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Large numbers of genetically stable, homozygous plants are needed for classical and molecular breeding programmes. In vitro anther culture has proved to be a useful tool for haploid/doubled haploid (DH) induction in pepper (Capsicum annuum L.) for more than twenty years. The present paper reports on a great improvement in the in vitro haploid induction and genome duplication methods routinely used for resistance breeding in sweet and spice peppers by two Hungarian research institutions, the Agricultural Biotechnology Center in Gödöllő and the Budapest Research Unit of the Vegetable Crops Research Institute. As a result of the colchicine-stimulated early genome induction method, the critically low (<0.1%) regeneration frequency of spice pepper types became ten times greater, reaching a value of around 1.0%, though this was still considerably lower than that achieved in pepper varieties for fresh consumption (5-10%). Moreover, the ratio of useful doubled haploids was far higher (H:DH = 1:2 or 1:4) in some cases after colchicine treatment than that of untreated control plants (H:DH = 2:1 or 3:1, depending on the genotype). An efficient method with good reproducibility, requiring less manual work, was elaborated for the in vitro genome duplication of pepper haploid regenerants using colchicine. When the haploid induction ability of plants conventionally cultured in the greenhouse was compared to that of plants raised under artificial conditions in phytotron chambers (satisfactory day and night temperatures, illumination, humidity), the responsiveness of the latter microspores (ratio of plant regeneration) was found to be almost twice as high. The application of 3% maltose for six days at 35°C resulted in a 1.45% increase in the ratio of responding anthers and a 0.34% increase in plant regeneration, averaged over all the variety types. Phenosafranin staining was used for the analysis of microspore viability. The reduction in viability during the induction period proved to be less pronounced in lines with better androgenetic responses than in those with poorer responsiveness.

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As locus-specific co-dominant PCR-based markers that allow semi-automated, high-throughput investigation technologies, microsatellites are ideal tools for genotype identification. Eleven of a set of 114 microsatellite markers available at the Agricultural Biotechnology Center proved to be suitable to distinguish between the parents of at least one of nine sweet pepper hybrid combinations. Markers with the highest information capacity were found to be capable of distinguishing between the parents of four different hybrid combinations and exhibited up to four different alleles in 18 haplotypes.

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Acta Agronomica Hungarica
Authors:
J. Pauk
,
C. Lantos
,
G. Somogyi
,
P. Vági
,
Z. Ábrahám Táborosi
,
A. Gémes Juhász
,
R. Mihály
,
Z. Kristóf
,
N. Somogyi
, and
Z. Tímár

Spice pepper production has a history of almost 300 years in the southern part of Hungary. In this study the results of two biotechnological improvements are summarized. Anther and isolated microspore culture techniques were improved to release haploid and doubled haploid (DH) lines for spice pepper breeding. Both the anther and isolated microspore culture methods were successfully used in spice pepper haploid production. Microspore culture-derived structures were analysed to identify their different parts. Green plantlets were regenerated from embryos derived from both anther and microspore cultures. Their doubled haploid analogues were integrated into Hungarian spice pepper hybrid seed breeding programmes. One hybrid, Sláger, was released as a new genotype for spice pepper production in 2008 and two hybrid candidates (Délibáb and Bolero) are now being tested in official trials.

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