Gene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9

Bananas are a staple food source and a major export commodity worldwide. The Cavendish dessert banana is a triploid AAA genome type and accounts for around 47% of global production. Being essentially sterile, genetic modification is perhaps the only pathway available to improve this cultivar. In thi...

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Main Authors: Eftekhari, Fatima, Dugdale, B., Kleidon, J., Brinin, A., Shand, K., Waterhouse, P., Dale, J.
Format: Journal Article
Language:English
Published: SPRINGER 2018
Subjects:
Online Access:http://hdl.handle.net/20.500.11937/78229
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author Eftekhari, Fatima
Dugdale, B.
Kleidon, J.
Brinin, A.
Shand, K.
Waterhouse, P.
Dale, J.
author_facet Eftekhari, Fatima
Dugdale, B.
Kleidon, J.
Brinin, A.
Shand, K.
Waterhouse, P.
Dale, J.
author_sort Eftekhari, Fatima
building Curtin Institutional Repository
collection Online Access
description Bananas are a staple food source and a major export commodity worldwide. The Cavendish dessert banana is a triploid AAA genome type and accounts for around 47% of global production. Being essentially sterile, genetic modification is perhaps the only pathway available to improve this cultivar. In this study, we used the CRISPR/Cas9 gene editing system to deliver a self-cleaving polycistronic guide RNA (gRNA) designed to target exon 1 of the Phytoene desaturase (PDS) gene in the Cavendish cultivar “Williams”. Genotyping of 19 independent events showed a 100% PDS modification rate primarily in the form of insertions (1–105 nt) or deletions (1–55 nt) (indels) at the predicted cleavage site. Tri-allelic disruptive modifications were observed in 63% of plants and resulted in both albinism and dwarfing. Pale green (16%) and wildtype green (21%) phenotypes generally correlated with in-frame indels in at least one of the three PDS alleles. Editing efficiency was dependent on both target site selection and Cas9 abundance. This is the first report of a highly effective CRISPR/Cas9 modification system using a polycistronic gRNA in Cavendish banana. Such an editing platform will be of considerable utility for the development of disease resistance and novel agro-traits in this commercially important cultivar into the future.
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spelling curtin-20.500.11937-782292020-06-08T00:23:56Z Gene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9 Eftekhari, Fatima Dugdale, B. Kleidon, J. Brinin, A. Shand, K. Waterhouse, P. Dale, J. Science & Technology Life Sciences & Biomedicine Biochemical Research Methods Biochemistry & Molecular Biology Biotechnology & Applied Microbiology Genome editing PDS Cavendish banana CRISPR/Cas9 SYSTEM GENOME PLANTS RNAS Bananas are a staple food source and a major export commodity worldwide. The Cavendish dessert banana is a triploid AAA genome type and accounts for around 47% of global production. Being essentially sterile, genetic modification is perhaps the only pathway available to improve this cultivar. In this study, we used the CRISPR/Cas9 gene editing system to deliver a self-cleaving polycistronic guide RNA (gRNA) designed to target exon 1 of the Phytoene desaturase (PDS) gene in the Cavendish cultivar “Williams”. Genotyping of 19 independent events showed a 100% PDS modification rate primarily in the form of insertions (1–105 nt) or deletions (1–55 nt) (indels) at the predicted cleavage site. Tri-allelic disruptive modifications were observed in 63% of plants and resulted in both albinism and dwarfing. Pale green (16%) and wildtype green (21%) phenotypes generally correlated with in-frame indels in at least one of the three PDS alleles. Editing efficiency was dependent on both target site selection and Cas9 abundance. This is the first report of a highly effective CRISPR/Cas9 modification system using a polycistronic gRNA in Cavendish banana. Such an editing platform will be of considerable utility for the development of disease resistance and novel agro-traits in this commercially important cultivar into the future. 2018 Journal Article http://hdl.handle.net/20.500.11937/78229 10.1007/s11248-018-0083-0 English http://creativecommons.org/licenses/by/4.0/ SPRINGER fulltext
spellingShingle Science & Technology
Life Sciences & Biomedicine
Biochemical Research Methods
Biochemistry & Molecular Biology
Biotechnology & Applied Microbiology
Genome editing
PDS
Cavendish banana
CRISPR/Cas9
SYSTEM
GENOME
PLANTS
RNAS
Eftekhari, Fatima
Dugdale, B.
Kleidon, J.
Brinin, A.
Shand, K.
Waterhouse, P.
Dale, J.
Gene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9
title Gene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9
title_full Gene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9
title_fullStr Gene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9
title_full_unstemmed Gene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9
title_short Gene editing the phytoene desaturase alleles of Cavendish banana using CRISPR/Cas9
title_sort gene editing the phytoene desaturase alleles of cavendish banana using crispr/cas9
topic Science & Technology
Life Sciences & Biomedicine
Biochemical Research Methods
Biochemistry & Molecular Biology
Biotechnology & Applied Microbiology
Genome editing
PDS
Cavendish banana
CRISPR/Cas9
SYSTEM
GENOME
PLANTS
RNAS
url http://hdl.handle.net/20.500.11937/78229