Characterization of a newly established potato tuber moth (Phthorimaea operculella Zeller) cell line.
OBJECTIVE
Potato tuber moth (PTM), Phthorimaea operculella Zeller is a widely distributed, devastating pest of potatoes attacking the foliage and infest the tubers in both field and store causing serious economic damage. As application of PTM granulovirus (PTM-GV) has shown significant reduction in damage, attempts were made to develop a new cell line from this insect to grow PTM-GV for use as a biopesticide.


METHODS
Approximately 100 mg of insect eggs were collected, surface sterilized and crushed gently in a boiling tube aseptically. The tissues were washed with physiological saline, suspended in growth medium and incubated stationary at 28 degrees C. Morphology of cells was studied after staining with Giemsa. Besides karyological and growth curve studies, PCR amplification was also done for rapid amplified polymorphic DNA pattern.


RESULTS
A new cell line from the embryonic tissue of PTM was maintained in Mitsuhashi Maramorosch medium supplemented with 10 per cent foetal bovine serum. It is in the 78th passage level and designated as NIV-PTM-1095. Random amplified polymorphic DNA profile analysis indicated this as a new cell line from potato tuber moth and differed from the profiles of two other lepidopteran cell lines maintained in the laboratory. Three different cell types were observed at the 40th passage level and comprised of epithelial-like cells (77%), fibroblast-like cells (20%) and giant cells (3%). The chromosome number varied from 54-176. The cell line had a cell doubling time of approximately 42 h during the logarithmic phase of growth. The cell line did not support the multiplication of any of the baculoviruses used in the study.


CONCLUSIONS
Since the new cell line is found to replicate PTM-GV, it may be useful for the propagation of PTM-GV in large scale. Studies to scale up the production of the GV in the cell line and field trials may lead to its widespread use as an eco-friendly biopesticide.
Sudeep
A B
AB
National Institute of Virology (ICMR), Pune, India.
Khushiramani
R
R
Athawale
S S
SS
Mishra
A C
AC
Mourya
D T
DT
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D004247:DNA / Q000235:genetics; D025722:Granulovirus / Q000502:physiology; D009036:Moths / Q000166:cytology* / Q000235:genetics / Q000472:pathogenicity / Q000821:virology; D010572:Pest Control, Biological; D010935:Plant Diseases / Q000469:parasitology; D019105:Random Amplified Polymorphic DNA Technique; D011198:Solanum tuberosum / Q000469:parasitology
D004247:DNA