Pollen Cryopreservation in Stevia rebaudiana Bertoni

Abstract

AbstractThis chapter focuses on pollen germination in vitro and cryopreservation of Stevia rebaudiana. At the time of anthesis, pollen were collected from the field and pre-storage germination assessment was carried by hanging drop technique using modified Brewbaker and Kwack medium containing different combination and concentration of sucrose and PEG (Polyethylene Glycol). Pollen were then packed in butter paper covers enclosed in sealed aluminum pouches which were rapidly plunged into the cryobiological system containing liquid nitrogen. After 1 week of cryopreservation, the germination test was carried out to know the percentage of germination of cryopreserved pollen. SEM (Scanning Electron Microscopy) analysis of pollen was carried out to detect any morphological changes in pollen structure. The results indicated that there is no significant decline in viability of cryopreserved pollen. Therefore, it can be concluded that cryopreservation of Stevia rebaudiana pollen can be achieved effectively without loss of its viability and can be conserved successfully to pollinate and use in breeding programs to broaden the genetic base through pollen.Key words Stevia rebaudiana Pollen cryopreservationIn vitro germination SEM studies