Creating yellow seed Camelina sativa with enhanced oil accumulation by CRISPR‐mediated disruption of Transparent Testa 8

Author:

Cai Yuanheng12ORCID,Liang Yuanxue1ORCID,Shi Hai1ORCID,Cui Jodie12ORCID,Prakash Shreyas1ORCID,Zhang Jianhui3ORCID,Anaokar Sanket1ORCID,Chai Jin1ORCID,Schwender Jorg1ORCID,Lu Chaofu3ORCID,Yu Xiao‐Hong12ORCID,Shanklin John1ORCID

Affiliation:

1. Department of Biology Brookhaven National Laboratory Upton NY USA

2. Department of Biochemistry and Cell Biology Stony Brook University Stony Brook NY USA

3. Department of Plant Sciences and Plant Pathology Montana State University Bozeman MT USA

Abstract

SummaryCamelina (Camelina sativa L.), a hexaploid member of the Brassicaceae family, is an emerging oilseed crop being developed to meet the increasing demand for plant oils as biofuel feedstocks. In other Brassicas, high oil content can be associated with a yellow seed phenotype, which is unknown for camelina. We sought to create yellow seed camelina using CRISPR/Cas9 technology to disrupt its Transparent Testa 8 (TT8) transcription factor genes and to evaluate the resulting seed phenotype. We identified three TT8 genes, one in each of the three camelina subgenomes, and obtained independent CsTT8 lines containing frameshift edits. Disruption of TT8 caused seed coat colour to change from brown to yellow reflecting their reduced flavonoid accumulation of up to 44%, and the loss of a well‐organized seed coat mucilage layer. Transcriptomic analysis of CsTT8‐edited seeds revealed significantly increased expression of the lipid‐related transcription factors LEC1, LEC2, FUS3, and WRI1 and their downstream fatty acid synthesis‐related targets. These changes caused metabolic remodelling with increased fatty acid synthesis rates and corresponding increases in total fatty acid (TFA) accumulation from 32.4% to as high as 38.0% of seed weight, and TAG yield by more than 21% without significant changes in starch or protein levels compared to parental line. These data highlight the effectiveness of CRISPR in creating novel enhanced‐oil germplasm in camelina. The resulting lines may directly contribute to future net‐zero carbon energy production or be combined with other traits to produce desired lipid‐derived bioproducts at high yields.

Funder

Basic Energy Sciences

Biological and Environmental Research

Publisher

Wiley

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