Skip to main content
OSTI.GOVU.S. Department of Energy Office of Scientific and Technical Information
U.S. Department of Energy
Office of Scientific and Technical Information
 

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

Journal Article · · Plant Biotechnology Journal
DOI:https://doi.org/10.1111/pbi.14403· OSTI ID:2446759
 [1];  [2];  [2];  [1];  [2];  [3];  [2];  [2];  [2];  [3];  [1];  [2]
  1. Department of Biology Brookhaven National Laboratory Upton NY USA, Department of Biochemistry and Cell Biology Stony Brook University Stony Brook NY USA
  2. Department of Biology Brookhaven National Laboratory Upton NY USA
  3. Department of Plant Sciences and Plant Pathology Montana State University Bozeman MT USA
+ Show Author Affiliations

Summary

Camelina ( 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.

Sponsoring Organization:
USDOE
Grant/Contract Number:
NONE; SC0021369; SC0018420; SC0012704
OSTI ID:
2446759
Alternate ID(s):
OSTI ID: 2396622
OSTI ID: 2426608
OSTI ID: 2477874
Journal Information:
Plant Biotechnology Journal, Journal Name: Plant Biotechnology Journal Journal Issue: 10 Vol. 22; ISSN 1467-7644
Publisher:
Wiley-BlackwellCopyright Statement
Country of Publication:
United Kingdom
Language:
English

References (56)

Histochemical characterisation of unextractable seed coat pigments and quantification of extractable lignin in the Brassicaceae journal January 2004
RNA isolation from recalcitrant plant tissue journal December 1994
New sources of water-soluble seed gums journal April 1965
Natural mutations in two homoeologous TT8 genes control yellow seed coat trait in allotetraploid Brassica juncea (AABB) journal November 2013
Generation of transgenic plants of a potential oilseed crop Camelina sativa by Agrobacterium-mediated transformation journal September 2007
Current progress in lipid-based biofuels: Feedstocks and production technologies journal May 2022
Camelina uses, genetics, genomics, production, and management journal December 2016
Mutagenesis of the FAE1 genes significantly changes fatty acid composition in seeds of Camelina sativa journal February 2018
TTG1 complex MYBs, MYB5 and TT2, control outer seed coat differentiation journal January 2009
Targeted Knockout of BnTT2 Homologues for Yellow-Seeded Brassica napus with Reduced Flavonoids and Improved Fatty Acid Composition journal May 2020
The production of yellow-seeded Brassica napus (AACC) through crossing interspecific hybrids of B. campestris (AA) and B. carinata (BBCC) with B. napus journal October 1998
The emerging biofuel crop Camelina sativa retains a highly undifferentiated hexaploid genome structure journal April 2014
Effective extraction of Arabidopsis adherent seed mucilage by ultrasonic treatment journal January 2017
Oil and protein content of the seeds of some pasture legumes journal January 1974
Modulating seed β-ketoacyl-acyl carrier protein synthase II level converts the composition of a temperate seed oil to that of a palm-like tropical oil journal March 2007
Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system journal March 2015
Desaturation and Hydroxylation: RESIDUES 148 AND 324 OF journal February 2002
Evolutionary studies of the bHLH transcription factors belonging to MBW complex: their role in seed development journal July 2023
Recent advances in the transcriptional regulation of the flavonoid biosynthetic pathway journal January 2011
CRISPR-P: A Web Tool for Synthetic Single-Guide RNA Design of CRISPR-System in Plants journal September 2014
Relative expression software tool (REST(C)) for group-wise comparison and statistical analysis of relative expression results in real-time PCR journal May 2002
Biotin attachment domain-containing proteins mediate hydroxy fatty acid-dependent inhibition of acetyl CoA carboxylase journal January 2021
The GLABRA2 gene encodes a homeo domain protein required for normal trichome development in Arabidopsis. journal June 1994
Participation of the Arabidopsis bHLH Factor GL3 in Trichome Initiation Regulatory Events journal September 2007
TRANSPARENT TESTA8 Inhibits Seed Fatty Acid Accumulation by Targeting Several Seed Development Regulators in Arabidopsis journal April 2014
Expression of a Lychee PHOSPHATIDYLCHOLINE:DIACYLGLYCEROL CHOLINEPHOSPHOTRANSFERASE with an Escherichia coli CYCLOPROPANE SYNTHASE Enhances Cyclopropane Fatty Acid Accumulation in Camelina Seeds journal May 2019
TRANSPARENT TESTA GLABRA2 , a Trichome and Seed Coat Development Gene of Arabidopsis, Encodes a WRKY Transcription Factor journal June 2002
The TT8 Gene Encodes a Basic Helix-Loop-Helix Domain Protein Required for Expression of DFR and BAN Genes in Arabidopsis Siliques journal October 2000
TT2, TT8, and TTG1 synergistically specify the expression of BANYULS and proanthocyanidin biosynthesis in Arabidopsis thaliana journal August 2004
TT8 controls its own expression in a feedback regulation involving TTG1 and homologous MYB and bHLH factors, allowing a strong and cell-specific accumulation of flavonoids in Arabidopsis thaliana journal June 2006
Genetic variation of yellow‐seeded rapeseed lines (Brassica napus L.) from different genetic sources journal October 1997
Plant carbon allocation in a changing world – challenges and progress: introduction to a Virtual Issue on carbon allocation: Introduction to a virtual issue on carbon allocation journal July 2020
AtMIF1 increases seed oil content by attenuating GL2 inhibition journal November 2020
Significant enhancement of fatty acid composition in seeds of the allohexaploid, Camelina sativa , using CRISPR/Cas9 gene editing journal January 2017
Selective gene dosage by CRISPR-Cas9 genome editing in hexaploid Camelina sativa journal April 2017
Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil journal January 2018
Targeted mutagenesis of BnTT8 homologs controls yellow seed coat development for effective oil production in Brassica napus L. journal May 2020
Enhancing the accumulation of eicosapentaenoic acid and docosahexaenoic acid in transgenic Camelina through the CRISPR‐Cas9 inactivation of the competing FAE1 pathway journal July 2022
Targeted mutagenesis of flavonoid biosynthesis pathway genes reveals functional divergence in seed coat colour, oil content and fatty acid composition in Brassica napus L. journal October 2023
The new frontier of genome engineering with CRISPR-Cas9 journal November 2014
Genetics and Biochemistry of seed Flavonoids journal June 2006
CRISPR/Cas9 editing of three CRUCIFERIN C homoeologues alters the seed protein profile in Camelina sativa journal July 2019
Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation journal July 2015
Intron-mediated enhancement of DIACYLGLYCEROL ACYLTRANSFERASE1 expression in energycane promotes a step change for lipid accumulation in vegetative tissues journal October 2023
A network of redundant bHLH proteins functions in all TTG1-dependent pathways of Arabidopsis journal October 2003
Assessing Diversity in the Camelina Genus Provides Insights into the Genome Structure of Camelina sativa journal April 2020
Arabidopsis Seed Coat Mucilage is a Specialized Cell Wall that Can be Used as a Model for Genetic Analysis of Plant Cell Wall Structure and Function journal January 2012
Regulation of Carbon Partitioning in the Seed of the Model Legume Medicago truncatula and Medicago orbicularis: A Comparative Approach journal December 2017
Metabolic Engineering a Model Oilseed Camelina sativa for the Sustainable Production of High-Value Designed Oils journal February 2020
Increasing Monounsaturated Fatty Acid Contents in Hexaploid Camelina sativa Seed Oil by FAD2 Gene Knockout Using CRISPR-Cas9 journal June 2021
An expanded role for the transcription factor WRINKLED1 in the biosynthesis of triacylglycerols during seed development journal August 2022
New Flowering and Architecture Traits Mediated by Multiplex CRISPR-Cas9 Gene Editing in Hexaploid Camelina sativa journal August 2022
Integument-Specific Transcriptional Regulation in the Mid-Stage of Flax Seed Development Influences the Release of Mucilage and the Seed Oil Content journal October 2021
Toluidine blue: A review of its chemistry and clinical utility journal January 2012
A review of Brassica seed color journal May 2011
Seed coat mucilage cells of Arabidopsis thaliana as a model for plant cell wall research journal July 2010

Similar Records

Related Subjects