PRELIMINARY STUDIES ON EFFECTS OF ACTIVATED CHARCOAL(AC) AND ANTHER DENSITY ON EFFICIENCY OF BARLEY ANTHER CULTURE AND GENE TRANSFER-BY PARTICLE BOMBARDMENT IN BARLEY(Hordeum vulgare L. )
Zhang Yuhua; Wu Huixial; Huang Jianhua; Lu Ruiju; Sun Yufang (Crop Breeding and Cultivation Research Institute, ShanghaiAcademy of Agricultural Sciences, Shanghai 201106 )(Institute of Genetics, Academia Sinica, Beijing 100101, China. )
Effects of AC and anther density on pollen embryo and callus production and green plant regeneration were studied in a filter-sterilized, liquid-medium culture system. AC could reduce much more embryo and callus production when applied to induction culture than to mannitol pre-culture, and obvious deterioration of pollen embryos and calli was observed, which resulted in a drop in green plant regeneration,while difference in anther density in mannitol pre-culture didn' t show any effects on anther culture, difference in anther density during induction culture did have an influence.That is, higher anther density (60 ￣ 90 anthers/ml) gave an adverse effect on the growth of pollen embryos and calli, causing a decrease both in pollen embryo and callus production and green plant regeneration ; lower anther density(15￣30 anthers/ml ) generated a favorable effect on the growth of pollen embryos and calli, resulting in an increase both in pollen embryo and callus production and green plant regeneration. Gene transfer into pollen embryos and calli was performed using particle bombardment.Abundant transient expression of reporter gene(GUS) occured on the bombarded pollen embryos and calli. However, we failed to obtain transgenic barley plants because the totipotency of transformed cells was probably lost during subsequent selection subculture.Some possible causes of negative effects of AC and anther density on anther culture in the present work were discussed, and the application of isolated microspores rather than microspore-derived embryos and calli to gene transfer experiments was suggested by in barley the authors.
【CateGory Index】： S512.305.5