Molecular characterization of an isoamylase 1-type starch debranching enzyme (DBEI) in grain amaranth (Amaranthus cruentus L.).

The purpose of this study was to characterize the molecular profile of a starch debranching enzyme (DBE) in grain amaranth. A cDNA clone that encodes a putative DBE was isolated from amaranth perisperm and then sequenced. This amaranth DBE appears to be an ISA1-type DBE (DBEI), based on its substrate specificity and the sequence similarity between the 2,391-bp cDNA clone and ISA1 s from potato and Arabidopsis. The mature DBEI of amaranth consists of 796 amino acids (90.5 kDa). We analyzed the transcript levels of the DBEI gene in amaranth seeds during various developmental stages and in plant tissues by qRT-PCR and RT-PCR analyses. The transcript levels of the DBEI gene rapidly increased at the middle stage of seed maturation. This result indicates that the enzyme encoded by the amaranth DBEI gene plays an important role in starch accumulation throughout the seed during the middle stage of seed development. We detected DBEI transcripts in storage and non-storage tissues. At the six-leaf stage, there were high levels of the DBEI transcripts in leaves, petioles, and the stem, and low levels in the root. Therefore, we suggest that the DBEI expression is not specific to non-storage and/or storage tissues. This summary of the basic characteristics of the DBEI gene will contribute to further studies on starch biosynthesis in Amaranthus.