Production of triploid plants of Japanese pear (Pyrus pyrifolia Nakai) by anther culture

Triploid plants were grown from the diploid pear scion cultivar (Pyrus pyrifolia Nakai) by anther culture. Androgenic callogenesis occurred at about 10 days after anther excision, and most calli formed within four weeks and stopped growing at less than 1-3 mm in diameter. Androgenic embryogenesis occurred within 8-12 weeks after anther culture. The formed embryos were generally white, globular or heart-shaped. To increase callogenesis and embryogenesis, the effects of cold pre-treatment, light and activated charcoal (AC) on anthers collected from buds were investigated. Cold pre-treatment, dark and lack of AC increased callogenesis more than no cold pre-treatment, light and adding AC. A decisive condition was not found for embryogenesis. The embryogenesis frequency was extremely low and only six embryos were grown. Two embryos expanded greenish cotyledons and developed foliage, and they proliferated when transferred monthly to fresh proliferation medium. The proliferated shoots were rooted and acclimated. Flow cytometric analysis and investigation of the number of chromosomes of regenerants showed that all cells were triploid.