The terminal flower buds of 6-yr-old Hanfu apple were used to study the ovule development, ovular characteristics, cell death of abortive ovules, and dynamic change of starch grain quantity in the embryo sac with paraffin slices and terminal deoxynucleotidyl transferase-mediated fluorescein deoxyuridine triphosphate nick-end labeling (TUNEL) system. Four anatropous ovules in each ventricle could be observed before flowering. With the developing of floral organ, the bulk of normal ovules enlarged in each ventricle, the mature embryo sac differentiated into nucellus, and the egg cell developed into zygote by double fertilization. A large number of starch grains were observed during pollen tube growth and double fertilization, which guaranteed basic nutrient supply in the normal development of ovules. Moreover, abortion phenomenon of runtish ovules emerged at the stages of mature embryo sac, double fertilization and zygote development. The abortion characteristics included deformity of ovule development, degradation of nucellus tissue, separation between funiculus and ovule, abnormality of four-nucleate embryo sac, as well as development interruption of mature embryo sac. TUNEL analysis proved that ovule abortion was programmed cell death.

In this study, we investigated the potential nitrification and community structure of soil-based ammonia-oxidizing bacteria (AOB) in apple orchard soil during different growth periods and explored the effects of environmental factors on nitrification activity and AOB community composition in the soil of a Hanfu apple orchard, using a culture-dependent technique and denaturing gradient gel electrophoresis (DGGE). We observed that nitrification activity and AOB abundance were the highest in November, lower in May, and the lowest in July. The results of statistical analysis indicated that total nitrogen (N) content, NH4 +-N content, NO3 --N content, and pH showed significant correlations with AOB abundance and nitrification activity in soil. The Shannon-Winner diversity, as well as species richness and evenness indices (determined by PCR-DGGE banding patterns) in soil samples were the highest in September, but the lowest in July, when compared to additional sampled dates. The DGGE fingerprints of soil-based 16S rRNA genes in November were apparently distinct from those observed in May, July, and September, possessing the lowest species richness indices and the highest dominance indices among all four growth periods. Fourteen DGGE bands were excised for sequencing. The resulting analysis indicated that all AOB communities belonged to the β-Proteobacteria phylum, with the dominant AOB showing high similarity to the Nitrosospira genus. Therefore, soil-based environmental factors, such as pH variation and content of NH4 +-N and NO3 --N, can substantially influence the abundance of AOB communities in soil, and play a critical role in soil-based nitrification kinetics.