Protoplast-Based Transient Expression and Gene Editing in Shrub Willow (Salix purpurea L.)

Hyden, Brennan; Yuan, Guoliang; Liu, Yang; Smart, Lawrence B.; Tuskan, Gerald A.; Yang, Xiaohan

doi:10.3390/plants11243490

Protoplast-Based Transient Expression and Gene Editing in Shrub Willow (Salix purpurea L.)

Journal Article · Mon Dec 12 23:00:00 EST 2022 · Plants

DOI:https://doi.org/10.3390/plants11243490· OSTI ID:1906582

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Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States); Cornell University, Ithaca, NY (United States)
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Cornell University, Ithaca, NY (United States)

Shrub willows (Salix section Vetrix) are grown as a bioenergy crop in multiple countries and as ornamentals across the northern hemisphere. To facilitate the breeding and genetic advancement of shrub willow, there is a strong interest in the characterization and functional validation of genes involved in plant growth and biomass production. While protocols for shoot regeneration in tissue culture and production of stably transformed lines have greatly advanced this research in the closely related genus Populus, a lack of efficient methods for regeneration and transformation has stymied similar advancements in willow functional genomics. Moreover, transient expression assays in willow have been limited to callus tissue and hairy root systems. Here we report an efficient method for protoplast isolation from S. purpurea leaf tissue, along with transient overexpression and CRISPR-Cas9 mediated mutations. This is the first such report of transient gene expression in Salix protoplasts as well as the first application of CRISPR technology in this genus. These new capabilities pave the way for future functional genomics studies in this important bioenergy and ornamental crop.

View Accepted Manuscript (DOE)

Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1906582

Journal Information:: Plants, Journal Name: Plants Journal Issue: 24 Vol. 11; ISSN 2223-7747

Publisher:: MDPICopyright Statement

Country of Publication:: United States

Language:: English

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