Brachypodium distachyon promoters as efficient building blocks for transgenic research in maize.

The biotechnological approach to improve performance or yield of crops or for engineering metabolic pathways requires the expression of a number of transgenes, each with a specific promoter to avoid induction of silencing mechanisms. In maize (Zea mays), used as a model for cereals, an efficient Agrobacterium tumefaciens-mediated transformation system has been established that is applied for translational research. In the current transformation vectors, the promoters of the 35S gene of the cauliflower mosaic virus and of the ubiquitin gene of maize are often used to drive the bialaphos-selectable marker and the transgene, respectively. To expand the number of promoters, genes with either constitutive or seed-specific expression were selected in Brachypodium distachyon, a model grass distantly related to maize. After the corresponding Brachypodium promoters had been fused to the β-glucuronidase reporter gene, their activity was followed throughout maize development and quantified in a fluorimetric assay with the 4-methylumbelliferyl β-D-glucuronide substrate. The promoters pBdEF1α and pBdUBI10 were constitutively and highly active in maize, whereas pBdGLU1 was clearly endosperm-specific, hence, expanding the toolbox for transgene analysis in maize. The data indicate that Brachypodium is an excellent resource for promoters for transgenic research in heterologous cereal species.

[1]  Dawn H. Nagel,et al.  The B73 Maize Genome: Complexity, Diversity, and Dynamics , 2009, Science.

[2]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[3]  P. Barceló,et al.  A facile method for screening for phosphinothricin (PPT)-resistant transgenic wheats , 1999, Molecular Breeding.

[4]  M. D. Block,et al.  The selection mechanism of phosphinothricin is influenced by the metabolic status of the tissue , 1995, Planta.

[5]  K. Lamkey,et al.  Registration of Five Inbred Lines of Maize: B102, B103, B104, B105, and B106 , 1997 .

[6]  Jukon Kim,et al.  Analysis of five novel putative constitutive gene promoters in transgenic rice plants , 2010, Journal of experimental botany.

[7]  Kan Wang,et al.  Genetic Transformation and Hybridization Improved Agrobacterium-mediated Transformation of Three Maize Inbred Lines Using Ms Salts , 2022 .

[8]  A. Furtado,et al.  Analysis of promoters in transgenic barley and wheat. , 2009, Plant biotechnology journal.

[9]  P. Quail,et al.  Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation , 1992, Plant Molecular Biology.

[10]  M. D. Block,et al.  Energy efficiency and energy homeostasis as genetic and epigenetic components of plant performance and crop productivity , 2011 .

[11]  Chung-Mo Park,et al.  Exploring valid reference genes for gene expression studies in Brachypodium distachyon by real-time PCR , 2008, BMC Plant Biology.

[12]  P. Lemaux,et al.  Transformation of Maize Cells and Regeneration of Fertile Transgenic Plants. , 1990, The Plant cell.

[13]  P. Waterhouse,et al.  RNA Silencing in Plants: Yesterday, Today, and Tomorrow , 2008, Plant Physiology.

[14]  D. Nettleton,et al.  Agrobacterium tumefaciens-Mediated Transformation of Maize Embryos Using a Standard Binary Vector System1 , 2002, Plant Physiology.

[15]  Carolyn J. Lawrence-Dill,et al.  Comparative Plant Genomics Resources at PlantGDB1 , 2005, Plant Physiology.

[16]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[17]  J. Draper,et al.  Brachypodium distachyon. A new model system for functional genomics in grasses. , 2001, Plant physiology.

[18]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[19]  M. Montagu,et al.  Quantitative kinetic analysis of β-glucuronidase activities using a computer-directed microtiter plate reader , 1993, Plant Molecular Biology Reporter.

[20]  T. Komari,et al.  High efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens , 1996, Nature Biotechnology.

[21]  P. Christou,et al.  When more is better: multigene engineering in plants. , 2010, Trends in plant science.

[22]  D. Douchkov,et al.  A Set of Modular Binary Vectors for Transformation of Cereals1[W][OA] , 2007, Plant Physiology.

[23]  J. Leemans,et al.  Transgenic maize plants by tissue electroporation. , 1992, The Plant cell.

[24]  R. Twyman,et al.  Promoter diversity in multigene transformation , 2010, Plant Molecular Biology.

[25]  M. Norman,et al.  The Ecology of Tropical Food Crops: Maize ( Zea mays ) , 1995 .

[26]  J. Giraudat,et al.  A Brassica napus transcript encoding a protein related to the Künitz protease inhibitor family accumulates upon water stress in leaves, not in seeds. , 1992, The Plant journal : for cell and molecular biology.

[27]  M. Montagu,et al.  Transgenic plants protected from insect attack , 1987, Nature.

[28]  W. V. Camp.  Yield enhancement genes: seeds for growth. , 2005 .

[29]  Sai Guna Ranjan Gurazada,et al.  Genome sequencing and analysis of the model grass Brachypodium distachyon , 2010, Nature.

[30]  P. Beyer,et al.  Engineering the provitamin A (beta-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. , 2000, Science.

[31]  Josh Strable,et al.  Maize (Zea mays): a model organism for basic and applied research in plant biology. , 2009, Cold Spring Harbor protocols.

[32]  P. Hilson,et al.  Somatic embryogenesis and plant regeneration of tropical maize genotypes , 2010, Plant Cell, Tissue and Organ Culture (PCTOC).

[33]  A. Depicker,et al.  Plants as bioreactors for protein production: avoiding the problem of transgene silencing , 2004, Plant Molecular Biology.

[34]  M. Bevan,et al.  GUS fusions: beta‐glucuronidase as a sensitive and versatile gene fusion marker in higher plants. , 1987, The EMBO journal.

[35]  M. Chilton,et al.  The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA , 1986, Journal of bacteriology.

[36]  K. Koch,et al.  Steady-state transposon mutagenesis in inbred maize. , 2005, The Plant journal : for cell and molecular biology.

[37]  Jürgen Breitenbach,et al.  Transgenic multivitamin corn through biofortification of endosperm with three vitamins representing three distinct metabolic pathways , 2009, Proceedings of the National Academy of Sciences.

[38]  M. Van Montagu,et al.  Engineering herbicide resistance in plants by expression of a detoxifying enzyme , 1987, The EMBO journal.

[39]  P. Hilson,et al.  Building Blocks for Plant Gene Assembly1[W][OA] , 2007, Plant Physiology.

[40]  Jürgen Breitenbach,et al.  Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize , 2008, Proceedings of the National Academy of Sciences.

[41]  F. Nagy,et al.  Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter , 1985, Nature.

[42]  A. Depicker,et al.  Single-copy T-DNAs integrated at different positions in the Arabidopsis genome display uniform and comparable β-glucuronidase accumulation levels , 2004, Cellular and Molecular Life Sciences CMLS.