[Embryogenesis of microspore derived multicells in Capsicum annuum L].

Abstract

Microspores and derived multicells were isolated and cultured in modified liquid CP medium after a 15d's preculture of anthers on solidified medium. Thirty days later in suspension culture, at 28 degrees C dark condition embryoids with different developmental stages were formed. Up to 22 embryoids could be formed from the cell suspension of 12 anthers, and about 23% of the embryoids were at the cotyledonary stage. Fluorescence and light microscope observations revealed that these embryoids derived from microspores. After several symmetrical division of the nuclei of uninucleated microspores, multi-nuclei cells or multi-cells were formed, and developed further into embryoids. There were white hairs on the surface of pepper embryoids, and some embryoids showed low vigor while others showed normal by TTC staining. Plants could be formed from torpedo and cotyledonary stage embryoids on solidified medium. Embryoids could be induced by 7 degrees C, 32 degrees C or 35 degrees C stress treatment on anthers, Higher embryogenesis frequencies were got at 7 degrees C and 35 degrees C condition in anther culture while 35 degrees C and 32 degrees C treatment showed a higher embryogenesis in isolated multicell culture. The reason of this result was discussed. There were obvious differences in embryogenesis frequency among different genotypes and different temperature stress conditions. Flow cytometric analysis revealed that there were haploidy, doubled haploidy and haploid-diploid chimera in the regenerated plants.