Function Analysis Of AtPLC2 Regulates Pollen Development In Arabidopsis Thaliana

Pollen development involves complex developmental control of gene expression by the haploid genome. Phosphoinositide-dependent phospholipase C (PI-PLC) is a major enzyme of inositol phospholipid signaling system. This signal system makes important contribution to transmembrane signal transduction, plant growth and development. These two messenger substance play crucial role in extracelluar stimuli and mediating various physiological processes.Here, we adopted a reverse genetics approach to investigate the function of AtPLC2 the Arabidopsis thaliana encoding AtPLC gene.In this study, PLC2/plc2 was the AtPLC2 heterozygous line and atplc2i was the AtPLC2 RNAi line. We attempted to isolate Arabidopsis plants that do not express the AtPLC2 gene, but could not recover homozygous mutant plants. The progeny of PLC2/plc2 heterozygous plants, harboring a T-DNA insertion, showed a segregation ratio of 1:1:0 for Wild-type, heterozygous and homozygous mutant plants, indicating a gametophytic defect. Genetic transmission analysis showed that the abnormal segregation ratio was due to failure to transmit the mutant allele through the male gametophyte. AtPLC2 gene homozygous mutant died with T-DNA insertion. The decreasing of pollen grains in anthers of the heterozygous PLC2/plc2-2 was detected and the number of mature nuclear pollen was less than the wild type. Microscopic observation revealed that many mature pollen grains from the heterozygous plants contained large vacuoles even until the mature pollen stage, whereas pollen from wild-type plants contained many small vacuoles beginning from the vacuolated pollen stage, which indicated that vacuoles in many of the heterozygous mutant pollen did not undergo normal fission after the first mitotic division. The AtPLC2 is important for pollen development. The quantity of silique on stem in WT, PLC2/plc2-1, PLC2/plc2-2 and atplc2i plants shown that PLC2/plc2-1, PLC2/plc2-2 and atplc2i plants have more defects silique than WT in the growth of the late. By biochemical staining of pollen, AtPLC2 lacking caused the pollen development interrupted.