Assessment of genetic and epigenetic changes in virus-free garlic (Allium sativum L.) plants obtained by meristem culture followed by in vitro propagation

KEY MESSAGE : This is the first report assessing epigenetic variation in garlic. High genetic and epigenetic polymorphism during in vitro culture was detected. Sequencing of MSAP fragments revealed homology with ESTs. Garlic (Allium sativum) is a worldwide crop of economic importance susceptible to...

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Bibliographic Details
Published in:	Plant cell reports Vol. 35; no. 1; pp. 129 - 141
Main Authors:	Gimenez, Magalí Diana, Yañez-Santos, Anahí Mara, Paz, Rosalía Cristina, Quiroga, Mariana Paola, Marfil, Carlos Federico, Conci, Vilma Cecilia, García-Lampasona, Sandra Claudia
Format:	Journal Article
Language:	English
Published:	Berlin/Heidelberg Springer Berlin Heidelberg 01-01-2016 Springer Nature B.V
Subjects:	Allium sativum Biomedical and Life Sciences Biotechnology Cell Biology Cloning Cluster Analysis Cultivars DNA Methylation Economic importance Epigenesis, Genetic epigenetics expressed sequence tags garlic Garlic - genetics Garlic - physiology Genetic Loci - genetics Genetic Variation Genomic Instability Life Sciences Meristem - genetics Meristem - physiology meristems Methylation micropropagation Original Article Phenotype phenotypic variation Plant Biochemistry Plant propagation Plant Sciences Reproduction Retroelements retrotransposons sanitizing Sequence Analysis, DNA somaclonal variation Specific Pathogen-Free Organisms tissue culture Tissue Culture Techniques virus-free plants MSAP Retrotransposon MSAP fragment cloning Somaclonal variation AFLP
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Summary:	KEY MESSAGE : This is the first report assessing epigenetic variation in garlic. High genetic and epigenetic polymorphism during in vitro culture was detected. Sequencing of MSAP fragments revealed homology with ESTs. Garlic (Allium sativum) is a worldwide crop of economic importance susceptible to viral infections that can cause significant yield losses. Meristem tissue culture is the most employed method to sanitize elite cultivars. Often the virus-free garlic plants obtained are multiplied in vitro (micro propagation). However, it was reported that micro-propagation frequently produces somaclonal variation at the phenotypic level, which is an undesirable trait when breeders are seeking to maintain varietal stability. We employed amplification fragment length polymorphism and methylation sensitive amplified polymorphism (MSAP) methodologies to assess genetic and epigenetic modifications in two culture systems: virus-free plants obtained by meristem culture followed by in vitro multiplication and field culture. Our results suggest that garlic exhibits genetic and epigenetic polymorphism under field growing conditions. However, during in vitro culture system both kinds of polymorphisms intensify indicating that this system induces somaclonal variation. Furthermore, while genetic changes accumulated along the time of in vitro culture, epigenetic polymorphism reached the major variation at 6 months and then stabilize, being demethylation and CG methylation the principal conversions. Cloning and sequencing differentially methylated MSAP fragments allowed us to identify coding and unknown sequences of A. sativum, including sequences belonging to LTR Gypsy retrotransposons. Together, our results highlight that main changes occur in the initial 6 months of micro propagation. For the best of our knowledge, this is the first report on epigenetic assessment in garlic.
Bibliography:	http://dx.doi.org/10.1007/s00299-015-1874-x ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0721-7714 1432-203X
DOI:	10.1007/s00299-015-1874-x