AFLP and AMP Fingerprints as Markers to Evaluate Genetic Differences between Medicago truncatula Line Jemalong and 2HA, a New Line Produced by in vitro Culture Selection

R. R. Irwanto, R. J. Rose

Abstract


A new line, Medicago truncatula cv. Jemalong 2HA (herein known as 2HA) has been developed via repetitive regeneration and selection of M. truncatula cv. Jemalong. During somatic embryogenesis, 2HA produces 500 times more embryos than its progenitor, Jemalong. It is interesting to study if those two lines are isogenic or has genetic differences. The main objectives of the study was to evaluate the genotypic differences between Jemalong and 2HA also to evaluate the methylation event in 2HA utilized two DNA fingerprinting techniques, i.e AFLP fingerprints (Amplified Length of Polymorphism) and AMP (Amplified Methylation Polymorphism). The results showed that AFLP analysis using eight primers combinations could not detect any differences between Jemalong and 2HA. However, using AMP methylation sensitive primers it could detect 15 polymorphisms out of 840 markers. These results lead to a conclusion that Jemalong and 2HA are isogenic lines. 2HA may have higher regeneration capacities due to methylation process which occurs during the production of 2HA through repetitive regeneration cycles.

Full Text:

PDF

References


Rose, R., Nolan, K. & Bicego, L., The development of the highly regenerable seed line Jemalong 2HA for transformation of Medicago truncatula: implications for regenerability via somatic embryogenesis, J. Plant. Physiol. 155, 788–791, 1999.

Evans, D., Sharp, W. & Medina-Filho, H., Somaclonal and gametoclonal variation, Amer. J. Bot. 71, 759-774, 1984.

Scowcroft, W. & Ryan, S., Tissue culture and plant breeding, Plant cell culture technology, M. Yeoman (ed), Blackwell Scientific Publications, pp. 67-94, 1986.

Phillips, R., Kaeppler, S. & Olhoft. P., Genetic instability of plant tissue cultures: Breakdown of normal controls, Proc. Natl. Acad. Sci. USA, 91, pp. 5222-5226, 1994.

Xu, Mingliang., Li, XiangQian. & Korban, S.S., AFLP-based Detection of DNA Methylation, Plant Molecular Biology Reporter, 18, 361–368, 2000.

Vos, P. & Hogers, R., AFLP: a new technique for DNA fingerprinting, Nucleic Acids Res., 23, 4407- 4414, 1995.

Murray, M. & Thompson, W., Rapid isolation of high-molecular-weight plant DNA, Nucleic Acids Res., 8, 4321–4325, 1980.

Carrol, B., Klimyuk, V., Thomas, C., Bishop, G., Harrison, K., Scofield, S. & Jones, J., Germinal transpositions of maize element dissociation from T-DNA loci in tomato, Genetics, 139, 407- 420, 1995.

Finnegan, E., Genger, R., Peacock,W. & Dennis, E., DNA methylation in plants, Plant Molecular Biology, 49, 223–2247, 1998.

Peraza-Echeverria S., Hen'era-Valencia V.A. & James-Kay A., Detection of DNA methylation changes in micropropagated banana plants using methylation-sensitive amplification polymorphism (MSAP), Plant Sci., 161, 359-367, 2001.

Raimondi, J., Masuelli, R. & Camadro, E., Assessment of somaclonal variation in asparagus by RAPD fingerprinting and cytogenetic analysis, Sci. Hort., 90, 19-29, 2001.

Chabaud, M., Larsonneau, C., Marmouget, C. & Huguet, T., Transformation of barrel medic (Medicago truncatula Gaertn) by Agrobacterium tumefaciens and regeneration via somatic embryogenesis of transgenic plants with the Mtenod12 nodulin promoter fused to the gus reporter gene, Plant Cell Rep., 15, 305–310, 1996.

Nolan, K., Irwanto, R. & Rose, R., Auxin Up-Regulates MtSERK1 Expression in Both Medicago truncatula Root-Forming and Embryogenic Cultures, Plant Physiol., 133, 218–230, 2003.

Hecht, V., Vielle-Calzada, J., Hartog, M., Schmidt, E., Boutilier, K., Grossniklaus, U. & de Vries, S., The Arabidopsis SOMATIC EMBRYOGENESIS RECEPTOR KINASE1 gene is expressed in developing ovules and embryos and enhances embryogenic competence in culture, Plant Physiol,. 127, 803–816, 2001.

Halperin, W., In vitro Embryogenesis: some historical issues and unresolved problems. In vitro embryogenesis in plants, (T. Thorpe, ed), Kluwer Academic Publishers, pp. 1-16, 1995.

Thrieffry, D. & Sanchez, L., Dynamic modelling of pattern formation during embryogenic development, Curr. Opin. Biotechnol., 13, 326-330, 2003.

Meyer, P., Introduction: epigenetic strategies in animals and plants,

Cell & Developmental Biology, 14, 51-52, 2003.

Santos D. & Fevereiro, P., Loss of DNA methylation affects somatic embryogenesis in Medicago truncatula, Plant Cell, Tissue and Organ Culture, 70, 155-161, 2002.

Chakrabarty, D., Yu, K. & Paek, K., Detection of DNA methylation changes during somatic embryogenesis of Siberian ginseng (Eleuterococcus senticosus), Plant Sci., 165, 61-68, 2003.




DOI: http://dx.doi.org/10.5614%2Fitbj.sci.2008.40.2.4

Refbacks

  • There are currently no refbacks.


View my Stats

Creative Commons License
This work is licensed under a Creative Commons Attribution-NoDerivatives 4.0 International License.