title: Knockdown of Polyphenol Oxidase Gene Expression in Potato (Solanum tuberosum L.) with Artificial MicroRNAs.
abstract: It is of great importance and interest to develop crop varieties with low polyphenol oxidase (PPO) activity for the food industry because PPO-mediated oxidative browning is a main cause of post-harvest deterioration and quality loss of fresh produce and processed foods. We recently demonstrated that potato tubers with reduced browning phenotypes can be produced by inhibition of the expression of several PPO gene isoforms using artificial microRNA (amiRNA) technology. The approach introduces a single type of 21-nucleotide RNA population to guide silencing of the PPO gene transcripts in potato tissues. Some advantages of the technology are: small RNA molecules are genetically transformed, off-target gene silencing can be avoided or minimized at the stage of amiRNA designs, and accuracy and efficiency of the processes can be detected at every step using molecular biological techniques. Here we describe the methods for transformation and regeneration of potatoes with amiRNA vectors, detection of the expression of amiRNAs, identification of the cleaved product of the target gene transcripts, and assay of the expression level of PPO gene isoforms in potatoes.
authors: Pacific Agri-Food Research Center, Agriculture and Agri-Food Canada, 4200 Highway 97, Box 5000, Summerland, BC, Canada, V0H 1Z0.|College of Forestry, Northwest A & F University, Yangling, Shaanxi, China.
authors: Ming
authors: M
authors: Chi
authors: Pacific Agri-Food Research Center, Agriculture and Agri-Food Canada, 4200 Highway 97, Box 5000, Summerland, BC, Canada, V0H 1Z0.
authors: Basdeo
authors: B
authors: Bhagwat
authors: Department of Biological Sciences, Michigan Technological University, Houghton, MI, 49931, USA.|Provincial State Key Laboratory of Wheat and Maize Crop Science, Henan Agricultural University, Zhengzhou, China.
authors: Guiliang
authors: G
authors: Tang
authors: Pacific Agri-Food Research Center, Agriculture and Agri-Food Canada, 4200 Highway 97, Box 5000, Summerland, BC, Canada, V0H 1Z0. yu.xiang@agr.gc.ca.
authors: Yu
authors: Y
authors: Xiang
mesh_terms: D004156:Catechol Oxidase / Q000235:genetics*; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D035683:MicroRNAs / Q000235:genetics*; D030821:Plants, Genetically Modified; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D011198:Solanum tuberosum / Q000235:genetics*
chemical_list: D035683:MicroRNAs; D004156:Catechol Oxidase
keywords: Artificial microRNA (amiRNA); Gene knockdown; Polyphenol oxidase (PPO); Potato genetic transformation; RNA silencing; Solanum tuberosum; StuPPO