Silencing of Aflatoxin Synthesis Through Rna Interference (Rnai) in Peanut Plants
Project Number: 6044-21000-004-01
Start Date: Sep 30, 2013
End Date: Sep 30, 2018
Prevent formation and accumulation of aflatoxins in peanut plants and peanut seeds by silencing several genes in the aflatoxin biosynthesis pathway using RNAi transgenic peanut plants”
1. Clone Aspergillus flavus gene fragments from the aflatoxin biosynthesis pathway to be used in molecular constructs for inducible and constitutive RNAi silencing.
2. Generate transgenic peanut plants with molecular constructs for silencing
3. Screen transgenic plants for presence of aflatoxins upon challenging with Aspergillus flavus and/or Aspergillus parasiticus.
4. Evaluate gene silencing by transcriptome sequencing of pathogen challenged infection sites, and confirm specific gene silencing by RT-PCR.
We will make molecular constructs for RNAi using fragments of the aflatoxin biosynthesis pathway. Genetically transform peanut plants with the RNAi constructs for constitutive and inducible silencing of the biosynthesis genes. Transgenic plants will be challenged with the pathogens Aspergillus flavus and Aspergillus parasiticus, and infected tissues will be analyzed using transcriptome sequencing, and by Real-time PCR of specific genes. The same tissues will be analyzed for presence of aflatoxin.