Modified diet assay for studying feeding RNAi in cotton leafhopper, Amrasca biguttula biguttula v1

Amrasca biguttula biguttula (Ishida) commonly known as cotton leafhopper is a serious pest of cotton and okra. Nutritional requirements or growth physiology for this insect has not been much known till date. We developed artificial membrane feeding method for leafhopper which can be further used for RNA interference studies. Main components of this diet were L-cysteine (5.0 mg), glycine (2.0 mg), nicotinic acid (1.0 mg), sucrose (500 mg), K2HPO4(50 mg), ZnCl2 (0.04 mg), Thiamine HCl (0.25 mg), Vitamin B6 (0.25 mg), Becosules capsule powder (Pfizer Limited, USA) (2.0 mg), Green food dye (GanpatiSyn Food Colour, India) (2.0 mg) per 10ml of autoclaved double distilled water. To make diet accessible to the cotton leafhoppers the 1.5 ml of Microcentrifuge tube’s (MCT) cap (Tarsons Products Pvt. Ltd. India) was removed and diet was filled in 2 layers of parafilm sachet (diet was poured on first layer of parafilm on cap and covered with another layer of stretched parafilm) into cap. The MCT’s was cut 1/4th from bottom end, which was covered with muslin cloth (40 count mesh size). This MCTs with leafhopper was kept in incubator at 65-70% RH, 14:10 h light and dark photoperiod and 27± 2°C.