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The Lack of Beta‐amylase Activity in Soybean Cultivar Altona sp 1 is Associated with a 1.2 kb Deletion in the 5′ Region of Beta‐amylase I Gene
Author(s) -
Kim WonSeok,
Krishnan Hari B.
Publication year - 2010
Publication title -
crop science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.76
H-Index - 147
eISSN - 1435-0653
pISSN - 0011-183X
DOI - 10.2135/cropsci2009.11.0660
Subject(s) - biology , amylase , gene , glycine soja , genetics , microbiology and biotechnology , glycine , biochemistry , enzyme , amino acid
Previous studies have identified near‐isogenic soybean [ Glycine max (L.) Merr.] lines, one containing normal β‐amylase [α‐1,4‐glucan maltohydrolase (EC 3.2.1.2)] activity (Altona Sp 1 b ) and the other with undetectable β‐amylase activity (Altona sp 1 ) 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 sp 1, was identified as β‐amylase by matrix‐assisted‐laser‐desorption‐time‐of‐flight mass spectrometry (MALDI‐TOF‐MS) and immunoblot analysis using antibodies generated against Arabidopsis β‐amylase. Southern blot analysis showed differences in the sizes of the DNA fragments hybridizing to β‐amylase probe between the near‐isogenic soybean lines. A search of the soybean genome database revealed the presence of nine β‐amylase genes in the soybean genome. We have isolated the β‐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 sp 1 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 β‐amylase activity in soybean cultivar Altona Altona sp 1.