Census of Candidatus Liberibacter asiaticus population inside the phloem of citrus trees

2020 ◽  
Author(s):  
Fernanda N.C. Vasconcelos ◽  
Jinyun Li ◽  
zhiqian pang ◽  
Christopher Vincent ◽  
Nian Wang
Keyword(s):  
Population Size ◽  
Sweet Orange ◽  
Bacterial Pathogens ◽  
Candidatus Liberibacter Asiaticus ◽  
Metabolic Burden ◽  
Whole Plant ◽  
Candidatus Liberibacter ◽  
Leaf Tissues ◽  
Liberibacter Asiaticus ◽  
Orange Trees

Candidatus Liberibacter asiaticus (CLas) is the predominant causal agent of citrus huanglongbing (HLB). The pathogen population size in local tissues and the whole plant are critical for the development of disease symptoms via pathogenicity factors and causing metabolic burden to the host. However, the total population size of CLas in a whole plant and the ratio of CLas vs. citrus cells in local tissues have not been addressed previously. The total CLas population size for 2.5-year-old Valencia sweet orange trees was quantified using quantitative PCR to be approximately 1.74 x 109, whereas that of 7 and 20-year-old sweet orange trees were estimated to be 4.3 x 1010, and 6.0 x 1010, respectively. The majority of CLas cells were distributed in the leaf tissues (55.58%), followed by that in the branch tissues (36.78%), feeder roots (4.75%), trunk (2.39%), and structural root (0.51%) tissues. The ratios of citrus cells vs. CLas cells for branch, leaf, trunk, feeder root, and structural root samples were approximately 39, 44, 153, 191, and 561, respectively, representing the metabolic burden of CLas in different organs. Approximately 0.01% of the total citrus phloem volume was estimated to be occupied by CLas. The CLas titer inside the leaf was estimated to be approximately 1.64 x 106 cells/leaf or 9.2 x 104 cells cm-2 in leaves, approximately 104 times less than that of typical apoplastic bacterial pathogens. This study provides quantitative estimates of phloem colonization by bacterial pathogens and further understands the biology and virulence mechanism of CLas.

scholarly journals Ultrastructural Changes and Putative Phage Particles Observed in Sweet Orange Leaves Infected with ‘Candidatus Liberibacter asiaticus’

Plant Disease ◽  
2015 ◽  
Vol 99 (3) ◽  
pp. 320-324 ◽  
Author(s):  
S. M. Fu ◽  
John Hartung ◽  
C. Y. Zhou ◽  
H. N. Su ◽  
J. Tan ◽  
...  
Keyword(s):  
Sweet Orange ◽  
Light And Electron Microscopy ◽  
Ultrastructural Changes ◽  
Candidatus Liberibacter Asiaticus ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Orange Trees ◽  
First Time ◽  
Citrus Disease ◽  
Putative Phage

Huanglongbing (HLB), also known as citrus greening, is currently the most destructive citrus disease. Anatomical analyses of HLB-affected sweet orange were carried out by light and electron microscopy. As compared with healthy citrus, the phloem plasmodesmata were plugged with callose, and in some samples the phloem was collapsed. Chloroplast structures were deformed. Prophage sequences occupy a significant portion of the genome of ‘Candidatus Liberibacter asiaticus’ and have been used to distinguish strains from Yunnan and Guangdong provinces in China and Florida. Interestingly, a large number of possible putative phage particles were observed attached on the surface of ‘Ca. L. asiaticus’ cells in plants inoculated with strain FJ3 from Fujian Province, China. Phage particles have been observed previously only in periwinkle plants artificially inoculated in Florida with ‘Ca. L. asiaticus’ that carried the SC1-type prophage. PCR assays verified the presence of the SC1-type prophage sequences previously described from this bacterium in Florida in the FJ3 isolate. This is the first time that suspected phage particles have been observed in sweet orange trees infected with ‘Ca. L. asiaticus.’

scholarly journals Liberibacters Associated with Citrus Huanglongbing in Brazil: ‘Candidatus Liberibacter asiaticus’ Is Heat Tolerant, ‘Ca. L. americanus’ Is Heat Sensitive

Plant Disease ◽  
2009 ◽  
Vol 93 (3) ◽  
pp. 257-262 ◽  
Author(s):  
S. A. Lopes ◽  
G. F. Frare ◽  
E. Bertolini ◽  
M. Cambra ◽  
N. G. Fernandes ◽  
...  
Keyword(s):  
Sweet Orange ◽  
Sao Paulo ◽  
São Paulo ◽  
Candidatus Liberibacter Asiaticus ◽  
Paulo State ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
São Paulo State ◽  
Heat Tolerant ◽  
Orange Trees

In São Paulo State, Brazil, ‘Candidatus Liberibacter americanus’ and ‘Candidatus Liberibacter asiaticus’ are associated with huanglongbing (HLB). Affected municipalities occur mainly in the central and southern regions, where the annual number of hours above 30°C is two to five times lower than that in the extreme northern and western regions. The influence of temperature on sweet orange trees infected with ‘Ca. L. asiaticus’ or ‘Ca. L. americanus’ was studied in temperature-controlled growth chambers. Symptom progression on new shoots of naturally infected and experimentally graft-inoculated symptomatic sweet orange trees was assessed. Mottled leaves developed on all infected trees at 22 to 24°C, but not on any ‘Ca. L. americanus’–infected trees at 27 to 32°C. Quantitative, real time-PCR was used to determine the liberibacter titers in the trees. After 90 days, ‘Ca. L. asiaticus’–infected trees had high titers at 32 and 35°C, but not at 38°C, while ‘Ca. L. americanus’–infected trees had high titers at 24°C, but at 32°C the titers were very low or the liberibacters could not be detected. Thus, the multiplication of ‘Ca. L. asiaticus’ is not yet affected at 35°C, while a temperature of 32°C is detrimental to ‘Ca. L. americanus’. Thus, ‘Ca. L. americanus’ is less heat tolerant than ‘Ca. L. asiaticus’. The uneven distribution of these two liberibacters in São Paulo State might be in relation with these results.

scholarly journals Evidence That ‘Candidatus Liberibacter asiaticus’ Moves Predominantly Toward New Tissue Growth in Citrus Plants

Plant Disease ◽  
2021 ◽  
Vol 105 (1) ◽  
pp. 34-42 ◽  
Author(s):  
Laudecir L. Raiol-Junior ◽  
Juan C. Cifuentes-Arenas ◽  
Everton V. de Carvalho ◽  
Eduardo A. Girardi ◽  
Silvio A. Lopes
Keyword(s):  
Sweet Orange ◽  
Force Production ◽  
Tissue Growth ◽  
Candidatus Liberibacter Asiaticus ◽  
In Planta ◽  
Potted Plants ◽  
Candidatus Liberibacter ◽  
Leaf Tissues ◽  
Basal Portion ◽  
Liberibacter Asiaticus

‘Candidatus Liberibacter asiaticus’ (Las) is an unculturable, phloem-limited, insect-transmitted bacterium associated with the Asiatic form of huanglongbing (HLB), the most destructive citrus disease. In Asia and the Americas, it is transmitted by the Asian citrus psyllid (Diaphorina citri Kuwavama). Despite considerable research, little is known about the processes involved in plant infection and colonization by Las. This study was conducted to determine whether the basal portion (below girdling) of the plant is an important route for Las to move laterally from a point of inoculation on a branch to pathogen-free branches elsewhere in the canopy, and to quantify the influence of actively growing tissues on vertical upward (acropetally) or downward (basipetally) movement of Las. Nongirdled and fully or partially girdled stems of potted plants of ‘Pera’ sweet orange, graft-inoculated above or below girdling, were sampled in distinct regions and assessed by qPCR, 6 months postinoculation. Las invaded all regions of partially and nongirdled plants but remained restricted to the inoculated regions of fully girdled plants, evidence that in planta bacterium movement is limited to the phloem. In fully girdled plants, starch accumulated above the girdling site, probably because of changes in flow of phloem sap. To study the influence of actively growing tissues, inoculated ‘Valencia’ sweet orange plants were kept intact or were top- or root-pruned to force production of new tissues, and sampled at 15-day intervals. Las migrated rapidly and most predominantly toward newly developing root and leaf tissues. The rapid and predominant movement of Las to newly developed shoots and roots would explain failures of canopy heat treatments and pruning to cure HLB-affected trees, and reinforces the need to protect rapidly growing new shoots from feeding by D. citri in order to minimize transmission and spread of the pathogen by the vector within and between orchards.

A Review of Huanglongbing (Citrus Greening) Management in Citrus Using Nutritional Approaches in China

2011 ◽  
Vol 12 (1) ◽  
pp. 24 ◽  
Author(s):  
Yulu Xia ◽  
Gecheng Ouyang ◽  
Ronald A. Sequeira ◽  
Yu Takeuchi ◽  
Ignacio Baez ◽  
...  
Keyword(s):  
Nutrient Management ◽  
Field Survey ◽  
Cultural Practices ◽  
Sweet Orange ◽  
Asian Citrus Psyllid ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
Candidatus Liberibacter ◽  
Citrus Maxima ◽  
Liberibacter Asiaticus

The Asian form of huanglongbing (HLB) is caused by ‘Candidatus Liberibacter asiaticus (Las),’ a phloem-limited bacterium transmitted by the Asian citrus psyllid, Diaphorina citri Kuwayama. Nutrient management, together with other cultural practices such as pruning and irrigation, for mitigation of the disease has been practiced in China for many years. Our literature review, field survey, and interviews with Chinese scientists and growers indicate that these cultural practices were generally ineffective for the disease management. However, a nutritional approach in conjunction with other cultural practices such as irrigation can maintain grove productivity for a certain time depending on the type of citrus species/cultivars, the age of the trees, the propagation method of the plants, the Asian citrus psyllid (ACP) (Diaphorina citri Kuwayama) population, and other factors. Symptomatic mature pommelo (Citrus maxima Merr) and sweet orange (C. sinensis L. Osbeck) plants can commonly survive and maintain a certain level of productivity for an additional 4 to 5 years, even longer assuming vigorous ACP control. Accepted for publication 27 June 2011. Published 3 October 2011.

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.

Comparative iTRAQ proteome and transcriptome analyses of sweet orange infected by “Candidatus Liberibacter asiaticus”

2011 ◽  
Vol 143 (3) ◽  
pp. 235-245 ◽  
Author(s):  
Jing Fan ◽  
Chunxian Chen ◽  
Qibin Yu ◽  
Ronald H. Brlansky ◽  
Zheng-Guo Li ◽  
...  
Keyword(s):  
Sweet Orange ◽  
Candidatus Liberibacter Asiaticus ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Transcriptome Analyses
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