The dynamic behavior of storage organelles in developing cereal seeds and its impact on the production of recombinant proteins

Cereal endosperm is a highly differentiated tissue containing specialized organelles for the accumulation of storage proteins, which are ultimately deposited either within protein bodies derived from the endoplasmic reticulum, or in protein storage vacuoles (PSVs). During seed maturation endosperm cells undergo a rapid sequence of developmental changes, including extensive reorganization and rearrangement of the endomembrane system and protein transport via several developmentally regulated trafficking routes. Storage organelles have been characterized in great detail by the histochemical analysis of fixed immature tissue samples. More recently, in vivo imaging and the use of tonoplast markers and fluorescent organelle tracers have provided further insight into the dynamic morphology of PSVs in different cell layers of the developing endosperm. This is relevant for biotechnological applications in the area of molecular farming because seed storage organelles in different cereal crops offer alternative subcellular destinations for the deposition of recombinant proteins that can reduce proteolytic degradation, allow control over glycan structures and increase the efficacy of oral delivery. We discuss how the specialized architecture and developmental changes of the endomembrane system in endosperm cells may influence the subcellular fate and post-translational modification of recombinant glycoproteins in different cereal species.

[1]  S. Hayashida,et al.  The Relationship of the Feces Protein Particles to Rice Protein Bodies , 1975 .

[2]  D. Wettstein,et al.  Protein body formation in the developing barley endosperm , 1980 .

[3]  Russell L. Jones,et al.  Ca2+-Stimulated Secretion of α-Amylase During Development in Barley Aleurone Protoplasts , 1986 .

[4]  M. Cornejo,et al.  Ca-stimulated secretion of alpha-amylase during development in barley aleurone protoplasts. , 1986, Plant physiology.

[5]  Kunisuke Tanaka,et al.  Purification of protein body-I of rice seed and its polypeptide composition , 1987 .

[6]  J. Jacobsen,et al.  Regulation of synthesis and transport of secreted proteins in cereal aleurone. , 1991, International review of cytology.

[7]  G. Galili,et al.  Evidence for a novel route of wheat storage proteins to vacuoles , 1992, The Journal of cell biology.

[8]  B. Priem,et al.  Mannosyl- and Xylosyl-Containing Glycans Promote Tomato (Lycopersicon esculentum Mill.) Fruit Ripening. , 1992, Plant physiology.

[9]  C. Damerval,et al.  Quantification of dominance for proteins pleiotropically affected by opaque-2 in maize , 1993, Heredity.

[10]  V. Cameron-Mills,et al.  A role for gamma 3 hordein in the transport and targeting of prolamin polypeptides to the vacuole of developing barley endosperm. , 1993, The Plant journal : for cell and molecular biology.

[11]  G. Leubner-Metzger,et al.  Class I [beta]-1,3-Glucanases in the Endosperm of Tobacco during Germination , 1995, Plant physiology.

[12]  D. Robinson,et al.  Protein storage vacuoles form de novo during pea cotyledon development. , 1995, Journal of cell science.

[13]  P. Shewry,et al.  Seed storage proteins: structures and biosynthesis. , 1995, The Plant cell.

[14]  R. L. Jones,et al.  Isolation of Intact Protein Storage Vacuoles from Barley Aleurone (Identification of Aspartic and Cysteine Proteases) , 1996, Plant physiology.

[15]  H. Nonogaki,et al.  An Endo-[beta]-Mannanase Develops Exclusively in the Micropylar Endosperm of Tomato Seeds Prior to Radicle Emergence , 1996, Plant physiology.

[16]  S. Swanson,et al.  Gibberellic Acid Induces Vacuolar Acidification in Barley Aleurone. , 1996, The Plant cell.

[17]  Russell L. Jones,et al.  From storage compartment to lytic organelle: the metamorphosis of the aleurone protein storage vacuole , 1998 .

[18]  Russell L. Jones,et al.  Barley Aleurone Cells Contain Two Types of Vacuoles: Characterization of Lytic Organelles by Use of Fluorescent Probes , 1998, Plant Cell.

[19]  M. Nishimura,et al.  Transport of Storage Proteins to Protein Storage Vacuoles Is Mediated by Large Precursor-Accumulating Vesicles , 1998, Plant Cell.

[20]  K. Müntz.  Deposition of storage proteins , 1998 .

[21]  F. Culiáñez-macià,et al.  The dormancy-related peroxiredoxin anti-oxidant, PER1, is localized to the nucleus of barley embryo and aleurone cells. , 1999, The Plant journal : for cell and molecular biology.

[22]  Jones,et al.  Barley aleurone cell death is not apoptotic: characterization of nuclease activities and DNA degradation , 1999, The Plant journal : for cell and molecular biology.

[23]  Nichols,et al.  Developmental biology of the cereal endosperm. , 1999, Trends in plant science.

[24]  James E. Bailey,et al.  Engineered glycoforms of an antineuroblastoma IgG1 with optimized antibody-dependent cellular cytotoxic activity , 1999, Nature Biotechnology.

[25]  S. Ritchie,et al.  Physiology of the aleurone layer and starchy endosperm during grain development and early seedling growth: new insights from cell and molecular biology , 2000, Seed Science Research.

[26]  Y. Karamanos,et al.  Evidence of two enzymes performing the de-N-glycosylation of proteins in barley: expression during germination, localization within the grain and set-up during grain formation. , 2000, Journal of experimental botany.

[27]  Y. Kimura,et al.  Structural Analysis of Free N-Glycans Occurring in Soybean Seedlings and Dry Seeds , 2000, Bioscience, biotechnology, and biochemistry.

[28]  T. Okamoto,et al.  Mass Transport of Proform of a Kdel-Tailed Cysteine Proteinase (Sh-EP) to Protein Storage Vacuoles by Endoplasmic Reticulum–Derived Vesicle Is Involved in Protein Mobilization in Germinating Seeds , 2000, The Journal of cell biology.

[29]  F. Chen,et al.  Expression of an expansin is associated with endosperm weakening during tomato seed germination. , 2000, Plant physiology.

[30]  F. Meins,et al.  Plant Physiol . ( 1995 ) 109 : 751-759 Class I ß-1 , 3-Glucanases in the Endosperm of Tobacco during Germination , 2000 .

[31]  B. Larkins,et al.  Genomics Analysis of Genes Expressed in Maize Endosperm Identifies Novel Seed Proteins and Clarifies Patterns of Zein Gene Expression , 2001, The Plant Cell Online.

[32]  R. L. Jones,et al.  Cell death of barley aleurone protoplasts is mediated by reactive oxygen species. , 2008, The Plant journal : for cell and molecular biology.

[33]  P. Shewry,et al.  Endosperm-specific activity of a storage protein gene promoter in transgenic wheat seed. , 2001, Journal of experimental botany.

[34]  E. Golovina,et al.  Mechanisms of plant desiccation tolerance. , 2001, Trends in plant science.

[35]  R. Twyman,et al.  Native and artificial reticuloplasmins co-accumulate in distinct domains of the endoplasmic reticulum and in post-endoplasmic reticulum compartments. , 2001, Plant physiology.

[36]  G. Bryant,et al.  Membrane behaviour in seeds and other systems at low water content: the various effects of solutes , 2001, Seed Science Research.

[37]  E. Herman,et al.  Expression patterns of genes encoding endomembrane proteins support a reduced function of the Golgi in wheat endosperm during the onset of storage protein deposition. , 2001, Journal of experimental botany.

[38]  R. L. Jones,et al.  Cell death of barley aleurone protoplasts is mediated by reactive oxygen species. , 2008, The Plant journal : for cell and molecular biology.

[39]  P. Shewry,et al.  Cereal seed storage proteins: structures, properties and role in grain utilization. , 2002, Journal of experimental botany.

[40]  B. Larkins,et al.  Maize Opaque Endosperm Mutations Create Extensive Changes in Patterns of Gene Expression Online version contains Web-only data. Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.003905. , 2002, The Plant Cell Online.

[41]  Y. Kimura,et al.  Subcellular localization of endo-beta-N-acetylglucosaminidase and high-mannose type free N-glycans in plant cell. , 2002, Biochimica et biophysica acta.

[42]  E. Sato,et al.  [Reactive oxygen]. , 2002, Nihon eiseigaku zasshi. Japanese journal of hygiene.

[43]  A. Depicker,et al.  Boosting heterologous protein production in transgenic dicotyledonous seeds using Phaseolus vulgaris regulatory sequences , 2002, Nature Biotechnology.

[44]  K. Yano,et al.  Endo-β-N-acetylglucosaminidase, an enzyme involved in processing of free oligosaccharides in the cytosol , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[45]  M. Nishimura,et al.  C-Terminal KDEL Sequence of A KDEL-Tailed Cysteine Proteinase (Sulfhydryl-Endopeptidase) Is Involved in Formation of KDEL Vesicle and in Efficient Vacuolar Transport of Sulfhydryl-Endopeptidase1 , 2003, Plant Physiology.

[46]  B. Coddeville,et al.  Comparative analysis of the site-specific N-glycosylation of human lactoferrin produced in maize and tobacco plants. , 2003, European journal of biochemistry.

[47]  B. Svensson,et al.  Feasibility study of a tissue-specific approach to barley proteome analysis: aleurone layer, endosperm, embryo and single seeds , 2003 .

[48]  V. Kauppinen,et al.  Expression of fungal thermotolerant endo-1,4-β-glucanase in transgenic barley seeds during germination , 1999, Plant Molecular Biology.

[49]  P. Lerouge,et al.  N-Glycoprotein biosynthesis in plants: recent developments and future trends , 1998, Plant Molecular Biology.

[50]  B. Svensson,et al.  Aspects of the barley seed proteome during development and germination. , 2004, Biochemical Society transactions.

[51]  M. Ogawa,et al.  The Accumulation of α-Zein in Transgenic Tobacco Endosperm is Stabilized by Co-expression of β-Zein , 2004 .

[52]  F. Takaiwa,et al.  Evaluation of tissue specificity and expression strength of rice seed component gene promoters in transgenic rice. , 2004, Plant biotechnology journal.

[53]  J. Jacobsen,et al.  Gibberellic-acid-responsive protoplasts from mature aleurone of Himalaya barley , 1985, Planta.

[54]  P. Christou,et al.  Unexpected Deposition Patterns of Recombinant Proteins in Post-Endoplasmic Reticulum Compartments of Wheat Endosperm1 , 2004, Plant Physiology.

[55]  K. Müntz.  Deposition of storage proteins , 1998, Plant Molecular Biology.

[56]  J. Jacobsen,et al.  The structure and composition of aleurone grains in the barley aleurone layer , 2004, Planta.

[57]  P. Bethke,et al.  Programmed cell death in cereal aleurone , 2000, Plant Molecular Biology.

[58]  Christer Jansson,et al.  The barley starch granule proteome—internalized granule polypeptides of the mature endosperm , 2004 .

[59]  Gad Galili,et al.  ER-Derived Compartments Are Formed by Highly Regulated Processes and Have Special Functions in Plants , 2004, Plant Physiology.

[60]  B. Larkins,et al.  Immunolocalization of avenin and globulin storage proteins in developing endosperm of Avena sativa L. , 1989, Planta.

[61]  T. Young,et al.  Programmed cell death during endosperm development , 2000, Plant Molecular Biology.

[62]  O. Langella,et al.  A two-dimensional proteome map of maize endosperm. , 2004, Phytochemistry.

[63]  J. Jacobsen,et al.  The structure and composition of Aleurone grains in the Barley Aleurone layer , 1971, Planta.

[64]  P. Christou,et al.  Recent progress in plantibody technology. , 2005, Current pharmaceutical design.

[65]  Liwen Jiang,et al.  Selective Membrane Protein Internalization Accompanies Movement from the Endoplasmic Reticulum to the Protein Storage Vacuole Pathway in Arabidopsis , 2005, The Plant Cell Online.

[66]  P. Christou,et al.  Sowing the seeds of success: pharmaceutical proteins from plants. , 2005, Current opinion in biotechnology.

[67]  Hur-Song Chang,et al.  A Gibberellin-Regulated Calcineurin B in Rice Localizes to the Tonoplast and Is Implicated in Vacuole Function1 , 2005, Plant Physiology.

[68]  T. Jomori,et al.  Genetically modified rice seeds accumulating GLP‐1 analogue stimulate insulin secretion from a mouse pancreatic beta‐cell line , 2005, FEBS letters.

[69]  A. Vitale,et al.  Sorting of proteins to storage vacuoles: how many mechanisms? , 2005, Trends in plant science.

[70]  Hideyuki Takahashi,et al.  A novel vesicle derived directly from endoplasmic reticulum is involved in the transport of vacuolar storage proteins in rice endosperm. , 2005, Plant & cell physiology.

[71]  M. Bardor,et al.  A KDEL-tagged monoclonal antibody is efficiently retained in the endoplasmic reticulum in leaves, but is both partially secreted and sorted to protein storage vacuoles in seeds. , 2006, Plant biotechnology journal.

[72]  P. Christou,et al.  The Intracellular Fate of a Recombinant Protein Is Tissue Dependent1 , 2006, Plant Physiology.

[73]  B. Svensson,et al.  Enrichment and identification of integral membrane proteins from barley aleurone layers by reversed-phase chromatography, SDS-PAGE, and LC-MS/MS. , 2006, Journal of proteome research.

[74]  Marc Strickert,et al.  Gene expression patterns reveal tissue-specific signaling networks controlling programmed cell death and ABA- regulated maturation in developing barley seeds. , 2006, The Plant journal : for cell and molecular biology.

[75]  Martin J. Mueller,et al.  Nonenzymatic Lipid Peroxidation Reprograms Gene Expression and Activates Defense Markers in Arabidopsis Tocopherol-Deficient Mutants[W] , 2006, The Plant Cell Online.

[76]  Pingfang Yang,et al.  Proteomic analysis of rice (Oryza sativa) seeds during germination , 2007, Proteomics.

[77]  M. Van Montagu,et al.  Aberrant localization and underglycosylation of highly accumulating single-chain Fv-Fc antibodies in transgenic Arabidopsis seeds , 2007, Proceedings of the National Academy of Sciences.

[78]  C. Stemmer,et al.  In vivo glyco‐engineered antibody with improved lytic potential produced by an innovative non‐mammalian expression system , 2007, Biotechnology journal.

[79]  H. Kiyono,et al.  Rice-based mucosal vaccine as a global strategy for cold-chain- and needle-free vaccination , 2007, Proceedings of the National Academy of Sciences.

[80]  P. Roepstorff,et al.  Spatio‐temporal changes in germination and radical elongation of barley seeds tracked by proteome analysis of dissected embryo, aleurone layer, and endosperm tissues , 2007, Proteomics.

[81]  B. Larkins,et al.  The Maize Floury1 Gene Encodes a Novel Endoplasmic Reticulum Protein Involved in Zein Protein Body Formation[W] , 2007, The Plant Cell Online.

[82]  Winfriede Weschke,et al.  Analysis of barley (Hordeum vulgare) grain development using three-dimensional digital models. , 2007, The Plant journal : for cell and molecular biology.

[83]  John L Cleveland,et al.  Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes , 2008, Autophagy.

[84]  R. Fischer,et al.  Recombinant antibody 2G12 produced in maize endosperm efficiently neutralizes HIV-1 and contains predominantly single-GlcNAc N-glycans. , 2008, Plant biotechnology journal.

[85]  P. Christou,et al.  Maize plants: An ideal production platform for effective and safe molecular pharming , 2008 .

[86]  P. Christou,et al.  Cost-effective production of a vaginal protein microbicide to prevent HIV transmission , 2008, Proceedings of the National Academy of Sciences.

[87]  Uwe Scholz,et al.  Barley Grain Maturation and Germination: Metabolic Pathway and Regulatory Network Commonalities and Differences Highlighted by New MapMan/PageMan Profiling Tools1[W][OA] , 2008, Plant Physiology.

[88]  A. Chan,et al.  Advancing Cell Biology and Functional Genomics in Maize Using Fluorescent Protein-Tagged Lines1 , 2008, Plant Physiology.

[89]  J. Scheller,et al.  Influence of elastin-like peptide fusions on the quantity and quality of a tobacco-derived human immunodeficiency virus-neutralizing antibody. , 2009, Plant biotechnology journal.

[90]  P. Shewry,et al.  Trafficking of storage proteins in developing grain of wheat , 2009, Journal of experimental botany.

[91]  S. Hirose,et al.  Deposition of a recombinant peptide in ER-derived protein bodies by retention with cysteine-rich prolamins in transgenic rice seed , 2009, Planta.

[92]  B. Svensson,et al.  Barley seed proteomics from spots to structures. , 2009, Journal of proteomics.

[93]  R A Knight,et al.  Guidelines for the use and interpretation of assays for monitoring cell death in higher eukaryotes , 2009, Cell Death and Differentiation.

[94]  Daniel J Klionsky,et al.  In search of an “autophagomometer” , 2009, Autophagy.

[95]  T. Kumamaru,et al.  ER membrane-localized oxidoreductase Ero1 is required for disulfide bond formation in the rice endosperm , 2009, Proceedings of the National Academy of Sciences.

[96]  F. Liu,et al.  OsRab5a regulates endomembrane organization and storage protein trafficking in rice endosperm cells. , 2010, The Plant journal : for cell and molecular biology.

[97]  R. Vierstra,et al.  ATG8 lipidation and ATG8-mediated autophagy in Arabidopsis require ATG12 expressed from the differentially controlled ATG12A AND ATG12B loci. , 2010, The Plant journal : for cell and molecular biology.

[98]  Wenfeng Li,et al.  iTRAQ Protein Profile Analysis of Arabidopsis Roots Reveals New Aspects Critical for Iron Homeostasis1[C][W] , 2010, Plant Physiology.

[99]  S. Hirose,et al.  Rice seed ER-derived protein body as an efficient delivery vehicle for oral tolerogenic peptides , 2010, Peptides.

[100]  Youko Oono,et al.  Analysis of ER stress in developing rice endosperm accumulating beta-amyloid peptide. , 2010, Plant biotechnology journal.

[101]  U. Hellman,et al.  Barley as a green factory for the production of functional Flt3 ligand , 2010, Biotechnology journal.

[102]  Lee-Anne S. Milburn,et al.  Sowing the Seeds of Success , 2010, Landscape Journal.

[103]  K. Shiraki,et al.  Extraction and purification of human interleukin-10 from transgenic rice seeds. , 2010, Protein expression and purification.

[104]  R. Fischer,et al.  The Changing Fate of a Secretory Glycoprotein in Developing Maize Endosperm1[C][W] , 2010, Plant Physiology.

[105]  P. Hussey,et al.  BODIPY probes to study peroxisome dynamics in vivo. , 2010, The Plant journal : for cell and molecular biology.

[106]  J. Stadlmann,et al.  The two endo-β-N-acetylglucosaminidase genes from Arabidopsis thaliana encode cytoplasmic enzymes controlling free N-glycan levels , 2011, Plant Molecular Biology.

[107]  Jialing Yao,et al.  Constructing the metabolic and regulatory pathways in germinating rice seeds through proteomic approach , 2011, Proteomics.

[108]  A. Depicker,et al.  Non-food/feed seeds as biofactories for the high-yield production of recombinant pharmaceuticals. , 2011, Plant biotechnology journal.

[109]  David Holding,et al.  Delivery of Prolamins to the Protein Storage Vacuole in Maize Aleurone Cells[W] , 2011, Plant Cell.

[110]  A. Depicker,et al.  Expression of Antibody Fragments with a Controlled N-Glycosylation Pattern and Induction of Endoplasmic Reticulum-Derived Vesicles in Seeds of Arabidopsis1[C][W][OA] , 2011, Plant Physiology.

[111]  W. Weschke,et al.  Laser-capture microdissection of developing barley seeds and cDNA array analysis of selected tissues. , 2011, Methods in molecular biology.

[112]  J. G. Contreras,et al.  Centrality Dependence of the Charged-Particle Multiplicity Density at Midrapidity in Pb-Pb Collisions at √sNN=2.76 TeV , 2010, 1012.1657.

[113]  H. Guo,et al.  Preparation of new 5-fluorouracil-loaded zein nanoparticles for liver targeting. , 2011, International journal of pharmaceutics.

[114]  M. Maeda,et al.  Double-Knockout of Putative Endo-β-N-acetylglucosaminidase (ENGase) Genes in Arabidopsis thaliana: Loss of ENGase Activity Induced Accumulation of High-Mannose Type Free N-Glycans Bearing N,N′-Acetylchitobiosyl Unit , 2011, Bioscience, biotechnology, and biochemistry.

[115]  T. B. Skaali,et al.  Centrality Dependence of the Charged-Particle Multiplicity Density at Midrapidity in Pb-Pb Collisions at ffiffiffiffiffiffiffiffi s NN p 1⁄4 2 : 76 TeV , 2011 .

[116]  J. Rogers.  Internal Membranes in Maize Aleurone Protein Storage Vacuoles: Beyond Autophagy , 2011, Plant Cell.

[117]  B. Svensson,et al.  Proteomes of the barley aleurone layer: A model system for plant signalling and protein secretion , 2011, Proteomics.

[118]  Lang Shen,et al.  Large-scale production of functional human serum albumin from transgenic rice seeds , 2011, Proceedings of the National Academy of Sciences.

[119]  A. Depicker,et al.  Production of monoclonal antibodies with a controlled N-glycosylation pattern in seeds of Arabidopsis thaliana. , 2011, Plant biotechnology journal.

[120]  W. Gruissem,et al.  iTRAQ-based analysis of changes in the cassava root proteome reveals pathways associated with post-harvest physiological deterioration. , 2011, The Plant journal : for cell and molecular biology.

[121]  S. Hirose,et al.  Expression of hypoallergenic Der f 2 derivatives with altered intramolecular disulphide bonds induces the formation of novel ER-derived protein bodies in transgenic rice seeds , 2012, Journal of experimental botany.

[122]  N. Packer,et al.  Production of α-L-iduronidase in maize for the potential treatment of a human lysosomal storage disease , 2012, Nature Communications.

[123]  T. Masumura,et al.  Accumulation of rice prolamin–GFP fusion proteins induces ER-derived protein bodies in transgenic rice calli , 2012, Plant Cell Reports.

[124]  Hideyuki Takahashi,et al.  Recombinant protein yield in rice seed is enhanced by specific suppression of endogenous seed proteins at the same deposit site. , 2012, Plant biotechnology journal.

[125]  Kunisuke Tanaka,et al.  Formation mechanism of the internal structure of type I protein bodies in rice endosperm: relationship between the localization of prolamin species and the expression of individual genes. , 2012, The Plant journal : for cell and molecular biology.

[126]  Mohammad Pessarakli,et al.  Reactive Oxygen Species, Oxidative Damage, and Antioxidative Defense Mechanism in Plants under Stressful Conditions , 2012 .

[127]  F. Takaiwa,et al.  Expression of OsBiP4 and OsBiP5 is highly correlated with the endoplasmic reticulum stress response in rice , 2012, Planta.

[128]  Siqi Liu,et al.  Stress Responsive Proteins Are Actively Regulated during Rice (Oryza sativa) Embryogenesis as Indicated by Quantitative Proteomics Analysis , 2013, PloS one.

[129]  Zhouxin Shen,et al.  Reconstruction of protein networks from an atlas of maize seed proteotypes , 2013, Proceedings of the National Academy of Sciences.

[130]  B. Svensson,et al.  Gibberellic Acid-Induced Aleurone Layers Responding to Heat Shock or Tunicamycin Provide Insight into the N-Glycoproteome, Protein Secretion, and Endoplasmic Reticulum Stress1[W] , 2013, Plant Physiology.

[131]  M. Grebe,et al.  The Endoplasmic Reticulum Is the Main Membrane Source for Biogenesis of the Lytic Vacuole in Arabidopsis[W] , 2013, Plant Cell.

[132]  Stuart K Johnson,et al.  Encapsulation of Hydrocortisone and Mesalazine in Zein Microparticles , 2013, Pharmaceutics.

[133]  Y. Ishino,et al.  A Guanine Nucleotide Exchange Factor for Rab5 Proteins Is Essential for Intracellular Transport of the Proglutelin from the Golgi Apparatus to the Protein Storage Vacuole in Rice Endosperm1[C][W][OA] , 2013, Plant Physiology.

[134]  J. Šamaj,et al.  Maize proteomics: An insight into the biology of an important cereal crop , 2013, Proteomics.

[135]  Z. Yin,et al.  Small GTPase Sar1 is crucial for proglutelin and α-globulin export from the endoplasmic reticulum in rice endosperm , 2013, Journal of experimental botany.

[136]  M. Sack,et al.  Plant species and organ influence the structure and subcellular localization of recombinant glycoproteins , 2013, Plant Molecular Biology.

[137]  Y. Onda,et al.  Oxidative Protein-Folding Systems in Plant Cells , 2013, International journal of cell biology.

[138]  Hideyuki Takahashi,et al.  Transgenic rice seeds accumulating recombinant hypoallergenic birch pollen allergen Bet v 1 generate giant protein bodies. , 2013, Plant & cell physiology.

[139]  F. Takaiwa,et al.  The use of rice seeds to produce human pharmaceuticals for oral therapy , 2013, Biotechnology journal.

[140]  P. Christou,et al.  Efficient recovery of recombinant proteins from cereal endosperm is affected by interaction with endogenous storage proteins. , 2013, Biotechnology journal.

[141]  C. Viotti.  ER and vacuoles: never been closer , 2014, Front. Plant Sci..

[142]  P. Cosette,et al.  Proteomic analysis. , 2014, Methods in molecular biology.

[143]  E. Arcalis,et al.  Fusion, rupture, and degeneration: the fate of in vivo-labelled PSVs in developing barley endosperm * , 2014, Journal of experimental botany.

[144]  M. Moloney,et al.  Plant molecular pharming for the treatment of chronic and infectious diseases. , 2014, Annual review of plant biology.

[145]  B. Schneider,et al.  Laser microdissection: a sample preparation technique for plant micrometabolic profiling. , 2014, Phytochemical analysis : PCA.

[146]  P. Shewry,et al.  The trafficking pathway of a wheat storage protein in transgenic rice endosperm , 2014, Annals of botany.

[147]  K. Hefferon,et al.  Plant Molecular Pharming , 2016 .