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Transformation and gene-disruption in the apple-pathogen, Neonectria ditissima

Vélëz, Heriberto and Skytte af Sätra, Jonas and Odilbekov, Firuz and Bourras, Salim and Gustavsson, Larisa and Dalman, Kerstin (2022). Transformation and gene-disruption in the apple-pathogen, Neonectria ditissima. HEREDITAS. 159 , 31
[Research article]

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Abstract

Background Apple production in Sweden and elsewhere is being threatened by the fungus, Neonectria ditissima, which causes a disease known as European canker. The disease can cause extensive damage and the removal of diseased wood and heavily infected trees can be laborious and expensive. Currently, there is no way to eradicate the fungus from infected trees and our knowledge of the infection process is limited. Thus, to target and modify genes efficiently, the genetic transformation technique developed for N. ditissima back in 2003 was modified. Results The original protocol from 2003 was upgraded to use enzymes currently available in the market for making protoplasts. The protoplasts were viable, able to uptake foreign DNA, and able to regenerate back into a mycelial colony, either as targeted gene-disruption mutants or as ectopic mutants expressing the green fluorescent protein (GFP). Conclusions A new genetic transformation protocol has been established and the inclusion of hydroxyurea in the buffer during the protoplast-generation step greatly increased the creation of knockout mutants via homologous recombination. Pathogenicity assays using the GFP-mutants showed that the mutants were able to infect the host and cause disease.

Authors/Creators:Vélëz, Heriberto and Skytte af Sätra, Jonas and Odilbekov, Firuz and Bourras, Salim and Gustavsson, Larisa and Dalman, Kerstin
Title:Transformation and gene-disruption in the apple-pathogen, Neonectria ditissima
Series Name/Journal:HEREDITAS
Year of publishing :2022
Volume:159
Article number:31
Number of Pages:10
Publisher:BMC
Associated Programs and Other Stakeholders:Z - SLU - Library > Odla mera
ISSN:1601-5223
Language:English
Publication Type:Research article
Article category:Scientific peer reviewed
Version:Published version
Copyright:Creative Commons: Attribution 4.0
Full Text Status:Public
Subjects:(A) Swedish standard research categories 2011 > 4 Agricultural Sciences > 404 Agricultural Biotechnology > Genetics and Breeding
(A) Swedish standard research categories 2011 > 4 Agricultural Sciences > 401 Agricultural, Forestry and Fisheries > Agricultural Science
Keywords:European canker, Fruit tree canker, Neonectria ditissima, Hydroxyurea, Fungal transformation, GFP
URN:NBN:urn:nbn:se:slu:epsilon-p-118749
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-p-118749
Additional ID:
Type of IDID
DOI10.1186/s41065-022-00244-x
Web of Science (WoS)000839650800001
ID Code:28809
Faculty:S - Faculty of Forest Sciences
LTV - Fakulteten för landskapsarkitektur, trädgårds- och växtproduktionsvetenskap
NJ - Fakulteten för naturresurser och jordbruksvetenskap
Department:(NL, NJ) > Dept. of Forest Mycology and Plant Pathology
(S) > Dept. of Forest Mycology and Plant Pathology

(LTJ, LTV) > Department of Plant Breeding (from 130101)
(NL, NJ) > Department of Molecular Sciences
Deposited By: SLUpub Connector
Deposited On:09 Sep 2022 07:43
Metadata Last Modified:10 Sep 2022 04:03

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