ADNAN RIAZ 05-ARID-22

SUPERVISORY COMMITTEE
Dr. Abid Riaz (Supervisor) Dr. Atiq Rattu (Co-supervisor) Dr.

INTRODUCTION
Wheat (Triticum aestivum) belongs to Gramineae family Worlds wheat production is 681.9 million metric tons (FAOSTAT, 2009). In 2010, wheat contributed about 14.4 percent to the total value added in Agriculture and 3.1 percent in GDP (GOP, 2010). During 2009-10, wheat was cultivated on an area of 9.042 mha with an annual production of about 23.864 million metric tons, thus averaging yield of 2.639 tons/hec (GOP, 2010).

DISEASES OF WHEAT
Fungal Diseases
Rusts
Leaf (brown) rust Stripe (yellow) rust Stem (black) rust

Powdery mildew Bunt Smut4 Spot blotch Tan spot Septoria leaf blotch Fusarium head blight

 Wheat rust fungi are highly specific obligate parasites Effects of rusts on wheat plant
Rusts often reduce root growth, foliage and yield by reducing photosynthetic rate increases the rate of respiration decreases translocation of photosynthates from infected tissue, The rusted plants yield grains that are poor in quantity and quality. The grains are either devoid of starch or of low nutritional quality for human consumption

 Leaf rust is an important disease in Pakistan and occurring through out the country. The disease is favored by moderate temperature and high humidity for long period of time When pathogen attacks plants, they defend themselves against such invasion with both passive and active defense mechanisms.

 Passive or preexisting defense mechanisms involves

certain structural barriers accumulation of antimicrobial compounds which prevent pathogen development and colonization of the tissue.

Active or induced defense responses includes:

hypersensitive response (HR) the production of phytoalexins and pathogenesis- related (PR) proteins, ion fluxes across the plasma membrane the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) (oxidative bursts), lignification and the reinforcement of the cell wall through both the cross-linking of cell wall structural proteins and callose deposition (Almagro et al., 2009).

 Phenylalanine Ammonia-Lyase (PAL) and Peroxidase are considered to be important in resistance and susceptible reaction of wheat against rust infections. Changes in the physical climate viz., temperature, rainfall and humidity, also the atmospheric composition affects the management of rusts of wheat.

OBJECTIVES
To study the effect of changing climate and find an appropriate relationship between disease severity with meteorological data (temperature, relative humidity and wind velocity) on disease development. To assess severity response of wheat cultivars against leaf rust at research institutes in Faisalabad, Bahawalpur and Sakrand. To analyse the activity of Phenylalanine AmmoniaLyase (PAL) and Peroxidase in the most resistant and susceptible wheat cultivars against leaf rust through spectrophotometer.

MATERIALS AND METHODS

COLLECTION OF DISEASE AND METEOROLOGICAL DATA Disease scale (Cobbs scale) (Peterson et al., 1948) ORMRNo visible infection. Resistant: visible chlorosis or necrosis, no uredia are present. Moderately resistant: small uredia, surrounded by either chlorotic or necrotic areas. Moderately susceptible: uredia medium size with no necrotic margins but possibly some distinct chlorosis. Susceptible: Large uredia with no necrosis and little or no chlorosis.

MSS-

Reaction Type No Disease Resistant Resistant-Moderately Resistant Moderately Resistant Moderately Resistant-Moderately Susceptible Moderately Susceptible Moderately Susceptible-Susceptible Susceptible

Observation 0 R R-MR MR

Response Value 0 0.2 0.3 0.4

0.6 MR-MS MS MS-S S

0.8
0.9

Response values given against each reaction and observation to calculate the co-efficient of infection for brown rust of wheat Loegering, (1959).

Varieties\Year Morroco Sarsabz Inqilab-91 Tandojam-83 Soughat-90 Kirin-95

2002-03 50S 20MS 20MRMS 10S 10S 10S

2003-04 60S 20S 30MRMS 20MSS 10S 30MR

2004-05 80S 0 0 0 0 0

2005-06 80S 0 0 0 0 0

2006-07 80S 5S 0 0 0 0

2007-08 0 0 0 0 0 0

2008-09 80S 5S 0 0 0 0

Table 1: Wheat leaf rust data for Faisalabad (2002-2009) as obtained from CDRP, NARC.

Varieties\Year Morroco Sarsabz Inqilab-91 Tandojam-83 Soughat-90 Kirin-95

2002-03 80S 20S 20MS 20S 10MSS 10S

2003-04 90S 30MSS 30MRMS 30S 20MSS 40MR

2004-05 100S 50MRMS 30MRMS 20S 20MS 20MRMS

2005-06 100S 30S 20MSS 30S 20MSS 20MS

2006-07 0 0 0 10S 0 0

2007-08 80S 0 0 0 0 0

2008-09 80S 10S 0 50S 0 0

Table 2: Wheat leaf rust data for Bahawalpur (2002-2009) as obtained from CDRP, NARC.

Varieties\Year Morroco Sarsabz Inqilab-91 Tandojam-83 Soughat-90 Kirin-95

2002-03 40S 20MSS 20MS 20MS 10S 10S

2003-04 90S 20S 20MSS 20MSS 10S 10S

2004-05 0 0 0 0 0 0

2005-06 60S 20S 20S 20S 10S 20S

2006-07 40S 0 0 0 0 0

2007-08 0 0 0 0 0 0

2008-09 70S 10MSS 0 5MSS 0 0

Table 3: Wheat leaf rust data for Sakrand (2002-2009) as obtained from CDRP, NARC.

Average Year 2003 2004 2005 2006 2007 2008 2009 Temperature C 16.74 20.11 16.47 21.31 15.83 15.82 17.4

Average Relative Humidity (%) 63.23 55.01 64.07 43.97 59.73 37.23 57.73

Average Wind Velocity (Km/hr) 5.26 4.77 3.72 3.46 2.74 6.93 2.62

Table 4: One month (30 days) average value of three environmental factors (temperature, relative humidity and wind velocity) of different years at Faisalabad.

Average Year 2003 2004 2005 2006 2007 2008 2009

Average

Average

Temperature C
17.45 20.44 17.41 20.7 17.57 16.85 18.25

Relative Humidity (%)

67.53 62.48 76.95 68.23 73.32 57.08 65.98

Wind Velocity (Km/hr)

3.95 2.47 4.07 2.9 1.98 3.02 3.89

Table 5: One month (30 days) average value of three environmental factors (temperature, relative humidity and wind velocity) of different years at Bahawalpur.

Average Temperature Year 2003 2004 2005 2006 2007 2008 2009 C 19.81 20.45 17.35 22.33 20.24 15.59 20.12

Average Relative Humidity (%) 58.48 54.13 58.43 52.78 60.87 53.55 58.9

Average Wind Velocity (Km/hr) 8.21 6.05 8.67 6.2 6.79 7.69 6.27

Table 6: One month (30 days) average value of three environmental factors (temperature, relative humidity and wind velocity) of different years at Sakrand.

 FIELD TRIAL DETERMINATION OF ACTIVITIES OF PHENYLALANINE AMMONIA LYASE (PAL) AND PEROXIDASE IN WHEAT CULTIVARS

Sowing of seedlings Spore germination test Inoculation Preparation of buffers Sampling and processing of samples
Phenylalanine Ammonia-Lyase (PAL) assay Peroxidase assay

RESULTS AND DISCUSSION

ANALYSIS OF DISEASE AND METEOROLOGICAL DATA

Figure 1a: Effect of temperature on co-efficient of infection of leaf rust on Morroco, Inqilab-91 and Sarsabz.

Figure 1b: Effect of temperature on co-efficient of infection of leaf rust on Soughat-90, Tandojam-83 and Kirin-95.

Figure 2a: Effect of relative humidity on co-efficient of infection of leaf rust on Morroco, Inqilab-91 and Sarsabz.

Figure 2b: Effect of relative humidity on co-efficient of infection of leaf rust on Soughat-90, Tandojam-83 and Kirin-95.

Figure 3a: Effect of wind velocity on co-efficient of infection of leaf rust on Morroco, Inqilab-91 and Sarsabz

Figure 3b: Effect of wind velocity on co-efficient of infection of leaf rust on Soughat-90, Tandojam-83 and Kirin-95

DISEASE SEVERITY OF FIELD TRIAL AT FAISALABAD AND BAHAWALPUR, 2010-2011

Disease severity on different cultivars of wheat during 2010-2011 field trials at Faisalabad and Bahawalpur

DETERMINATION OF ACTIVITIES OF PHENYLALANINE AMMONIA LYASE (PAL) AND PEROXIDASE IN WHEAT CULTIVARS

PAL activity (IU/ml/min) at 270 nm after 8hr, 24hr, 48hr and 72hr after inoculation in resistant cultivar, Inqilab-91.

Peroxidase activity (IU/ml/min) at 270 nm after 8hr, 24hr, 48hr and 72hr after inoculation in resistant cultivar, Inqilab-91.

PAL activity (IU/ml/min) at 270 nm after 8hr, 24hr, 48hr and 72hr after inoculation in susceptible cultivar, Kirin-95

Peroxidase activity (IU/ml/min) at 470 nm after 8hr, 24hr, 48hr and 72hr after inoculation in susceptible cultivar, Kirin-95

Absorbance of standards for cinnamic acid used in PAL assay observed at 270nm wavelength

Absorbance of standards for guaiacol used in peroxidase assay observed at 470nm wavelength.