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United States Department of Agriculture

Agricultural Research Service

Title: Remarkable Physiological Variability among Isolates of the Mite Pathogenic Fungus Neozygites Floridana: Cryopreservation and Nutritional Requirements

item Delalibera, Jr, I - CORNELL UNIVERSITY
item Humber, Richard

Submitted to: Entomology International Congress Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: August 7, 2000
Publication Date: N/A

Technical Abstract: Neozygites floridana is a fungal pathogen that is currently being released in Africa for the control of the introduced cassava green mite (CGM). The inability in producing N. floridana in vitro is one factor that has limited the initiations of molecular studies to monitor the exotic isolates in regions where the pathogen already exists. This study reports culture media and cryoprotection treatments for isolates specific to CGM. The growth in different culture media and ability to be cryopreserved of a CGM isolate (CNPMF 8), was compared to two other isolates with wider host ranges (ARSEF 662 and ARSEF 5376). The CGM isolate, is more fastidious than the two other isolates and only grows in a limited number of media. IPL-41 insect cell culture medium supplemented with 5% fetal bovine serum (FBS), 0.3% lactalbumin hydrolyzate and 0.3% yeastolate affords growth of greater than 107 hyphal bodies/ml of all isolates. FBS, the most expensive eingredient of the media can be replaced by lipids concentrate and plurioni F-68. Cultures of the ARSEF isolates can be stored in standard procedures for ultra-low temperature (cooling at 1 deg C/min to -30C followed by rapid uncontrolled cooling to -80C or -196C) with 5% glycerol or 7% polydextrose. These methods are not suitable for the CGM isolate. Trehalose and Sorbitol are appropriate cryoprotectans for CGM isolates and best results are obtained using directly rapid uncontrolled cooling.

Last Modified: 4/22/2015
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