scholarly journals Quantitative Distribution of Candidatus Liberibacter asiaticus in the Aerial Parts of the Huanglongbing-infected Citrus Trees in Texas

HortScience ◽  
2014 ◽  
Vol 49 (1) ◽  
pp. 65-68 ◽  
Author(s):  
Madhurababu Kunta ◽  
John V. da Graça ◽  
Nasir S.A. Malik ◽  
Eliezer S. Louzada ◽  
Mamoudou Sétamou
Keyword(s):  
Sweet Orange ◽  
Plant Parts ◽  
Citrus Greening Disease ◽  
Bacterial Titer ◽  
Citrus Groves ◽  
Significant Difference ◽  
Bark Tissue ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Orange Trees

The Asian citrus psyllid, Diaphorina citri Kuwayama, one of the known vectors for citrus greening disease or Huanglongbing (HLB) pathogens, has been present in Texas for over a decade, but the detection of the disease is recent. HLB has been confirmed in only two adjacent commercial citrus groves of grapefruit and sweet orange. A study was conducted to compare the population of Candidatus Liberibacter asiaticus (CLas) cells in different plant parts including peduncle, columella, leaves, seeds, young shoots, flower buds, flowers, and bark of 6-year-old known infected grapefruit and sweet orange trees. The bacterial population was estimated using a previously described grand universal regression equation Y = 13.82 – 0.2866X, where Y is the log of the target copy number and X is the Ct (threshold cycle) of the assay. Except for bark tissue, there was no significant difference in the concentration of CLas cells in other plant parts between the two cultivars. Within the cultivar, the bacterial concentration also varied with the plant part, with peduncle, columella, midrib having significantly higher titer of CLas compared with other plant parts. The obtained results here are in agreement with previous studies conducted on Florida samples, but the consistently lowest bacterial titer recorded in young shoots, leaf blade, and especially leaf margins relative to the midrib has never been previously reported.

scholarly journals Identification and Characterization of the Huanglongbing Bacterium in Pummelo from Multiple Locations in Guangdong, P. R. China

Plant Disease ◽  
2008 ◽  
Vol 92 (4) ◽  
pp. 513-518 ◽  
Author(s):  
X. Deng ◽  
J. Chen ◽  
Z. Feng ◽  
Z. Shan ◽  
H. Guo ◽  
...  
Keyword(s):  
Sweet Orange ◽  
Nucleotide Polymorphisms ◽  
Sequence Comparisons ◽  
Research Attention ◽  
Republic Of China ◽  
Citrus Greening Disease ◽  
Citrus Production ◽  
History Of ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

Huanglongbing (HLB, yellow shoot disease, ex. citrus greening disease), caused by Candidatus Liberibacter spp., is highly destructive to citrus production in Asia, Africa, and South America. Although primarily affecting sweet orange and mandarin, HLB has long been observed in pummelo in Guangdong Province, People's Republic of China; however, the disease in pummelo has received little research attention. Accordingly, it remains unclear how closely related the strains of Ca. Liberibacter in pummelo are to those in other citrus species. In this study, the loci of 16S rDNA, rplAJ (β-operon of ribosomal protein), and an outer membrane protein (omp) gene were analyzed and characterized among strains of Ca. Liberibacter in pummelo samples from six different locations in Guangdong. Sequence comparisons indicated that ‘Candidatus Liberibacter asiaticus’, but not ‘Ca. Liberibacter africanus’ or ‘Ca. Liberibacter americanus’, was exclusively associated with HLB symptoms in pummelo. The pummelo strains of ‘Ca. Liberibacter asiaticus’ from Guangdong were highly homogeneous. Analyses of single-nucleotide polymorphisms in the omp locus showed that the Guangdong pummelo strains grouped with ‘Ca. Liberibacter asiaticus’ strains from Thailand, Nepal, and an unspecified location in China but differed from the Philippine and China-Behai strains. Based on the sequence homogeneity at the omp locus, the history of pummelo culture and the means by which HLB is known to be spread, we believe that, likely, the pummelo strain of ‘Ca. Liberibacter asiaticus’ recently was spread to pummelo in the study areas from infected sweet orange or mandarin trees by insect vectors or by propagation of pummelo infected elsewhere.

scholarly journals Comparative Ability of Six Fungicides to Inhibit Development of Phytophthora Gummosis on Citrus

Plant Disease ◽  
2002 ◽  
Vol 86 (6) ◽  
pp. 687-690 ◽  
Author(s):  
M. E. Matheron ◽  
M. Porchas
Keyword(s):  
Sweet Orange ◽  
Lesion Size ◽  
Lesion Development ◽  
Outer Bark ◽  
Significant Difference ◽  
Bark Tissue ◽  
Lesion Growth ◽  
Average Size ◽  
Canker Development ◽  
Orange Trees

The activity of the registered fungicides fosetyl-Al and metalaxyl (subsequently replaced with mefenoxam by the manufacturer) was compared with other potentially useful compounds, azoxystrobin, dimethomorph, fluazinam, and zoxamide, for suppression of canker development on citrus bark after inoculation with Phytophthora citrophthora or P. nicotianae. The number of sweet orange trees on which cankers developed after inoculation with P. citrophthora and the average size of cankers when present were lower on plants treated with dimethomorph, fosetyl-Al, or metalaxyl compared with nontreated trees and those treated with azoxystrobin or fluazinam. When bark removed from treated trees was inoculated with P. citrophthora on the cambium surface at 5, 30, or 60 days after treatment (DAT), inhibition of lesion development on bark strips treated with dimethomorph, fosetyl-Al, or metalaxyl was significantly greater than that detected on bark treated with azoxystrobin, fluazinam, or zoxamide. When inoculated with P. nicotianae at 5 or 30 DAT, reduction of lesion size on bark strips treated with dimethomorph, fosetyl-Al, or metalaxyl was significantly greater than that detected on bark treated with azoxystrobin or fluazinam. Inhibition of lesion development by zoxamide was significantly less than that observed with metalaxyl at 5 DAT on bark inoculated with P. nicotianae; however, at 30, 60, and 90 DAT there was no significant difference in the performance of either fungicide. Reduction of lesion growth on the cambium surface compared with outer bark surface, when inoculated with P. citrophthora, did not differ significantly from 5 to 30 DAT for bark tissue treated with azoxystrobin, dimethomorph, fosetyl-Al, or metalaxyl. Among the nonregistered fungicides tested, dimethomorph provided the best level of Phytophthora gummosis control on citrus.

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 Canopy health, but not Candidatus Liberibacter asiaticus Ct values, are correlated with fruit yield in Huanglongbing affected sweet orange trees

2021 ◽  
Author(s):  
Amit Levy ◽  
Taylor Livingston ◽  
Chunxia Wang ◽  
Diann Achor ◽  
Tripti Vashisth
Keyword(s):  
Sweet Orange ◽  
Field Trials ◽  
Fruit Yield ◽  
Ct Value ◽  
Independent Field ◽  
Canopy Volume ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Almost All ◽  
Orange Trees

AbstractIn Florida, almost all citrus trees are infected with Huanglongbing (HLB), caused by the gram-negative, intracellular phloem limited bacteria Candidatus liberibacter asiaticus (CLas). Distinguishing between the severely and mildly sick trees is important for managing the groves and testing new HLB therapies. A mildly sick tree is one that produces higher fruit yield, compared to a severely sick tree, but measuring yields is laborious and time consuming. Here we characterized HLB affected sweet orange trees in the field in order to identify the specific traits that are correlated with the yields. We found that canopy volume, fruit detachment force (FDF) and the percentage of photosynthetically active radiation interception in the canopy (%INT) were positively correlated with fruit yields. Specifically, %INT measurements accurately distinguished between mild and severe trees in independent field trials. We could not find a difference in the Ct value between high and low producing HLB trees. Moreover, Ct values did not always agree with the number of CLas in the phloem that were visualized by transmission electron microscopy. Overall, our work identified an efficient way to distinguish between severe and mild HLB trees in Florida by measuring %INT and suggests that health of the canopy is more important for yields than the Ct value.

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 Quantitative Distribution of ‘Candidatus Liberibacter asiaticus’ in Citrus Plants with Citrus Huanglongbing

2009 ◽  
Vol 99 (2) ◽  
pp. 139-144 ◽  
Author(s):  
Wenbin Li ◽  
Laurene Levy ◽  
John S. Hartung
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Citrus Greening Disease ◽  
Rdna Sequences ◽  
16S Rdna Sequences ◽  
Candidatus Liberibacter ◽  
Citrus Huanglongbing ◽  
Liberibacter Asiaticus ◽  
Greening Disease ◽  
Individual Trees ◽  
Orange Trees

Citrus huanglongbing (HLB), or greening disease, is strongly associated with any of three nonculturable gram-negative bacteria belonging to ‘Candidatus Liberibacter spp.’ ‘Ca. Liberibacter spp.’ are transmitted by citrus psyllids to all commercial cultivars of citrus. The diseases can be lethal to citrus and have recently become widespread in both São Paulo, Brazil, and Florida, United States, the locations of the largest citrus industries in the world. Asiatic HLB, the form of the disease found in Florida, is associated with ‘Ca. Liberibacter asiaticus’ and is the subject of this report. The nonculturable nature of the pathogen has hampered research and little is known about the distribution of ‘Ca. L. asiaticus’ in infected trees. In this study, we have used a quantitative polymerase chain reaction assay to systematically quantify the distribution of ‘Ca. L. asiaticus’ genomes in tissues of six species of citrus either identified in the field during survey efforts in Florida or propagated in a greenhouse in Beltsville, MD. The populations of ‘Ca. L. asiaticus’ inferred from the distribution of 16S rDNA sequences specific for ‘Ca. L. asiaticus’ in leaf midribs, leaf blades, and bark samples varied by a factor of 1,000 among samples prepared from the six citrus species tested and by a factor of 100 between two sweet orange trees tested. In naturally infected trees, above-ground portions of the tree averaged 1010 ‘Ca. L. asiaticus’ genomes per gram of tissue. Similar levels of ‘Ca. L. asiaticus’ genomes were observed in some but not all root samples from the same plants. In samples taken from greenhouse-inoculated trees, levels of ‘Ca. L. asiaticus’ genomes varied systematically from 104 genomes/g at the graft inoculation site to 1010 genomes/g in some leaf petioles. Root samples from these trees also contained ‘Ca. L. asiaticus’ at 107 genomes/g. In symptomatic fruit tissues, ‘Ca. L. asiaticus’ genomes were also readily detected and quantified. The highest levels of ‘Ca. L. asiaticus’ in fruit tissues were found in the locular membranes and septa (108 genomes/g), with 100-fold lower levels of ‘Ca. L. asiaticus’ in the meso and pericarp of such fruit. Our results demonstrate both the ubiquitous presence of ‘Ca. L. asiaticus’ in symptomatic citrus trees as well as great variation between individual trees and among samples of different tissues from the same trees. Our methods will be useful in both the management and scientific study of citrus HLB, also known as citrus greening disease.

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.

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