BIOCHEMICAL AND PHYSIOLOGICAL ANALYSIS AND PROTEIN PROFILING IN WHEAT (Triticum aestivum L.) GENOTYPES UNDER HEAT STRESS
KrishiKosh
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Title |
BIOCHEMICAL AND PHYSIOLOGICAL ANALYSIS AND PROTEIN PROFILING IN WHEAT (Triticum aestivum L.) GENOTYPES UNDER HEAT STRESS
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Creator |
HITESH RANCHHODBHAI RAMANI
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Contributor |
Mandavia M. K.
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Subject |
WHEAT
BIOCHEMISTRY |
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Description |
The present experiment on ?Biochemical and Physiological Analysis and Protein Profiling in Wheat (Triticum aestivum L.) Genotypes under Heat Stress? was conducted at Department of Biotechnology, Junagadh Agricultural University, Junagadh. The experiment-1 was carried out using fourteen wheat genotypes and four heat treatments using Factorial CRD design, where seeds were grown in germination bag filled with soil for 10 days. The seedlings were subjected to control and heat treatments at 35?C, 40?C and 45?C for four hour and samples were analysed for relative water content, membrane stability and injury, lipid peroxidation, hydrogen peroxide content and chlorophyll stability index. Heat tolerant genotype GW-190 genotype showed highest membrane stability and relative water content and lowest membrane injury, lipid peroxidation and hydrogen peroxide compared to other genotype so it was selected as best heat tolerant genotype. Heat susceptible genotype J-2010-11 showed lowest membrane stability (MS) and relative water content and highest membrane injury, lipid peroxidation and hydrogen peroxide content so it was selected as highly susceptible genotype. Above physiological and biochemical parameters may be used for screening the susceptible and tolerant wheat genotypes against heat stress. H2O2 and MS are more effective indicators for screening heat tolerant genotypes under stress condition. From results of the experiment-1, one heat tolerant (GW-190) and heat susceptible (J-2010-11) wheat genotypes were selected and the plants were subjected to two groups: control and heat treatments where 40?C and 45?C heat treatments given for 2 h and 4 h of duration and physiological, biochemical, antioxidant enzyme activities, Isoenzymes and Protein profiling by 2D electrophoresis analysis were performed. Relative water content and membrane stability were found to be higher in heat tolerant genotype GW-190 compared to heat susceptible genotype J-2010-11 at tillering and grain filling stages. As heat stress and duration of heat stress increased the relative water content and membrane stability of heat tolerant and heat susceptible genotypes were decreased at both the stages of wheat development. Compared to tillering stage, relative water content and membrane stability were found lower in grain filling stage because of increased temperature. Protein, Proline and glycine betaine, Glutathione reductase, Peroxidase and Superoxide dismutase acitivities were found to be higher in heat tolerant genotype GW-190 compared to heat susceptible genotype J-2010-11 at tillering and grain filling stages. As heat stress and duration of heat stress increased, the biochemical constitutes and antioxidant enzymes activities of heat tolerant and heat susceptible genotypes also increased at both the stages of wheat development. As well all the biochemical constitutes and all three antioxidant enzymes activities were found to be higher in tillering stage compared to grain filling stage. At tillering stage, in case of Peroxidase, band No. 5 (Rm= 0.289) and band No. 6 (Rm=0.495) were present only in heat tolerant genotype while it was absent in heat susceptible genotype. At grain filling stage, band No. 3 (Rm=0.160) was present only in heat tolerant genotype. In case of Superoxide dismutase at tillering stage all the bands were present in heat tolerant as well in heat susceptible genotypes. At graing filling stage, band No. 2 (Rm=0.072) was present only in heat tolerant genotype. So isoenzymes may be useful for screening the heat tolerant and heat susceptible genotypes. At tillering stage, more total protein spots (1207) were recorded compared to that of (972) spots at grain filling stage. Compared to control, in heat stress condition expression of spots were increased. This was not true for grain filling stage. At tillering stage, highest numbers of protein spots (207) were found at 45?C for 4h duration in heat tolerant genotype GW-190 while it was true (148) spots at 40?C for 2h duration in heat tolerant genotype GW-190 at grain filling stage. The protein spots showed the differential expression pattern in treated heat tolerant genotype might be responsible for the stronger heat tolerance. Scanning electron microscopy of wheat leaves showed that analysis of variance indicated significant differences for stomatal length existed among heat tolerant and susceptible genotype as well significant differences were found for stomatal width among heat tolerant and susceptible genotype. Total 12 Operon series RAPD primers were amplified to generate the 105 fragments. The percent polymorphism obtained for RAPD primers were ranged from 71.4% to 100% with an average value of 92.33% per primer. Subcluster A1 (b) of cluster- I consisted of only one heat tolerant genotype J-2010-06 with more than 66% of similarity. Subcluster A2 of cluster-I consisted of only one heat susceptible genotype J-2010-13 similarity of more than 85%. Cluster-II consisted of two genotypes J-2010-05 and GW-190 showing similarity of more than 85% that belongs to heat tolerant groups. These primers may be used to screen the genotypes against heat stress. |
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Date |
2016-09-21T14:59:36Z
2016-09-21T14:59:36Z 2014-10 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/78098
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Language |
en
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Format |
application/pdf
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