scholarly journals Rapid molecular detection of Candidatus Liberibacter asiaticus, the bacterium associated with citrus huanglongbing (greening) disease of Mandarin using polymerase chain reaction

2013 ◽  
Vol 12 (29) ◽  
pp. 4623-4629
Author(s):  
V Mahajan M ◽  
P Jagtap G ◽  
Dey Utpal
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Detection ◽  
Candidatus Liberibacter Asiaticus ◽  
Chain Reaction ◽  
Candidatus Liberibacter ◽  
Citrus Huanglongbing ◽  
Liberibacter Asiaticus ◽  
Polymerase Chain ◽  
Greening Disease

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 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 Live Population Dynamics of ‘Candidatus Liberibacter asiaticus’, the Bacterial Agent Associated with Citrus Huanglongbing, in Citrus and Non-Citrus Hosts

Plant Disease ◽  
2014 ◽  
Vol 98 (7) ◽  
pp. 876-884 ◽  
Author(s):  
H. Hu ◽  
Avijit Roy ◽  
R. H. Brlansky
Keyword(s):  
Bacterial Population ◽  
Candidatus Liberibacter Asiaticus ◽  
Bacterial Populations ◽  
Bacterial Agent ◽  
New Findings ◽  
Candidatus Liberibacter ◽  
Citrus Huanglongbing ◽  
Liberibacter Asiaticus ◽  
Polymerase Chain ◽  
Severinia Buxifolia

Citrus huanglongbing (HLB) is a century-old destructive disease which presents an unprecedented challenge to citrus industries worldwide. In Florida, HLB is associated with the phloem-limited bacterium ‘Candidatus Liberibacter asiaticus’ and is mainly transmitted by Asian citrus psyllid (Diaphorina citri). Quantification of the pathogen population in a host aids in investigation of virulence mechanisms and disease management. Recently a procedure was developed to detect live bacterial populations using a novel DNA-binding dye, propidium monoazide, in conjunction with real-time polymerase chain reaction (PMA-qPCR). Chinese box orange (Severinia buxifolia) is a common ornamental present in Florida which could host D. citri and ‘Ca. L. asiaticus’. For 20 months, the change of the live ‘Ca. L. asiaticus’ populations in graft- and psyllid-transmitted Valencia sweet orange (Citrus sinensis ‘Valencia’) and S. buxifolia plants was monitored by PMA-qPCR. Our results showed that the live ‘Ca. L. asiaticus’ population was significantly lower in the months of December, January, and February than the rest of the year in both hosts. No statistically significant pattern in the total bacterial population was observed in either host. This pattern may indicate a seasonal growth of ‘Ca. L. asiaticus’ along with the growth of both plants. These new findings should provide useful information on HLB management.

scholarly journals Detection of ‘Candidatus Liberibacter asiaticus’ in Diaphorina citri and Its Importance in the Management of Citrus Huanglongbing in Florida

2008 ◽  
Vol 98 (4) ◽  
pp. 387-396 ◽  
Author(s):  
K. L. Manjunath ◽  
S. E. Halbert ◽  
C. Ramadugu ◽  
S. Webb ◽  
R. F. Lee
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
Citrus Production ◽  
Candidatus Liberibacter ◽  
Citrus Huanglongbing ◽  
Liberibacter Asiaticus ◽  
Polymerase Chain ◽  
Polymerase Chain Reaction Methodology

Citrus huanglongbing (HLB or citrus greening), is a highly destructive disease that has been spreading in both Florida and Brazil. Its psyllid vector, Diaphorina citri Kuwayama, has spread to Texas and Mexico, thus threatening the future of citrus production elsewhere in mainland North America. Even though sensitive diagnostic methods have been developed for detection of the causal organisms, Candidatus Liberibacter spp., the pathogen cannot be detected consistently in plants until symptoms develop, presumably because of low titer and uneven distribution of the causal bacteria in nonsymptomatic tissues. In the present study, TaqMan based real-time quantitative polymerase chain reaction methodology was developed for detection of ‘Ca. L. asiaticus’ in D. citri. Over 1,200 samples of psyllid adults and nymphs, collected from various locations in Florida, from visually healthy and HLB symptomatic trees at different times of the year were analyzed to monitor the incidence and spread of HLB. The results showed that spread of ‘Ca. L. asiaticus’ in an area may be detected one to several years before the development of HLB symptoms in plants. The study suggests that discount garden centers and retail nurseries may have played a significant role in the widespread distribution of psyllids and plants carrying HLB pathogens in Florida.

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