Knockdown of Polyphenol Oxidase Gene Expression in Potato (Solanum tuberosum L.) with Artificial MicroRNAs

  • Ming Chi
  • Basdeo Bhagwat
  • Guiliang Tang
  • Yu XiangEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1405)


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.

Key words

Artificial microRNA (amiRNA) Gene knockdown Polyphenol oxidase (PPO) Potato genetic transformation RNA silencing Solanum tuberosum StuPPO 


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Ming Chi
    • 1
    • 2
  • Basdeo Bhagwat
    • 1
  • Guiliang Tang
    • 3
    • 4
  • Yu Xiang
    • 1
    Email author
  1. 1.Pacific Agri-Food Research CenterAgriculture and Agri-Food CanadaSummerlandCanada
  2. 2.College of ForestryNorthwest A & F UniversityYanglingChina
  3. 3.Department of Biological SciencesMichigan Technological UniversityHoughtonUSA
  4. 4.Provincial State Key Laboratory of Wheat and Maize Crop ScienceHenan Agricultural UniversityZhengzhouChina

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