SpringerLink
Log in

Association between sheath blight resistance and chitinase activity in transgenic rice plants expressing McCHIT1 from bitter melon

  • Original Paper
  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

No usable resources with high-level resistance to sheath blight (SB) have yet been found in rice germplasm resources worldwide. Therefore, creating and breeding new disease-resistant rice resources with sheath blight resistance (SBR) are imperative. In this study, we inoculated rice plants with hyphae of the highly pathogenic strain RH-9 of rice SB fungus Rhizoctonia solani to obtain eight stable transgenic rice lines harbouring the chitinase gene (McCHIT1) of bitter melon with good SBR in the T5 generation. The mean disease index for SB of wild-type plants was 92% and 37–44% in transgenic lines. From 24 h before until 120 h after inoculation with R. solani, chitinase activity in stable transgenic plants with increased SBR was 2.0–5.5 and 1.8–2.7 times that of wild-type plants and plants of a disease-susceptible stable transgenic line, respectively. The correlation between SBR and chitinase activity in McCHIT1-transgenic rice line plants was significant. This work stresses how McCHIT1 from bitter melon can be used to protect rice plants from SB infection.

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

Access this article

Log in via an institution

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.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Abbreviations

ANOVA:

Analysis of variance

BSA:

Bovine serum albumin

GlcNAc:

N-acetylglucosamine

GUS:

β-Glucuronidase

NaAc:

Sodium acetate

PMSF:

Pheylmethylsulfonyl fluoride

PR:

Pathogenesis-related gene

SB:

Sheath blight

SBR:

Sheath blight resistance

SSR:

Shortest significant ranges

X-Gluc:

5-Bromo-4-chloro-3-indolyl glucuronide

References

  • Boller T, Mauch F (1988) Colorimetric assay for chitinase. Methods Enzymol 161:430–435

    Article  CAS  Google Scholar 

  • Boller T, Gehri A, Mauch F, Vögeli U (1983) Chitinase in bean leaves: induction by ethylene, purification, properties, and possible function. Planta 157:22–31

    Article  CAS  PubMed  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  Google Scholar 

  • Chen H, Lin YJ, Zhang QF (2009) Review and prospect of transgenic rice research. Chin Sci Bull 54:4049–4068

    Article  CAS  Google Scholar 

  • Chen B, Li LW, Lin YJ, Wang ZH, Lu GD (2014) Evaluation of seedling resistance of rice germplasm. Cereal Res Commun 42:495–502

    Article  Google Scholar 

  • Datta K, Tu J, Oliva N, Ona I, Velazhahan R, Mew TW, Muthukrishnan S, Datta SK (2001) Enhanced resistance to sheath blight by constitutive expression of infection-related rice chitinase in transgenic elite indica rice cultivars. Plant Sci 160:405–414

    Article  CAS  PubMed  Google Scholar 

  • Groth DE, Bond JA (2006) Initiation of rice sheath blight epidemics and effect of application timing of azoxystrobin on disease incidence, severity, yield, and milling quality. Plant Dis 90:1073–1076

    Article  CAS  PubMed  Google Scholar 

  • Hedrick SA, Bell JN, Boller TS, Lamb CJ (1988) Chitinase cDNA cloning and mRNA induction by fungal elicitor, wounding, and infection. Plant Physiol 86:182–186

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ignacimuthu S, Ceasar SA (2012) Development of transgenic finger millet (Eleusine coracana (L.) Gaertn.) resistant to leaf blast disease. Biosci J 37:135–147

    Article  CAS  Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405

    Article  CAS  Google Scholar 

  • Kalpana K, Maruthasalam S, Rajesh T, Poovannan K, Kumar KK, Kokiladevi E, Raja JAJ, Sudhakar D, Velazhahan R, Samiyappan R, Balasubramanian P (2006) Engineering sheath blight resistance in elite indica rice cultivars using genes encoding defense proteins. Plant Sci 170:203–215

    Article  CAS  Google Scholar 

  • Kim JK, Jang IC, Wu R, Zuo WN, Boston RS, Lee YH, Ahn IP, Nahm BH (2003) Co-expression of a modified maize ribosome-inactivating protein and a rice basic chitinase gene in transgenic rice plants confers enhanced resistance to sheath blight. Transgenic Res 12:475–484

    Article  CAS  PubMed  Google Scholar 

  • Kumar KK, Poovannan K, Nandakumar R, Thamilarasi K, Geetha C, Jayashree N, Kokiladevi E, Raja JAJ, Samiyappan R, Sudhakar D, Balasubramanian P (2003) A high throughput functional expression assay system for a defence gene conferring transgenic resistance on rice against the sheath blight pathogen, Rhizoctonia solani. Plant Sci 165:969–976

    Article  CAS  Google Scholar 

  • Lee FN, Rush MC (1983) Rice sheath blight: a major rice disease. Plant Dis 67:829–832

    Article  Google Scholar 

  • Li P, Pei Y, Sang XC, Ling YH, Yang ZL, He GH (2009) Transgenic indica rice expressing a bitter melon (Momordica charantia) class I chitinase gene (McCHIT1) confers enhanced resistance to Magnaporthe grisea and Rhizoctonia solani. Eur J Plant Pathol 125:533–543

    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. Nat Biotech 13:686–691

    Article  CAS  Google Scholar 

  • Liu M, Sun ZX, Zhu J, Xu T, Harman GE, Lorito M (2004) Enhancing rice resistance to fungal pathogens by transformation with cell wall degrading enzyme genes from Trichoderma atroviride. Zhejiang Univ Sci 5:133–136

    Article  CAS  Google Scholar 

  • Maximova SN, Marelli JP, Young A, Pishak S, Verica JA, Guiltinan MJ (2006) Over-expression of a cacao class I chitinase gene in Theobroma cacao L. enhances resistance against the pathogen, Colletotrichum gloeosporioides. Planta 224:740–749

    Article  CAS  PubMed  Google Scholar 

  • Mew TW, Cottyn B, Pamplona R, Barrios H, Li XM, Chen ZY, Lu F, Nilpanit N, Arunyanart P, Kim PV, Du PV (2004a) Applying rice seed-associated antagonistic bacteria to manage rice sheath blight in develo** countries. Plant Dis 88:557–564

    Article  PubMed  Google Scholar 

  • Mew TW, Leung H, Savary S, Cruz CMV, Leach JE (2004b) Looking ahead in rice disease research and management. Crit Rev Plant Sci 23:103–127

    Article  Google Scholar 

  • Molla KA, Karmakar S, Chanda PK, Ghosh S, Sarkar SN, Datta SK, Datta K (2013) Rice oxalate oxidase gene driven by green tissue-specific promoter increases tolerance to sheath blight pathogen (Rhizoctonia solani) in transgenic rice. Mol plant pathol 14:910–922

    Article  CAS  PubMed  Google Scholar 

  • Nandakumar R, Babu S, Kalpana K, Raguchander T, Balasubramanian P, Samiyappan R (2007) Agrobacterium-mediated transformation of indica rice with chitinase gene for enhanced sheath blight resistance. Biol Plant 51:142–148

    Article  CAS  Google Scholar 

  • Neuhaus JM, Sticker L, Meins F, Boller T (1991) A short C-terminal sequence is necessary and sufficient for the targeting of chitinases to the plant vacuole. Proc Natl Acad Sci USA 88:10362–10366

    Article  CAS  PubMed  Google Scholar 

  • Park DS, Sayler RJ, Hong YG, Nam MH, Yang Y (2008) A method for inoculation and evaluation of rice sheath blight disease. Plant Dis 92:25–29

    Article  PubMed  Google Scholar 

  • Pasonen HL, Seppänen SK, Degefu Y, Rytkönen A, von Weissenberg K, Pappinen A (2004) Field performance of chitinase transgenic silver birches (Betula pendula): resistance to fungal diseases. Theor Appl Genet 109:562–570

    Article  CAS  PubMed  Google Scholar 

  • Peng D, Li SD, Wang JX, Chen CJ, Zhou MG (2014) Integrated biological and chemical control of rice sheath blight by Bacillus subtilis NJ-18 and **ggangmycin. Pest Manag Sci 70:258–263

    Article  CAS  PubMed  Google Scholar 

  • Punja ZK (2001) Genetic engineering of plants to enhance resistance to fungal pathogens—a review of progress and future prospects. Can J Plant Pathol 23:216–235

    Article  CAS  Google Scholar 

  • Rassouli H, Tohidfar M, Haghnazari A, Ghareyazie B, Najafi J (2008) Evaluation of stability of chitinase gene in transgenic offspring of cotton (Gossypium hirsutum). Iran J Biotechnol 6:208–214

    Google Scholar 

  • Selitrennikoff CP (2001) Antifungal proteins. Appl Environ Microb 67:2883–2894

    Article  CAS  Google Scholar 

  • Shah JM, Raghupathy V, Veluthambi K (2009) Enhanced sheath blight resistance in transgenic rice expressing an endochitinase gene from Trichoderma virens. Biotechnol Lett 31:239–244

    Article  CAS  PubMed  Google Scholar 

  • Shah JM, Singh R, Veluthambi K (2013) Transgenic rice lines constitutively co-expressing tlp-D34 and chi11 display enhancement of sheath blight resistance. Biol Plant 57:351–358

    Article  CAS  Google Scholar 

  • Sharma KK, Bhatnagar-Mathur P, Thorpe TA (2005) Genetic transformation technology: status and problems. In Vitro Cell Dev Biol Plant 41:102–112

    Article  CAS  Google Scholar 

  • Shimahara K, Takiguchi Y (1988) Preparation of crustacean chitin. Methods Enzymol 161:417–423

    Article  CAS  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 Bot 59:2371–2378

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sripriya R, Raghupathy V, Veluthambi K (2008) Generation of selectable marker-free sheath blight resistant transgenic rice plants by efficient co-transformation of a cointegrate vector T-DNA and a binary vector T-DNA in one Agrobacterium tumefaciens strain. Plant Cell Rep 27:1635–1644

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Pinson SRM, Fjellstrom RG, Tabien RE (2012) Phenotypic gain from introgression of two QTL, qSB9-2 and qSB12-1, for rice sheath blight resistance. Mol Breed 30:293–303

    Article  CAS  Google Scholar 

  • **an HQ, Li JR, Zhang LQ, Li DC (2012) Cloning and functional analysis of a novel chitinase gene Trchi1 from Trichothecium roseum. Biotechnol Lett 34:1921–1928

    Article  PubMed  Google Scholar 

  • **ao YH, Li XB, Yang XY, Luo M, Hou L, Guo SH, Luo XY, Pei Y (2007) Cloning and characterization of a balsam pear class I chitinase gene (Mcchit1) and its ectopic expression enhances fungal resistance in transgenic plants. Biosci Biotechnol Biochem 71:1211–1219

    Article  CAS  PubMed  Google Scholar 

  • Zeng XF, Li L, Li JR, Zhao DG (2016) Constitutive expression of McCHIT1–PAT enhances resistance to rice blast and herbicide, but does not affect grain yield in transgenic glutinous rice. Biotechnol Appl Biochem 63:77–85

    Article  CAS  PubMed  Google Scholar 

  • Zhu YY, Zhao FL, Zhao DG (2011) Regeneration and transformation of a maize elite inbred line via immature embryo culture and enhanced tolerance to a fungal pathogen Exserohilum turcicum with a balsam pear class I chitinase gene. Afr J Agr Res 6:1923–1930

    Google Scholar 

  • Zou JH, Pan XB, Chen ZX, Xu JY, Lu JF, Zhai WX, Zhu LH (2000) Map** quantitative trait loci controlling sheath blight resistance in two rice cultivars (Oryza sativa L.). Theor Appl Genet 101:569–573

    Article  CAS  Google Scholar 

  • Zuo SM, Zhang L, Wang H, Yin YJ, Zhang YF, Chen ZX, Ma YY, Pan XB (2008) Prospect of the QTL-qSB-9Tq utilized in molecular breeding program of japonica rice against sheath blight. J Genet Genomics 35:499–505

    Article  CAS  PubMed  Google Scholar 

  • Zuo SM, Zhang YF, Chen ZX, Chen XJ, Pan XB (2010) Current progress in genetics and breeding in resistance to rice sheath blight. Scientia Sinica Vitae 40:1014–1023. https://doi.org/10.1360/052010-323 (in Chinese)

    Article  Google Scholar 

  • Zuo SM, Yin YJ, Pan CH, Chen ZX, Zhang YF, Gu SL, Zhu LH, Pan XB (2013) Fine map** of qSB-11 LE, the QTL that confers partial resistance to rice sheath blight. Theor Appl Genet 126:1257–1272

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

The study was funded by Major Projects of National New Transgenic Species Breeding (2016ZX08001-002), General Projects of Technological Innovation and Application Demonstration (Social Livelihood) in Chongqing City (cstc2018jscx-msybX0250), and PhD Funds (including the Talent Introduction Plan) of Southwest University (Project SWU112077).

Author information

Author notes

    Authors and Affiliations

    1. Rice Research Institute, Key Laboratory of Application and Safety Control of Genetically Modified Crops, Academy of Agricultural Sciences, Southwest University, Chongqing, 400715, China

      Changwei Zhang, Manyu Huang, **anchun Sang, ** Li, Yinghua Ling, Fangming Zhao, Dan Du, Yunfeng Li, Zhenglin Yang & Guanghua He

    Authors
    1. Changwei Zhang
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Manyu Huang
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. **anchun Sang
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. ** Li
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Yinghua Ling
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Fangming Zhao
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Dan Du
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Yunfeng Li
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. Zhenglin Yang
      View author publications

      You can also search for this author in PubMed Google Scholar

    10. Guanghua He
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Guanghua He.

    Additional information

    Publisher's Note

    Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

    Changwei Zhang and Manyu Huang have equally contributed

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Zhang, C., Huang, M., Sang, X. et al. Association between sheath blight resistance and chitinase activity in transgenic rice plants expressing McCHIT1 from bitter melon. Transgenic Res 28, 381–390 (2019). https://doi.org/10.1007/s11248-019-00158-x

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s11248-019-00158-x

    Keywords

    Search

    Navigation