Inconsistent transmission of banana bunchy top virus in micropropagated bananas and its implication for germplasm screening

Banana bunchy top virus (BBTV) was readily transmitted through tissue culture in banana (Musa sp.) cv. Lady finger (AAB) and Cavendish cv. Williams (AAA). Lines derived from infected and healthy field plants had similar in vitro multiplication rates. BBTV-infected in vitro cultures displayed symptoms of stunting, leaf curling, chlorotic and green flecks, and poor root growth. Symptoms became milder with time, and were often difficult to discern in older, rapidly multiplying cultures. A triple antibody sandwich ELISA using polyclonal and monoclonal antibodies was very efficient for detecting BBTV in vitro. Symptomless, ELISA-negative plants arose in 10 out of 11 lines derived from BBTV-infected field plants and first appeared after 9 months continuous in vitro culture at a constant 28�C. Meristem tip culture or heat therapy was not used. These plants remained symptomless and ELISA-negative after planting out in the glasshouse (individual plants checked for up to 16 months). The implications of this inconsistent transmission of BBTV for germplasm indexing and exchange are discussed.

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Two Monoclonal Antibodies That Specifically Recognize Aspergillus Cell Wall Antigens and Can Detect Circulating Antigens in Infected Mice

International Journal of Molecular Sciences ◽

10.3390/ijms23010252 ◽

2021 ◽

Vol 23 (1) ◽

pp. 252

Author(s):

Xihua Lian ◽

Stephen Chambers ◽

John G. Lewis ◽

Amy Scott-Thomas ◽

Madhav Bhatia

Keyword(s):

Cell Wall ◽

Monoclonal Antibodies ◽

Invasive Aspergillosis ◽

Candida Species ◽

Sandwich Elisa ◽

Diagnostic Techniques ◽

Patient Treatment ◽

Diagnostic Tools ◽

Mouse Tissues

Invasive aspergillosis (IA) is a life-threatening disease mainly caused by Aspergillus fumigatus and Aspergillus flavus. Early diagnosis of this condition is crucial for patient treatment and survival. As current diagnostic techniques for IA lack sufficient accuracy, we have raised two monoclonal antibodies (1D2 and 4E4) against A. fumigatus cell wall fragments that may provide a platform for a new diagnostic approach. The immunoreactivity of these antibodies was tested by immunofluorescence and ELISA against various Aspergillus and Candida species in vitro and by immunohistochemistry in A. fumigatus infected mouse tissues. Both monoclonal antibodies (mAbs) showed intensive fluorescence with the hyphae wall of A. fumigatus and A. flavus, but there was no staining with other Aspergillus species or Candida species. Both mAbs also showed strong immunoreactivity to the cell wall of A. fumigatus hyphae in the infected liver, spleen and kidney of mice with IA. The antigens identified by 1D2 and 4E4 might be glycoproteins and the epitopes are most likely a protein or peptide rather than a carbohydrate. An antibody-based antigen capture ELISA detected the extracellular antigens released by A. fumigatus, A. flavus, A. niger and A. terreus, but not in Candida species. The antigen could be detected in the plasma of mice after 48 h of infection by double-sandwich ELISA. In conclusion, both 1D2 and 4E4 mAbs are potentially promising diagnostic tools to investigate invasive aspergillosis.

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Inhibition of bovine sperm–zona binding by bovine herpesvirus-1

Reproduction ◽

10.1530/rep.1.00636 ◽

2005 ◽

Vol 130 (2) ◽

pp. 251-259 ◽

Cited By ~ 9

Author(s):

S Tanghe ◽

G Vanroose ◽

A Van Soom ◽

L Duchateau ◽

M T Ysebaert ◽

...

Keyword(s):

Tissue Culture ◽

Monoclonal Antibodies ◽

Bovine Herpesvirus ◽

Inhibitory Effect ◽

Bovine Herpesvirus 1 ◽

Bovine Sperm ◽

Viral Glycoproteins ◽

Herpesvirus 1 ◽

Dose Dependent

The purpose of the present study was to identify a potential interference of bovine herpesvirus-1 (BoHV-1) with sperm–oocyte interactions during bovinein vitrofertilization. An inhibition of almost 70% of sperm–zona binding was observed when bovine cumulus-denuded oocytes were inseminated in the presence of 10750% tissue culture infective dose/ml BoHV-1. The inhibitory effect of BoHV-1 on sperm–zona binding was mediated by an interaction of the virus with spermatozoa, but not with oocytes. Treatment of spermatozoa with BoHV-1, however, did not affect sperm motility and acrosomal status. Antiserum against BoHV-1 prevented the virus-induced inhibition of sperm–zona binding, indicating that BoHV-1 itself affects the fertilization process. In order to investigate which BoHV-1 glycoprotein(s) are responsible for the virus–sperm interaction, BoHV-1 was treated with monoclonal antibodies against the viral glycoproteins gB, gC, gD and gH prior to insemination. Anti-gC completely prevented the inhibitory effect of BoHV-1 on sperm–zona binding, while anti-gD caused a reduction of this inhibition. Further evidence for the involvement of gC and gD in the virus–sperm interaction was provided by the fact that purified gC and gD decreased sperm–zona binding in a dose-dependent way with gC being more effective than gD. These results indicated that BoHV-1 inhibits bovine sperm–zona binding by interacting with spermatozoa. The binding of BoHV-1 to a spermatozoon is mediated by the viral glycoproteins gC and gD, and therefore seems to be comparable with the mechanisms of BoHV-1 attachment to its natural host cell.

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DELAY OF ONSET OF LEAFROLL SYMPTOM EXPRESSION IN Vitis vinifera ’LIEMBERGER’ FROM RIBAVIRIN-TREATED IN VITRO CULTURES

Canadian Journal of Plant Science ◽

10.4141/cjps83-068 ◽

1983 ◽

Vol 63 (2) ◽

pp. 557-560 ◽

Cited By ~ 6

Author(s):

J. H. STEVENSON ◽

P. L. MONETTE

Keyword(s):

Tissue Culture ◽

Vitis Vinifera ◽

Drug Concentration ◽

Day Treatment ◽

In Vitro Cultures ◽

Symptom Expression ◽

Leafroll Symptom

Ribavirin added at a concentration of 10 mg/L or more to in vitro cultures of Vitis vinifera ’Liemberger’ infected with grapevine leafroll delayed the onset of symptom expression by an average of 50 days. At the concentrations tested, the effect was independent of drug concentration and a 30-day treatment was as effective as a 90-day treatment.Key words: Ribavirin, tissue culture, grapevine leafroll

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Use of in vitro Cultures of Daphne cneorum L. for the Western Detection of Daphne Virus X

Journal of Environmental Horticulture ◽

10.24266/0738-2898-10.3.153 ◽

1992 ◽

Vol 10 (3) ◽

pp. 153-156

Author(s):

C. Wei ◽

M.J. Green ◽

S.E. Godkin ◽

P.L. Monette

Keyword(s):

In Vitro Cultures ◽

Early Screening ◽

Virus Elimination ◽

Viral Coat Protein ◽

Western Analysis ◽

Heat Therapy ◽

Viral Coat ◽

Ornamental Shrub ◽

Commercial Micropropagation

Abstract In vitro cultures of Daphne cneorum L. were successfully used as test samples for the detection of the viral coat protein of daphne virus X (DVX) by Western analysis. This powerful serological technique could thus be used where viral antigen concentrations may be very low, such as in the early screening of in vitro-cultured “heat therapy” tips from virus elimination programs. Western analysis was conducted on randomly selected D. cneorum plants from a commercial nursery and on in vitro cultures from a commercial micropropagation lab. All the plants and cultures tested were DVX-infected, supporting the view that this ornamental shrub may be universally infected with DVX.

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In vitro Plant Regeneration in Banana (Musa sp.) cv. Grand Nain

Asian Journal of Advances in Agricultural Research ◽

10.9734/ajaar/2021/v16i430180 ◽

2021 ◽

pp. 9-17

Author(s):

Aiman Yousif ◽

Muhammad Ijaz ◽

Hamza Iftikhar ◽

Aqsa Mustafa ◽

Nerum Tasneem ◽

...

Keyword(s):

Tissue Culture ◽

Acetic Acid ◽

Plant Regeneration ◽

Ms Medium ◽

In Vitro Plant Regeneration ◽

Green Mass ◽

Musa Sp ◽

Hardening Process ◽

Indole 3 Acetic Acid

This study was conducted at the tissue culture laboratory in Pakistan. Suckers obtained from the banana cv. Grand Nain to explore the effects of different concentrations of BAP and IAA. After inoculation with Murashige and Skoog (MS) medium supplemented with different concentrations of 6-Benzylaminopurine (BAP) and the Indole-3- acetic acid (IAA) the suckers first turn into the brown after 5-10 days then turned into a green mass-like structure after 30 days. From this mass-like structure shoots and roots were developed. Among the different concentrations, BAP 7.5 mg/l and IAA 0.5 mg/l showed the highest number of shoots 0.75, 2.75 and 6.25 per explant and shows the highest shoot lengths which 1.03 cm, 2.45 cm and 3.40 cm respectively. IAA is essential for only roots proliferation. The roots of different lengths were produced when the concentration of IAA 0.5 mg/l was used and 2.93 cm, 4.63 cm and 5.88 cm roots length produced at 15, 30 and 45 days after occultation respectively. Then these plants were transferred to the greenhouse for hardening acclimatization with the environment, the hardening process took place, and the established plantlets are ready for planting.

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Obtaining and Characterization of Hybridomas Producing Monoclonal Antibodies to Shiga-Like Toxins of I and II Types

Problems of Particularly Dangerous Infections ◽

10.21055/0370-1069-2021-3-83-88 ◽

2021 ◽

pp. 83-88

Author(s):

G. V. Kuklina ◽

S. S. Ipatov ◽

D. V. Pechenkin ◽

A. V. Eremkin ◽

A. A. Kytmanov ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Cell Lines ◽

Hybrid Cell ◽

Sandwich Elisa ◽

Minimum Detectable Concentration ◽

Myeloma Cells ◽

Hybrid Cell Lines

Objective – obtaining and characterization of hybrid cell lines producing monoclonal antibodies against I and II types of shiga-like toxins.Materials and methods. Shiga-like toxins obtained in “48thCentral Research Institute” of Ministry of Defense of Russian Federation (Kirov), BALB/c mice, myeloma cells SP2/0-Ag14 were used in research. Immune splenocytes and SP2/0-Ag14 myeloma cells were fused according to G. Kohler and C. Milstein method in De St. Fazekas and D. Scheidegger modifcation using 50 % polyethylene glycol. Hybrid cell lines producing specifc monoclonal antibodies were cloned by limited dilutions. Hybridomas growth and producing properties were studied in vitro and in vivo. Specifc activity of immune sera, culture and ascitic ﬂuids were studied by indirect ELISA. Monoclonal antibodies from ascitic ﬂuids were precipitated by saturated ammonium sulfate, followed by ion exchange chromatographyResults and discussion. 8 hybridomas producing monoclonal antibodies against I and II types shiga-like toxins were obtained. Hybridomas are characterized by stable proliferation and antibody-producing activity during 10 passages in vitro and 3 passages in vivo (observation period). Obtained monoclonal antibodies can be used for ELISA detection of I and II types shiga-like toxins. Minimum detectable concentration of shiga-like toxins in sandwich ELISA is 1 ng/ml. The possibility of detecting shiga-like toxins without typical diﬀerentiation was shown when using in the enzyme immunoassay a polyreceptor mixture of monoclonal antibodies for sensitizing the plate and a polyspecifc mixture of immunoperoxidase conjugates.

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INDUKSI MUTASI PADA PISANG (Musa sp. - ABB) cv. KEPOK DENGAN IRADIASI GAMMA SECARA IN VITRO

Jurnal BIOMA ◽

10.21009/bioma12(1).3 ◽

2017 ◽

Vol 12 (1) ◽

pp. 25

Author(s):

Luthfia Masykuroh ◽

Adisyahputra Adisyahputra ◽

Reni Indrayanti

Keyword(s):

Tissue Culture ◽

Gamma Irradiation ◽

Plant Tissue Culture ◽

Plant Tissue ◽

Irradiation Dose ◽

Induced Mutation ◽

Musa Sp ◽

Randomized Design ◽

Design Factors

Banana (Musa sp. - ABB) cv. Kepok is one of type banana processed that have a very potential commodities fruit developed to support food survival. The purpose of this study was to knowing the effect of gamma irradiation on the growth of banana plants cv. Kepok in vitro. This study was conducted in October 2014 – October 2015 in Plant Tissue Culture Laboratory, Biological – Science UNJ. The methods used was experiment with fully randomized design. Factors that tested was 6 gamma irradiation doses (0, 20, 30, 40, 50, 60 Gy) with 10 repetition. Observation of phenotypic generate diverse characters on the growth of the number of shoots and leaves. Gamma irradiation dose of 50 Gy is doses most inhibits the growth of character. Mutations that occur in banana plantlets cv. Kepok generated by the treatment doses gamma irradiation induced mutation is random.

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