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Direct electron microscopy and serology with plant viruses in leaf material dried and stored over calciumchloride

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Abstract

Most of 43 viruses could easily be detected directly in 53 out of 66 leaf samples dried and stored over CaCl2 for varying periods of time up to 20 1/2 years. Detection usually was with PTA pH 6.5, but alfalfa mosaic, cucumber mosaic and tomato aspermy viruses required PTA pH 3.0 to 4.0. Bean common mosaic, cowpea aphid-borne mosaic and cowpea mosaic viruses were also easily observed in newly dehydrated samples obtained for diagnosis from Morocco and Tanzania.

Broad bean wilt virus, cowpea mosaic virus and cucumber mosaic virus were detected with agar gel-diffusion tests in dry leaf material ground in buffer. This serological assay demonstrated a high concentration of cucumber mosaic virus in leaf material dried over CaCl2 20 years ago. This paper further coroborates the value of the CaCl2 method of dehydration and storage of plant viruses in leaf material.

Samenvatting

Zeer uiteenlopende plantevirussen kunnen goed worden bewaard in bladmateriaal van geïnfecteerde planten dat is gedroogd en bewaard boven CaCl2. Bij rechtstreekse elektronenmicroscopische toetsing van 66 monsters met verschillende isolaten van 43 virussen konden in 53 monsters gemakkelijk virusdeeltjes worden waargenomen (Tabel 1 en Fig. 1 en 2). Meestal lukte dit met fosforwolfraamzuur pH 6,5 dat doorgaans voor in ruw plantesap voorkomende virussen wordt gebruikt. Bij luzernemozaïekvirus en komkommermozaïek virus gelukte dit alleen maar bij lagere pH (3,0 en 4,0). Ook het tomate-aspermievirus was dan veel gemakkelijker aantoombaar. Methylaminewolfraamzuur gaf geen beter resultaat.

Draden van bonerolmozaïekvirus en van ‘cowpea aphid-borne mosaic virus’ en bolletjes van ‘cowpea mosaic virus’ waren snel waarneembaar in elektronenmicroscopische preparaten gemaakt van bladmateriaal van recent uit Marokko en Tanzania ontvangen monsters.

‘Cowpea mosaic virus’, tuinboneverwelkingsvirus en komkommermozaïekvirus konden eveneens gemakkelijk en snel serologisch worden aangetoond in met bufferoplossing vermalen droog blad van respectievelijk ‘cowpea’, erwt en tabak. Het laatstgenoemde, reeds 20 1/2 jaar geleden gedroogde bladmateriaal, bleek in vergelijking met pas geïnoculeerd vers blad van komkommer enChenopodium quinoa zelfs zeer veel serologisch actief virusmateriaal te bevatten (Fig. 3).

De beschreven waarnemingen bevestigen nogmaals de waarde van de toegepaste methode van virusbewaring en tonen aan dat het mogelijk is vele virussen te herkennen in van elders ontvangen gedroogde bladmonsters, zonder het risico te lopen van virusontsnapping, zoals altijd aanwezig bij werk met toetsplanten in de kas.

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Author notes

  1. M. P. Benetti

    Present address: Istituto Sperimentale per la Patologia Vegetale, Via Casal de'Pazzi 250, 00156, Rome, Italy

Authors and Affiliations

  1. Research Institute for Plant Protection (IPO), P.O. Box 42, 6700 AA, Wageningen, the Netherlands

    L. Bos &  M. P. Benetti

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  1. L. Bos
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  2. M. P. Benetti
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Guestworker from Istituto Sperimentale per la Patologia Vegetale, Rome, Italy, participating in the investigations from September 21 to December 21, 1977.

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Bos, L., Benetti, M.P. Direct electron microscopy and serology with plant viruses in leaf material dried and stored over calciumchloride. Netherlands Journal of Plant Pathology 85, 241–251 (1979). https://doi.org/10.1007/BF01977596

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  • DOI: https://doi.org/10.1007/BF01977596

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