Protocol

Centre : Central Tuber Crops Research Institute(CTCRI),TVM
Name of the Protocol
A rapid and efficient method for in vitro production of zoospores
Nature : Novel
Crop / Pathogen : Taro/Phytophthora colocasiae

Inventors

1. Nath V. S.
2. Shyni,
3. Hegde V. M.
4. Misra R. S.
5. Veena S. S.
6. Raj M.
7. Jeeva M. L.

Objectives

1. To mass produce zoospores for artificial inoculation

Procedure / Summary

Mycelial plugs (5 mm diameter) were excised from actively growing cultures of P. colocasiae on carrot agar (CA) medium. The plugs were placed on the adaxial side of the taro leaf discs (5x5 cm; cv Sree Kiran, leaf blight susceptible) floating on 50 ml sterile water in 200 mm sterile petri dishes. The lids of the petri plates were lined with moistened filter paper to maintain high humidity and incubated at 25°C in the dark for 3–4 days. Following incubation, the infected leaf portions were excised, placed with adaxial side upwards in 6-well plates with 2 ml sterile water and slowly immersed in water. Then, the plates were chilled at 4°C for 15 min and thereafter at 37°C for another 30 min.

Applications

1. The method could produce mass no. of zoospores in limited time.

Publications

1. Nath V. S., Shyni, Hegde V. M., Misra R. S., Veena S. S., Raj M., Jeeva M. L. (2016). A rapid and efficient method for in vitro screening of taro for leaf blight disease. Journal of Phytopathology. doi: 10.1111/jph.12477.