Skip to main content
Log in

RETRACTED ARTICLE: Expression of a rice chitinase gene enhances antifungal response in transgenic litchi (cv. Bedana)

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

This article was retracted on 02 December 2016

Abstract

To enhance the antifungal response of litchi (Litchi chinensis Sonn.), transgenic plants were generated by transferring rice chitinase gene driven by a maize-ubiquitin promoter along with its first intron into the zygotic embryos via Agrobacterium tumefaciens-mediated transformation. After co-cultivation for 2 days, zygotic embryos were transferred onto Murashige and Skoog (MS) modified media supplemented with 25 mgl−1 hygromycin and 400 mgl−1 cefotaxime. Consequently embryos were selected and the antibiotic resistant transgenic plantlets were regenerated. The culture time from zygotic embryo to transgenic plants was 14 months. The integration of the transgene was confirmed by PCR, RT-PCR, Southern and western blot analyses. The transgenic plants exhibited higher chitinase activity than the non-transformed plants. The chitinase activity when examined using the native polyacrylamide in-gel assay, indicated that the foreign gene expression resulting in the protein of expected molecular weight that showed chitinase activity. The transgenic plants showed delayed onset of the disease and smaller lesions following in vitro inoculation of die-back, leaf spots and blight pathogen (Phomopsis sp.). The transgenic plants were adapted to the greenhouse and did not show any phenotypic alterations.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

B5:

Gamborg et al. (1968)

CPW salts:

Frearson et al. (1973)

2,-4-d :

2,-4-dichlorophenoxyacetic acid

GA3 :

Gibberellic acid

HPT:

Hygromycin phospotransferase gene

IBA:

Indole3-butyric acid

MS:

Murashige and Skoog (1962)

NAA:

α-naphthalene acetic acid

8P:

Kao and Michayluk (1975)

STS:

Silver thiosulphate

SDS-PAGE:

Sodium dodecylsulphate polyacrylamide gel electrophoresis

WPM:

Woody plant medium

References

  • Asao H, Nishizawa Y, Arai S, Sato T, Hirai M, Yoshida K, Shinmyo A, Hibi T (1997) Enhanced resistance against a fungal pathogen Sphaerotheca humuli in transgenic strawberry expressing a rice chitinase gene. Plant Biotechnol 14:145–149

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Broglie K, Chet I, Holliday M, Cressman R, Biddle P, Knowlton S, Mauvais CJ, Broglie R (1991) Transgenic plants with enhanced resistance to the fungal pathogen Rhizoctonia solani. Science 254:1194–1197

    Article  CAS  PubMed  Google Scholar 

  • Chen WB, Xu X, Liang GH, Muthukrishnan S, Chen PD, Liu DJ, Gill BS (1998) Introduction and constitutive expression of a rice chitinase gene in bread wheat using biolistic bombardment and bar gene as a selectable marker. Theor Appl Genet 97:1296–1306

    Article  CAS  Google Scholar 

  • Chen A, Yang J, Niu Y, Yang D, Liu CP, Yu CY, Li CH (2010) High frequency somatic embryogenesis from germinated zygotic embryos of Schisandra chinensis and evaluation of the effects of medium, strength sucrose, GA3 and BA in on somatic embryo development. Plant Cell Tiss Org Cult 100:49–58

    Article  Google Scholar 

  • Chhikara S, Choudhary D, Dhankhar OP, Jaiwal PK (2011) Combined expression of a barley Class II Chitinase and type I ribosome inactivating protein in transgenic Brassica juncea provides protection against Alternaria brassica. Plant Cell Tiss Org Cult 108(1):83–89

  • Das DK, Rahman A (2010) Expression of a bacterial chitinase (ChiB) gene enhances antifungal potential in transgenic Litchi chinensis Sonn. (cv. Bedana). Curr Trends Biotenol Pharm 41:820–833

    Google Scholar 

  • Das DK, Shiva Prakash N, Sarin NB (1996) Regeneration and Transformation of Litchi (Litchi chinensis Sonn.). In Vitro Cell Dev Biol Plant 32:63A

  • Das DK, Reddy MK, Upadhyaya KC, Sopory SK (2002) An efficient leaf disc culture method for the regeneration via somatic embryogenesis and transformation of grape (Vitis vinifera L.). Plant Cell Rep 20:999–1005

    Article  CAS  Google Scholar 

  • Emani C, Garcia JM, Lopata-Finch E, Pozo MJ, Uribe P, Kim DJ, Sunikumar G, Cook DR, Kenerley CM, Rathore KS (2003) Enhanced fungal resistance in transgenic cotton expressing an endo chitinase gene from Trichoderma virens. Plant Biotech J 1:321–326

    Article  CAS  Google Scholar 

  • Frearson EM, Power JB, Cocking EC (1973) The isolation, culture and regeneration of Petunia leaf protoplasts. Dev Biol 33:130–137

    Article  CAS  PubMed  Google Scholar 

  • Fu L, Tang D (1983) Introduction pollen plants of litchi tree (Litchi chinensis Sonn.). Acta Genetica Sinica 10:369–374

    Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Plant Cell Cultures 1. Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  CAS  PubMed  Google Scholar 

  • Girhepuje PV, Shinde GB (2011) Transgenic tomato plants expressing a wheat endochitin gene demonstrate enhanced resistance to Fusarium oxysporum f. Sp. lycopersici. Plant Cell Tiss Org Cult 105:242–253

    Article  Google Scholar 

  • Hahlbrock K, Scheel D (1989) Physiology and molecular biology of phenylpropanoid metabolism. Ann Rev Plant Physiol Plant Mol Biol 40:347–369

    Article  CAS  Google Scholar 

  • He X, Miyasaka SC, Fitch MM, Moore PH, Zhu YJ (2008) Agrobacterium mediated-transformation of taro (Colocasia esculenta (L).Schott) with a rice chitinase gene for improved tolerance to a fungal pathogen Sclerotium rolfsii. Plant Cell Rep 27:903–909

    Article  CAS  PubMed  Google Scholar 

  • Hoagland DR, Arnon DI (1950) The water culture method of growing plants without soil, Calif Agric Exp Stn Bull No. 347, Berkely, Calif

  • Jayaraj J, Punja ZK (2007) Combined expression of chitinase and lipid transfer protein genes in transgenic carrot plants enhances resistance to foliar fungal pathogens. Plant Cell Rep 26:1539–1546

    Article  CAS  PubMed  Google Scholar 

  • Jayraj J, Anand A, Muthukrishnan S (2004) Pathogenesis-related proteins and their roles in resistance to fungal pathogen. In: Punja ZK (ed) Fungal disease resistance in plants-biochemistry, molecular biology and genetic engineering. Food Products Press (Haworth Press), New York, pp 139–178

    Google Scholar 

  • Jonathan HC, Robert ES, Robert TM (1997) Control of lychee anthracnose by foliar application of Tebuconazole, Mancozeb and copper hydroxide a ‘Mauritius’ Lychee fruit under South Florida conditions. Proc Fla State Hort Soc 110:149–152

    Google Scholar 

  • Kao KM, Michayluk MR (1975) Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126:105–110

    Article  CAS  PubMed  Google Scholar 

  • Konieczny R, Pilarska M, Tuleja M, Salai T (2009) Somatic embryogenesis and plant regeneration in zygotic embryos of Trifolium nigrescens (Viv). Plant Cell Tiss Org Cult 11:36–47

    Google Scholar 

  • Kumar SM, Kumar BK, Sharma KK, Devi P (2004) Genetic transformation of Pigeon pea with rice chitinase gene. Plant Breeding 123:485–489

    Article  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriphage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  • Lagrimi LM (1992) Wound-induced deposition of polyphenols in transgenic plants over expressing peroxidase. Plant Physiol 96:577–583

    Article  Google Scholar 

  • Legrand M, Kauffmann S, Geoffroy P, Fritig B (1987) Biological function of pathogenesis-related proteins: four tobacco pathogenesis-related proteins are chitinases. Proc Natl Acad Sci USA 84:6750–6754

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li M, Li H, Hu X, Pan X, Niu G (2010) An Agrobacterium tumefaciens mediated transformation system using callus of Zoysia tenuifolia Willd ex Trin. Plant Cell Tiss Org Cult 102:321–327

    Article  CAS  Google Scholar 

  • Lin W, Anuratha CS, Datta K, Potrykus I, Muthukrishnan S, Datta SK (1995) Genetic engineering of rice for resistance to sheath blight. Bio Technol 13:685–691

    Article  Google Scholar 

  • Lloyd G, McCown B (1981) Commercialy feasible micropropagation of mountain laurel, Kalmia latifolia, by the use of shoot tip culture. Comb Int Plant Prop Soc 30:421–427

    Google Scholar 

  • Lodhi Ma, Ye GN, Weeden NF, Reisch BI (1994) A simple and efficient method for DNA extraction from grapevine cultivars and Vitis species. Plant Mol Biol Rep 12:6–13

    Article  CAS  Google Scholar 

  • Manju Y, Darsana C, Manish S (2010) Agrobacterium tumefaciense-mediated transformation of Sesame (Sesamum indicum L.). Plant Cell Tiss Org Cult 103:377–386

    Article  Google Scholar 

  • Menzel CM, Waife GK (2005) Litchi and longan:botany, production and uses. CABI, Wallingford, UK, p 305. ISBN 0-85199-696-5

  • Miller GL (1959) Use of dinitrosalicylic acid reagent for the determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  • Morton JF (1987) Litchi. In: Morton JF (ed) Fruits of warm climates. Lewis S. Maxwell, Miami, pp 249–259

  • Murashige T, Skoog F (1962) A revised medium rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nishizawa Y, Hibi T (1991) Rice chitinase gene: cDNA cloning and stress-induced expression. Plant Sci 76:211–218

    Article  CAS  Google Scholar 

  • Nishizawa Y, Nishio Z, Nakazono K, Soma M, Nakajima E, Ugaki M, Hibi T (1999) Enhanced resistance to blast (Magnaporthe grisea) in transgenic Japonica rice by constitutive expression of rice chitinase. Theor Appl Genet 99:383–390

    Article  CAS  PubMed  Google Scholar 

  • Peña-Ramírez YJ, García-Sheseña I, Hernández-Espinoza Á, Domínguez-Hernández A, Barredo-Pool FA, González-Rodríguez JA, Robert ML (2011) Induction of somatic embryogenesis and plant regeneration in the tropical timber tree Spanish red cedar (Cedrata oderata L. (Mediaceae). Plant Cell Tiss Org Cult 105:203–209

    Google Scholar 

  • Puchooa D (2004) Expression of green fluorescent protein gene in litchi (Litchi chinensis Sonn.) tissues. J Appl Hort 6:11–15

    CAS  Google Scholar 

  • Punja ZK (2006) Recent developments towards achieving fungal disease resistance in transgenic plants. Can J Plant Pathol 28:S298–S308

    Article  CAS  Google Scholar 

  • Rangan P, Akshatha V, Parvatan G, Ravishankar GA (2010) Somatic embryogenesis and Agrobacterium-mediated transformation in Bixa orellana L. Plant Cell Tiss Org Cult 16:1–12

    Google Scholar 

  • Rolle RS (eds) (2004) Reports of the APO seminar on “Reduction of postharvest losses of fruits and vegetables”, held in India, New Delhi from 5th–11th October

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Article  CAS  PubMed  Google Scholar 

  • Sridevi G, Sabapathi N, Meena P, Nandakumar R, Samiyappan R, Muthukrishnan S, Veluthambi K (2003) Transgenic indica rice variety Pusa Basmati 1 constitutively expressing a rice chitinase gene exhibits enhanced resistance to Rhizoctonia solani. J Plant Biochem Biotechnol 12:93–101

    Article  CAS  Google Scholar 

  • Stehpan JW, Wolf AG (1990) Dye-labelled substrates for the assay and detection of chitinase and lysozyme activity. J Microbial Methods 12:197–205

    Article  Google Scholar 

  • Tanveer K, Vanga R, Leelavathi S (2010) High-frequency regeneration via somatic embryogenesis of an elite recalcitrant cotton genotype (Gossypium hirsutum L.) and efficient Agrobacterium mediated transformation. Plant Cell Tiss Org Cult 101:323–330

    Article  Google Scholar 

  • Thiago S, Lucana C, Fatima A, Julio C, Marcio (2009) Studies on genetic transformation of Theobroma cacao L: evaluation of different polyamines and antibiotics on somatic embryogenesis and the efficiency of UidA gene transfer by Agrobacterium tumefaciens. Plant Cell Tiss Org Cult 99:281–298

    Google Scholar 

  • Trudel J, Asselin A (1989) Detection of chitinase activity after polyacrylamide gel electrophoresis. Anal Biochem 178:362–366

    Article  CAS  PubMed  Google Scholar 

  • Trudel J, Asselin A (1990) Detection of chitin deacetylase activity after polyacrylamide gel electrophoresis. Anal Biochem 189:249–253

    Article  CAS  PubMed  Google Scholar 

  • Yamamoto T, Iketani H, Ieki H, Nishizawa Y, Notsuka K, Hibi T, Hayashi T, Matsuta N (2000) Transgenic grapevine plants expressing a rice chitinase with enhanced resistance to fungal pathogens. Plant Cell Rep 19:639–646

    Article  CAS  Google Scholar 

  • Yu CH, Chen ZG, Lu LX, Lin JW (2000) Somatic embryogenesis and plant regeneration from litchi protoplasts isolated from embryogenic suspensions. Plant Cell Tiss Org Cult 61:51–58

    Article  CAS  Google Scholar 

  • Zhou LN, Kuang ZS, Ma XJ, Chen JQ, Tang XJ, Ling X (1993) Preliminary studies on culture of immature embryos and embryogenesis of somatic cells of litchi (Litchi chinensis Sonn.). Guangdong Agric Sci 5:14–15

    Google Scholar 

Download references

Acknowledgments

Authors are grateful to the University Grants Commission, Govt. of India, New Delhi for providing financial assistance. We are highly thankful to Dr. M.V.Razam, Department of Genetics, South Campus, University of Delhi, New Delhi, India, for providing rice chitinase 11 cDNA and antibody, Mr. S.C.B. Sharma, Advanced Instrumentation Research Facility, JNU, New Delhi for providing Electron micrography and Dr. Shiva Prakash, Senior Scientist, Monsanto company, Bangalore, India for manuscript updating.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to D. K. Das.

Additional information

An erratum to this article is available at http://dx.doi.org/10.1007/s11240-016-1143-x.

This article has been retracted. The retraction was undertaken at the request of Dr. Das, the corresponding author of this paper, as it was found that this paper included duplicated information already published in Current Trends in Biotechnology and Pharmacy 4 (2010), 820-833, by the same corresponding author, and data that did not correspond to the species studied in this publication.

About this article

Cite this article

Das, D.K., Rahman, A. RETRACTED ARTICLE: Expression of a rice chitinase gene enhances antifungal response in transgenic litchi (cv. Bedana). Plant Cell Tiss Organ Cult 109, 315–325 (2012). https://doi.org/10.1007/s11240-011-0097-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-011-0097-2

Keywords

Navigation