Assessment of natural and induced genetic variation in Alstroemeria using random amplified polymorphic DNA (RAPD) markers

Abstract

We have used random amplified polymorphic DNA (RAPD) markers to study genetic variation in Alstroemeria. The first objective was to examine the discriminatory power of RAPD markers in different genotypes of Alstroemeria obtained by traditional breeding. All genotypes examined, including commercial Alstroemeria varieties, could be distinguished on the basis of their RAPD profiles. Progeny plants could be distinguished from their parents. A second objective of this study was to investigate whether RAPD markers can be used as a routine tool to detect mutant plants, as an alternative to glasshouse testing. To address this objective, we analysed AZstroemeria plants that carried phenotypically visible mutations that either were induced by irradiation using X-rays or were the result of somaclonal variation. In eight out of a total of 13 mutant Alstroemeria plants obtained after irradiation or tissue culture we detected no polymorphisms when compared to control plants that were considered to be non-mutated. Only in five of the mutant plants analysed we detected one to two polymorphisms. These results suggest that frequent genome rearrangements had not occurred in the mutant plants analysed. These results also demonstrate that the RAPD technique is an inappropriate tool for the rapid screening of Alstroemeria for induced variation. It seems probable that this conclusion would be equally applicable in other plant genera in which induced variation has occurred. However, the RAPD technique is a simple and effective tool for genetic fingerprinting of Alstroemeria varieties, provided their differences are due to sexual propagation.

Somaclonal variation in plants: causes and detection methods

“…Nevertheless, RAPD technique has been inconclusive or ineffective in some species. For example, RAPD markers did not detect somaclonal variation in begonia plants regenerated from leaf explants treated with increasing concentrations of nitrosomethylurea (Bouman and De Klerk 2001) as well as X-ray-induced garlic mutants (Anastassopoulos and Keil 1996). The technique also has a lower reproducibility and reliability (Jones et al 1997) as well as less informative compared to amplified fragment length polymorphism (AFLP) markers thereby limiting its application in some species (Mulcahy et al 1993;Vos et al 1995).…”

“…Genetic variation in mutants can be induced either by specific treatments with physical and chemical mutagens or by tissue culture, see Predieri (2001) and Roux (2004) for detailed reviews. Anastassopoulos and Keil (1996) referred to somaclonal variation as mutations in plants regenerated from tissue culture. Remarkably, it has also been observed that the frequency of expression of dominance is higher in somaclonal variation than in other forms of mutation (Larkin 1998;Yang et al 2010).…”

Divergência genética entre cultivares de gérbera utilizando marcadores RAPD

“…Em plantas ornamentais, já foram utilizados marcadores RAPD para o estudo da variabilidade genética de diversas espécies, como, por exemplo: Alstroemeria (ANASTASSOPOULOS & KEIL, 1996), crisântemo (WOLFF & PETERS- VAN RIJN, 1993), rosa (TORRES et al, 1993), lírio (YAMAGISHI, 1995), gérbera (DA MATTA et al, 2009), entre outras.…”

Efficient production of transgenic Alstroemeria plants by using Agrobacterium tumefaciens

“…Alstroemeria is a monocotyledonous plant and has its centres of origin in Chile and Brazil (Aker & Healy, 1990). Breeding activities have resulted in modern hybrids of Alstroemeria with large and beautiful flowers (Anastassopoulos & Keil, 1996). Biotechnological strategies such as recombinant DNA technologies and genetic transformation could provide efficient approaches in Alstroemeria breeding, but the lack of a reliable regeneration system to generate transgenic plants has hindered the use of genetic transformation in Alstroemeria .…”