Over expression of anti-apoptotic gene in banana cv Rasthali enhances resistance against Fusarium oxysporum f. sp. cubense Race 1

M Umesha; C Sunisha; T R Usharani; H D Sowmya; S Sriram

doi:10.24154/jhs.v18i1.2169

Authors

M Umesha entre for Post-Graduate Studies Jain University, Bengaluru - 560069, Karnataka, India Author
C Sunisha ICAR-Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bengaluru - 560089, Karnataka, India Author
T R Usharani ICAR-Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bengaluru - 560089, Karnataka, India Author
H D Sowmya ICAR-Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bengaluru - 560089, Karnataka, India Author
S Sriram ICAR-Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bengaluru - 560089, Karnataka, India Author

DOI:

https://doi.org/10.24154/jhs.v18i1.2169

Keywords:

Anti-apoptotic gene, banana, constitutive expression, fusarium wilt, rasthali

Abstract

The most popular banana cv Rasthali was transformed with anti-apoptotic gene, AtBAG4 regulated with two different promoters viz., ZmBgl and ubiquitin to enhance the tolerance levels to Fusarium oxysporum f. sp. cubense Race 1 (FOC1). The differences in gene expression driven by two promoters revealed that stronger expression of AtBAG4 gene under the ubiquitin promoter suppressed the infection and spreading processes of FOC1 in transgenic banana under standard bioassay systems. Analysis using the real time PCR showed the varying levels of AtBAG4 gene expression under two promoters. It was evident that ZmBgl driven AtBAG4 lead to lower gene expression in leaves which correlated with lesser levels of resistance to FOC1. Constitutive expression of AtBAG4 under the control of ubiquitin promoter showed increased transgene transcripts which directly correlated with the enhanced tolerance against FOC1 from seedlings stage to active vegetative phases. This study reveals the importance of constitutive expression of anti-apoptotic gene showing enhanced tolerance against the most dreaded FOC1 in highly susceptible variety Rasthali.

References

Dale, J. L, James, A, Paul, J. Y, Khanna, H, Smith, M, Echeverria, S. P, Bastidas, F. G, Kema, G, Waterhouse, P, Mengersen, K. and Harding, R. 2017. Transgenic Cavendish bananas with resistance to Fusarium wilt tropical race 4. Nature Comm. 8:1496. doi: 10.1038/s41467- 017-01670-6. DOI: https://doi.org/10.1038/s41467-017-01670-6

Dickman, M. B. and de Figueiredo, P. 2013. Death Be Not Proud—Cell Death Control in Plant Fungal Interactions. PLoS Pathog. 9: e1003542. https://doi.org/10.1371/journal. ppat.1003542 DOI: https://doi.org/10.1371/journal.ppat.1003542

Doukhanina, E. V., Chen, S., van der Zalm, E., Godzik, A., Reed, J. and Dickman, M. B. 2006. Identification and functional characterization of the BAG protein family in Arabidopsis thaliana. J. Bio. Chem., 281: 18793-18801. DOI: https://doi.org/10.1074/jbc.M511794200

Ge, S., Kang, Z., Li, Y., Zhang, F., Shen, Y., Ge, R. and Huang, Z. 2016. Cloning and function analysis of BAG family genes in wheat. Func. Pl. Biol., 43: 393-402. DOI: https://doi.org/10.1071/FP15317

Ghag, S. B, Shekhawat, U. K. and Ganapathi, T. R. 2014. Native cell-death genes as candidates for developing wilt resistance in transgenic banana plants. AoB Plants. 1:6 https:// doi:10.1093/ aobpla/plu037. DOI: https://doi.org/10.1093/aobpla/plu037

Gu, R., Zhao, L., Zhang, Y., Chen, X., Bao, J., Zhao, J. and Wang, G. 2006. Isolation of a maize beta- glucosidase gene promoter and characterization of its activity in transgenic tobacco. Plant Cell Rep., 25: 1157-1165. DOI: https://doi.org/10.1007/s00299-006-0177-7

Jacobs, A. T. and Marnett, L. J. 2009. HSF1-mediated BAG3 expression attenuates apoptosis in 4- hydroxynonenal-treated colon cancer cells via stabilization of anti-apoptotic Bcl-2 proteins. J. Biol. Chem., 284: 9176-9183. DOI: https://doi.org/10.1074/jbc.M808656200

Jiang, P., Zhang, K., Ding, Z., He, Q., Li, W., Zhu, S., Cheng, W., Zhang, K. and Li, K. 2018. Characterization of a strong and constitutive promoter from the Arabidopsis serine carboxypeptidase-like gene AtSCPL30 as a potential tool for crop transgenic breeding. BMC Biotech., 18:1-13 DOI: https://doi.org/10.1186/s12896-018-0470-x

Khanna, H.K., Becker, D., Kleidon, J. and Dale, J. 2004. Centrifugation assisted Agrobacterium- mediated transformation (CAAT) of embryogenic cell suspension of banana (Musa spp. Cavendish AAA and Lady finger AAB). DOI: https://doi.org/10.1023/B:MOLB.0000047771.34186.e8

Mol. Breed., 14: 239–252.

Li, C. Y, Chen, S, Zuo, C. W, Sun, Q. M, Ye, Q, Yi, G. J. and Huang, B. Z. 2011. The use of GFP– transformed isolates to study infection of banana with Fusarium oxysporum f. sp. cubense race 4. Euro. J. Pl. Pathol., 131: 327-340. DOI: https://doi.org/10.1007/s10658-011-9811-5

Linclon, J. E, Richael, C, Overduin, B, Smith, K, Bostock, R. and Gilchrist, D. G. 2002. Expression of the antiapoptotic baculovirus p. 53 gene in tomato blocks programmed cell death and provides broad-spectrum resistance to disease. PNAS, USA., 99: 15217-15221. DOI: https://doi.org/10.1073/pnas.232579799

Li, W. and Dickman, M. B. 2004. Abiotic stress induces apoptotic-like features in tobacco that is inhibited by expression of human Bcl-2. Biotechnol Lett., 26: 8. DOI: https://doi.org/10.1023/B:BILE.0000012896.76432.ba

Magambo, B. 2012. Generating transgenic banana (cv. Sukali Ndizi) resistant to Fusarium wilt (Doctoral dissertation, Queensland University of Technology).

Moore, N. Y., Pegg, K., Allen, A. R. and Irvin, J. A. G. 1993. Vegetative compatibility and distribution of Fusarium oxysporum f. sp. cubense in Australia. Aus. J. Expt. Agric., 33: 797-802. DOI: https://doi.org/10.1071/EA9930797

Onyango, S. O., Roderick, H., Tripathi, J. N., Collins, R., Atkinson, H. J., Oduor, R. O. and Tripathi, L. 2016. The ZmRCP-1 promoter of maize provides root tip specific expression of transgenes in plantain. J. Biol. Res.- Thessaloniki., 23: 1-9. DOI: https://doi.org/10.1186/s40709-016-0041-z

Paul, J.Y., Becker, D.K., Dickman, M.B., Harding, R.M., Khanna, H.K. and Dale, J.L. 2011. Apoptosis-related genes confer resistance to Fusarium wilt in transgenic ‘Lady Finger’ bananas. Plant Biotech. J., 9: 1141-1149. DOI: https://doi.org/10.1111/j.1467-7652.2011.00639.x

Ploetz, R. C. and Pegg, K. G. 2000. Fusarium wilt. In: Diseases of Banana, Abacá and Enset. D. R. Jones, (ed.) pp.143-159.CABI Publishing, Wallingford, UK.

Ploetz, R. C. 2015. Management of Fusarium wilt of banana: a review with special reference to tropical race 4. Crop Prot., 73: 7–15. DOI: https://doi.org/10.1016/j.cropro.2015.01.007

Schmitz, R. J., Grotewold, E. and Stam, M. 2022. Cis- regulatory sequences in plants: their importance, discovery, and future challenges. Plant Cell. 34: 718-741. DOI: https://doi.org/10.1093/plcell/koab281

Singhal, V. 1999. Bananas. In: Indian agriculture 1999. Indian Economic Data Research Centre, New Delhi. pp. 150–152.

Sondermann, H., Scheufler, C., Schneider, C., Hohfeld, J., Hartl, F. U., and Moarefi, I. 2001. Structure of a Bag/Hsc70 complex: convergent functional evolution of Hsp70 nucleotide exchange factors. Science. 291: 1553-1557. DOI: https://doi.org/10.1126/science.1057268

Sunisha, C., Sowmya, H.D., Usharani, T.R., Umesha, M., Gopalkrishna, H.R. and Saxena, A., 2020a. Deployment of stacked antimicrobial genes in banana for stable tolerance against Fusarium oxysporum f. sp. cubense through genetic transformation. Mol. Biotech., 62: 8-17. DOI: https://doi.org/10.1007/s12033-019-00219-w

Sunisha, C., Sowmya, H. D., Usharani, T. R., Umesha, M., Gopalkrishna, H. R., and Sriram, S. 2020b. Induction of Ced9 mediated anti- apoptosis in commercial banana cultivar Rasthali for stable resistance against Fusarium wilt. 3 Biotech., 10: 1-8. DOI: https://doi.org/10.1007/s13205-020-02357-4

Thaler, J. S, Owen, B. and Higgins, V. J. 2004. The role of the jasmonate response in plant susceptibility to diverse pathogens with a range of lifestyles. Plant Physio., 135: 530-538. DOI: https://doi.org/10.1104/pp.104.041566

Thangavelu, R., Sundararaju, P., Sathiamoorthy, S., Raghuchander, T., Velazhahan, R., Nakkeeran, S. and Palaniswami, A. 2001. Status of Fusarium wilt of banana in India. p.58-63. In: A.B. Molina, N.H. Nikmasdek, K.W. Liew (eds.), Banana Fusarium Wilt Management: Towards Sustainable Cultivation. INIBAP- ASPNET, Los Banos, Philippines.