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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:
; ; ; ; ORCiD logo;
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)
OSTI Identifier:
1618915
Alternate Identifier(s):
OSTI ID: 1608281
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Published Article
Journal Name:
BMC Research Notes
Additional Journal Information:
Journal Name: BMC Research Notes Journal Volume: 13 Journal Issue: 1; Journal ID: ISSN 1756-0500
Publisher:
BioMed Central
Country of Publication:
United Kingdom
Language:
English
Subject:
09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; Agrobacterium; CRISPR; sgRNA; Sorghum; Transformation; Transient

Citation Formats

Sharma, Rita, Liang, Yan, Lee, Mi Yeon, Pidatala, Venkataramana R., Mortimer, Jenny C., and Scheller, Henrik V. Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies. United Kingdom: N. p., 2020. Web. doi:10.1186/s13104-020-04968-9.
Sharma, Rita, Liang, Yan, Lee, Mi Yeon, Pidatala, Venkataramana R., Mortimer, Jenny C., & Scheller, Henrik V. Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies. United Kingdom. doi:https://doi.org/10.1186/s13104-020-04968-9
Sharma, Rita, Liang, Yan, Lee, Mi Yeon, Pidatala, Venkataramana R., Mortimer, Jenny C., and Scheller, Henrik V. Thu . "Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies". United Kingdom. doi:https://doi.org/10.1186/s13104-020-04968-9.
@article{osti_1618915,
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 R. and Mortimer, Jenny C. and Scheller, Henrik V.},
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 = {10.1186/s13104-020-04968-9},
journal = {BMC Research Notes},
number = 1,
volume = 13,
place = {United Kingdom},
year = {2020},
month = {2}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: https://doi.org/10.1186/s13104-020-04968-9

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