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Genomic instability in phenotypically normal regenerants of medicinal plant Codonopsis lanceolata Benth. et Hook. f., as revealed by ISSR and RAPD markers

  • Cell Biology and Morphogenesis
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Abstract

Codonopsis lanceolata Benth. et Hook. f., commonly known as bonnet bellflower, is a high-valued herb medicine and vegetable. In this study, a large number of plants were regenerated via organogenesis from immature seed-derived calli in C. lanceolata by a simple and efficient method. Compared with the mother donor plant, the regenerated plants did not exhibit visible phenotypic variations in six major morphological traits examined at the stage of one-season-maturity under field conditions. To gain insight into the genomic stability of these regenerated plants, 63 individuals were randomly tagged among a population of more than 2,000 regenerants, and were compared with the single mother donor plant by two molecular markers, the inter-simple sequence repeats (ISSR) and randomly amplified polymorphic DNA (RAPD). Apparent genomic variation was detected in the 63 regenerants, whereas preexisting heterozygosiy in the donor plant was deemed minimal by testing 30 seedlings germinated from selfed seeds of the same donor plant. The percentages of polymorphic bands (PPB) in the ISSR and RAPD analysis were respectively 15.7 and 24.9% for the 63 regenerated plants. Cluster analysis indicates that the genetic similarity values calculated on the basis of RAPD and ISSR data among the 64 plants (63 regenerated and one donor) were respectively 0.894 and 0.933, which allow classification of the plants into distinct groups. Nineteen randomly isolated bands underlying the changed RAPD or ISSR patterns were sequenced, and three of them showed significant homology to known-function genes. Detailed pairwise sequence comparison at one locus between the donor plant and a regenerant revealed that insertion of two short (24 and 19 bp) stretches of nucleotides in the regenerated plant relative to the donor plant occurred in an apparently stochastic manner.

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Abbreviations

6-BA :

6-Benzylaminopurine

CTAB :

Cetyl trimethylammonium bromide

GA 3 :

Gibberellic acid

IBA :

Indole-butyric acid

ISSR :

Inter-simple sequence repeat

MS :

Murashige and Skoog's medium

NAA :

α-Naphthalene acetic acid

RAPD :

Randomly amplified polymorphic DNA

UPGMA :

Unweighted pair group method with arithmetic averaging

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Acknowledgements

This study was supported by the Program for Changjiang Scholars and Innovative Research Team (PCSIRT) in University, the National Science Award for Distinguished Young Investigators in China (30225003), and by the National Natural Science Foundation of China (30370768)

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Authors and Affiliations

  1. Laboratory of Plant Molecular Epigenetics, Institute of Genetics and Cytology, Northeast Normal University, Changchun, 130024, China

    W. L. Guo, L. Gong, Z. F. Ding, Y. D. Li, F. X. Li, S. P. Zhao & B. Liu

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  1. W. L. Guo
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  2. L. Gong
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  3. Z. F. Ding
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  4. Y. D. Li
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  5. F. X. Li
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  6. S. P. Zhao
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  7. B. Liu
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Correspondence to B. Liu.

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Communicated by F. Sato

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299_2006_131_MOESEM1_ESM.doc

Somaclonal variation of 63 regenerated Codonopsis lanceolata plants (designated as C1 to C63) as compared to the single donor mother plant, revealed by ISSR and RAPD markers. NB novel bands; DB disappeared bands; PB polymorphic bands; TB total bands; PPB percentage of polymorphic bands.

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Guo, W.L., Gong, L., Ding, Z.F. et al. Genomic instability in phenotypically normal regenerants of medicinal plant Codonopsis lanceolata Benth. et Hook. f., as revealed by ISSR and RAPD markers. Plant Cell Rep 25, 896–906 (2006). https://doi.org/10.1007/s00299-006-0131-8

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  • DOI: https://doi.org/10.1007/s00299-006-0131-8

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