Abstract
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.
Similar content being viewed by others
Abbreviations
- ADP:
-
Adenosine diphosphate
- BE:
-
Branching enzymes
- DBE:
-
Debranching enzyme
- DBEI:
-
ISA1-type DBE
- Glc:
-
Glucose
- GBSS:
-
Granule-bound starch synthase
- ISA:
-
Isoamylase
- RACE:
-
Rapid amplification of cDNA end
- qRT-PCR:
-
Quantitative real-time PCR
- RT-PCR:
-
Reverse transcriptase polymerase chain reaction
- SSS:
-
Soluble starch synthases
- UTRs:
-
Untranslated regions
References
Saunders RM, Becker R (1984) Amaranthus: a potential food and feed resource. Adv Cereal Sci Technol 6:357–396
Marcone MF (2001) Starch properties of Amaranthus pumilus (seabeach amaranth): a threatened plant species with potential benefits for the breeding/amelioration of present Amaranthus cultivars. Food Chem 73:61–66
Inouchi N, Nishi K, Tanaka S, Asai M, Kawase Y, Hata Y, Konishi Y, Yue S, Fuwa H (1999) Characterization of amaranth and quinoa starches. J Appl Glycosci 46:233–240
Stone LA, Lorenz K (1984) The starch of amaranthus—physico-chemical properties and functional characteristics. Starch-Stärke 36:232–237
Smith A, Denyer K, Martin C (1997) The synthesis of the starch granule. Annu Rev Plant Biol 48:67–87
Martin C, Smith AM (1995) Starch biosynthesis. Plant Cell 7:971
Kubo A, Rahman S, Utsumi Y, Li Z, Mukai Y (2005) Complementation of sugary-1 phenotype in rice endosperm with the wheat isoamylase1 gene supports a direct role for isoamylase1 in amylopectin biosynthesis. Plant Physiol 137:43–56
van der Maarel MJEC, van der Veen B, Uitdehaag JCM, Leemhuis H, Dijkhuizen L (2002) Properties and applications of starch-converting enzymes of the alpha-amylase family. J Biotechnol 94:137–155
Burton RA, Jenner H, Carrangis L, Fahy B (2002) Starch granule initiation and growth are altered in barley mutants that lack isoamylase activity. Plant J 31:97–112
James MG, Robertson DS, Myers AM (1995) Characterization of the maize gene sugary1, a determinant of starch composition in kernels. Plant Cell 7:417–429
Nakamura Y, Kubo A, Shimamune T, Matsuda T, Harada K, Satoh H (1997) Correlation between activities of starch debranching enzyme and alpha-polyglucan structure in endosperms of sugary-1 mutants of rice. Plant J 12:143–153
Dauvillée D, Colleoni C, Mouille G, Morell MK, d’Hulst C, Wattebled F, Liénard L, Delvallé D, Ral J-P, Myers AM, Ball SG (2001) Biochemical characterization of wild-type and mutant isoamylases of Chlamydomonas reinhardtii supports a function of the multimeric enzyme organization in amylopectin maturation. Plant Physiol 125:1723–1731
Posewitz MC, Smolinski SL, Kanakagiri S, Melis A, Seibert M, Ghirardi ML (2004) Hydrogen photoproduction is attenuated by disruption of an isoamylase gene in Chlamydomonas reinhardtii. Plant Cell 16:2151–2163
Delatte T, Trevisan M, Parker ML, Zeeman SC (2005) Arabidopsis mutants Atisa1 and Atisa2 have identical phenotypes and lack the same multimeric isoamylase, which influences the branch point distribution of amylopectin during starch synthesis. Plant J 41:815–830
Bustos R (2004) Starch granule initiation is controlled by a heteromultimeric isoamylase in potato tubers. Proc Natl Acad Sci USA 101:2215–2220
Bierhals J, Lajolo F, Cordenunsi B (2004) Activity, cloning, and expression of an isoamylase-type starch-debranching enzyme from banana fruit. J Agric Food Chem 52:7412–7418
Park Y-J, Nemoto K, Nishikawa T, Matsushima K, Minami M, Kawase M (2011) Genetic diversity and expression analysis of granule bound starch synthase I gene in the new world grain amaranth (Amaranthus cruentus L.). J Cereal Sci 53:298–305
Murray M, Thompson W (1980) Rapid isolation of high molecular weight plant DNA. Nucl Acids Res 8:4321–4325
Toguri T (1991) Changes of a rice debranching enzyme during seed formation and germination. J Plant Physiol 137:541–546
Nakamura Y, Umemoto T, Takahata Y (1996) Changes in structure of starch and enzyme activities affected by sugary mutations in developing rice endosperm. Possible role of starch debranching enzyme (R-enzyme) in amylopectin biosynthesis. Physiol Plantarum 97:491–498
Kim S, Hamada T, Otani M (2005) Cloning and characterization of sweetpotato isoamylase gene (IbIsa1) isolated from tuberous root. Breed Sci 55:453–458
Kubo A, Colleoni C, Dinges JR, Lin Q, Lappe RR, Rivenbark JG, Meyer AJ, Ball SG, James MG, Hennen-Bierwagen TA, Myers AM (2010) Functions of heteromeric and homomeric isoamylase-type starch-debranching enzymes in developing maize endosperm. Plant Physiol 153:956–969
Joshi C (1987) Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucl Acids Res 15:9627–9640
Ball SG, Morell MK (2003) From bacterial glycogen to starch: understanding the biogenesis of the plant starch granule. Annu Rev Plant Biol 54:207–233
Hussain H, Mant A, Seale R, Zeeman S, Hinchliffe E, Edwards A, Hylton C, Bornemann S, Smith AM, Martin C, Bustos R (2003) Three isoforms of isoamylase contribute different catalytic properties for the debranching of potato glucans. Plant Cell 15:133–149
Ohdan T, Francisco PB, Sawada T, Hirose T, Terao T, Satoh H, Nakamura Y (2005) Expression profiling of genes involved in starch synthesis in sink and source organs of rice. J Exp Bot 56:3229–3244
Park Y-J, Nishikawa T (2012) Characterization and expression analysis of the starch synthase gene family in grain amaranth (Amaranthus cruentus L.). Genes Genet Syst 87:281–289
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Park, YJ., Nishikawa, T., Tomooka, N. et al. Molecular characterization of an isoamylase 1-type starch debranching enzyme (DBEI) in grain amaranth (Amaranthus cruentus L.). Mol Biol Rep 41, 7857–7864 (2014). https://doi.org/10.1007/s11033-014-3679-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-014-3679-3