Morphology and lipid body and vacuole dynamics during secondary conidia formation inColletotrichum acutatum: laser scanning confocal analysis

2006 ◽  
Vol 52 (2) ◽  
pp. 117-124 ◽  
Author(s):  
Ariani Corrêa Barbosa ◽  
Anousca Evelyn do Carmo ◽  
Letícia Graf ◽  
Roberto Tomaz ◽  
Caroline Fogaça de Souza ◽  
...  

Colletotrichum acutatum may develop one or more secondary conidia after conidial germination and before mycelial growth. Secondary conidia formation and germination were influenced by conidia concentration. Concentrations greater than 1 × 105conidia/mL were associated with germination decrease and with secondary conidia emergence. Secondary conidia can form either alone or simultaneously with germ tubes and appressoria. Confocal analysis showed numerous lipid bodies stored inside ungerminated conidia, which diminished during germ tube and appressoria formation, with or without secondary conidia formation. They were also reduced during secondary conidia formation alone. While there was a decrease inside germinated conidia, lipid bodies appeared inside secondary conidia since the initial stages. Intense vacuolization inside primary germinated conidia occurred at the same time as the decrease in lipid bodies, which were internalized and digested by vacuoles. During these events, small acidic vesicles inside secondary conidia were formed. Considering that the conidia were maintained in distilled water, with no exogenous nutrients, it is clear that ungerminated conidia contain enough stored lipids to form germ tubes, appressoria, and the additional secondary conidia replete with lipid reserves. These results suggested a very complex and well-balanced regulation that makes possible the catabolic and anabolic pathways of these lipid bodies.Key words: secondary conidia, lipid bodies, vacuoles, confocal microscopy, Colletotrichum.


1994 ◽  
Vol 72 (12) ◽  
pp. 1758-1765 ◽  
Author(s):  
G. H. Egley

Substrates influenced germination of conidia of the mycoherbicide, Colletotrichum truncatum. Very few (< 10%) conidia germinated when incubated while suspended in water or when incubated on or in partially liquefied agar. Many (> 70%) were induced to germinate when incubated on firm agar. The percentage of germinated conidia increased as agar firmness increased. Not all solid substrates equally influenced germination. Over 50% of the conidia germinated on chromatography paper, cellulose acetate filter, or on hard (plastic cover slip) substrates. In contrast, germination was relatively poor on cellulose nitrate filter and glass cover slips. Some natural substrates of dissimilar texture (wood, live plant leaf) produced good germination. Lower O2 levels or limited gas exchange for submerged conidia did not prevent germination because many conidia germinated while submerged on or between firm substrates. Subjecting conidia to motion during incubation between glass cover slips favored germ tube production rather than appressoria formation. Evidence suggests a positive germination response of C. truncatum conidia to solid substrates that occurs prior to the well-documented thigmotropic response of germ tubes of germinated conidia. Key words: Colletotrichum truncatum, conidia germination, thigmotropism, substrate.



1978 ◽  
Vol 56 (22) ◽  
pp. 2865-2872 ◽  
Author(s):  
Ichiko Tsuneda ◽  
Lorene L. Kennedy

Germination of basidiospores in Fomes fomentarius (Fries) Kickx is bipolar with germ tubes emerging at both ends. Ungerminated spores are smooth with a thick cell wall consisting of two layers: an outer thin, electron-dense layer and an inner thick, electron-light layer. During the early stage of germination, two additional cell wall layers are formed: a very thin, electron-dense layer and a relatively thick, electron-light layer. Germ tube walls originate from these newly formed, inner layers. Ungerminated spores are uninucleate and contain numerous lipid bodies, ribosomes, and cisternae of endoplasmic reticulum. Germinated spores have distinct mitochondria and an invaginated plasma membrane and are usually devoid of endoplasmic reticulum.



Plant Disease ◽  
2016 ◽  
Vol 100 (10) ◽  
pp. 2050-2056 ◽  
Author(s):  
Bruna B. Forcelini ◽  
Teresa E. Seijo ◽  
Achour Amiri ◽  
Natalia A. Peres

Anthracnose fruit rot of strawberry, caused by Colletotrichum acutatum, is a major disease in Florida and frequent quinone-outside inhibitor (QoI) fungicide applications are needed for disease control. From 1994 to 2014, 181 C. acutatum isolates were collected from multiple strawberry fields in Florida with or without QoI spray history. Sensitivity to azoxystrobin and pyraclostrobin was tested based upon mycelial growth and germ tube elongation inhibition. Mean effective concentration where growth was reduced by 50% (EC50) values for isolates collected prior to 2013 based upon mycelial growth were 0.22 and 0.013 μg/ml and upon germ tube elongation were 0.57 and 0.03 μg/ml for azoxystrobin and pyraclostrobin, respectively. Mycelial growth and germ tube elongation of 48 isolates collected in 2013 and 2014 were not inhibited with azoxystrobin at 3 μg/ml and pyraclostrobin at 0.110 μg/ml. A fungicide discriminatory dose assay indicated that 43 of the 48 isolates had EC50 values higher than 100 and 10 μg/ml for azoxystrobin and pyraclostrobin, respectively. Azoxystrobin and pyraclostrobin sprayed preventively on strawberry fruit inoculated with C. acutatum failed to control resistant isolates. Sequencing of the cytochrome b gene of sensitive and resistant isolates showed that QoI-resistant isolates contained either G143A or F129L amino acid substitutions.



1976 ◽  
Vol 22 (1) ◽  
pp. 92-100 ◽  
Author(s):  
Gordon M. Murray ◽  
Douglas P. Maxwell

Multicelled conidia of Stemphylium sarcinaeforme germinate in water forming several germ tubes. Individual cells within conidia are connected by pores which are plugged in ungerminated conidia and open in germinated ones. During germination, vacuoles enlarge, endoplasmic reticulum profiles increase in number, and mitochondria change from spherical to elongate. The germ tube wall is laid down at the site of emergence from the conidium. Shortly after germination, a septum with a central pore forms where the germ tube emerged. The germ tube wall is surrounded by a fibrillar sheath. Lipid bodies are closely associated with vacuoles during germination. The ultrastructural location of lipid was found by extraction of conidia with lipid solvents. Total lipid decreases from 14.4% of the dry weight of ungerminated conidia to 13.4% of the dry weight of conidia germinated for 10 h. No qualitative changes occurred in the major lipid classes of conidia during germination. The activities of lipase and acid phosphatase were detected in ungerminated and germinated conidia.



2021 ◽  
Vol 9 (4) ◽  
pp. 839
Author(s):  
Muhammad Rafiullah Khan ◽  
Vanee Chonhenchob ◽  
Chongxing Huang ◽  
Panitee Suwanamornlert

Microorganisms causing anthracnose diseases have a medium to a high level of resistance to the existing fungicides. This study aimed to investigate neem plant extract (propyl disulfide, PD) as an alternative to the current fungicides against mango’s anthracnose. Microorganisms were isolated from decayed mango and identified as Colletotrichum gloeosporioides and Colletotrichum acutatum. Next, a pathogenicity test was conducted and after fulfilling Koch’s postulates, fungi were reisolated from these symptomatic fruits and we thus obtained pure cultures. Then, different concentrations of PD were used against these fungi in vapor and agar diffusion assays. Ethanol and distilled water were served as control treatments. PD significantly (p ≤ 0.05) inhibited more of the mycelial growth of these fungi than both controls. The antifungal activity of PD increased with increasing concentrations. The vapor diffusion assay was more effective in inhibiting the mycelial growth of these fungi than the agar diffusion assay. A good fit (R2, 0.950) of the experimental data in the Gompertz growth model and a significant difference in the model parameters, i.e., lag phase (λ), stationary phase (A) and mycelial growth rate, further showed the antifungal efficacy of PD. Therefore, PD could be the best antimicrobial compound against a wide range of microorganisms.



2010 ◽  
Vol 76 (21) ◽  
pp. 7217-7225 ◽  
Author(s):  
Daniel P. MacEachran ◽  
M. E. Prophete ◽  
A. J. Sinskey

ABSTRACT Generally, prokaryotes store carbon as polyhydroxyalkanoate, starch, or glycogen. The Gram-positive actinomycete Rhodococcus opacus strain PD630 is noteworthy in that it stores carbon in the form of triacylglycerol (TAG). Several studies have demonstrated that R. opacus PD630 can accumulate up to 76% of its cell dry weight as TAG when grown under nitrogen-limiting conditions. While this process is well studied, the underlying molecular and biochemical mechanisms leading to TAG biosynthesis and subsequent storage are poorly understood. We designed a high-throughput genetic screening to identify genes and their products required for TAG biosynthesis and storage in R. opacus PD630. We identified a gene predicted to encode a putative heparin-binding hemagglutinin homolog, which we have termed tadA (triacylglycerol accumulation deficient), as being important for TAG accumulation. Kinetic studies of TAG accumulation in both the wild-type (WT) and mutant strains demonstrated that the tadA mutant accumulates 30 to 40% less TAG than the parental strain (WT). We observed that lipid bodies formed by the mutant strain were of a different size and shape than those of the WT. Characterization of TadA demonstrated that the protein is capable of binding heparin and of agglutinating purified lipid bodies. Finally, we observed that the TadA protein localizes to lipid bodies in R. opacus PD630 both in vivo and in vitro. Based on these data, we hypothesize that the TadA protein acts to aggregate small lipid bodies, found in cells during early stages of lipid storage, into larger lipid bodies and thus plays a key role in lipid body maturation in R. opacus PD630.



2001 ◽  
Vol 41 (5) ◽  
pp. 697 ◽  
Author(s):  
D. R. Beasley ◽  
D. C. Joyce ◽  
L. M. Coates ◽  
A. H. Wearing

Saprophytic bacteria, yeasts and filamentous fungi were isolated from Geraldton waxflower flowers and screened to identify potential antagonism towards Botrytis cinerea. Isolates from other sources (e.g. avocado) were also tested. Isolates were initially screened in vitro for inhibition of B. cinerea conidial germination, germ tube elongation and mycelial growth. The most antagonistic bacteria, yeasts and fungi were selected for further testing on detached waxflower flowers. Conidia of the pathogen were mixed with conidia or cells of the selected antagonists, co-inoculated onto waxflower flowers, and the flowers were sealed in glass jars and incubated at 20˚C. The number of days required for the pathogen to cause flower abscission was determined. The most antagonistic bacterial isolate, Pseudomonas sp. 677, significantly reduced conidial germination and retarded germ tube elongation of B. cinerea. None of the yeast or fungal isolates tested was found to significantly reduce conidial germination or retard germ tube elongation, but several significantly inhibited growth of B. cinerea. Fusarium sp., Epicoccum sp. and Trichoderma spp. were the most antagonistic of these isolates. Of the isolates tested on waxflower, Pseudomonas sp. 677 was highly antagonistic towards B. cinerea and delayed waxflower abscission by about 3 days. Trichoderma harzianum also significantly delayed flower abscission. However, as with most of the fungal antagonists used, inoculation of waxflower flowers with this isolate resulted in unsightly mycelial growth.



2010 ◽  
Vol 25 (2) ◽  
pp. 151-156 ◽  
Author(s):  
Natasa Duduk ◽  
Aleksa Obradovic ◽  
Mirko Ivanovic

Effects of the volatile phase of thyme, cinnamon and clove essential oils on Colletotrichum acutatum were investigated. Mycelial disc was placed in the center of the Petri dish (V=66 ml) containing PDA. Different volumes of either non- or ethanol-diluted essential oils were placed on the inner side of the dish cover to obtain final concentrations of 153, 107, 76, 46, 15, 14, 12, 11, 7.6, 3.82, 1.53, 0.153 and 0.0153 ?l/L of air. The dishes were sealed with Parafilm and incubated in up-side-down position. After 7 days of incubation, mycelial growth was recorded by measuring the colony diameter. If no mycelial growth was recorded, the disc was transferred to a new PDA plate in order to evaluate whether the activity was either fungistatic or fungicidal. Mean growth values were obtained and then converted to inhibition percentage of mycelial growth compared with the control treatment. All the tested essential oils inhibited mycelial growth of C. acutatum in the dose dependent manner. Mycelial growth was totally inhibited by thyme oil in the concentration of 76 ?l/L of air. The same results were obtained by cinnamon and clove oil in the concentration of 107 ?l/L of air. Thyme and cinnamon oil had fungicidal effect in concentrations of 107 and 153 ?l/L respectively. The results obtained provide evidence on the antifungal in vitro effect of the tested essential oils as potential means for the control of C. acutatum.





2018 ◽  
Vol 16 (02) ◽  
pp. 15-24
Author(s):  
Zaker M ◽  
Zaker L

The efficacy of a large number of plant extracts and essential oils in controlling plant diseases has been proven worldwide. Botrytis cinerea has attacked a wide host range causing severe loss in the field and at storage. In this study the antifungal efficacy of essential oils of three medicinal plants namely wild marjoram (Zataria multifolia), wild savory (perennial) (Satureja mutica) and savory (annual) (Satureja hortensis) possessing these compounds at three concentrations: 50, 100 and 200 ppm were evaluated in controlling the mycelial growth, spore germination and germ tube elongation of B. cinerea. All treatments except savory (annual) essential oil at 50 ppm showed significant differences with the control in inhibiting the mycelial growth as well as spore germination and germ tube elongation of B. cinerea (p=0.01). It was also noted that wild marjoram at 100 ppm and wild savory (perennial) essential oils at 200 ppm could completely (100%) inhibit the growth of the fungus. Essential oils from wild marjoram and wild savory had higher antifungal activity than annual savory. Their suitable formulations could be prepared and used as safe alternatives for controlling moulds of horticultural products during storage. The Agriculturists 2018; 16(2) 15-24



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