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Application of pollen for studies on genetic diversity with the using of the Andean lupin (Lupinus mutabilis Sweet) as example

100%

Sawicka-Sienkiewicz E., Augustyniak H., Pietkiewicz G., Bartkowiak S.

The cultivated American species Lupinus mutabilis was selected in order to assess its possibilities of being introduced as an alternative high protein crop in Polish conditions. The realized programme involved identification of earlier maturing and higher yielding genotypes as well as interspecific hybridization with lupins belonging to section Albus, which includes the oldest cultivated species Lupinus albus. Recent reports (1) on determination of DNA polymorphism by the PCR method indicate usefulness of direct application of small amounts of pollen. The advantage of this method is the precisely determined and physiologically stable material used for the analysis. Furthermore, pollen can be preserved above the hygroscopic substance at ?20oC for a long time and small amounts of pollen (20-100 grains) can be used for the analysis without any damage to the plant material. For PCR reaction, ten selected primers were used. A polyacrylamide gel comparison of the PCR products revealed that random primers as well as primers which amplified a fragment of the whole gene can be applied to analyse the DNA of polymorphism of L. mutabilis and L. albus. The conclusion is that it is possible to differentiate within both groups of species by application of pollen to DNA analysis.

Screening of cytoplasmic DNA diversity between and within Lupinus mutabilis Sweet and Lupinus albus sensu lato by restriction fragment length polymorphism (RFLP)

88%

Olczak T., Rurek M., Janska H., Augustyniak H., Sawicka-Sienkiewicz E.

Journal of Applied Genetics

2001

vol. 42

issue 2

127-137

Seven populations and five mutant lines of the Andean lupin and four species from the section Albus were screened for their mitochondrial and chloroplast polymorphisms. For this purpose the RFLP method with EcoRI as a restriction enzyme was used. Lupinus luteus, Lupinus albus and Phaseolus vulgaris organellar clones as well as amplified fragments were used as probes. We found that mitochondrial probes were more suitable than chloroplast probes for identification of inter- and intra-specific variations within the examined material. Most mitochondrial probes differentiate the two species investigated. A high level of mitochondrial polymorphism was observed among the populations of L. mutabilis in contrast to monomorphism among the species in the section Albus. A limited polymorphism was detected between the mutant lines of L. mutabilis. We conclude from this study that the mitochondrial RFLP analysis is a valuable tool for identification of variability among Andean lupin populations.

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1 Biotechnologia

1 Journal of Applied Genetics

2 2001

2 Augustyniak H.

2 Sawicka-Sienkiewicz E.

1 Bartkowiak S.

1 Janska H.

1 Olczak T.

1 Pietkiewicz G.

1 Rurek M.

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Application of pollen for studies on genetic diversity with the using of the Andean lupin (Lupinus mutabilis Sweet) as example

Screening of cytoplasmic DNA diversity between and within Lupinus mutabilis Sweet and Lupinus albus sensu lato by restriction fragment length polymorphism (RFLP)