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Materials and methods

Seed of five improved wheat cultivars i.e., HD 2135, HD 2160, HD 2189, HD 2285 and Vaishali (test cultivars), was obtained from Division of Genetics, IARI, New Delhi. Cultivar, Agra Local (AL), was used as a susceptible parent in making crosses.

The above test cultivars were crossed with AL to get F1 seed. Reciprocal crosses were also attempted to study the role of cytoplasm in inheritance of resistance. HD 2189 and HD 2285 were also crossed with isogenic wheat lines Sr11 and Sr30 for test of allelism. After emergence of ear heads crossing was attempted following emasculation procedure. A few F1 seeds were kept in reserve while others were multiplied to raise F2 seed. A part of F2 seed was further advanced to get F3 seed for testing. To identify whether resistance is similar or different in test cultivars, they were crossed among themselves except in reciprocal manner, to make diallele crosses.

For testing, parents, F1, F2 and F3 seedlings were raised in aluminium trays (11" x 4" x 3") filled with soil and farmyard manure. The see&as in trays were ready for inoculation after 10 days of sowing. A set of differentials (Bahadur et al. 1985) was also sown along with each set for ascertaining the purity of the pathotype. The urediospore inoculum of various pathotypes was obtained from Directorate of Wheat Research, Regional Station, Flowerdale, Shimla. The urediospore-inoculum of each pathotype was multiplied on AL, following standard procedures (Josh) et al. 1988). The urediospore suspension of the pathotype was prepared in a clean petri plate by mixing spore dust with a few drops of water and a pinch of tween 20 to break the surface tension. Adequate water was added to make the spore suspension and filled in an atomizer and sprayed uniformly on the seedlings. Pots/trays were sprayed with tap water and kept in moist chambers for 48h for incubation at a temperature 20-25C. The above cultivars were also grown along with isogenic lines and inoculated with 12 pathotypes 11(79G31), 21(9G5), 21A-2(75G5), 34-1(l0G13), 40-A(62G29), 40-1(62G29-1), 42(19G35), 117- 1(166G2), 117-A(36G2), 117A-1(38G18), 122(7G11), 295(7G43) for matching Sr genes following Bahadur et al. (1993).

The differential sets were recorded after 15 days of inoculation when disease developed. The infection types were noted according to the classification of Stakman et al. (1962). Further minor variations in infection types were recorded by putting + and - signs after the number, where - sign indicated infection type lower and + sign higher than normal categories. Symbols represent 0= immune, 0;=nearly immune, 1 = very resistant, 2 = resistant, 3 = moderately susceptible, 4 = susceptible, and N = Necrosis.

F2 seedlings, showing different infection types were grouped separately and counted to determine F2 ratios. F3 seedlings of each family, were also recorded for their segregation into resistant, segregating and susceptible families. The chi square test (chi2) for goodness of fit, described by Panse and Sukhatme (1967) was used for testing validity of observations in relation to expected one in segregating population on the basis of Mendelian segregation.

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