Reference Report for RTN20170303.1
| Title: | The Lack of Beta-amylase Activity in Soybean Cultivar Altona sp is Associated with a 1.2 kb Deletion in the 5' Region of Beta-amylase I Gene |
| Authors: | Kim, S.K., Krishnan, H.B. |
| Source: | Crop Sci. 2010 50:1942-1949 |
| Abstract: | Previous studies have identified near-isogenic soybean [Glycine max (L.) Merr.] lines, one containing normal _-amylase [_-1,4 - glucan malto -hydrolase (EC 3.2.1.2)] activity (Altona Sp1b) and the other with undetectable _-amylase activity (Altona sp1) in seeds. SDS-PAGE analysis of 50% isopropanol extracted proteins from soybean seeds revealed a prominent 56 kDa protein. This protein, which was absent in Altona sp1, was identifi ed as _-amylase by matrix-assisted-laser-desorption-time-of-flight mass spectrometry (MALDI-TOF-MS) and immunoblot analysis using antibodies generated against Arabidopsis _-amylase. Southern blot analysis showed differences in the sizes of the DNA fragments hybridizing to _-amylase probe between the near-isogenic soybean lines. A search of the soybean genome database revealed the presence of nine _-amylase genes in the soybean genome. We have isolated the _-amylase gene (GmBAM 1) by screening a genomic library of wild-type soybean and determined its nucleotide sequence. Analysis of the nucleotide sequence of the GmBAM 1 revealed a complete open reading frame that was interrupted by six introns. In contrast, the GmBAM 1 from Altona sp1 had a 1207 bp deletion near the 5' region that included the second and third exon regions. Our results suggest that this deletion may be responsible for the lack of _-amylase activity in soybean cultivar Altona Altona sp |
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