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Conferred resistance to Botrytis cinerea in Lilium by overexpression of the RCH10 chitinase gene

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Transgenic Lilium lines have been generated by Agrobacterium -mediated transformation that have enhanced resistance to Botrytis cinerea as a consequence of ectopic expression of a rice chitinase gene.

Abstract

The production of ornamentals is an important global industry, with Lilium being one of the six major bulb crops in the world. The international trade in ornamentals is in the order of £60–75 billion and is expected to increase worldwide by 2–4 % per annum. The continued success of the floriculture industry depends on the introduction of new species/cultivars with major alterations in key agronomic characteristics, such as resistance to pathogens. Fungal diseases are the cause of reduced yields and marketable quality of cultivated plants, including ornamental species. The fungal pathogen Botrytis causes extreme economic losses to a wide range of crop species, including ornamentals such as Lilium. Agrobacterium-mediated transformation was used to develop Lilium oriental cv. ‘Star Gazer’ plants that ectopically overexpress the Rice Chitinase 10 gene (RCH10), under control of the CaMV35S promoter. Levels of conferred resistance linked to chitinase expression were evaluated by infection with Botrytis cinerea; sporulation was reduced in an in vitro assay and the relative expression of the RCH10 gene was determined by quantitative reverse transcriptase-PCR. The extent of resistance to Botrytis, compared to that of the wild type plants, showed a direct correlation with the level of chitinase gene expression. Transgenic plants grown to flowering showed no detrimental phenotypic effects associated with transgene expression. This is the first report of Lilium plants with resistance to Botrytis cinerea generated by a transgenic approach.

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References

  • Anand A, Zhou T, Trick HN, Gill BS, Bockus WW, Muthukrishnan S (2003) Greenhouse and field testing of transgenic wheat plants stably expressing genes for thaumatin-like protein, chitinase and glucanase against Fusarium graminearum. J Exp Bot 54:1101–1111

    Article  CAS  PubMed  Google Scholar 

  • Balconi C, Lanzanova C, Conti E, Triulzi T, Forlani F, Cattaneo M, Lupotto E (2007) Fusarium head blight evaluation in wheat transgenic plants expressing the maize b-32 antifungal gene. Eur J Plant Pathol 117:129–140

    Article  CAS  Google Scholar 

  • Balode A (2009) Breeding for resistance against Botrytis in lily. Acta Hort (ISHS) 836:143–148

    Google Scholar 

  • Chandler S, Tanaka Y (2007) Genetic modification in floriculture. Crit Rev Plant Sci 26:169–197

    Article  CAS  Google Scholar 

  • Conijn CGM (2014) Developments in the control of lily diseases. Acta Hort (ISHS) 1027:213–229

    Google Scholar 

  • Daughtrey ML, Bridgen MP (2013) Evaluating resistance to Botrytis elliptica in field-grown lilies. Acta Hort (ISHS) 1002:313–318

    Google Scholar 

  • Distefano G, La Malfa S, Vitale A, Lorito M, Deng ZN, Gentile A (2008) Defence-related gene expression in transgenic lemon plants producing an antimicrobial Trichoderma harzianum endochitinase during fungal infection. Transgenic Res 17:873–879

    Article  CAS  PubMed  Google Scholar 

  • Edwards K, Johnstone C, Thompson C (1991) A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucl Acid Res 19:1349

    Article  CAS  Google Scholar 

  • EFSA (2006) Opinion of the scientific panel on genetically modified organisms on a request from the commission related to the notification (Reference C/NL/04/02) for the placing on the market of the genetically modified carnation Moonlite 123.2.38 with a modified colour, for import of cut flowers for ornamental use, under Part C of directive 2001/18/EC from Florigene. EFSA J 362:1–19

    Google Scholar 

  • Esposito S, Colucci MG, Frusciante L, Filippone E, Lorito M, Bressan RA (2000) Antifungal transgenes expression in Petunia hybrida. In: Proceedings of the nineteenth international symposium on improvement of ornamental plants. Acta Hort (ISHS), vol 508, pp 157–161

  • Fiddaman PJ, O’Neill TM, Rossall S (2000) Screening of bacteria for the suppression of Botrytis cinerea and Rhizoctonia solani on lettuce (Lactuca sativa) using leaf disc bioassays. Ann Appl Biol 137:223–235

    Article  Google Scholar 

  • Grasotti A, Gimelli F (2011) Bulb and cut flower production in the genus Lilium: current status and the future. Acta Hortic 900:21–42

    Google Scholar 

  • Hou PF, Chen CY (2003) Early stages of infection of lily leaves by Botrytis elliptica and Botrytis cinerea. Plant Pathol Bull 12:103–108

    Google Scholar 

  • Inglis GD, Kawchuk LM (2002) Comparative degradation of oomycete, ascomycete, and basidiomycete cell walls by microparasitic and biocontrol fungi. Can J Microbiol 48:60–70

    Article  CAS  PubMed  Google Scholar 

  • Katsumoto Y, Fukuchi-Mitzutani M, Fukui Y, Brugliera F, Holton TA, Karan M, Nakamura N, Yonekura-Sakakibara K, Togami J, Pigeaire A, Tao GQ, Nehra NS, Lu CY, Dyson BK, Tsuda S, Ashikari T, Kusumi T, Mason JG, Tanaka Y (2007) Engineering of the rose flavonoid biosynthetic pathway successfully generated blue-hued flowers accumulating delphinidin. Plant Cell Physiol 48:1589–1600

    Article  CAS  PubMed  Google Scholar 

  • Kishimoto K, Nishizawa Y, Tabei Y, Hibi T, Nakajima M, Akutsu K (2002) Detailed analysis of rice chitinase gene expression in transgenic cucumber plants showing different levels of disease resistance to gray mold (Botrytis cinerea). Plant Sci 162:655–662

    Article  CAS  Google Scholar 

  • Le Nard M, De Hertogh AA (1993) Tulipa. In: De Hertogh AA, Le Nard M (eds) The Physiology of Flower Bulbs. Elsevier, Amsterdam, pp 617–682

    Google Scholar 

  • Li X, Wang C, Sun H, Li T (2011) Establishment of the total RNA extraction system for lily bulbs with abundant polysaccharides. African J Biotechnol 10:17907–17915

    CAS  Google Scholar 

  • Marchant R, Power JB, Lucas JA, Davey MR (1998) Biolistic transformation of rose (Rosa hybrida L.). Ann Bot 81:109–114

    Article  Google Scholar 

  • Mohandas S, Kempaiah P, Choudhary ML, Murthy BNS, Gowda TKS (2009) Somatic embryogenesis and Agrobacterium-mediated genetic transformation of geranium with chitinase-glucanase encoding genes. Indian J Horticult 66:1–6

    Google Scholar 

  • Nishikawa T, Okazaki K, Uchino T, Arakawa K, Nagamine T (1999) A molecular phylogeny of Lilium in the internal transcribed spacer region of nuclear ribosomal DNA. J Mol Evol 49:238–249

    Article  CAS  PubMed  Google Scholar 

  • Núñez de Cáceres FF, Davey MR, Wilson ZA (2011) A rapid and efficient Agrobacterium-mediated transformation protocol for Lilium. In: Proceedings of the II international symposium on the genus Lilium. Acta Hort (ISHS) vol 900, pp 161–167

  • Palli SR, Retnakaran A (1999) Molecular and biochemical aspects of chitin synthesis inhibition. In: Jollès P, Muzzarelli RAA (eds) Chitin and chitinases. Birkhäuser, Switzerland, pp 85–98

    Chapter  Google Scholar 

  • Pierpoint WS, Hargreaves JA, Shewry PR (1996) Modifying resistance to plant-pathogenic fungi. Genetic Engineering of crop plants for resistance to pests and diseases. British Crop Protection Council, UK, pp 66–83

    Google Scholar 

  • Ram MSN, Mohandas S (2003) Transformation of African violet (Saintpaulia ionantha) with glucanase-chitinase genes using Agrobacterium tumefaciens. Elegant Sci Floricult 624:471–478

    CAS  Google Scholar 

  • Rohini VK, Rao KS (2001) Transformation of peanut (Arachis hypogaea L.) with tobacco chitinase gene: variable response of transformants to leaf spot disease. Plant Sci 160:889–898

    Article  CAS  PubMed  Google Scholar 

  • Rommens CM, Kishore GM (2000) Exploiting the full potential of disease-resistance genes for agricultural use. Curr Opin Biotech 11:120–125

    Article  CAS  PubMed  Google Scholar 

  • Shin S, Mackintosh CA, Lewis J, Heinen SJ, Radmer L, Dill-Macky R, Baldridge GD, Zeyen RJ, Muehlbauer GJ (2008) Transgenic wheat expressing a barley class II chitinase gene has enhanced resistance against Fusarium graminearum. J Exp Botany 59:2371–2378

    Article  CAS  Google Scholar 

  • Staats M, van Baarlen P, van Kan JAL (2005) Molecular phylogeny of the plant pathogenic genus Botrytis and the evolution of host specificity. Mol Biol Evol 22:333–346

    Article  CAS  PubMed  Google Scholar 

  • Takatsu Y, Nishizawa Y, Hibi T, Akutsu K (1999) Transgenic chrysanthemum [Dendranthema grandiflorum (Ramat.) Kitamura] expressing a rice chitinase gene shows enhanced resistance to gray mold (Botrytis cinerea). Sci Hortic 82:113–123

    Article  CAS  Google Scholar 

  • Tanaka Y, Katsumoto Y, Brugliera F, Mason J (2005) Genetic engineering in floriculture. Plant Cell Tiss Org Cult 80:1–24

    Article  CAS  Google Scholar 

  • Tanaka Y, Brugliera F, Chandler S (2009) Recent progress of flower colour modification by biotechnology. Int J Mol Sci 10:5350–5369

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Vellicce GR, Ricci JCD, Hernandez L, Castagnaro AP (2006) Enhanced resistance to Botrytis cinerea mediated by the transgenic expression of the chitinase gene ch5B in strawberry. Transgenic Res 15:57–68

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Kausch AP, Chandlee JM, Luo H, Ruemmele BA, Browning M, Jackson N, Goldsmith MR (2003) Co-transfer and expression of chitinase, glucanase and bar genes in creeping bent grass for conferring fungal disease resistance. Plant Sci 165:497–506

    Article  CAS  Google Scholar 

  • Wu G, Shortt BJ, Lawrence EB, Levine EB, Fitzsimmons KC, Shah DM (1995) Disease resistance conferred by expression of a gene encoding H2O2-generating glucose oxidase in transgenic potato plants. Plant Cell 7:1357–1368

    CAS  PubMed Central  PubMed  Google Scholar 

  • Zhu Q, Lamb CJ (1990) Isolation and characterization of a rice gene encoding a basic chitinase. Mol General Genetics 226:289–296

    Google Scholar 

  • Zhu Q, Maher EA, Masoud S, Dixon RA, Lamb CJ (1994) Enhanced protection against fungal attack by constitutive co-expression of chitinase and glucanase genes in transgenic tobacco. Nature Biotechnol 12:807–812

    Article  CAS  Google Scholar 

  • Zuker A, Tzfira T, Vainstein A (1998) Genetic engineering for cut-flower improvement. Biotechnol Adv 16:33–79

    Article  CAS  PubMed  Google Scholar 

  • Zuker A, Shklarman E, Scovel G, Ben-Meir H, Ovadis M, Neta-Sharir I (2001) Genetic engineering of agronomic and ornamental traits in carnation. Plant Sci 560:91–94

    CAS  Google Scholar 

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Acknowledgments

The authors would like to thank Dr. S. Rossall and Prof. M. Dickinson for their valuable suggestions; Dr. J. Fernandez Gomez and Louise Cheetham for technical assistance. This work was supported by a scholarship to FFNCG by the Consejo Nacional de Ciencia y Tecnología (CONACYT) of Mexico.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, LE12 5RD, UK

    Francisco F. Núñez de Cáceres González, Michael R. Davey, Ester Cancho Sanchez & Zoe A. Wilson

  2. Ciudad del Conocimiento, Centro de Investigaciones Biológicas, Universidad Autónoma del Estado de Hidalgo, Km. 4.5 Carr. Pachuca-Tulancingo, 42184, Pachuca, Hidalgo, Mexico

    Francisco F. Núñez de Cáceres González

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  1. Francisco F. Núñez de Cáceres González
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  2. Michael R. Davey
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  3. Ester Cancho Sanchez
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  4. Zoe A. Wilson
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Correspondence to Zoe A. Wilson.

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Communicated by W. Harwood.

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Núñez de Cáceres González, F.F., Davey, M.R., Cancho Sanchez, E. et al. Conferred resistance to Botrytis cinerea in Lilium by overexpression of the RCH10 chitinase gene. Plant Cell Rep 34, 1201–1209 (2015). https://doi.org/10.1007/s00299-015-1778-9

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  • DOI: https://doi.org/10.1007/s00299-015-1778-9

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