Transforming, Genome Editing and Phenotyping the Nitrogen-fixing Tropical Cannabaceae Tree Parasponia andersoniiTitis A.K. Wardhani 1,2, Yuda Purwana Roswanjaya 1,2, Simon Dupin 1,3, Huchen Li 1,4, Sidney Linders 1, Marijke Hartog 1, Rene Geurts 1, Arjan van Zeijl 1
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.