New Cell Lines from Freshly Hatched Larvae, Embryos and Hemocytes of the Tarnished Plant Bug Lygus lineolaris

摘要

The tarnished plant bug, Lygus lineolaris, is a major threat to both the agri-food sector and to coniferous nursery stocks in Canada, the USA, and Mexico. It is an extremely polyphagous insect feeding on the developing reproductive organs resulting in shriveled seeds and seeds without embryos. Although L. lineolaris has been reared in laboratories, there are no literature reports of a cell line from this insidious pests of seed crops. In this report, we describe the establishment of 24 continuous cell lines of L. lineolaris. The cell lines designated NRcan-Ll-1030 consist of 22 neonate lines, with 12 freely suspended and 10 attached, 10 embryonic lines, with 8 freely suspended and 2 attached, and 8 hemocyte lines with 4 freely suspended and 4 attached. They were initiated in three different media, namely, Grace's medium, Ex-Cell 401, and Mitsuhashi and Maramorosch insect medium. Concentration of FBS was started at 10% and gradually reduced to 3%. These cells are heterogeneous in nature with cells growing predominantly in large 'soccer ball' like aggregates. These aggregates, visible with the naked eye, resist vigorous pipetting and even enzymatic treatment for dispersal. Other morphologies present include small and large individual round cells, epithelial and fibroblast-like attached cells, and vacuolated cells. Initial cell growth was very slow with initial passages requiring 6 mo intervals. This interval was reduced to 1 mo by the 10th passage and to 7 d by the 13th. The freely suspended cells grow well in 250 ml shaker flasks. All cultures can be cryopreserved and revived successfully.