Abstract
The establishment of intraspecific hybrid plant between two varieties of common buckwheat plant was examined by in vitro flowering, in vitro pollination, fertilized flower/ovule culture, subsequent backcrosses and possible line establishment by micropropagation procedure. Varieties Kitawasesoba and Fukuizairaisoba were selected as mother and father plants, respectively. The fertilization and subsequent plant regeneration in vitro were significantly improved by the application of coconut water. About 40% of pollinated flowers cultured in coconut water medium grew into fertilized ovules and about 35% of pollinated flowers grew into immature embryos. All the embryos grew spontaneously into young plants.
In vitro born plants were identified as hybrid plants by zymogram, RAPD, and morphological and agriculture traits.
After the first in vitro cross, 3 backcrosses in vitro were sufficient to produce hybrid plants that had 94% of maternal genes. These plants were obtained in 240 days. One life cycle took about 55-60days in vitro in the present research. In vitro born plants were micropropagated in vitro to establish potentially stable lines. Possibility of cross breeding of plants in vitro to develop breeding factory is discussed.
