipe ◽  
Francisco de Assis Alves Mourão Filho ◽  
Silvio Aparecido Lopes ◽  
Beatriz Madalena Januzzi Mendes ◽  
Maurel Behling ◽  
...  
Keyword(s):  
Sweet Orange ◽  
Pcr Analysis ◽  
Candidatus Liberibacter Asiaticus ◽  
Randomized Design ◽  
Transgenic Lines ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Polymerase Chain ◽  
Attacin A ◽  
Leaf Symptoms

The objective of this work was to evaluate the reaction of four sweet orange cultivars expressing the attacin A gene to 'Candidatus Liberibacter asiaticus' (Las) infection, a bacterium associated to huanglongbing (HLB) disease. Transgenic sweet orange plants of Hamlin, Natal, Pêra, and Valência cultivars, as well as nontransgenic controls received inocula by grafting budwood sections of HLB-infected branches. Disease progression was evaluated through observations of leaf symptoms and by polymerase chain reaction (PCR) analysis, eight months after inoculation. A completely randomized design was used, with four experiments (one for each cultivar) performed simultaneously. Bacteria title was estimated by quantitative PCR (qPCR). HLB symptoms and Las titers were present in nontransgenic and transgenic plants expressing the attacin A gene of the four sweet orange cultivars, eight months after bacteria inoculation. Five transgenic lines (transformation events) of 'Pêra' sweet orange expressing the attacin A gene have significantly lower Las titers in comparison with nontransgenic plants of this cultivar.

scholarly journals A New Diagnostic System for Detection of ‘Candidatus Liberibacter asiaticus’ Infection in Citrus

Plant Disease ◽  
2013 ◽  
Vol 97 (10) ◽  
pp. 1295-1300 ◽  
Author(s):  
Lianming Lu ◽  
Baoping Cheng ◽  
Jinai Yao ◽  
Aitian Peng ◽  
Danchao Du ◽  
...  
Keyword(s):  
Affinity Chromatography ◽  
Polyclonal Antibodies ◽  
Diagnostic System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Candidatus Liberibacter Asiaticus ◽  
Negative Results ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Polymerase Chain

In this study, two polyclonal antibodies were produced against the Omp protein of ‘Candidatus Liberibacter asiaticus’. First, omp genes were sequenced to exhibit 99.9% identity among 137 isolates collected from different geographical origins. Then, two peptides containing the hydrophobic polypeptide-transport-associated (POTRA) domain and β-barrel domain, respectively, were identified on Omp protein. After that, these two peptides were overexpressed in Escherichia coli and purified by affinity chromatography to immunize the white rabbits. Finally, the antiserum was purified by affinity chromatography. The two Omp antibodies gave positive results (0.454 to 0.633, 1:1,600 dilution) in enzyme-linked immunosorbent assay against ‘Ca. L. asiaticus’-infected samples collected from different geographical origins but revealed negative results against other pathogen-infected, nutrient-deficient and healthy samples. The antibody against the POTRA domain of Omp protein could detect ‘Ca. L. asiaticus’ in 45.7% of the symptomatic samples compared with a 56.2% detection rate with a polymerase chain reaction assay. These new antibodies will provide a very useful supplement to the current approaches to ‘Ca. L. asiaticus’ detection and also provide powerful research tools for tracking distribution of this pathogen in vivo.

scholarly journals Deleções do DNA Mitocondrial no Envelhecimento: Efeito da Disfunção na Fosforilação Oxida tiva

1999 ◽  
Vol 5 (3) ◽  
pp. 65-66
Author(s):  
Celia Harumi Tengan
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Pcr Polymerase Chain Reaction ◽  
Polymerase Chain

Vários estudos descreveram o acúmulo de uma deleção do DNA mitocondrial (DNAmt), denominada de deleção comum, em tecidos pós-mitóticos durante o processo de envelhecimento. Esses achados levaram A hipótese de que radicais livres, gerados dentro da mitocandria, poderiam lesar o DNAmt durante a vida normal. Acredita-se que um defeito no funciomento da cadeia respirat6ra, decorrente da lesão do DNAmt, levaria a um aumento na produção de radicais livres, que por sua vez, lesariam o DNAmt, criando um ciclo vicioso é um fator importante no acúmulo de deleções do DNAmt, pacientes com deficiência da função oxidativa (independente do defeito primário) deveriam apresentar um acúmulo acelerado de deleções do DNAmt. Nós testamos esta hipótese através de três analises: (a) comparação dos níveis de deleção comum em controles normais e pacientes com doenças mitocondriais geneticamente caracterizadas e associadas com uma mutação do DNAmt; (b) análise da cosegregação da deleção comum (associada com o envelhecimento) com uma mutação de ponto patogênica do DNAmt; e (c) detecção de deleções múltiplas do DNAmt através de PCR (polymerase chain reaction) longo em controles e pacientes com doenças mitocondriais. Observamos uma correlação positiva entre a idade e MN/6s de deleção comum em controles (r = 0,80) e pacientes (r = 0,69). As inclinações das curvas eram semelhantes, sugerindo que a taxa de acúmulo da deleção comum associada com a idade era a mesma em ambos os grupos. Não conseguimos observar a co-segregação das moléculas de DNAmt contendo a mutação de ponto com a deleção comum e nem aumento no número de deleções em pacientes. Nossos resultados não suportam a hipótese de que o ciclo vicioso (lesão do DNAmt afeta a função da cadeia respiratória, levando a uma maior produção de radicais livres que, por sua vez, provocaria mais lesão do DNAmt) é um fator importante no acúmulo de deleções do DNAmt no processo de envelhecimento.

scholarly journals Prevalence of Adhesion and Regulation of Biofilm-Related Genes in Different Clones ofStaphylococcus aureus

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Salman Sahab Atshan ◽  
Mariana Nor Shamsudin ◽  
Zamberi Sekawi ◽  
Leslie Than Thian Lung ◽  
Rukman Awang Hamat ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Biofilm Formation ◽  
Clinical Information ◽  
Microtiter Plate ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Microtiter Plate Assay ◽  
High Prevalence ◽  
Different Types ◽  
Polymerase Chain

Clinical information about genotypically different clones of biofilm-producingStaphylococcus aureusis largely unknown. We examined whether different clones of methicillin-sensitive and methicillin-resistantS. aureus(MSSA and MRSA) differ with respect to staphylococcal microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) in biofilm formation. The study used 60 different types ofspaand determined the phenotypes, the prevalence of the 13 MSCRAMM, and biofilm genes for each clone. The current investigation was carried out using a modified Congo red agar (MCRA), a microtiter plate assay (MPA), polymerase chain reaction (PCR), and reverse transcriptase polymerase chain reaction (RT-PCR). Clones belonging to the samespatype were found to have similar properties in adheringto thepolystyrene microtiter plate surface. However, their ability to produce slime on MCRA medium was different. PCR experiments showed that 60 clones of MSSA and MRSA were positive for 5 genes (out of 9 MSCRAMM genes).icaADBCgenes were found to be present in all the 60 clones tested indicating a high prevalence, and these genes were equally distributed among the clones associated with MSSA and those with MRSA. The prevalence of other MSCRAMM genes among MSSA and MRSA clones was found to be variable. MRSA and MSSA gene expression (MSCRAMM andicaADBC) was confirmed by RT-PCR.

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