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An Efficient Microinoculation Procedure to Study Plant Virus Multiplication at Predetermined Individual Infection Sites on the Leaves
Author(s) -
Konate G.,
Fritig B.
Publication year - 1984
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/j.1439-0434.1984.tb00700.x
Subject(s) - virus , inoculation , biology , tobacco mosaic virus , epidermis (zoology) , virology , horticulture , botany , anatomy
The microinoculation procedure (inoculated are 0.02—0.05 mm 2 was carried out using the very thin tip of a stretched Pasteur pipette. It was dipped in a 500 μg/ml suspension of purified TMV, brought into contact with the upper epidermis of the leaf which had been dusted With an abrasive and rotated twice without actually pricking the leaf tissue. Five minutes later, the excess abrasive and virus were removed by rinsing the leaves under water. The procedure was applied to several tobacco‐TMV combinations. It induced the formation of single local lesions with over 90% efficiency on hypersensitively reacting hosts. Comparable efficiencies were obtained with systemically reacting hosts, as evidenced by a radiochemical procedure coupled with indirect ELISA (KONATE and FRITIG 1983). It was found that residual virus originating from the inoculum was negligible and could casily be distinguished form newly synthesized virus, even shortly after the microinoculation. This makes it possible to measure the rates of virus cell‐to‐cell spread and of virus multiplication at various times after inoculation and at various distances form the points of virus entry. These approadies can be extended to the comparison between differently reacting hosts and to the study of interference between different viruses or different virus strains.