Establishment of Asian citrus psyllid (Diaphorina citri) primary cultures

• Published in: In vitro cellular & developmental biology. Animal 2009-07, Vol.45 (7), p.317-320
• Main Authors: Marutani-Hert, Mizuri, Hunter, Wayne B, Hall, David G
• Format: Article
• Language: English
• Subjects: Animal Genetics and Genomics; Animals; Biomedical and Life Sciences; Cell Adhesion; Cell Biology; Cell Culture; Cell Culture Techniques; Cell growth; Cell lines; Cell migration; Cell Movement; Citrus; Culture Media; Cultured cells; Developmental Biology; Diaphorina citri; Disease vectors; Embryo, Nonmammalian - cytology; Embryos; Fruits; Genomics; Greening; Hemiptera; Hemiptera - cytology; Hemiptera - embryology; Hemiptera - microbiology; Insect cells; Insect eggs; Insect genetics; Insect vectors; Kuwayama; Life Sciences; Life span; Limiting factors; Media (culture); Nutrients; Polymerase chain reaction; Psyllidae; Pure culture; Rhizobiaceae; Stem Cells; Trees
• ISSN: 1071-2690; 1543-706X
• DOI: 10.1007/s11626-009-9188-3

The Asian citrus psyllid (AsCP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is a highly competent vector of the phloem-inhabiting bacterium Candidatus Liberibacter asiaticus associated with the citrus disease huanglongbing (HLB). Commonly referred to as citrus greening disease in the USA, HLB causes reduced fruit yields, quality, and ultimately tree death and is considered the most serious citrus disease. HLB has become a major limiting factor to the production of citrus worldwide. Studies of HLB have been impeded by the fact that C. Liberibacter has not yet been cultured on artificial nutrient media. After being acquired by a psyllid, C. Liberibacter asiaticus is reported to replicate within the psyllid and is retained by the psyllid throughout its life span. We therefore hypothesized that C. Liberibacter asiaticus could be cultured in vitro using psyllid cell cultures as the medium and investigated the establishment of a pure culture for AsCP cells. Several commercially available insect cell culture media along with some media we developed were screened for viability to culture cells from AsCP embryos. Cells from psyllid tissues adhered to the plate and migration was observed within 24 h. Cells were maintained at 20°C. We successfully established primary psyllid cell cultures, referred to as DcHH-1, for D. citri Hert-Hunter-1, with a new media, Hert-Hunter-70.