Aradhika TRIPATHI, Rashmi AGGARWAL, Anita YADAV

Differential expression analysis of defense-related genes responsive to Tilletia indica infection in wheat

Karnal bunt caused by Tilletia indica is a quarantine disease responsible for qualitative and quantitative losses in wheat. The present study aimed to determine the expression of defense-related genes involved in early stages of infection in response to T. indica by using the differential display reverse transcriptase-PCR method (DDRT-PCR) in susceptible (WL 711) and resistant (HD 29) cultivars of wheat. The DDRT-PCR banding profiles generated by the random primers in combination with oligo-dT primers of resistant and susceptible wheat plants were evaluated at 0 h, 12 h, 24 h, and 48 h after inoculation. Out of 9 sets of primers, 7 sets of primers produced remarkable induced transcripts. The highest up-regulated transcripts were observed in the resistant cultivar (HD 29), whereas the down-regulated transcripts were found in the susceptible (WL 711) cultivar. Several sets of primers were designed from the defense-related gene sequences available in the GenBank database, and of these, 4 primer sets amplified the desired specific bands. Based on the putative functions, 3 defense-related genes, namely puroindoline protein PINB (AT- KB2), \Beta-1,4-glucanase (AT- KB3), and chitinase (AT- KB7), and 1 housekeeping gene, actin (AT- KB1), were characterized.

Anahtar Kelimeler:

Key words: Defense-related genes, expression, DDRT-PCR, Tilletia indica, wheat

Differential expression analysis of defense-related genes responsive to Tilletia indica infection in wheat

Keywords:

Key words: Defense-related genes, expression, DDRT-PCR, Tilletia indica, wheat,

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Aggarwal R, Kharbikar LL, Renu (2011). Identification of Bipolaris sorokiniana- responsive differential transcripts in wheat (Triticum aestivum L.). Indian Phytopath 64: 24–27.
Aggarwal R, Tripathi A, Yadav A (2010). Pathogenic and genetic variability in Tilletia indica monosporidial culture lines using universal rice primer-PCR. Eur J Plant Pathol 128: 333–342.
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1997). Basic local alignment search tool. J Mol Biol 215: 403–410.
Benito EP, Prins T, Van Kan JAL (1996). Application of differential display RT-PCR to the analysis of gene expression in a plantfungus interaction. Plant Mol Biol 32: 947–957.
Bonde MR, Berner DK, Peterson GL, Olsen M, Cunfer B, Sim T (2004). Survival of Tilletia indica teliospores in different soils. Plant Dis 88: 316–324.
Bonde MR, Peterson GL, Schaad NW, Smilanick JL (1997). Karnal bunt of wheat. Plant Dis 81: 1370–1377.
Bozkurt TO, McGrann GR, MacCormack R, Boyd LA, Akkaya MS (2010). Cellular and transcriptional responses of wheat during compatible and incompatible race-specific interactions with Puccinia striiformis f. sp. tritici. Mol Plant Pathol 11: 625–640.
Bozkurt TO, Unver T, Akkaya MS (2007). Genes associated with resistance to wheat yellow rust disease identified by differential display analysis. Physiol Mol Plant Pathol 71: 251–259.
Carris LM, Castlebury LA, Goates BJ (2006). Nonsystemic bunt fungi Tilletia indica and T. horrida: a review of history, systematics, and biology. Annu Rev Phytopathol 44: 113–133.
Collinge DB, Kragh KM, Mikkelsen JD, Nielsen KK, Rasmussen U, Vad K (1993). Plant chitinases. Plant J 3: 31–40.
Collinge M, Boller T (2001). Differential induction of two potato genes, Stprx2 and StNAC, in response to infection by Phytophthora infestans and wounding. Plant Mol Biol 46: 521– 5
Directorate of Economics and Statistics (2010). Agricultural Statistics at a Glance. New Delhi: Department of Agriculture, Ministry of Agriculture, Government of India.
Ge X, Chen J, Li N, Lin Y, Sun C, Cao K (2003). Resistance function of rice lipid transfer protein LTP110. J Biochem Mol Biol 36: 603–607.
Giroux MJ, Morris CF (1998). Wheat grain hardness results from highly conserved mutations in the friabilin components puroindoline a and b. Proc Natl Acad Sci USA 95: 6262–6266.
Goswami RS, Xu JR, Trail F, Hilburn K, Kistler HC (2006). Genomic analysis of host–pathogen interaction between Fusarium graminearum and wheat during early stages of disease development. Microbiology 152: 1877–1890.
Hall TA (1999). BIOEDIT: a user-friendly biological sequences alignment editor and analysis of program for Windows 95/98/ NT. Nucleic Acids Symposium Series 41: 95–98.
Johnk N, Hietala AM, Fossdal CG, Collinge DB, Newman MA (2005). Defense-related genes expressed in Norway spruce roots after infection with the root rot pathogen Ceratobasidium bicorne (anamorph: Rhizoctonia sp.). Tree Physiol 25: 1533–1543.
Kong L, Anderson JM, Ohm HW (2005). Induction of wheat defense and stress-related genes in response to Fusarium graminearum. Genome 48: 29–40.
Kumar J, Saharan MS, Sharma AK, Sharma S, Somvir, Tiwari R, Nagarajan S (2004). Pathogenic and molecular variation among Indian isolates of Tilletia indica causing Karnal bunt of wheat. Indian Phytopath 57: 144–149.
Landjeva S, Korzun V, Borner A (2007). Molecular markers: actual and potential contributions to wheat genome characterization and breeding. Euphytica 156: 121–296.
Li WL, Faris JD, Chittoor JM, Leach JE, Hulbert SH, Liu DJ, Chen PD, Gill BS (1999). Genomic mapping of defense response genes in wheat. Theor Appl Genet 98: 226–233.
Liang P, Averboukh L, Pardee AB (1993). Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization. Nucleic Acids Res 21: 3269–3125.
Lu ZX, Gaudet DA, Frick M, Puchalski B, Genswein B, Laroche A (2005). Identification and characterization of genes differentially expressed in the resistance reaction in wheat infected with Tilletia tritici, the common bunt pathogen. J Biochem Mol Biol 38: 420–431.
Mauch F, Mauch-Mani B, Boller T (1988). Antifungal hydrolases in pea tissue II. Inhibition of fungal growth by combinations of chitinase and β-1,3-glucanase. Plant Physiology 88: 936–942.
Mitra M (1931). A new bunt on wheat in India. Ann Appl Biol 18: 178–179.
Munir F, Naqvi SMS, Mahmood T (2013). In vitro and in silico characterization of Solanum lycopersicum wound-inducible proteinase inhibitor-II gene. Turk J Biol 37: 1–10.
Munoz CI, Bailey AM (1998). A cutinase-encoding gene from Phytophthora capsici isolated by differential-display RT-PCR. Curr Genet 33: 225–230.
Oh BJ, Ko MK, Kim KS, Kim YS, Lee HH, Jeon WB, Im KH (2003). Isolation of defense-related genes differentially expressed in the resistance interaction between pepper fruits and the anthracnose fungus Colletotrichum gloeosporioides. Mol Cells 15: 349–355.
Okay S, Özcengiz G (2011). Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. Turk J Biol 35: 1–7.
Reza KA, Sharmin S, Moosa MM, Mahmood N, Ghosh A, Begum RA, Khan H (2013). Identification and molecular characterization of a receptor-like protein kinase gene from Corchorus capsularis. Turk J Biol 37: 11–17.
Sambrook JE, Fritsch F, Maniatis T (2001). Molecular Cloning: A Laboratory Manual, 3rd ed. Cold Spring Harbor, NY, USA: Cold Spring Harbor Laboratory Press.
Vasanthaiah HKN, Basha SM, Katam R (2010). Differential expression of chitinase and stilbene synthase genes in Florida hybrid bunch grapes to Elsinoe ampelina infection. Plant Growth Regul 61:127–134.
Warham EJ (1987). Studies on Karnal bunt of wheat. PhD, University of Wales, Aberystwyth, UK.
Yoshikawa M, Yamaoka N, Rakeuchi Y (1993). Elicitors: their significance and primary modes of action in the induction of plant defense reactions. Plant Cell Physiol 34: 1163–1173.
Zhang J, Martin JM, Balint-Kurti P, Huang L, Girouz M J (2011). The wheat puroindoline genes confer fungal resistance in transgenic corn. J Phytopathol 159: 188–190.