Functional expression of MAP kinase TiHOG1 gene in Tilletia indica inciting Karnal bunt of wheat
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Tilletia indica is an important quarantined fungal pathogen inciting Karnal bunt disease of wheat. In fungal pathogens, the Hog1-type MAPK is involved in response to various stresses and virulence. In the present study,we identified, characterized and analyzed functional expression of the TiHog1-type gene. As HOG1 homologous genes are salt-inducible, the TiHOG1 gene (1104 bp) was amplified in 1 M NaCl stressed condition in T. indica; cloned in pJET2.0 cloning vector. The TiHOG1 gene was subcloned in expression vector (pRS426Gpd plasmid) and sequences were analyzed. The TiHOG1 in pRS426GPD (pRS426GPD-TiHOG1) was expressed functionally in heterologous system (Saccaromyces cerevisiae). TiHOG1 protein exhibits high amino acid similarity with the homologous protein sequences in Tilletia caries, Ustilago maydis and Puccinia graminis f. sp. tritici. Functional expression of TiHOG1 in S. cerevisiae ∆hog1 mutant showed that TiHOG1 was restored salinity stress tolerance up to 1 M NaCl, 1 M KCl, 0.25 CsCl2, 0.5 M MgCl2 and 0.5 M CaCl2 and osmostress tolerance up to 1 M sorbitol and 1 M sucrose. The expression of TiHOG1 was elicited up to 19.81-fold in response to a susceptible host, whereas resistant host factors showed 9.65-fold transcripts. This first study revealed involvement of a TiHOG1 gene in stress tolerance and may have a role in pathogenesis of T. indica (Karnal bunt of wheat).
KeywordsTilletia indica Karnal bunt Wheat HOG1-type mitogen-activated protein kinase Stress tolerance Functional expression
This work is financially supported by the Indian Council of Agricultural Research (ICAR) -Consortium Research Platform (CRP) on Genomics (ICAR-G/CRP-Genomics/2015-2720/IARI-12-151). We are highly thankful to the Director, Joint Director (Research), ICAR-IARI, New Delhi for providing guidance and facilities for carrying out this study.
- Gietz RD, Woods RA (1994) High efficiency transformation with lithium acetate. In Johnston JR (ed) Molecular genetics of yeast: a practical approach. Oxford University Press, Oxford, pp 121–134Google Scholar
- Gurjar MS, Jogawat A, Kulshresta D, Sharma S, Aggarwal R (2016) Intraspecific variation of Tilletia indica isolates causing Karnal bunt of wheat in India. Indian Phytopathol 69(4):352–356Google Scholar
- Gurjar MS, Aggarwal R, Sharma S, Kulshreshtha D, Gupta A, Gogoi R, Thirumalaisamy PP, Saini A (2017) Development of real time PCR assay for the detection and quantification of teliospores of Tilletia indica causing Wheat Karnal bunt in soil. Indian J Exp Biol 55(6):549–554Google Scholar
- Po LI, Gong X, Hui JIA, Fan Y, Zhang Y, Cao Z, Hao Z, Han J, Gu S, Dong J (2016) MAP kinase gene STK1 is required for hyphal, conidial, and appressorial development, toxin biosynthesis, pathogenicity, and hypertonic stress response in the plant pathogenic fungus Setosphaeria turcica. J Integr Agric 15(12):2786–2794CrossRefGoogle Scholar
- Rana M, Arora C, Ram B, Kumar A (2001) Floret specificity of Karnal bunt infection due to presence of fungal growth-promotory activity in wheat spikes. J Plant Biol 28(3):283–290Google Scholar
- Singh DV, Gogoi R (2011) Karnal bunt of wheat (Triticum sp.): a global scenario. Indian J Agric Sci 81(1):3–14Google Scholar