Diversity of abnormal embryo sacs in indica/japonica hybrids in rice demonstrated by confocal microscopy of ovaries

Abstract Embryo sac abortion occurs frequently in the spikelet of indica/japonica hybrids in rice. To provide a comprehensive understanding of embryo sac sterility in typical indica/japonica hybrids, a simple eosin B staining procedure for use with confocal microscopy was developed to observe the mature embryo sacs of 35 indica/japonica F 1 s making up an incomplete diallel cross design (5 × 7). A diversity of abnormal embryo sacs existed in the hybrids. The main types were classified into five categories: (i) embryo sac degeneration (ESD), (ii) embryo sac without egg apparatus, (iii) embryo sac without female germ unit, (iv) embryo sac with abnormal polar nuclei and (v) abnormal small embryo sac (ASES). The frequencies of abnormal embryo sacs in the 35 hybrids ranged from 26.0% to 76.3%, with an average of 48.6%. The highest frequency of abnormality was 31.3% ESD. The frequencies of various abnormalities varied with parental genotype and environment.