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Title: Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies

Abstract

Objectives Sorghum is one of the most recalcitrant species for transformation. Considering the time and effort required for stable transformation in sorghum, establishing a transient system to screen the efficiency and full functionality of vector constructs is highly desirable. Results Here, we report an Agrobacterium -mediated transient transformation assay with intact sorghum leaves using green fluorescent protein as marker. It also provides a good monocot alternative to tobacco and protoplast assays with a direct, native and more reliable system for testing single guide RNA (sgRNA) expression construct efficiency. Given the simplicity and ease of transformation, high reproducibility, and ability to test large constructs, this method can be widely adopted to speed up functional genomic and genome editing studies.

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1572061
DOE Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article
Country of Publication:
United States
Language:
English

Citation Formats

Sharma, Rita, Liang, Yan, Lee, Mi Yeon, Pidatala, Venkataramana, Mortimer, Jenny, and Scheller, Henrik. Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies. United States: N. p., 2019. Web. doi:10.1101/779918.
Sharma, Rita, Liang, Yan, Lee, Mi Yeon, Pidatala, Venkataramana, Mortimer, Jenny, & Scheller, Henrik. Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies. United States. doi:10.1101/779918.
Sharma, Rita, Liang, Yan, Lee, Mi Yeon, Pidatala, Venkataramana, Mortimer, Jenny, and Scheller, Henrik. Tue . "Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies". United States. doi:10.1101/779918.
@article{osti_1572061,
title = {Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies},
author = {Sharma, Rita and Liang, Yan and Lee, Mi Yeon and Pidatala, Venkataramana and Mortimer, Jenny and Scheller, Henrik},
abstractNote = {Objectives Sorghum is one of the most recalcitrant species for transformation. Considering the time and effort required for stable transformation in sorghum, establishing a transient system to screen the efficiency and full functionality of vector constructs is highly desirable. Results Here, we report an Agrobacterium -mediated transient transformation assay with intact sorghum leaves using green fluorescent protein as marker. It also provides a good monocot alternative to tobacco and protoplast assays with a direct, native and more reliable system for testing single guide RNA (sgRNA) expression construct efficiency. Given the simplicity and ease of transformation, high reproducibility, and ability to test large constructs, this method can be widely adopted to speed up functional genomic and genome editing studies.},
doi = {10.1101/779918},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2019},
month = {9}
}

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