scholarly journals Early Events of Citrus Greening (Huanglongbing) Disease Development at the Ultrastructural Level

2010 ◽  
Vol 100 (9) ◽  
pp. 949-958 ◽  
Author(s):  
Svetlana Y. Folimonova ◽  
Diann S. Achor

Citrus greening (Huanglongbing [HLB]) is one of the most destructive diseases of citrus worldwide. The causal agent of HLB in Florida is thought to be ‘Candidatus Liberibacter asiaticus’. Understanding of the early events in HLB infection is critical for the development of effective measures to control the disease. In this work, we conducted cytopathological studies by following the development of the disease in citrus trees graft inoculated with ‘Ca. L. asiaticus’-containing material under greenhouse conditions to examine the correlation between ultrastructural changes and symptom production, with the main objective of characterizing the early events of infection. Based on our observations, one of the first degenerative changes induced upon invasion of the pathogen appears to be swelling of middle lamella between cell walls surrounding sieve elements. This anatomical aberration was often observed in samples from newly growing flushes in inoculated sweet orange and grapefruit trees at the early “presymptomatic” stage of HLB infection. Development of symptoms and their progression correlated with an increasing degree of microscopic aberrations. Remarkably, the ability to observe the bacterium in the infected tissue also correlated with the degree of the disease progression. Large numbers of bacterial cells were found in phloem sieve tubes in tissue samples from presymptomatic young flushes. In contrast, we did not observe the bacteria in highly symptomatic leaf samples, suggesting a possibility that, at more advanced stages of the disease, a major proportion of ‘Ca. L. asiaticus’ is present in a nonviable state. We trust that observations reported here advance our understanding of how ‘Ca. L. asiaticus’ causes disease. Furthermore, they may be an important aid in answering a question: when and where within an infected tree the tissue serves as a better inoculum source for acquisition and transmission of the bacterium by its psyllid vector.

Insects ◽  
2019 ◽  
Vol 10 (12) ◽  
pp. 422 ◽  
Author(s):  
El-Desouky Ammar ◽  
Diann Achor ◽  
Amit Levy

Huanglongbing, the most destructive citrus disease worldwide, is caused by the bacterium ‘Candidatus Liberibacter asiaticus’ (CLas) and is vectored by the Asian citrus psyllid (ACP). Very little is known about the form and distribution of CLas in infected psyllids, especially at the ultrastructural level. Here, we examined these aspects by transmission electron microscopy, combined with immunogold labeling. In CLas-exposed ACP adults, the CLas bacterial cells were found to be pleomorphic taking tubular, spherical, or flask-shaped forms, some of which seemed to divide further. Small or large aggregates of CLas were found in vacuolated cytoplasmic pockets of most ACP organs and tissues examined, including the midgut, filter chamber, hindgut, Malpighian tubules, and secretory cells of the salivary glands, in addition to fat tissues, epidermis, muscle, hemocytes, neural tissues, bacteriome, and walls of the female spermatheca and oviduct. Large aggregates of CLas were found outside the midgut within the filter chamber and between the sublayers of the basal lamina of the hindgut and Malpighian tubules. Novel intracytoplasmic structures that we hypothesized as ‘putative CLas multiplication sites’ were found in the cells of the midgut, salivary glands, and other tissues in CLas-exposed ACP. These structures, characterized by containing a granular matrix and closely packed bacterial cells, were unbound by membranes and were frequently associated with rough endoplasmic reticulum. Our results point to the close association between CLas and its psyllid vector, and provide support for a circulative-propagative mode of transmission.


2010 ◽  
Vol 100 (8) ◽  
pp. 756-762 ◽  
Author(s):  
John S. Hartung ◽  
Cristina Paul ◽  
Diann Achor ◽  
R. H. Brlansky

Huanglongbing, or citrus greening, threatens the global citrus industry. The presumptive pathogens, ‘Candidatus Liberibacter asiaticus’ and ‘Ca. L. americanus’ can be transferred from citrus to more easily studied experimental hosts by using holoparasitic dodder plants. However, the interaction between ‘Candidatus Liberibacter’ spp. and the dodder has not been studied. We combined quantitative polymerase chain reaction with electron microscopy to show that only 65% of tendrils of Cuscuta indecora grown on ‘Ca. Liberibacter’ spp.-infected host plants had detectable levels of the pathogen. Among tendrils that were colonized by Liberibacter in at least one 2 cm segment, most were not colonized in all segments. Furthermore, the estimated population levels of the pathogen present in serial 2 cm segments of dodder tendrils varied widely and without any consistent pattern. Thus, there was generally not a concentration gradient of the pathogen from the source plant towards the recipient and populations of the pathogen were sometimes found in the distal segments of the dodder plant but not in the proximal or middle segments. Populations of the pathogens ranged from 2 × 102 to 3.0 × 108 cells per 2 cm segment. On a fresh weight basis, populations as high as 1.4 × 1010 cells per g of tissue were observed demonstrating that ‘Ca. Liberibacter’ spp. multiplies well in Cuscuta indecora. However, 55% of individual stem segments did not contain detectable levels of the pathogen, consistent with a pattern of nonuniform colonization similar to that observed in the much more anatomically complex citrus tree. Colonization of dodder by the pathogen is also nonuniform at the ultrastructural level, with adjacent phloem vessel elements being completely full of the pathogen or free of the pathogen. We also observed bacteria in the phloem vessels that belonged to two distinct size classes based on the diameters of cross sections of cells. In other sections from the same tendrils we observed single bacterial cells that were apparently in the process of differentiating between the large and round forms to the long and thin forms (or vice versa). The process controlling this morphological differentiation of the pathogen is not known. The highly reduced and simplified anatomy of the dodder plant as well as its rapid growth rate compared with citrus, and the ability of the plant to support multiplication of the pathogen to high levels, makes it an interesting host plant for further studies of host–pathogen interactions.


Weed Science ◽  
1968 ◽  
Vol 16 (3) ◽  
pp. 377-380 ◽  
Author(s):  
E. R. Hill ◽  
E. C. Putala ◽  
J. Vengris

Electronmicrographs of barnyardgrass (Echinochloa crusgalli (L.) Beauv.) grown under controlled conditions and treated with 2-chloro-4-ethylamino-6-isopropylamino-s-triazine (atrazine) at the one to three-leaf stages show that degradation of the chloroplast starts as a swelling of the fret system followed by swelling and disruption of the granal discs. In advanced stages of breakdown, the membranes of the grana and chloroplast envelope were ruptured. These changes were initiated 2 hr after treatment with rates of 2, 5, 10, and 20 ppm atrazine. The incidence of starch grains greatly decreased as the duration of treatment exceeded 4 hr. The mitochondria appeared normal throughout, not being affected by concentration or duration of treatment. The stage of growth at which plants were treated was insignificant in determining changes at the ultrastructural level. The ultrastructural changes preceded any macroscopically discernable symptoms of tissue breakdown.


Author(s):  
K.W. Lee ◽  
R.H. Meints ◽  
D. Kuczmarski ◽  
J.L. Van Etten

The physiological, biochemical, and ultrastructural aspects of the symbiotic relationship between the Chlorella-like algae and the hydra have been intensively investigated. Reciprocal cross-transfer of the Chlorellalike algae between different strains of green hydra provide a system for the study of cell recognition. However, our attempts to culture the algae free of the host hydra of the Florida strain, Hydra viridis, have been consistently unsuccessful. We were, therefore, prompted to examine the isolated algae at the ultrastructural level on a time course.


Author(s):  
Amankwah K.S. ◽  
A.D. Weberg ◽  
R.C. Kaufmann

Previous research has revealed that passive (involuntary inhalation) tobacco smoking during gestation can have adverse effects upon the developing fetus. These prior investigations did not concentrate on changes in fetal morphology. This study was undertaken to delineate fetal neural abnormalities at the ultrastructural level in mice pups exposed in utero to passive maternal smoking.Pregnant study animals, housed in a special chamber, were subjected to cigarette smoke daily from conception until delivery. Blood tests for determination of carbon monoxide levels were run at 15-18 days gestation. Sciatic nerve tissue from experimental and control animals were obtained following spontaneous delivery and fixed in 2.5% gluteraldehyde in 0.1M cacodylate buffer pH 7.3. The samples were post-fixed in osmium ferrocyanide (1:1 mixture of 1.5% aqueous OSO4 and 2.5% K4 Fe(CN)6). Following dehydration, the tissues were infiltrated with and embedded in Spurr. Sections were stained with uranyl acetate and lead citrate.


Author(s):  
H. J. Finol ◽  
M. E. Correa ◽  
L.A. Sosa ◽  
A. Márquez ◽  
N.L. Díaz

In classical oncological literature two mechanisms for tissue aggression in patients with cancer have been described. The first is the progressive invasion, infiltration and destruction of tissues surrounding primary malignant tumor or their metastases; the other includes alterations produced in remote sites that are not directly affected by any focus of disease, the so called paraneoplastic phenomenon. The non-invaded tissue which surrounds a primary malignant tumor or its metastases has been usually considered a normal tissue . In this work we describe the ultrastructural changes observed in hepatocytes located next to metastases from diverse malignant tumors.Hepatic biopsies were obtained surgically in patients with different malignant tumors which metatastized in liver. Biopsies included tumor mass, the zone of macroscopic contact between the tumor and the surrounding tissue, and the tissue adjacent to the tumor but outside the macroscopic area of infiltration. The patients (n = 5), 36–75 years old, presented different tumors including rhabdomyosarcoma, leiomyosarcoma, pancreas carcinoma, biliar duct carcinoma and colon carcinoma. Tissue samples were processed with routine techniques for transmission electron microscopy and observed in a Hitachi H-500 electron microscope.


Author(s):  
E. M. B. Sorensen ◽  
R. R. Mitchell ◽  
L. L. Graham

Endemic freshwater teleosts were collected from a portion of the Navosota River drainage system which had been inadvertently contaminated with arsenic wastes from a firm manufacturing arsenical pesticides and herbicides. At the time of collection these fish were exposed to a concentration of 13.6 ppm arsenic in the water; levels ranged from 1.0 to 20.0 ppm during the four-month period prior. Scale annuli counts and prior water analyses indicated that these fish had been exposed for a lifetime. Neutron activation data showed that Lepomis cyanellus (green sunfish) had accumulated from 6.1 to 64.2 ppm arsenic in the liver, which is the major detoxification organ in arsenic poisoning. Examination of livers for ultrastructural changes revealed the presence of electron dense bodies and large numbers of autophagic vacuoles (AV) and necrotic bodies (NB) (1), as previously observed in this same species following laboratory exposures to sodium arsenate (2). In addition, abnormal lysosomes (AL), necrotic areas (NA), proliferated rough endoplasmic reticulum (RER), and fibrous bodies (FB) were observed. In order to assess whether the extent of these cellular changes was related to the concentration of arsenic in the liver, stereological measurements of the volume and surface densities of changes were compared with levels of arsenic in the livers of fish from both Municipal Lake and an area known to contain no detectable level of arsenic.


2020 ◽  
Vol 22 (99) ◽  
pp. 63-68
Author(s):  
O. М. Shchebentovska ◽  
A. K. Kostyniuk

Liver problems of various etiologies in turkeys have been reported in many countries for the last 20 years. Poultry dies having no clinical manifestations of the disease, and at pathological autopsy, diffuse haemorrhages and marked dystrophic changes of the organ are noted. To date, there are several factors that can cause such changes, these are unbalanced amino-acid feed, insufficient calcium, biotin, selenium, the very high energy content of feed; zootechnical factors – limited mobility of birds due to cage density violations, high temperature; genetic factors – the influence of estrogens; infectious factors – E. coli, Clostridium, and viruses of Picornaviridae family. The article describes the histopathological and ultrastructural changes in the turkey liver under the influence of various factors. The material for the research was obtained from a farm where turkeys of the “Hybrid Converter” cross are grown, same age, fed with a standard diet that changed according to the technological map of cultivation. On the 50th day of life, a pathological autopsy of the dead poultry was performed, pieces of liver were selected for histological and ultrastructural examination. The visual assessment revealed significantly enlarged liver, the colour from dark red to light brown, flabby consistency. In some cases, diffuse fatty infiltrations of hepatocytes were histologically revealed, in other cases, focal necrosis with the growth of the connective tissue and the formation of massive perivascular couplings were registered. Large vacuolar fatty degeneration of hepatocytes with subsequent development of fibrosis indicates chronic intoxication, probably caused by slow breakdown of fatty acids in cells due to insufficient oxidative phosphorylation, as well as reduced levels of lipotropic factors: choline, methionine and the vitamins. At the ultrastructural level, a large number of lipid inclusions of various sizes, dystrophic changes in mitochondria were observed, which indicates a decrease in the synthetic activity of cells.


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