Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application. Issue 7 (6th February 2018)

• Record Type: Journal Article
• Authors: Che, Ping; Anand, Ajith; Wu, Emily; Sander, Jeffry D.; Simon, Marissa K.; Zhu, Weiwei; Sigmund, Amy L.; Zastrow‐Hayes, Gina; Miller, Michael; Liu, Donglong; Lawit, Shai J.; Zhao, Zuo‐Yu; Albertsen, Marc C.; Jones, Todd J.
• Is Part Of: Plant biotechnology journal. Volume 16:Issue 7(2018)
• Page Start: 1388
• Page End: 1395
• Publication Date: 2018-02-06
• Subjects: sorghum transformation -- Africa sorghum varieties -- Agrobacterium -- genome modification -- ternary vector -- CRISPR/Cas9; Plant biotechnology -- Periodicals; Plant genetic engineering -- Periodicals; 630.272
• DOI: 10.1111/pbi.12879
• Languages: English
• ISSNs: 1467-7644

Abstract

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.