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Transgenic banana plants expressing a Stellaria media defensin gene (Sm-AMP-D1) demonstrate improved resistance to Fusarium oxysporum

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Abstract

Plant defensins are group of small, cysteine stabilized antimicrobial peptides rich in basic amino acids which inhibit growth of a multitude of phytopathogens. These defensins have been explored widely to generate transgenic crop plants resistant to varied fungal and bacterial diseases. In the present study, gene sequence coding for a seed defensin (Sm-AMP-D1) of common chickweed Stellaria media was synthesized artificially and cloned downstream of a strong constitutive promoter in pCAMBIA-1301 plant expression vector. Transgenic banana plants expressing the Sm-AMP-D1 gene were subsequently generated via Agrobacterium-mediated genetic transformation. Transgenic nature of the regenerated banana plants was confirmed by genomic DNA PCR and Southern blotting analysis. Northern blots demonstrated efficient expression of Sm-AMP-D1 mRNA in transgenic banana plants. Further, two selected transgenic lines challenged with a pathogenic isolate of Fusarium oxysporum f. sp. cubense race 1 showed improved resistance as compared to untransformed control banana plants. These transgenic lines continued to show resistance against Foc race 1 6 months post-infection. This study demonstrates that overexpression of potent plant defensins such as Sm-AMP-D1 in important food crops like banana can lead to development of durable resistance against fungal pathogens.

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References

  • Allen A, Snyder AK, Preuss M, Nielsen EE, Shah DM, Smith TJ (2008) Plant defensins and virally encoded fungal toxin KP4 inhibit plant root growth. Planta 227:331–339

    Article  PubMed  CAS  Google Scholar 

  • Anderson MA, Heath RL, Lay FT, Poon S (2009) US patent application 2009; application 20090083880. Modified plant defensin

  • Bakry F, Carreel F, Jenny C, Horry JP (2009) Genetic improvement of banana. In: Jain SM, Priyadarshan PM (eds) Breeding plantation tree crops: tropical species. Springer, Berlin, pp 3–51

    Chapter  Google Scholar 

  • Butler D (2013) Fungus threatens top banana: fears rise for Latin American industry as devastating disease hits leading variety in Africa and Middle East. Nature 504:195–196. doi:10.1038/504195a

    Article  PubMed  CAS  Google Scholar 

  • Cary JW, Rajasekaran K, Jaynes JM, Cleveland TE (2000) Transgenic expression of a gene encoding a synthetic antimicrobial peptide results in inhibition of fungal growth in vitro and in planta. Plant Sci 154:171–181

    Article  PubMed  CAS  Google Scholar 

  • Chakrabarti A, Ganapathi TR, Mukherjee PK, Bapat VA (2003) MSI-99, a magainin analogue, imparts enhanced disease resistance in transgenic tobacco and banana. Planta 216:587–596

    PubMed  CAS  Google Scholar 

  • Cote FX, Domergue R, Monmarson S, Schwendiman J, Teisson C, Escalant JV (1996) Embryogenic cell suspensions from the male flower of Musa AAA cv. Grand Nain. Physiol Plant 97:285–290

    Article  CAS  Google Scholar 

  • D’Hont A, Denoeud F, Aury JM, Baurens FC, Carreel F, Garsmeur O, Noel B, Bocs S, Droc G, Rouard M, Da Silva C, Jabbari K, Cardi C, Poulain J, Souquet M, Labadie K, Jourda C, Lengellé J, Rodier-Goud M, Alberti A, Bernard M, Correa M, Ayyampalayam S, Mckain MR, Leebens-Mack J, Burgess D, Freeling M, Mbéguié-A-Mbéguié D, Chabannes M, Wicker T, Panaud O, Barbosa J, Hribova E, Heslop-Harrison P, Habas R, Rivallan R, Francois P, Poiron C, Kilian A, Burthia D, Jenny C, Bakry F, Brown S, Guignon V, Kema G, Dita M, Waalwijk C, Joseph S, Dievart A, Jaillon O, Leclercq J, Argout X, Lyons E, Almeida A, Jeridi M, Dolezel J, Roux N, Risterucci AM, Weissenbach J, Ruiz M, Glaszmann JC, Quétier F, Yahiaoui N, Wincker P (2012) The banana (Musa acuminata) genome and the evolution of monocotyledonous plants. Nature 488:213–217

    Article  PubMed  Google Scholar 

  • Ganapathi TR, Higgs NS, Balint-Kurti PJ, Arntzen CJ, May GD, Van Eck JM (2001) Agrobacterium-mediated transformation of the embryogenic cell suspensions of the banana cultivar Rasthali (AAB). Plant Cell Rep 20:157–162

    Article  CAS  Google Scholar 

  • Gao AG, Hakimi SM, Mittanck CA, Wu Y, Woerner BM, Stark DM, Shah DM, Liang J, Rommens CM (2000) Fungal pathogen protection in potato by expression of a plant defensin peptide. Nat Biotechnol 18:1307–1310

    Article  PubMed  CAS  Google Scholar 

  • Ghag SB, Shekhawat UKS, Ganapathi TR (2012) Petunia floral defensins with unique prodomains as novel candidates for development of Fusarium wilt resistance in transgenic banana plants. PLoS One 7(6):e39557

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Ghag SB, Shekhawat UKS, Ganapathi TR (2013) Expression of C-terminal prodomain truncated Petunia floral defensins inhibit the growth of transgenic banana plants. Curr Trends Biotechnol Pharm 7:505–510

    CAS  Google Scholar 

  • Ghag SB, Shekhawat UKS, Ganapathi TR (2014) Host induced post-transcriptional hairpin RNA-mediated gene silencing of vital fungal genes confers efficient resistance against Fusarium wilt in banana. Plant Biotechnol J. doi:10.1111/pbi.12158

    PubMed  Google Scholar 

  • Ghosh A, Ganapathi TR, Nath P, Bapat VA (2009) Establishment of embryogenic cell suspension cultures and Agrobacterium-mediated transformation in an important Cavendish banana cv. Robusta (AAA). Plant Cell Tissue Organ Cult 97:131–139

    Article  Google Scholar 

  • Jha S, Chattoo BB (2009) Transgene stacking and coordinated expression of plant defensins confer fungal resistance in rice. Rice 2:143–154

    Article  Google Scholar 

  • Jha S, Chattoo BB (2010) Expression of a plant defensin in rice confers resistance to fungal phytopathogens. Transgenic Res 19:373–384

    Article  PubMed  CAS  Google Scholar 

  • Kanzaki H, Nirasawa S, Saitoh H, Ito M, Nishihara M, Terauchi R, Nakamura I (2002) Overexpression of the wasabi defensin gene confers enhanced resistance to blast fungus (Magnaporthe oryzae) in transgenic rice. Theor Appl Genet 105:809–814

    Article  PubMed  CAS  Google Scholar 

  • Lay FT, Anderson MA (2005) Defensins—components of the innate immune system in plants. Curr Protein Pept Sci 6:85–101

    Article  PubMed  CAS  Google Scholar 

  • Lay FT, Brugliera F, Anderson MA (2003) Isolation and properties of floral defensins from ornamental tobacco and Petunia. Plant Physiol 131:1283–1293

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Li C, Shao J, Wang Y, Li W, Guo D, Yan B, Xia Y, Peng M (2013) Analysis of banana transcriptome and global gene expression profiles in banana roots in response to infection by race 1 and tropical race 4 of Fusarium oxysporum f. sp. cubense. BMC Genomics 14:851

    Article  PubMed  PubMed Central  Google Scholar 

  • Mohandas S, Sowmya HD, Saxena AK, Meenakshia S, Rania RT, Mahmood R (2013) Transgenic banana cv. Rasthali (AAB, Silk gp) harboring Ace-AMP1 gene imparts enhanced resistance to Fusarium oxysporum f. sp. cubense race 1. Sci Hortic 164:392–399

    Article  CAS  Google Scholar 

  • Ploetz RC (2006) Fusarium wilt of banana is caused by several pathogens referred to as Fusarium oxysporum f. sp. cubense. Phytopathology 96:653–656

    Article  PubMed  Google Scholar 

  • Portieles R, Ayra C, Gonzalez E, Gallo A, Rodriguez R, Chacón O, López Y, Rodriguez M, Castillo J, Pujol M, Enriquez G, Borroto C, Trujillo L, Thomma BP, Borrás-Hidalgo O (2010) NmDef02, a novel antimicrobial gene isolated from Nicotiana megalosiphon confers high-level pathogen resistance under greenhouse and field conditions. Plant Biotechnol J 8:678–690

    Article  PubMed  CAS  Google Scholar 

  • Sagaram US, Pandurangi R, Kaur J, Smith TJ, Shah DM (2011) Structure-activity determinants in antifungal plant defensins MsDef1 and MtDef4 with different modes of action against Fusarium graminearum. PLoS One 6(4):e18550

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Sagaram US, El-Mounadi K, Buchko GW, Berg HR, Kaur J, Pandurangi RS, Smith TJ, Shah DM (2013) Structural and functional studies of a phosphatidic acid-binding antifungal plant defensin MtDef4: identification of an RGFRRR motif governing fungal cell entry. PLoS One 8(12):e82485

    Article  PubMed  PubMed Central  Google Scholar 

  • Schouten HJ, Krens FA, Jacobsen E (2006) Cisgenic plants are similar to traditionally bred plants: international regulations for genetically modified organisms should be altered to exempt cisgenesis. EMBO Rep 7:750–753

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Shekhawat UKS, Srinivas L, Ganapathi TR (2011) MusaDHN-1, a novel multiple stress-inducible SK(3)-type dehydrin gene, contributes affirmatively to drought- and salt-stress tolerance in banana. Planta 234:915–932

    Article  PubMed  CAS  Google Scholar 

  • Slavokhotova AA, Odintsova TI, Rogozhin EA, Musolyamov AK, Andreev YA, Grishin EV, Egorov TA (2010) Isolation, molecular cloning and antimicrobial activity of novel defensins from common chickweed (Stellaria media L.) seeds. Biochimie 93:450–456

    Article  PubMed  Google Scholar 

  • Stotz HU, Thomson JG, Wang Y (2009) Plant defensins: defense, development and application. Plant Signal Behav 4:1010–1012

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Stover RH (1962) Fusarial wilt (Panama disease) of bananas and other Musa species. Commonwealth Mycological Institute, Surrey

    Google Scholar 

  • Swathi Anuradha T, Divya K, Jami SK, Kirti PB (2008) Transgenic tobacco and peanut plants expressing a mustard defensin show resistance to fungal pathogens. Plant Cell Rep 27:1777–1786

    Article  PubMed  CAS  Google Scholar 

  • Thomma BPHJ, Cammue BPA, Thevissen K (2002) Plant defensins. Planta 216:193–202

    Article  PubMed  CAS  Google Scholar 

  • Zasloff M (2002) Antimicrobial peptides of multicellular organisms. Nature 415:389–395

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

Authors would like to thank Dr. S. P. Kale, Head, Nuclear Agriculture and Biotechnology Division, BARC for his support and encouragement and Dr. A K Saxena, IIHR, Bangalore for kindly providing the Foc cultures used in this study.

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Authors and Affiliations

  1. Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400 085, India

    Siddhesh B. Ghag, Upendra K. Singh Shekhawat & Thumballi R. Ganapathi

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  1. Siddhesh B. Ghag
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  2. Upendra K. Singh Shekhawat
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  3. Thumballi R. Ganapathi
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Correspondence to Thumballi R. Ganapathi.

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Ghag, S.B., Shekhawat, U.K.S. & Ganapathi, T.R. Transgenic banana plants expressing a Stellaria media defensin gene (Sm-AMP-D1) demonstrate improved resistance to Fusarium oxysporum . Plant Cell Tiss Organ Cult 119, 247–255 (2014). https://doi.org/10.1007/s11240-014-0529-x

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  • DOI: https://doi.org/10.1007/s11240-014-0529-x

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