Cloning and expression analysis of cDNAs encoding ADP-glucose pyrophosphorylase large and small subunits from hulless barley (Hordeum vulgare L. var. nudum)

Dongmei Li 1 , 2 , Zhimin Yang 1 , Xinchun Liu 1 , Zhen Song 2 , Zongyun Feng 1 ,  and Yang He 2
  • 1 College of Agronomy, Sichuan Agricultural University, Chengdu 611130, P.R. China
  • 2 College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, P.R. China
Dongmei Li
  • College of Agronomy, Sichuan Agricultural University, Chengdu 611130, P.R. China
  • College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, P.R. China
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, Zhimin Yang, Xinchun Liu, Zhen Song
  • College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, P.R. China
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, Zongyun Feng and Yang He
  • Corresponding author
  • College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, P.R. China
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Abstract

As an important plateau cereal crop, hulless barley is the principal food for the Tibetan people in China. ADP-glucose pyrophosphorylase (AGPase) is considered as the key enzyme for starch biosynthesis in plants. In this study, cDNAs encoding the small subunit (SSU I) and large subunit (LSU I) of AGPase were isolated from hulless barley. The results showed that SSU I and LSU I were 1438 and 1786 bp in length with a complete open reading frame (ORF) of 1419 and 1572 bp. The ORF-encoded polypeptides of 472 and 523 amino acids were having calculated molecular masses of 52.01 and 58.23 kDa, and the pI values were 5.59 and 6.30. In addition, phylogenetic analysis showed that SSU I and LSU I had the same phylogenetic trends with some species. Furthermore, expression levels in different growth periods and tissues of two hulless barley varieties were analyzed by quantitative reverse transcription-polymerase chain reaction. Gene expression levels of SSU I and LSU I were consistent with the total starch accumulation rate in endosperm. In conclusion, our data confirmed that SSU I and LSU I played an important role in hulless barley starch synthesis.

  • 1.

    Fu DX, Ruan RW, Dai XM, Liu YM. A study on ancient barley, wheat and millet discovered at Changguo of Tibet. Acta Agron Sin 2002;6:392–8.

  • 2.

    Zeng XQ, Bai Lj, Wei ZX, Yuan HJ, Wang YL, Qijun Xu, et al. Transcriptome analysis revealed the drought-responsive genes in Tibetan hulless barley. BMC Genomics 2016;17:386.

    • Crossref
    • PubMed
    • Export Citation
  • 3.

    Zeng XQ, Long H, Wang Z, Zhao SC, Tang YW, Huang ZY, et al. The draft genome of Tibetan hulless barley reveals adaptive patterns to the high stressful Tibetan Plateau. Proc Natl Acad Sci USA 2015;114:1095–100.

  • 4.

    Tang YW, Zeng XQ, Wang YL, Bai LJ, Xu QJ, Wei ZX, et al. Transcriptomics analysis of hulless barley during grain development with a focus on starch biosynthesis. Funct Integr Genomics 2017;17:107–17.

    • Crossref
    • PubMed
    • Export Citation
  • 5.

    Zheng XG, Qi JC, Hui HS, Lin LH, Wang F. Starch accumulation in hulless barley during grain filling. Bot Stu 2017;58:30.

    • Crossref
    • Export Citation
  • 6.

    Kang GZ, Liu GQ, Peng XQ, Wei LT, Wang CY, Zhu YJ, et al. Increasing the starch content and grain weight of common wheat by overexpression of the cytosolic AGPase large subunit gene. Plant Physiol Biochem 2013;73:93–8.

    • Crossref
    • PubMed
    • Export Citation
  • 7.

    Seng SS, Wu J, Sui JJ, Wu CY, Zhong XH, Liu C, et al. ADP-glucose pyrophosphorylase gene plays a key role in the quality of corm and yield of cormels in gladiolus. Biochem Bioph Res Co 2016;474:206–12.

    • Crossref
    • Export Citation
  • 8.

    Martin C, Smith AM. Starch biosynthesis. Plant Cell 1995;7:971–85.

  • 9.

    Bhave MR, Lawrence S, Barton C, Hannah LC. Identification and molecular characterization of shrunken-2 cDNA clones of maize. Plant Cell 1990;2:581–8.

    • PubMed
    • Export Citation
  • 10.

    Patron NJ, Keeling PJ. Common evolutionary origin of starch biosynthetic enzymes in green and red algae. J Phycol 2005;41:1131–41.

    • Crossref
    • Export Citation
  • 11.

    Hwang SK, Hamada S, Okita TW. Catalytic implications of the higher plant ADP-glucose pyrophosphorylase large subunit. Phytochemistry 2007;68:464–77.

    • Crossref
    • PubMed
    • Export Citation
  • 12.

    Boehlein SK, Shaw JR, Hannah LC, Stewart JD. Probing allosteric binding sites of the maize endosperm ADP-glucose pyrophosphorylase. Plant Physiol Bioch 2010;152:85–95.

    • Crossref
    • Export Citation
  • 13.

    Rösti S, Rudi H, Rudi K, Opsahl-Sorteberg HG, Fahy B, Denyer K. The gene encoding the cytosilic small subunit of ADP-glucose pyrophosphorylase in barley endosperm also encodes the major plastidial small subunit in the leaves. J Exp Bot 2006;57:3619–26.

    • Crossref
    • PubMed
    • Export Citation
  • 14.

    Ainsworth C, Hosein F, Tarvis M, Weir F, Burrell MM, Devos KM, et al. Adenosine diphosphate glucose pyrophosphorylase genes in wheat: differential expression and gene mapping. Planta 1995;197:1–10.

    • PubMed
    • Export Citation
  • 15.

    Lee SK, Hwang SK, Han M, Eom JS, Kang HG, Han Y, et al. Identification of the ADP-glucose pyrophosphorylase isoforms essential for starch synthesis in the leaf and seed endosperm of rice (Oryza sativa L.). Plant Mol Biol 2007;65:531–46.

    • Crossref
    • PubMed
    • Export Citation
  • 16.

    Corbi J, Duthiel JY, Damerval C, Tennailon MI, Manicacci D. Accelerated evolution and coevolution drove the evolutionary history of AGPase sub-units during angiosperm radiation. Ann Bot London 2012;109:693–708.

    • Crossref
    • Export Citation
  • 17.

    Chen X, Long H, Gao P, Deng GB, Pan ZF, Liang JJ, et al. Transcriptome Assembly and analysis of Tibetan Hulless barley (Hordeum vulgare L. var. nudum): developing grains, with emphasis on quality properties. PLoS One 2014;9:e98144.

    • Crossref
    • PubMed
    • Export Citation
  • 18.

    Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar M. MEGA5:molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 2011;28:2731–9.

    • Crossref
    • PubMed
    • Export Citation
  • 19.

    Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 1980;16:111–20. BRAC.

    • Crossref
    • PubMed
    • Export Citation
  • 20.

    Bao J, Corke H, Sun M. Microsatellites in starch-—synthesizing genes in relation to starch physicochemical properties in waxy rice (Oryza sativa L.). Theor Appl Genet 2002;105:898–905.

    • Crossref
    • PubMed
    • Export Citation
  • 21.

    Saripalli G, Gupta PK. AGPase: its role in crop productivity with emphasis on heat tolerance in cereals. Theor Appl Genet 2015;128:1893–916.

    • Crossref
    • PubMed
    • Export Citation
  • 22.

    Hwang SK, Hamada S, Okita TW. ATP binding site in the plant ADP-glucose pyrophosphorylase large subunit. Febs Lett 2006;580:6741–8.

    • Crossref
    • PubMed
    • Export Citation
  • 23.

    Fu YB, Ballicora MA, Preiss J. Mutagenesis of the Glucose-1-Phosphate-Binding Site of Potato Tuber ADP-glucose pyrophosphorylase. Plant Physiol 1998;117:989–96.

    • Crossref
    • PubMed
    • Export Citation
  • 24.

    La Cognata U, Willmitzer L, Muller RB. Molecular cloning and characterization of novel isoforms of potato ADP-glucose pyrophosphorylase. Mol Gen Genet 1995;246:538–48.

    • Crossref
    • PubMed
    • Export Citation
  • 25.

    Nakamura Y, Kawaguchi K. Multiple forms of ADP glucose pyrophosphorylase of rice endosperm. Physiol Plantarum 1992;84:336–42.

    • Crossref
    • Export Citation
  • 26.

    Cakira B, Tuncel AR, Green A, Koper K, Hwang Seon-Kap W, Okita T, et al. Substrate binding properties of potato tuber ADP-glucose pyrophosphorylase as determined by isothermal titration calorimetry. Febs Lett 2015;589:1444–9.

    • Crossref
    • PubMed
    • Export Citation
  • 27.

    Burton RA, Johnson PE, Beckles DM, Fincher GB, Jenner HL, Naldrett MJ, et al. Characterization of the genes encoding the cytosolic and plastidial forms of ADPglucose pyrophosphorylase in wheat endosperm. Plant Physiol 2002;130:1464–75.

    • Crossref
    • PubMed
    • Export Citation
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A Journal of Biosciences: Zeitschrift für Naturforschung C (ZNC) is an international scientific journal for the emerging field of natural and natural-like products. ZNC publishes original research on the isolation, bio-chemical synthesis and bioactivities of natural products, their biochemistry, pharmacology, biotechnology, and biological activity and innovative developed computational methods for predicting their structure and/or function.

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