Vol. 5, Issue 1 (2020)
Expression of small interfering RNAs in wheat plant specific to gluten hydrolyzing proteinase (GHP) transcripts of sunnpest (Eurygaster Integriceps)
Author(s): Siva Prasad Venkata Satya Chekkara, Alex Gaponenko
Abstract: Many times alternative approaches are not available against chemical pesticides for controlling pest of economic importance in an ecofriendly manner. Sunnpest is one such pest against which till now no biological approach is available for its radical control. After coming across Bacillus thuringensis (Bt) proteins resistance in the insect pest, the latest dsRNA based siRNA approach found to be more promising for the insect control alongwith no adverse effect on the environment. The present approach involves expressing multiple synthetic siRNAs to target vital transcript or multiple transcripts of a pest. To accomplish this, siRNAs were designed by taking into consideration of avoiding the perfect off targets problem. Sunnpest (Eurygaster integriceps), Gluten hydrolyzing proteinase (GHP) gene has been selected as a target gene and designed multiple shRNAs sequences and cloned in a single binary vector. Agrobacterium based transformation was done in Wheat embryos and screened through kanamycin resistance selectivity, positive plants evaluated for shRNA expression and quantified through Real-Time PCR. The shRNA was expressed in transgenic wheat plants. Once Sunnpest feeds upon these transgenic wheat, shRNAs enters through cell sap into the insect body and converted into functional siRNAs inside the insect and specifically knock down GPH mRNA which further protects wheat seeds from GPH induced external digestion resulting in to better dough making quality wheat seeds.