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An improved selective medium for the isolation of Verticillium dahliae Kleb

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Title An improved selective medium for the isolation of Verticillium dahliae Kleb
Names Zehsazian, Hassan (creator)
Horner, Chester E. (advisor)
Date Issued 1965-08-11 (iso8601)
Note Graduation date: 1966
Abstract Sodium polypectate (SPP) was added to an ethanol-streptomycin
agar (ESA) medium to determine if it would provide a selective
substrate for the growth of Verticillium dahliae and for isolation of
the fungus from soil and plant tissues in the presence of saprophytic
fungi. Sodium polypectate improved greatly the formation and pigmentation of microscierotia in four different isolates of V. dahliae.
Adding SPP to ESA medium also improved the recovery rate of the
fungus from soil and from infected plants. At high inoculum
densities, SPP did not increase colony numbers, and ESA without
SPP apparently allowed the recovery of all viable fungus propagules.
However, adding SPP to the medium, even at levels as low as 0.25
g/l, increased the size, microsclerotial formation, and pigmentation
of the colonies in every case. All the colonies on ESA-SPP medium
were very distinct and they could be observed and counted easily
and rapidly with the unaided eye. The time required for formation and pigmentation of microsclerotia was shortened by five to seven
days when SPP was added to the medium. The results suggest that
V. dahliae utilizes SPP primarily to form pigmented resting structures rather than vegetative mycelium.
In soil dilutions, addition of SPP up to 1.0 g/l of ESA increased
the recovery of V. dahliae. At higher concentrations of SPP,
vigorous growth of saprophytic soil fungi inhibited the growth and
masked the presence of V. dahliae.
Microsclerotial production by V. dahliae was obtained in shake
cultures by adding SPP to a 0.1 percent water agar or a 0.1 percent
Czapek Dox solution, both containing 0.76 percent ethanol and 101
ppm streptomycin. The intensity of production was always related
to the level of SPP in the media. Water agar containing SPP yielded
many more microsclerotia than Czapek Dox containing SPP, at all
tested levels.
For best microsclerotial production the combined effect of
SPP and ethanol was necessary in all experiments.
Improvement of ESA medium by addition of appropriate amounts
of SPP will allow more refined studies in the biology of V. dahliae
and the knowledge gained from such experiments should enhance our
understanding of this important plant pathogen and eventually aid
in its control.
Genre Thesis/Dissertation
Topic Verticillium
Identifier http://hdl.handle.net/1957/47763

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