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A D N A N R I A Z0 5 - A R I D - 2 2
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SUPERVISORY COMMITTEE Dr. Abid Riaz (Supervisor)
Dr. Atiq Rattu (Co-supervisor)
Dr.
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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).
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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
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Wheat rust fungi are highly specific obligateparasites
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 infectedtissue,
The rusted plants yield grains that are poor in quantity andquality.
The grains are either devoid of starch or of low nutritional
quality for human consumption
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Leaf rust is an important disease in Pakistan andoccurring through out the country.
The disease is favored by moderate temperatureand high humidity for long period of time
When pathogen attacks plants, they defend
themselves against such invasion with both passiveand active defense mechanisms.
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Passive or preexisting defense mechanisms involves certain structural barriers
accumulation of antimicrobial compounds which preventpathogen 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 throughboth the cross-linking of cell wall structural proteins and
callose deposition (Almagro et al., 2009).
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Phenylalanine Ammonia-Lyase (PAL) andPeroxidase are considered to be important inresistance 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 ofwheat.
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OBJECTIVES To study the effect of changing climate and find an
appropriate relationship between disease severitywith meteorological data (temperature, relativehumidity and wind velocity) on disease
development.
To assess severity response of wheat cultivars againstleaf rust at research institutes in Faisalabad,Bahawalpur and Sakrand.
To analyse the activity of Phenylalanine Ammonia-Lyase (PAL) and Peroxidase in the most resistant andsusceptible wheat cultivars against leaf rust through
spectrophotometer.
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MATERIALS AND METHODS COLLECTION OF DISEASE AND METEOROLOGICAL DATA
Disease scale (Cobbs scale) (Peterson et al., 1948)
O- No visible infection.
R- Resistant: visible chlorosis or necrosis, nouredia are present.
MR- Moderately resistant: small uredia, surrounded
by either chlorotic or necrotic areas.MS- Moderately susceptible: uredia medium size
with no necrotic margins but possibly somedistinct chlorosis.
S- Susceptible: Large uredia with no necrosis
and little or no chlorosis.
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Reaction Type Observation Response Value
No Disease 0 0
Resistant R 0.2
Resistant-Moderately Resistant R-MR 0.3
Moderately Resistant MR 0.4
Moderately Resistant-Moderately
Susceptible MR-MS
0.6
Moderately Susceptible MS 0.8
Moderately Susceptible-Susceptible MS-S 0.9
Susceptible S 1
Response values given against each reaction and observation to calculate the co-efficient ofinfection for brown rust of wheat Loegering, (1959).
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Varieties\Year 2002-03 2003-04 2004-05 2005-06 2006-07 2007-08 2008-09
Morroco 50S 60S 80S 80S 80S 0 80S
Sarsabz 20MS 20S 0 0 5S 0 5S
Inqilab-91 20MRMS 30MRMS 0 0 0 0 0
Tandojam-83 10S 20MSS 0 0 0 0 0
Soughat-90 10S 10S 0 0 0 0 0
Kirin-95 10S 30MR 0 0 0 0 0
Table 1: Wheat leaf rust data for Faisalabad (2002-2009) as obtained from CDRP, NARC.
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Varieties\Year 2002-03 2003-04 2004-05 2005-06 2006-07 2007-08 2008-09
Morroco 80S 90S 100S 100S 0 80S 80S
Sarsabz 20S 30MSS 50MRMS 30S 0 0 10S
Inqilab-91 20MS 30MRMS 30MRMS 20MSS 0 0 0
Tandojam-83 20S 30S 20S 30S 10S 0 50S
Soughat-90 10MSS 20MSS 20MS 20MSS 0 0 0
Kirin-95 10S 40MR 20MRMS 20MS 0 0 0
Table 2: Wheat leaf rust data for Bahawalpur (2002-2009) as obtained from CDRP, NARC.
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Varieties\Year 2002-03 2003-04 2004-05 2005-06 2006-07 2007-08 2008-09
Morroco 40S 90S 0 60S 40S 0 70S
Sarsabz 20MSS 20S 0 20S 0 0 10MSS
Inqilab-91 20MS 20MSS 0 20S 0 0 0
Tandojam-83 20MS 20MSS 0 20S 0 0 5MSS
Soughat-90 10S 10S 0 10S 0 0 0
Kirin-95 10S 10S 0 20S 0 0 0
Table 3: Wheat leaf rust data for Sakrand (2002-2009) as obtained from CDRP, NARC.
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Year
Average
Temperature C
Average
Relative Humidity (%)
Average
Wind Velocity (Km/hr)
2003 16.74 63.23 5.26
2004 20.11 55.01 4.77
2005 16.47 64.07 3.72
2006 21.31 43.97 3.46
2007 15.83 59.73 2.74
2008 15.82 37.23 6.93
2009 17.4 57.73 2.62
Table 4: One month (30 days) average value of three environmental factors (temperature, relativehumidity and wind velocity) of different years at Faisalabad.
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Year
Average
Temperature C
Average
Relative Humidity (%)
Average
Wind Velocity (Km/hr)
2003 17.45 67.53 3.95
2004 20.44 62.48 2.47
2005 17.41 76.95 4.07
2006 20.7 68.23 2.9
2007 17.57 73.32 1.98
2008 16.85 57.08 3.02
2009 18.25 65.98 3.89
Table 5: One month (30 days) average value of three environmental factors (temperature, relativehumidity and wind velocity) of different years at Bahawalpur.
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Year
Average Temperature
C
Average
Relative Humidity (%)
Average
Wind Velocity (Km/hr)
2003 19.81 58.48 8.21
2004 20.45 54.13 6.05
2005 17.35 58.43 8.67
2006 22.33 52.78 6.2
2007 20.24 60.87 6.79
2008 15.59 53.55 7.69
2009 20.12 58.9 6.27
Table 6: One month (30 days) average value of three environmentalfactors (temperature, relativehumidity and wind velocity) ofdifferent years at Sakrand.
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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
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RESULTS AND DISCUSSION ANALYSIS OF DISEASE AND METEOROLOGICAL DATA
Figure 1a: Effect of temperature on co-efficient of infection of leaf rust onMorroco, Inqilab-91 and Sarsabz.
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Figure 1b: Effect of temperature on co-efficient of infection of leaf rust onSoughat-90, Tandojam-83 and Kirin-95.
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Figure 2a: Effect of relative humidity on co-efficient of infection of leaf rust onMorroco, Inqilab-91 and Sarsabz.
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Figure 2b: Effect of relative humidity on co-efficient of infection of leaf rust onSoughat-90, Tandojam-83 and Kirin-95.
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Figure 3a: Effect of wind velocity on co-efficient of infection of leaf rust onMorroco, Inqilab-91 and Sarsabz
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Figure 3b: Effect of wind velocity on co-efficient of infection of leaf rust onSoughat-90, Tandojam-83 and Kirin-95
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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
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DETERMINATION OF ACTIVITIES OF PHENYLALANINEAMMONIA LYASE (PAL) AND PEROXIDASE IN WHEAT
CULTIVARS
PAL activity (IU/ml/min) at 270 nm after 8hr, 24hr, 48hr and 72hr after inoculation inresistant cultivar, Inqilab-91.
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Peroxidase activity (IU/ml/min) at 270 nm after 8hr, 24hr, 48hr and 72hr after inoculationin resistant cultivar, Inqilab-91.
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PAL activity (IU/ml/min) at 270 nm after 8hr, 24hr, 48hr and 72hr after inoculation insusceptible cultivar, Kirin-95
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Peroxidase activity (IU/ml/min) at 470 nm after 8hr, 24hr, 48hr and 72hr after inoculation insusceptible cultivar, Kirin-95
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Absorbance of standards for cinnamic acid used in PAL assay observed at 270nmwavelength
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Absorbance of standards for guaiacol used in peroxidase assay observed at470nm wavelength.
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