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ABSTRACT
A technique has been devised for maintaining the virulence of the endophyte of Alnus glutinosa L. during in vitro cultivation. Essential to the technique are the inclusion of an ether extract of Alnus roots and cycloheximide and chloramphenicol to the artificial medium. Using the devised medium, endophyte virulence has been maintained for 14 days whereas in the absence of the ether extract and antibiotics, virulence was lost within 3 days. Nodules that are subsequently formed after inoculation of plants with the incubated endophyte are normal. Visible nodules were formed within 7 to 12 weeks after inoculation, a period which was comparable to the nodulation time of control plants inoculated with fresh crushed nodules. In addition, the nodules were capable of reducing acetylene which is indicative of nitrogen fixing activity. Also, during this study it was found that freeze-drying nodules does not reduce the virulence of the nodule-inducing endophyte.
1 Paper number 4290 of the Journal Series of the North Carolina Agr. Exp. Sta., North Carolina State Univ., Raleigh 27607. Portions of this paper were presented before Div. S-3, Soil Sci. Soc. Amer., Las Vegas, Nev., 15 Nov. 1973.
2 Graduate Research Assistant and Associate Professor, Soil Sci. Dep., North Carolina State Univ., Raleigh.
Received for publication February 2, 1974. Accepted for publication May 13, 1974.
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