Open AccessDeveloping a flexible, high‐efficiency Agrobacterium ‐mediated sorghum transformation system with broad application
Author(s) -
Che Ping,
Anand Ajith,
Wu Emily,
Sander Jeffry D.,
Simon Marissa K.,
Zhu Weiwei,
Sigmund Amy L.,
ZastrowHayes Gina,
Miller Michael,
Liu Donglong,
Lawit Shai J.,
Zhao ZuoYu,
Albertsen Marc C.,
Jones Todd J.
Publication year - 2018
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12879
Subject(s) - biology , sorghum , agrobacterium , transformation (genetics) , crispr , selectable marker , microbiology and biotechnology , crop , genetically modified crops , cas9 , plasmid , transgene , gene , computational biology , agronomy , genetics
Summary Sorghum is the fifth most widely planted cereal crop in the world and is commonly cultivated in arid and semi‐arid regions such as Africa. Despite its importance as a food source, sorghum genetic improvement through transgenic approaches has been limited because of an inefficient transformation system. Here, we report a ternary vector (also known as cohabitating vector) system using a recently described pVIR accessory plasmid that facilitates efficient Agrobacterium ‐mediated transformation of sorghum. We report regeneration frequencies ranging from 6% to 29% in Tx430 using different selectable markers and single copy, backbone free ‘quality events’ ranging from 45% to 66% of the total events produced. Furthermore, we successfully applied this ternary system to develop transformation protocols for popular but recalcitrant African varieties including Macia , Malisor 84‐7 and Tegemeo . In addition, we report the use of this technology to develop the first stable CRISPR /Cas9‐mediated gene knockouts in Tx430.