In vitro Cultivation and Sporulation of Neozygites parvispora (Zygomycetes: Entomophthorales)

Abstract

Summary Neozygites parvispora was isolated from Tbrips tabaci infesting leek plants. In vitro cultivation of two isolates was achieved using supplemented Grace's insect tissue culture medium complemented with fetal bovine serum (FBS) and pre-treated lepidopteran hemolymph. This medium composition enabled exponential cell growth as rod-shaped hyphal bodies with a length over diameter ratio of 4.5-5.0. Both FBS and hemolymph were mandatory for sustained growth. The growth requirements indicate the presence of one or several growth factors in both the hemolymph and FBS. Absence of FBS resulted in growth arrest with subsequent cell lysis. Omission of hemolymph led to rounding and size decrease of the hyphal bodies, some of which formed septated germ tubes in a later stage. Eventually, cell death occurred under such conditions. Hyphal bodies from in vitro culture were capable of capilliconidia formation after entrapment in alginate pellets. This suggests the requirement of a liquid/air interface, which triggers primary sporulation and subsequent capilliconidium formation.