NDLI: Synonymous mutation gene design to overexpress ACCase in creeping bentgrass to obtain resistance to ACCase-inhibiting herbicides

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Synonymous mutation gene design to overexpress ACCase in creeping bentgrass to obtain resistance to ACCase-inhibiting herbicides

Content Provider	Springer Nature Link
Author	Heckart, Douglas L. Schwartz, Brian M. Raymer, Paul L. Parrott, Wayne A.
Copyright Year	2016
Abstract	Overexpression of a native gene can cause expression of both introduced and native genes to be silenced by posttranscriptional gene silencing (PTGS) mechanisms. PTGS mechanisms rely on sequence identity between the transgene and native genes; therefore, designing genes with mutations that do not cause amino acid changes, known as synonymous mutations, may avoid PTGS. For proof of concept, the sequence of acetyl-coA carboxylase (ACCase) from creeping bentgrass (Agrostis stolonifera L.) was altered with synonymous mutations. A native bentgrass ACCase was cloned and used as a template for the modified gene. Wild-type (WT) and modified genes were further modified with a non-synonymous mutation, coding for an isoleucine to leucine substitution at position 1781, known to confer resistance to ACCase-inhibiting herbicides. Five-hundred calli of creeping bentgrass ‘Penn A-4’ were inoculated with Agrobacterium containing either the WT or modified genes, with or without the herbicide-resistance mutation. Six herbicide-resistant-transgenic events containing the modified gene with the 1781 mutation were obtained. Transcription of the modified ACCase was confirmed in transgenic plants, showing that gene-silencing mechanisms were avoided. Transgenic plants were confirmed to be resistant to the ACCase-inhibiting herbicide, sethoxydim, providing evidence that the modified gene was functional. The result is a novel herbicide-resistance trait and shows that overexpression of a native enzyme with a gene designed with synonymous mutations is possible.
Starting Page	465
Ending Page	476
Page Count	12
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	25
Issue Number	4
e-ISSN	15739368
Language	English
Publisher	Springer International Publishing
Publisher Date	2016-04-26
Publisher Place	Cham
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Acetyl coenzyme A carboxylase (ACCase) Aryloxyphenoxypropionate (APP) Cyclohexandione (CHD) Herbicide-resistance Posttranscriptional gene silencing Sethoxydim resistant Synonymous mutation Synthetic gene design Animal Genetics and Genomics Plant Genetics & Genomics Transgenics Biomedical Engineering/Biotechnology Genetic Engineering Molecular Medicine
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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