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Generation of chemically induced mutations using in vitro propagated shoot tip tissues for genetic improvement of fruit trees

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Abstract

Mutagenesis via treatment of seeds with chemical mutagens such as ethyl methanesulfonate (EMS) has been widely used for crop improvement. However, this approach is not suitable for some crop species such as clonally propagated crops and allogamous perennial plants characteristically with high levels of genome heterozygosity and a long life cycle. Here, we report direct treatment of in vitro-induced peach shoot tip tissues with EMS for generation of mutations in peach, a perennial, woody fruit tree. We optimized EMS dosage and exposure time and found that treatment of the explants with 0.2 % EMS for 16 h was optimal for generation of genetic variations. So far we have generated nearly 2000 peach seedlings. Typical EMS-induced phenotypic variations were present in the seedlings. Next generation sequencing and subsequent data analyses were performed to monitor the efficiency of mutagenesis. The mutation rate was estimated to be one mutation per 150 kilobase pairs in the mutagenized population, suggesting effectiveness of this method in inducing mutagenesis in peach. Taken together, our data open an avenue for the generation of mutant populations suitable for crop improvement in allogamous perennial plants including fruit trees and clonally propagated plants.

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Abbreviations

BAQ:

Base alignment quality

BRM:

Bud reactivation medium

DD:

Damage degree

DMSO:

Dimethyl sulfoxide

EMS:

Ethyl methanesulfonate

ENU:

Ethylnitrosourea

IBA:

Indol butyric acid

LR:

Lethal rate

NGS:

Next generation sequencing

qPCR:

Quantitative polymerase chain reaction

RPM:

Revolutions per minute

SIM:

Shoot induction medium

SNP:

Single nucleotide polymorphism

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Acknowledgments

We are very grateful to Anna Xu for sharing peach tissue culture work, to Jamie McNeil for technical assistance, and to Brad Arbon for plant care in orchard. This work was supported by the Plum Pox Management and Monitoring Program (PPMMP), the Government of Canada.

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Author notes

  1. Mohamed Elhiti

    Present address: Department of Botany, Faculty of Science, Tanta University, Tanta, 31527, Egypt

Authors and Affiliations

  1. Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, N5V 4T3, Canada

    Mohamed Elhiti, Huaiyu Wang, Ryan S. Austin, Bin Chen, Dan Brown & Aiming Wang

  2. Canadian Centre for Agri-Food Research in Health and Medicine, 351 Tache Avenue, Winnipeg, MB, R2H 2A6, Canada

    Dan Brown

Authors
  1. Mohamed Elhiti
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  2. Huaiyu Wang
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  3. Ryan S. Austin
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  4. Bin Chen
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  5. Dan Brown
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  6. Aiming Wang
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Corresponding author

Correspondence to Aiming Wang.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 20 kb)

Supplementary Figure S1

An overview of the steps involved in generation of an ethyl methanesulfonate (EMS) mutagenized population of peach trees (TIFF 893 kb)

Supplementary material 3 (DOCX 23 kb)

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Elhiti, M., Wang, H., Austin, R.S. et al. Generation of chemically induced mutations using in vitro propagated shoot tip tissues for genetic improvement of fruit trees. Plant Cell Tiss Organ Cult 124, 447–452 (2016). https://doi.org/10.1007/s11240-015-0894-0

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  • DOI: https://doi.org/10.1007/s11240-015-0894-0

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