Development of Normalized Cdna Library from Fusarium Wilt Infected Roots of a Tolerant Banana Genotype 'Calcutta-4' Musa acuminata ssp. burmannicoides

Authors

  • V Swarupa Author
  • A Rekha Author
  • K V Ravishankar Author

DOI:

https://doi.org/10.24154/jhs.v9i1.223

Keywords:

Fusarium oxysporum f.sp. cubense, Banana, cDNA Library, Defense Response, Normalization

Abstract

Management of the most devasting disease, Fusarium wilt of banana, caused by the fungus Fusarium oxysporum f. sp., cubense, is a challenge to the plant pathologist and the banana grower. Currently, genomics is providing the way for understanding plant defense mechanism, having acquired an important place in crop improvement. To identify the relevant genes and to understand the defense mechanism induced during Fusarium wilt infection, a normalized cDNA library was constructed from infected root samples of a tolerant banana genotype, Musa acuminata spp. burmannicoides 'Calcutta-4', by duplex specific nuclease (DSN) based normalization, using the SMART (switching mechanism at 5' end of RNA transcript) full-length cDNA construction method. Sequencing and analysis of 600 clones revealed 392 non-redundant clones. In all, of 88% of the sequences were annotated using Musa genome database, and the remaining 12% were identified as novel loci not annotated. We observed several resistance genes, ROS scavenging genes and genes involved in ubiquitin-proteosome pathway in this study. These genes may have a possible role against Foc infection. These sequences would enrich the EST data developed against specific stress, which is an indispensable tool for predicting functional genes and understanding the defense mechanism.

References

Bai, T.T., Xie, W.B., Zhou, P.P., Wu, Z.L., Xiao, W.C., Zhou, L., Sun, J., Ruan, X.L., and Li, H.P. 2013. Transcriptome and expression profile analysis of highly resistant and susceptible banana roots challenged with Fusarium oxysporum f. sp. cubense Tropical Race 4. PLoS ONE:8:e73945. doi:10.1371/journal.pone.0073945

Bonaldo, M.F., Lennon, G. and Soares, M.B. 1996. Normalization and subtraction: Two approaches to facilitate gene discovery. Genome Res., 6:791–806

Craig, A., Ewan, E., Mesmar, J., Gudipati, G. and Sadanandom, A. 2009. E3 ubiquitin ligases and plant innate immunity. J. Exptl. Bot., 60:1123–1132

Dhawan, R., Luo. H., Foerster, A.M., Abuqamar, S., Du, H.N., Briggs, S.D., Mittelsten Scheid, O. and Mengiste, T. 2009. HISTONE MONOUBIQUITINATION 1 interacts with a subunit of the mediator complex and regulates defense against necrotrophic fungal pathogens in Arabidopsis. Pl. Cell, 21:1000–1019

Droc, G., Lariviere, D., Guignon, V., Yahiaoui, N., This, D., Garsmeur, O., Dereeper, A., Hamelin, C., Argout, X., Dufayard, J.F. et al. 2013. The banana genome hub. Database (Oxford) 2013:doi:10.1093/database/bat035

Garg, R., Patel, R.K., Tyagi, A.K. and Jain, M. 2011. De novo assembly of chickpea transcriptome using short reads for gene discovery and marker identification. DNA Res. 18:53–63

Li, C.Y., Deng, G.M., Yang, J., Viljoen, A., Jin, Y., Kuang, R.B., Zuo, C.W., Lv, Z.C., Yang, Q.S., Sheng, O., Wei, Y.R., Hu, C.H., Dong, T. and Yi, G.J. 2012. Transcriptome profiling of resistant and susceptible Cavendish banana roots following inoculation with Fusarium oxysporum f. sp. cubense Tropical Race 4. BMC Genomics, 13:374

Li, C., Shao, J., Wang, Y., Li, W., Guo, D., Yan, B., Xia, Y. and Peng, M. 2013a. 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

Li, X., Bai, T., Li, Y., Ruan, X. and Li, H. 2013b. Proteomic analysis of Fusarium oxysporum f. sp. cubense Tropical Race 4-inoculated response to Fusarium wilts in the banana root cells. Proteome Sci., 11:41

Liu, J.J., Goh, C.J., Loh, C.S., Liu, P. and Pua, E.C. 1998. A method for isolation of total RNA from fruit tissues of banana. Pl. Mol. Biol. Rep., 16:1-6

Marino, D., Peeters, N. and Rivas, S. 2012. Ubiquitination during plant immune signaling. Pl. Physiol., 160:15-27

Mittler, R., Vanderauwera, S., Gollery, M. and Van Breusegem, F. 2004. The reactive oxygen gene network in plants. Trends Pl. Sci., 9:490–498

Passos, M.A.N., de Cruz1, V.O., Emediato, F.L., de Teixeira, C., Azevedo, V.C.R., Brasileiro, A.C.M., Amorim, E.P., Ferreira, C.F., Martins, N.F., Togawa, R.C., Junior, G.J.P., da Silva Jr O.B. and Miller, R.N.G. 2013. Analysis of the leaf transcriptome of Musa acuminata during interaction with Mycosphaerella musicola: gene assembly, annotation and marker development. BMC Genomics, 14:78

Ploetz, R.C. 2000. Panama disease: A classic and destructive disease of banana. Online. Plant Health Progress doi:10.1094/PHP-2000-1204-01-HM.

Ravishankar, K.V., Rekha, A., Swarupa, V. and Savitha, G. 2011. Gene expression analysis in roots of Musa acuminata ssp. burmannicoides ‘Calcutta-4’, a banana genotype tolerant to Fusarium wilt. Acta Hort., 897:363–370

Schmelzer, E. 2002. Cell polarization, a crucial process in fungal defencse. Trends Plant Sci., 7:411–415

Swarupa, V., Ravishankar K.V. and Rekha, A. 2013. Characterization of tolerance to Fusarium oxysporum f. sp. cubense infection in banana using suppression subtractive hybridization and gene expression analysis. Physiol. Mol. Pl. Pathol., 83:1–7

Swarupa, V., Ravishankar K.V. and Rekha, A. 2014. Plant defense response against Fusarium oxysporum and strategies to develop tolerant genotypes in banana. Planta, DOI 10.1007/s00425-013-2024-8

Thakker, J.N., Shah, K. and Kothari, I.L. 2009. Elicitation, partial purification and antifungal activity of â- 1, 3 glucanse from banana plants. PRAJNA-Journal of Pure and Applied Science, 17:10–16

Torres, M.A., Jones, J.D.G. and Dangl, J.L. 2006. Reactive oxygen species signaling in response to pathogens. Pl. Physiol., 141:373–378

Van den Berg, N., Berger, D.K., Hein, I., Birch, P.R.J., Wingfield, M.J. and Viljoen, A. 2007. Tolerance in banana to Fusarium wilt is associated with early up- regulation of cell wall-strengthening genes in the roots. Mol. Pl. Pathol., 8:333–341

Young, B.D. and Anderson, M. 1985. Quantitative analysis of solution hybridization. In: Hames, B.D. and Higgins,

S.J. (eds) Nucleic Acid Hybridization: A practical approach. IRL Press, Oxford, pp. 47-71

Zhulidov, P.A., Bogdanova, E.A., Shcheglov, A.S., Vagner, L.L., Khaspekov, G.L., Kozhemyako, V.B., Mikhail V. Matz, M.V., Meleshkevitch, E., Moroz, L.L., Lukyanov, S.A. and Shagin, D.A. 2004. Simple cDNA normalization using kamchatka crab duplex-specific nuclease. Nucleic Acids Res., 32:1-8

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Published

30-06-2014

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Section

Original Research Papers

How to Cite

Swarupa, V., Rekha, A., & Ravishankar, K. V. (2014). Development of Normalized Cdna Library from Fusarium Wilt Infected Roots of a Tolerant Banana Genotype ’Calcutta-4’ Musa acuminata ssp. burmannicoides. Journal of Horticultural Sciences, 9(1), 55-60. https://doi.org/10.24154/jhs.v9i1.223

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