Approaches for the Isolation of Arabidopsis adh1 Regulatory Mutants Using Allyl Alcohol Selection

Approaches for the Isolation of Arabidopsis adh1 Regulatory Mutants Using Allyl Alcohol Selection Allyl alcohol, a suicide substrate for the alcohol dehydrogenase enzyme (EC.1.1.1.1), has been frequently used as a negative selection method for the isolation of alcohol dehydrogenase mutants in plants, animals and microorganisms. This approach led to the isolation of mutants that mapped to the ADH gene itself. We attempted to use allyl alcohol selection for the isolation of adh1 regulatory mutants in Arabidopsis. First we selected at plantlet level on ADH1–GUS transgenic plants. This enabled us to use GUS staining to discriminate between structural and regulatory mutants. Allyl alcohol selection of 50000 EMS-treated seeds did not yield any potential mutants. Secondly we selected EMS and γ-ray-treated seeds of a transgenic line transformed with an additional copy of the ADH1 gene including its own promoter. Fifteen allyl alcohol-resistant plants were selected from the mutagenized seed. Genetic analysis of three putative mutants (adr8, adr10, and adr15) indicated that the ADH1-null phenotype was due to monogenic trans-recessive mutations. But treatment with the demethylating agent 5-azacytidine and analysis of methylation levels of the ADH1 gene indicated that these mutant candidates have increased levels of methylation in the promoter and coding region of ADH1. These results suggest that the allyl alcohol resistance of adr8, adr10, and adr15 is due to silencing of ADH1 rather than to a mutation of a regulatory locus. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Plant Physiology Springer Journals

Approaches for the Isolation of Arabidopsis adh1 Regulatory Mutants Using Allyl Alcohol Selection

Loading next page...
 
/lp/springer_journal/approaches-for-the-isolation-of-arabidopsis-adh1-regulatory-mutants-57TCJ40aYK
Publisher
Kluwer Academic Publishers-Plenum Publishers
Copyright
Copyright © 2003 by MAIK “Nauka/Interperiodica”
Subject
Life Sciences; Plant Sciences
ISSN
1021-4437
eISSN
1608-3407
D.O.I.
10.1023/B:RUPP.0000003274.04439.d4
Publisher site
See Article on Publisher Site

Abstract

Allyl alcohol, a suicide substrate for the alcohol dehydrogenase enzyme (EC.1.1.1.1), has been frequently used as a negative selection method for the isolation of alcohol dehydrogenase mutants in plants, animals and microorganisms. This approach led to the isolation of mutants that mapped to the ADH gene itself. We attempted to use allyl alcohol selection for the isolation of adh1 regulatory mutants in Arabidopsis. First we selected at plantlet level on ADH1–GUS transgenic plants. This enabled us to use GUS staining to discriminate between structural and regulatory mutants. Allyl alcohol selection of 50000 EMS-treated seeds did not yield any potential mutants. Secondly we selected EMS and γ-ray-treated seeds of a transgenic line transformed with an additional copy of the ADH1 gene including its own promoter. Fifteen allyl alcohol-resistant plants were selected from the mutagenized seed. Genetic analysis of three putative mutants (adr8, adr10, and adr15) indicated that the ADH1-null phenotype was due to monogenic trans-recessive mutations. But treatment with the demethylating agent 5-azacytidine and analysis of methylation levels of the ADH1 gene indicated that these mutant candidates have increased levels of methylation in the promoter and coding region of ADH1. These results suggest that the allyl alcohol resistance of adr8, adr10, and adr15 is due to silencing of ADH1 rather than to a mutation of a regulatory locus.

Journal

Russian Journal of Plant PhysiologySpringer Journals

Published: Oct 17, 2004

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create lists to
organize your research

Export lists, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off