Increased Resistance to Penicillium Seed Rot in Transgenic Wheat Over‐expressing Puroindolines

Puroindolines (PINs) are the main components of the wheat grain hardness locus (Ha) and have in vitro antimicrobial activity against bacteria and fungi. Here, we examined the effect of variation in PINA and/or PINB content upon Penicillium sp. seed fungal growth inhibition. The Penicillium sp. assays were germination assays performed after incubating seeds in Penicillium sp. contaminated soil. The first set of wheat genotypes consisted of two sets of transgenic isolines created in the varieties ‘Bobwhite’ and ‘Hi‐Line’ having over‐expression of PINA and/or PINB. The second set of genotypes consisted of near‐isogenic lines (NILs) varying for mutations in PINA or PINB created in the varieties ‘Explorer’ and ‘Hank’. After incubation in Penicillium sp.‐infected soil, transgenic wheat seeds over‐expressing PINA in both ‘Hi‐Line’ and ‘Bobwhite’ and both PINs in ‘Hi‐Line’ exhibited significantly reduced fungal infection and increased germination. No significant differences in Penicillium sp. infection or germination rates were observed in seeds of the NILs. The results indicate that puroindolines native role in seeds is to increase seed viability and that when over‐expressed as transgenes, the puroindolines are effective antifungal proteins.

[1]  D. Marion,et al.  Spatial and temporal distribution of the major isoforms of puroindolines (puroindoline-a and puroindoline-b) and non specific lipid transfer protein (ns-LTP1e1) of Triticum aestivum seeds. Relationships with their in vitro antifungal properties , 1998 .

[2]  M. Gautier,et al.  Puroindoline genes are highly conserved in diploid ancestor wheats and related species but absent in tetraploid Triticum species. , 2000 .

[3]  M. Giroux,et al.  Complementation of the pina (null) allele with the wild type Pina sequence restores a soft phenotype in transgenic wheat , 2006, Theoretical and Applied Genetics.

[4]  J. Kader.  LIPID-TRANSFER PROTEINS IN PLANTS. , 1996, Annual review of plant physiology and plant molecular biology.

[5]  M. Giroux,et al.  Puroindolines: Their Role in Grain Hardness and Plant Defence , 2003, Biotechnology & genetic engineering reviews.

[6]  D. Mathre,et al.  Control of dry seed decay of wheat , 1991 .

[7]  M. Giroux,et al.  Hard Wheat Milling and Bread Baking Traits Affected by the Seed‐Specific Overexpression of Puroindolines , 2005 .

[8]  M. Giroux,et al.  Wheat puroindolines enhance fungal disease resistance in transgenic rice. , 2001, Molecular plant-microbe interactions : MPMI.

[9]  O. Anderson,et al.  Rapid Production of Multiple Independent Lines of Fertile Transgenic Wheat (Triticum aestivum) , 1993, Plant physiology.

[10]  Yang Li,et al.  Expression of puroindoline a enhances leaf rust resistance in transgenic tetraploid wheat , 2008, Molecular Biology Reports.

[11]  D. Marion,et al.  Triticum aestivum puroindolines, two basic cystine-rich seed proteins: cDNA sequence analysis and developmental gene expression , 1994, Plant Molecular Biology.

[12]  S. V. Oard,et al.  Expression of the antimicrobial peptides in plants to control phytopathogenic bacteria and fungi , 2006, Plant Cell Reports.

[13]  M. Zasloff.  Antimicrobial peptides of multicellular organisms , 2002, Nature.

[14]  E. Conti,et al.  Fusarium head blight evaluation in wheat transgenic plants expressing the maize b-32 antifungal gene , 2007, European Journal of Plant Pathology.

[15]  R. Hancock,et al.  Cationic peptides: a new source of antibiotics. , 1998, Trends in biotechnology.

[16]  C. Faleri,et al.  Two Plant Puroindolines Colocalize in Wheat Seed and in vitro Synergistically Fight Against Pathogens , 2005, Plant Molecular Biology.

[17]  W. Broekaert,et al.  Antimicrobial Peptides from Plants , 1997 .

[18]  F. Leuven,et al.  Antimicrobial peptides fromMirabilis jalapa andAmaranthus caudatus: expression, processing, localization and biological activity in transgenic tobacco , 1996, Plant Molecular Biology.

[19]  V. R. Stewart,et al.  Registration of “Hi‐Line” Wheat , 1992 .

[20]  G. Carlson,et al.  Registration of ‘Choteau’ Wheat , 2004 .

[21]  R. Bressan,et al.  In Defense against Pathogens. Both Plant Sentinels and Foot Soldiers Need to Know the Enemy1,212 , 2003, Plant Physiology.

[22]  A. Lovegrove,et al.  Promoter analysis and immunolocalisation show that puroindoline genes are exclusively expressed in starchy endosperm cells of wheat grain , 2007, Plant Molecular Biology.

[23]  C. Morris,et al.  Wheat grain hardness results from highly conserved mutations in the friabilin components puroindoline a and b. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[24]  H. Vogel,et al.  Tryptophan- and arginine-rich antimicrobial peptides: structures and mechanisms of action. , 2006, Biochimica et biophysica acta.

[25]  R. Ferreira,et al.  The role of plant defence proteins in fungal pathogenesis. , 2007, Molecular plant pathology.

[26]  Z. Punja.  Genetic engineering of plants to enhance resistance to fungal pathogens—a review of progress and future prospects , 2001 .

[27]  Rosalinda D'Amore,et al.  Constitutive over-expression of two wheat pathogenesis-related genes enhances resistance of tobacco plants to Phytophthora nicotianae , 2007, Plant Cell, Tissue and Organ Culture.

[28]  D. Marion,et al.  Puroindoline‐a and α1‐purothionin form ion channels in giant liposomes but exert different toxic actions on murine cells , 2006, The FEBS journal.

[29]  M. Giroux,et al.  The Wheat Puroindoline Genes Confer Fungal Resistance in Transgenic Corn , 2011 .

[30]  P. Carbonero,et al.  Susceptibility of phytopathogenic bacteria to wheat purothionins in vitro. , 1972, Applied microbiology.

[31]  M. Gautier,et al.  Expression of wheat puroindoline-b reduces scab susceptibility in transgenic apple (Malus × domestica Borkh.) , 2004 .

[32]  M. Giroux,et al.  Wheat puroindolines interact to form friabilin and control wheat grain hardness , 2004, Theoretical and Applied Genetics.

[33]  G. Carlson,et al.  Registration of 'Explorer' wheat , 2002 .

[34]  N. Sitaram.  Antimicrobial peptides with unusual amino acid compositions and unusual structures. , 2006, Current medicinal chemistry.

[35]  D. Marion,et al.  Puroindolines form ion channels in biological membranes. , 2003, Biophysical journal.