Developmental and environmental regulation of soybean SE60 gene expression during embryogenesis and germination.
Soybean SE60 belongs to the gamma-thionin family of proteins. We recently demonstrated that SE60 plays a role in defense during soybean development. Here, we show that SE60 is expressed in a tissue-specific and developmentally regulated manner. The expression of SE60 is distinct from that of the glycinin (Gy2) and extensin (SbHRGP3) genes of soybean during embryogenesis and germination. A SE60::GUS(-809) transgene, comprising -809 bp of the 5'-flanking region of SE60 fused to the GUS reporter gene, was expressed specifically in developing embryos, but not in the endosperms, from the globular stage of transgenic tobacco and Arabidopsis seeds. Furthermore, light affected the SE60::GUS(-809) expression pattern in germinating seedlings. Electrophoretic mobility shift assay (EMSA) revealed that soybean nuclear proteins as well as E. coli-expressed SB16, a high mobility group protein (HMG), were bound sequence-specifically to the fragment containing AT-rich motifs identified in the SE60 promoter. Interestingly, the soybean nuclear proteins binding to the two G-boxes and RY repeat were prevalent in seeds of 2-4 mm in size. In contrast, the nuclear proteins binding to the AT-rich motif and SE60 RNA expression were more prominent in seeds of 4-6 mm in size. Therefore, we propose that factors binding to the G-boxes or RY repeat initiate SE60 expression during embryogenesis.
Choi
Yeonhee
Y
School of Biological Sciences, Seoul National University, Seoul 151-747, Korea. yhc@snu.ac.kr
Jeong
Cheol Woong
CW
Ohr
Hyonhwa
H
Song
Sang-Kee
SK
Choi
Yang Do
YD
Lee
Jong Seob
JS
D020856:AT Rich Sequence / Q000235:genetics; D001483:Base Sequence; D015342:DNA Probes / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D047108:Embryonic Development / Q000235:genetics*; D004777:Environment; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D018525:Germination / Q000235:genetics*; D005966:Glucuronidase / Q000378:metabolism; D006609:High Mobility Group Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D020133:Reverse Transcriptase Polymerase Chain Reaction; D036226:Seedlings / Q000235:genetics; D012639:Seeds / Q000196:embryology / Q000235:genetics; D030262:Soybean Proteins / Q000235:genetics* / Q000378:metabolism; D013025:Glycine max / Q000196:embryology* / Q000235:genetics*; D013997:Time Factors
D015342:DNA Probes; D006609:High Mobility Group Proteins; D009687:Nuclear Proteins; C085202:SE60 protein, Glycine max; D030262:Soybean Proteins; D005966:Glucuronidase
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