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Changes in Variable Number of Tandem Repeats in 'Candidatus Liberibacter asiaticus' through Insect Transmission.

Katoh H, Inoue H, Iwanami T - PLoS ONE (2015)

Bottom Line: The disease is associated with three species of 'Candidatus Liberibacter' among which 'Ca.The results showed that some tandem repeats showed detectable changes after insect transmission.In contrast, changes in the number of tandem repeats increased with longer acquisition and inoculation access periods, whereas changes were not observed through psyllid transmission after relatively short acquisition and inoculation access periods, up to 20 and 19 days, respectively.

View Article: PubMed Central - PubMed

Affiliation: NARO Institute of Fruit Tree Science, Fujimoto 2-1, Tsukuba, Ibaraki 305-8605, Japan.

ABSTRACT
Citrus greening (huanglongbing) is the most destructive citrus disease worldwide. The disease is associated with three species of 'Candidatus Liberibacter' among which 'Ca. Liberibacter asiaticus' has the widest distribution. 'Ca. L. asiaticus' is commonly transmitted by a phloem-feeding insect vector, the Asian citrus psyllid Diaphorina citri. A previous study showed that isolates of 'Ca. L. asiaticus' were clearly differentiated by variable number of tandem repeat (VNTR) profiles at four loci in the genome. In this study, the VNTR analysis was further validated by assessing the stability of these repeats after multiplication of the pathogen upon host-to-host transmission using a 'Ca. L. asiaticus' strain from Japan. The results showed that some tandem repeats showed detectable changes after insect transmission. To our knowledge, this is the first report to demonstrate that the repeat numbers VNTR 002 and 077 of 'Ca. L. asiaticus' change through psyllid transmission. VNTRs in the recipient plant were apparently unrelated to the growing phase of the vector. In contrast, changes in the number of tandem repeats increased with longer acquisition and inoculation access periods, whereas changes were not observed through psyllid transmission after relatively short acquisition and inoculation access periods, up to 20 and 19 days, respectively.

No MeSH data available.


Related in: MedlinePlus

Schematic model of strategy of Table 2.The black circle indicates the area used for DNA extraction. Numbers at the lower right, which are x n, indicate the number of samples in Table 2.
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pone.0138699.g001: Schematic model of strategy of Table 2.The black circle indicates the area used for DNA extraction. Numbers at the lower right, which are x n, indicate the number of samples in Table 2.

Mentions: As for nymphal acquisition–adult transmission tests, healthy fifth instars maintained on ‘Ca. L. asiaticus’-negative M. paniculata were transferred to an infected rough lemon tree with a fine brush. These nymphs became adults during the acquisition feeding. After an AAP of 20–98 days, each adult individual was transferred to a healthy yuzu, tankan mandarin, unzoki, or orange jessamine seedling for an IAP of 4–23 days. The number of insect used for each IAP is dictate in Fig 1. Recipient plants were maintained for 3–4 months in a temperature-controlled greenhouse until DNA exrtraction. After inoculation feeding, all psyllids were collected and preserved at –50°C until DNA extraction. For all psyllid transmission experiment, single psyllid was used in IAP per plant. Twenty-seven of 144 inoculative psyllids died during IAPs (mortality rate = 18.8%) and the pathogen was successfully transmitted to 42 of 144 recipient plants by psyllids (transmission rate = 29.2%). Inoculative psyllid DNA samples from which sufficient copies of ‘Ca. L. asiaticus’ genome were detected by quantitative real-time PCR assays [24] were used for the subsequent analysis of VNTRs of the “Ca, L, asiaticus” genome in psyllids.


Changes in Variable Number of Tandem Repeats in 'Candidatus Liberibacter asiaticus' through Insect Transmission.

Katoh H, Inoue H, Iwanami T - PLoS ONE (2015)

Schematic model of strategy of Table 2.The black circle indicates the area used for DNA extraction. Numbers at the lower right, which are x n, indicate the number of samples in Table 2.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581716&req=5

pone.0138699.g001: Schematic model of strategy of Table 2.The black circle indicates the area used for DNA extraction. Numbers at the lower right, which are x n, indicate the number of samples in Table 2.
Mentions: As for nymphal acquisition–adult transmission tests, healthy fifth instars maintained on ‘Ca. L. asiaticus’-negative M. paniculata were transferred to an infected rough lemon tree with a fine brush. These nymphs became adults during the acquisition feeding. After an AAP of 20–98 days, each adult individual was transferred to a healthy yuzu, tankan mandarin, unzoki, or orange jessamine seedling for an IAP of 4–23 days. The number of insect used for each IAP is dictate in Fig 1. Recipient plants were maintained for 3–4 months in a temperature-controlled greenhouse until DNA exrtraction. After inoculation feeding, all psyllids were collected and preserved at –50°C until DNA extraction. For all psyllid transmission experiment, single psyllid was used in IAP per plant. Twenty-seven of 144 inoculative psyllids died during IAPs (mortality rate = 18.8%) and the pathogen was successfully transmitted to 42 of 144 recipient plants by psyllids (transmission rate = 29.2%). Inoculative psyllid DNA samples from which sufficient copies of ‘Ca. L. asiaticus’ genome were detected by quantitative real-time PCR assays [24] were used for the subsequent analysis of VNTRs of the “Ca, L, asiaticus” genome in psyllids.

Bottom Line: The disease is associated with three species of 'Candidatus Liberibacter' among which 'Ca.The results showed that some tandem repeats showed detectable changes after insect transmission.In contrast, changes in the number of tandem repeats increased with longer acquisition and inoculation access periods, whereas changes were not observed through psyllid transmission after relatively short acquisition and inoculation access periods, up to 20 and 19 days, respectively.

View Article: PubMed Central - PubMed

Affiliation: NARO Institute of Fruit Tree Science, Fujimoto 2-1, Tsukuba, Ibaraki 305-8605, Japan.

ABSTRACT
Citrus greening (huanglongbing) is the most destructive citrus disease worldwide. The disease is associated with three species of 'Candidatus Liberibacter' among which 'Ca. Liberibacter asiaticus' has the widest distribution. 'Ca. L. asiaticus' is commonly transmitted by a phloem-feeding insect vector, the Asian citrus psyllid Diaphorina citri. A previous study showed that isolates of 'Ca. L. asiaticus' were clearly differentiated by variable number of tandem repeat (VNTR) profiles at four loci in the genome. In this study, the VNTR analysis was further validated by assessing the stability of these repeats after multiplication of the pathogen upon host-to-host transmission using a 'Ca. L. asiaticus' strain from Japan. The results showed that some tandem repeats showed detectable changes after insect transmission. To our knowledge, this is the first report to demonstrate that the repeat numbers VNTR 002 and 077 of 'Ca. L. asiaticus' change through psyllid transmission. VNTRs in the recipient plant were apparently unrelated to the growing phase of the vector. In contrast, changes in the number of tandem repeats increased with longer acquisition and inoculation access periods, whereas changes were not observed through psyllid transmission after relatively short acquisition and inoculation access periods, up to 20 and 19 days, respectively.

No MeSH data available.


Related in: MedlinePlus