Enzyme fingerprint analyses in tissue regenerated from anther culture of maize

Abstract

The objectives of our studies were to investigate the effect of cold pre-treatment duration and the effect of two different culture media (YP and N6) on maize anther culture response in two maize genotypes (A 18 and A 19) and to identify the gametic origin of the maize regenerants. Androgenic induction and callus formation was compared in anther cultures following pre-treatment applied to both media tested and with both maize genotypes. Higher plant regeneration was observed in case of YP media independently of the genotype used. The best results were achieved when 12 days (genotype A 18) or 14 days (in case of genotype A 19) cold pre-treatment at 10°C was applied. We have tested the possibility of using enzyme isoform analyses to identify the microspore origin of calli and plants derived from anther cultures. The 11 enzymes tested in our experiments were acid phosphatase, alcohol dehydrogenase, catalase, diaphorase, b-glucosidase, glutamate oxaloacetate transaminase, isocitrate dehydrogenase, malate dehydrogenase, 6-phosphogluconate dehydrogenase, phosphoglucomutase and phosphoglucoisomerase. Analysis of malate dehydrogenase proved the gametic origin of the calli initiated and of the DH plants regenerated from anther culture, when the coleoptile of the donor plant material showed two forms of enzyme 3/6 and the analysed calli showed only one of the two forms (3 or 6).

Methodological Approaches to Simple Enzyme Polymorphism Analyses of Amaranth Species (Amaranthus SP.)

“…According to our and other laboratories experimental experiences, slightly modified methodology of horizontal starch gel electrophoresis published by Stuber et al (1988) and Bourgoin-Greneche et al (1998) for coleoptile section of maize seedling can be successfully utilised for study of enzyme multiplicity in pea, soybeen, grass pea, chickpea seeds and cotyledons of sunflower (Bourgoin- Greneche & Lallemand 1993;, 1999a, coleoptiles, leaves and roots of maize seedlings (Múdry & Dragúň 2007), leaf and root tissues regenerated from anther culture of maize (Uváčková et al 2008) or leaf tissues of sugar beet (Engelhardt & Múdry 1999;Engelhardt & Bežo 2000). The results on influence of some modifications of experimental conditions on amaranth isozymograms and convenience of methodology published by Stuber et al (1988) for enzyme polymorphism analysis of amaranths are presented by the electrophoretic phenotypes ( Fig.…”

The Bioindicative Role of Enzyme Polymorphism of Seven Enzyme Systems as Reaction on High Cadmium Ions Concentration in Maize (Zea Mays L.)

“…The standard technique (for coleoptilar section) of horizontal starch gel electrophoresis described by Stuber with co-workers (Stuber et al 1988) and recently published in our works (Múdry & Juráček 2001;Múdry & Kraic, 2007;Uváčková et al 2008) was used for analysis of polymorphism in acid phosphatase (aCP, E.C. 3.1.3.2), alcohol dehydrogenase (aDh, E.C.…”