Polyclonal antisera to epacrid mycorrhizae and to Hymenoscyphus ericae display specificity

2000 ◽  
Vol 78 (7) ◽  
pp. 841-850
Author(s):  
R A Parry ◽  
C B McLean ◽  
M R Alderton ◽  
P J Coloe ◽  
A C Lawrie
Keyword(s):  
Mycorrhizal Fungi ◽  
Polyclonal Antibodies ◽  
Calluna Vulgaris ◽  
Immunogold Labelling ◽  
Fungal Endophyte ◽  
Cross Reactivity ◽  
Plant Roots ◽  
Hymenoscyphus Ericae ◽  
Ericoid Mycorrhizae ◽  
Polyclonal Antisera

Three polyclonal antisera produced in mice were used to investigate specificity and cross-reactivity between ericaceous and epacridaceous mycorrhizal fungi. One antiserum was to a culture of Hymenoscyphus ericae (Read) Korf and Kernan, the fungal endophyte of Calluna vulgaris (L.) Hull (Ericaceae). The other two were to peloton preparations from roots of Epacris impressa Labill. (Epacridaceae) from two sites (Cranbourne and Grampians) in Victoria, Australia. By immunofluorescence, all three antisera recognised H. ericae but not Oidiodendron griseum Roback, suggesting a serological relationship with the former endophyte. They also recognised 10 of the 12 fungal isolates tested, from mycorrhizal roots of E. impressa (Cranbourne), and all 4 isolates from Astroloma pinifolium (R. Br.) Benth. (Epacridaceae) (Grampians). Furthermore, none of the antisera recognised any of the nine common soil-inhabiting fungi selected for screening. Antisera recognised only unmelanized hyphae on epacrid and other plant roots taken from the wild. With plants from Cranbourne, all antisera except the Grampians antiserum recognised hyphae only on epacrid roots, demonstrating specificity. Hyphae on other plant roots were not recognised by any of the antisera. With plants from the Grampians, all antisera recognised some hyphae on both epacrid and other plant roots, except in two instances. The immunogold labelling indicates that the antisera are specific for fungi and do not recognise the plant. Since the fungal isolate forms true mycorrhizal structures, this suggests that there is a serological similarity between fungi forming epacrid mycorrhiza and those (H. ericae) forming ericoid mycorrhiza.Key words: ericoid mycorrhizae, Epacridaceae, polyclonal antibodies, immunofluorescence, immunogold.

Growth of micropropagated lowbush blueberry with defined fungi in irradiated peat mix

1992 ◽  
Vol 70 (11) ◽  
pp. 2202-2206 ◽  
Author(s):  
Walter Litten ◽  
John M. Smagula ◽  
Yolande Dalpé
Keyword(s):  
Mycorrhizal Fungi ◽  
Stem Tissue ◽  
Γ Irradiation ◽  
Vaccinium Angustifolium ◽  
Hymenoscyphus Ericae ◽  
Ericoid Mycorrhizae ◽  
Lowbush Blueberry ◽  
Vegetative Multiplication ◽  
Mycorrhizal Inoculation

There is an interest in vegetative multiplication of high-yielding clones of Vaccinium angustifolium Ait. to establish or enhance blueberry production. This study evaluates mycorrhizal inoculation as an aid in such propagation from microcuttings. Shoots of Vaccinium angustifolium (clone 7062) generated in vitro were rooted in a peat–vermiculite–perlite substrate with or without ericoid mycorrhizal fungi fortification by Hymenoscyphus ericae or Scytalidium vaccinii and with or without peat sterilization by γ irradiation. Both in irradiated peat mix inoculated with S. vaccinii and in unirradiated peat mix with H. ericae, microcuttings grew taller and branched more than with the four other treatments. The profusely rooted plantlets available from all treatments of the cuttings put on significantly more total length of stems and branches after 167 days in the greenhouse when growing with either inoculant in unirradiated peat than in the unirradiated peat without inoculation. However, the magnitude of difference might be of borderline importance in commercial nursery operations. A higher level of copper and zinc in stem tissue was observed in stem tissue of plants grown with H. ericae with or without irradiation but not with S. vaccinii. Key words: ericoid mycorrhizae, micropropagation, Hymenoscyphus ericae, Scytalidium vaccinii, Vaccinium angustifolium.

scholarly journals Host Range of a Select Isolate of the Beneficial Ericoid Mycorrhizal Fungus Hymenoscyphus ericae

HortScience ◽  
2000 ◽  
Vol 35 (5) ◽  
pp. 827C-827
Author(s):  
Nicole R. Gorman ◽  
Mark C. Starrett
Keyword(s):  
Host Range ◽  
Mycorrhizal Fungus ◽  
Calluna Vulgaris ◽  
Vaccinium Corymbosum ◽  
Negative Control ◽  
Culture Collection ◽  
Hymenoscyphus Ericae ◽  
Ericoid Mycorrhizae ◽  
Commercial Applications ◽  
Ericoid Mycorrhizal Fungus

Studies were conducted to examine the host range of a select isolate of the ericoid mycorrhizal fungus, Hymenoscyphus ericae (Read) Korf and Kernan [American Type Culture Collection (ATCC) #32985]. Host status was tested for 15 ericaceous species, including Calluna vulgaris, Enkianthus campanulatus, Gaultheria procumbens, Kalmia latifolia, Leucothoe fontanesiana, Oxydendrum arboreum, Pieris floribunda, Rhododendron calendulaceum, Rhododendron carolinianum, Rhododendron catawbiense, Rhododendron maximum, Rhododendron mucronulatum, Vaccinium corymbosum, and Vaccinium macrocarpon. Arbutus unedo, an ericaceous species that forms arbutoid, not ericoid, mycorrhizae was used as a negative control. All of the species were colonized by the ericoid isolate with the exception of Enkianthus campanulatus and the negative control. The benefits of the association and possible commercial applications are discussed.

scholarly journals In Vitro Colonization of Micropropagated Pieris floribunda by Ericoid Mycorrhizae. II. Effects on Acclimatization and Growth

HortScience ◽  
2001 ◽  
Vol 36 (2) ◽  
pp. 357-359 ◽  
Author(s):  
Mark C. Starrett ◽  
Frank A. Blazich ◽  
Steven R. Shafer ◽  
Larry F. Grand
Keyword(s):  
Mycorrhizal Fungi ◽  
Hymenoscyphus Ericae ◽  
Ericoid Mycorrhizal Fungi ◽  
Ericoid Mycorrhizae ◽  
In Vitro Inoculation

Inoculation of microshoots of Pieris floribunda (Pursh ex Sims) Benth. and Hook. (mountain andromeda) with isolates of Hymenoscyphus ericae (Read) Korf and Kernan ericoid mycorrhizal fungi stimulated growth during 1 month in vitro. However, no benefits were apparent after 3 months in a greenhouse. Acclimatization of plantlets of P. floribunda to greenhouse conditions following in vitro inoculation improved survival (42% vs. 16% for controls). The protocol reported herein is similar to procedures utilized currently for micropropagation of various ericaceous species and has potential to improve plantlet survival during acclimatization.

Use of monoclonal and polyclonal antibodies to detect prostatic acid phosphatase by immunoblotting.

1986 ◽  
Vol 32 (10) ◽  
pp. 1832-1835 ◽  
Author(s):  
P C Patel ◽  
L Aubin ◽  
J Côte
Keyword(s):  
Monoclonal Antibodies ◽  
Acid Phosphatase ◽  
Western Blot ◽  
Polyclonal Antibodies ◽  
Prostatic Acid Phosphatase ◽  
The Other ◽  
Dot Blot ◽  
Western Blots ◽  
Dot Blot Assay ◽  
Polyclonal Antisera

Abstract We investigated two techniques of immunoblotting--the Western blot and the dot blot--for use in detecting prostatic acid phosphatase (PAP, EC 3.1.3.2). We used polyclonal antisera to human PAP, produced in rabbits by hyperimmunization with purified PAP, and PAP-specific monoclonal antibodies in the immunoenzymatic protocols. We conclude that PAP can be readily detected by Western blots with use of polyclonal antisera, but not with monoclonal antibodies. On the other hand, using a dot blot assay, we could easily detect PAP with both polyclonal and monoclonal antibodies.

scholarly journals Examination of the Efficacy and Cross-Reactivity of a Novel Polyclonal Antibody Targeting the Disintegrin Domain in SVMPs to Neutralize Snake Venom

Toxins ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 254
Author(s):  
Shelby S. Szteiter ◽  
Ilse N. Diego ◽  
Jonathan Ortegon ◽  
Eliana Salinas ◽  
Abcde Cirilo ◽  
...  
Keyword(s):  
Snake Venom ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Cross Reactivity ◽  
Snake Envenomation ◽  
Disintegrin Domain ◽  
The Common ◽  
Neutralization Ability ◽  
New Strategies

Snake envenomation can result in hemorrhage, local necrosis, swelling, and if not treated properly can lead to adverse systemic effects such as coagulopathy, nephrotoxicity, neurotoxicity, and cardiotoxicity, which can result in death. As such, snake venom metalloproteinases (SVMPs) and disintegrins are two toxic components that contribute to hemorrhage and interfere with the hemostatic system. Administration of a commercial antivenom is the common antidote to treat snake envenomation, but the high-cost, lack of efficacy, side effects, and limited availability, necessitates the development of new strategies and approaches for therapeutic treatments. Herein, we describe the neutralization ability of anti-disintegrin polyclonal antibody on the activities of isolated disintegrins, P-II/P-III SVMPs, and crude venoms. Our results show disintegrin activity on platelet aggregation in whole blood and the migration of the SK-Mel-28 cells that can be neutralized with anti-disintegrin polyclonal antibody. We characterized a SVMP and found that anti-disintegrin was also able to inhibit its activity in an in vitro proteolytic assay. Moreover, we found that anti-disintegrin could neutralize the proteolytic and hemorrhagic activities from crude Crotalus atrox venom. Our results suggest that anti-disintegrin polyclonal antibodies have the potential for a targeted approach to neutralize SVMPs in the treatment of snakebite envenomations.

Synaptonemal complex proteins: occurrence, epitope mapping and chromosome disjunction

1994 ◽  
Vol 107 (10) ◽  
pp. 2749-2760 ◽  
Author(s):  
M.J. Dobson ◽  
R.E. Pearlman ◽  
A. Karaiskakis ◽  
B. Spyropoulos ◽  
P.B. Moens
Keyword(s):  
Synaptonemal Complex ◽  
Fusion Proteins ◽  
Epitope Mapping ◽  
Core Protein ◽  
Binding Capacity ◽  
Polyclonal Antibodies ◽  
Meiotic Chromosome ◽  
Immunogold Labelling ◽  
Chromosome Disjunction ◽  
Metaphase I

We have used polyclonal antibodies against fusion proteins produced from cDNA fragments of a meiotic chromosome core protein, Cor1, and a protein present only in the synapsed portions of the cores, Syn1, to detect the occurrence and the locations of these proteins in rodent meiotic prophase chromosomes. The 234 amino acid Cor1 protein is present in early unpaired cores, in the lateral domains of the synaptonemal complex and in the chromosome cores when they separate at diplotene. A novel observation showed the presence of Cor1 axial to the metaphase I chromosomes and substantial amounts of Cor1 in association with pairs of sister centromeres. The centromere-associated Cor1 protein becomes dissociated from the centromeres at anaphase II and it is not found in mitotic metaphase centromeres. The extended presence of Cor1 suggests that it may have a role in chromosome disjunction by fastening chiasmata at metaphase I and by joining sister kinetochores, which ensures co-segregation at anaphase I. Two-colour immunofluorescence of Cor1 and Syn1 demonstrates that synapsis between homologous cores is initiated at few sites but advances rapidly relative to the establishment of new initiation sites. If the rapid advance of synapsis deters additional initiation sites between pairs of homologues, it may provide a mechanism for positive recombination interference. Immunogold epitope mapping of antibodies to four Syn1 fusion proteins places the amino terminus of Syn1 towards the centre of the synaptonemal complex while the carboxyl terminus extends well into the lateral domain of the synaptonemal complex. The Syn1 fusion proteins have a non-specific DNA binding capacity. Immunogold labelling of Cor1 antigens indicates that the lateral domain of the synaptonemal complex is about twice as wide as the apparent width of lateral elements when stained with electron-dense metal ions. Electron microscopy of shadow-cast surface-spread SCs confirms the greater width of the lateral domain. The implication of these dimensions is that the proteins that comprise the synaptic domain overlap with the protein constituents of the lateral domains of the synaptonemal complex more than was apparent from earlier observations. This arrangement suggests that direct interactions might be expected between some of the synaptonemal complex proteins.

scholarly journals Immunochemical characterization of NADPH-cytochrome P-450 reductase from Jerusalem artichoke and other higher plants

1989 ◽  
Vol 259 (3) ◽  
pp. 847-853 ◽  
Author(s):  
I Benveniste ◽  
A Lesot ◽  
M P Hasenfratz ◽  
F Durst
Keyword(s):  
Cytochrome C ◽  
Rat Liver ◽  
Higher Plants ◽  
Polyclonal Antibodies ◽  
Reductase Activity ◽  
Jerusalem Artichoke ◽  
Cross Reactivity ◽  
Cytochrome C Reductase ◽  
Nadph Cytochrome ◽  
Cytochrome P 450

Polyclonal antibodies were prepared against NADPH-cytochrome P-450 reductase purified from Jerusalem artichoke. These antibodies inhibited efficiently the NADPH-cytochrome c reductase activity of the purified enzyme, as well as of Jerusalem artichoke microsomes. Likewise, microsomal NADPH-dependent cytochrome P-450 mono-oxygenases (cinnamate and laurate hydroxylases) were efficiently inhibited. The antibodies were only slightly inhibitory toward microsomal NADH-cytochrome c reductase activity, but lowered NADH-dependent cytochrome P-450 mono-oxygenase activities. The Jerusalem artichoke NADPH-cytochrome P-450 reductase is characterized by its high Mr (82,000) as compared with the enzyme from animals (76,000-78,000). Western blot analysis revealed cross-reactivity of the Jerusalem artichoke reductase antibodies with microsomes from plants belonging to different families (monocotyledons and dicotyledons). All of the proteins recognized by the antibodies had an Mr of approx. 82,000. No cross-reaction was observed with microsomes from rat liver or Locusta migratoria midgut. The cross-reactivity generally paralleled well the inhibition of reductase activity: the enzyme from most higher plants tested was inhibited by the antibodies; whereas Gingko biloba, Euglena gracilis, yeast, rat liver and insect midgut activities were insensitive to the antibodies. These results point to structural differences, particularly at the active site, between the reductases from higher plants and the enzymes from phylogenetically distant plants and from animals.

scholarly journals Rapid immunoassays for detection of anabolic nortestosterone in dietary supplements

2013 ◽  
Vol 31 (No. 5) ◽  
pp. 514-519 ◽  
Author(s):  
B. Holubová ◽  
S. Göselová ◽  
L. Ševčíková ◽  
M. Vlach ◽  
M. Blažková ◽  
...  
Keyword(s):  
Dietary Supplements ◽  
Rapid Detection ◽  
Visual Detection ◽  
Limit Of Detection ◽  
Polyclonal Antibodies ◽  
Cross Reactivity ◽  
Rapid Analysis ◽  
Experimental Conditions ◽  
Immunochromatographic Strip ◽  
Strip Test

An enzyme immunoassay (ELISA) and an immunochromatographic strip were designed for a rapid detection of nortestosterone in dietary supplements. Two polyclonal antibodies and two types of nortestosterone-protein coating conjugates were tested to develop the most appropriate method. Under optimal experimental conditions, the most sensitive ELISA achieved the IC<sub>50 </sub>and the limit of detection values of 6.41 and 0.09 ng/ml, respectively. The assay specificity was tested measuring cross-reactivity of several steroids. The interference with the assay was negligible (&lt; 0.1%), except for cross-reactivity with another frequently abused steroid testosterone (23%). The optimised gold particle-based immunochromatographic strip provided in semi-quantitative test a visual detection limit of 1&nbsp;ng/ml. None of these methods showed the interference using a filtrate of the suspension of non-contaminated sample. After the validation for particular matrices, the ELISA and the strip test could be useful tools for a rapid analysis of nortestosterone in crude extracts of dietary supplements.

Isolation and ultrastructural localization of a soluble protein from Ophiostoma ulmi

1990 ◽  
Vol 68 (11) ◽  
pp. 2517-2524 ◽  
Author(s):  
R. S. Jeng ◽  
A. M. Svircev
Keyword(s):  
Polyclonal Antibodies ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein A ◽  
Leading Edge ◽  
Ultrastructural Localization ◽  
Immunogold Labelling ◽  
Two Dimensional ◽  
Thin Sections ◽  
Ophiostoma Ulmi ◽  
Gold Label

Two-dimensional polyacrylamide gel electrophoresis was used to identify and isolate a soluble polypeptide, the QP1 protein, which is characteristic of the vegetative hyphae of nonaggressive isolate Q412 of Ophiostoma ulmi. Individual QP1 spots were excised from 16 two-dimensional gels. Polypeptides were eluted from the gel spots by electroelution and lyophilized. The protein was injected into rabbits for the production of polyclonal antibodies. Antiserum specificity was tested by transferring polypeptides from a two-dimensional gel onto nitrocellulose and treating with QP1 serum. The resulting immunoblot contained a single spot that corresponded in shape and location to that of the QP1 polypeptide. Thin sections of fungal mycelia, from nonaggressive isolate Q412 and the aggressive isolate VA of O. ulmi, were treated with QP1 antibodies and protein A – gold. The gold label was localized in thin sections over conidial and hyphal cell walls of the nonaggressive isolate. The aggressive isolate was nonreactive. Mycelia from nonaggressive isolates Q412 and Q311 and aggressive isolates VA and CESS16K of O. ulmi were grown on solid medium, treated with QP1 antibodies, labelled with protein A – gold, and prepared for scanning electron microscopy. The gold-labelled QP1 polypeptide was detected on the leading edge of a small number of hyphae from nonaggressive isolates Q412 and Q311. Key words: immunogold labelling, Ophiostoma ulmi, soluble proteins.

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