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Agency for the Assessment and Application of Technology (BPPT)

1 ARTICLES PUBLISHED IN JoVE

JoVE Journal

Transforming, Genome Editing and Phenotyping the Nitrogen-fixing Tropical Cannabaceae Tree Parasponia andersonii

Titis A.K. Wardhani^1,2, Yuda Purwana Roswanjaya^1,2, Simon Dupin^1,3, Huchen Li^1,4, Sidney Linders¹, Marijke Hartog¹, Rene Geurts¹, Arjan van Zeijl¹

¹Laboratory of Molecular Biology, Department of Plant Sciences, Wageningen University & Research, ²Center of Technology for Agricultural Production, Agency for the Assessment and Application of Technology (BPPT), ³Department of Ecological Science, Faculty of Earth and Life Sciences, Vrije Universiteit Amsterdam, ⁴Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, Beijing University of Agriculture

Parasponia andersonii is a fast-growing tropical tree that belongs to the Cannabis family (Cannabaceae) and can form nitrogen-fixing root nodules in association with the rhizobium. Here, we describe a detailed protocol for reverse genetic analyses in P. andersonii based on Agrobacterium tumefaciens-mediated stable transformation and CRISPR/Cas9-based genome editing.