Tilletia indica, commonly called Karnal bunt, is an internationally quarantined wheat fungal path... more Tilletia indica, commonly called Karnal bunt, is an internationally quarantined wheat fungal pathogen which affects commercial seed trading as well as the quality of wheat grain for consumption. The teliospores of Tilletia indica surviving in soil serve as the primary source of inoculum and play a major role in disease development. Proper identification and detection of T. indica teliospores based on morphological features and germination of teliospores is time consuming and tedious. In this study, we validated PCR based species-specific primer which amplified 570 bp fragments using ITSKB primers. Further, the real time PCR assay was developed and standardized for detection and quantification of teliospores in soil. The (R) correlation coefficient (0.994) between CT values and DNA concentrations showed the accuracy of qPCR based quantification. The sensitivity of qPCR marker was 100 fg. Thirteen field soil samples were assessed by qPCR for quantification of teliospore DNA. Low funga...
Molecular characterization of 18 Chaetomium isolates collected from India based on the internal t... more Molecular characterization of 18 Chaetomium isolates collected from India based on the internal transcribed spacer (ITS) region of the rRNA gene sequences was done. Phylogenetic analysis of full length ITS region showed that Chaetomium globosum isolates, Cg1, Cg2, Cg6, Cg11 and Cg15, Chaetomium spp. isolates, C16, C17 and a Chaetomium perlucidum isolate formed a group with American isolates of Chaetomium spp., SW287, SW271 and CL024, thereby supporting the close relationships among these isolates. Other C. globosum isolates, Cg3, Cg4, Cg5, Cg10, Cg12, Cg13 and Cg14 clustered with European isolates, UOA/HCPF 9215 and UOA/HCPF 9860 and an Australian isolate NC1. Isolates Cg7, Cg8, and Cg9 were closely related to the Australian isolates but distantly related to the isolates from New Zealand. However, all these isolates clustered in the same Australia group as evident in the evolutionary history analysis using parsimony method. European isolate MU-2009 and Australian isolates NA26 were ...
Genetic variability among the isolates of Ustilago segetum f.sp. tritici collected from north wes... more Genetic variability among the isolates of Ustilago segetum f.sp. tritici collected from north west plain zone was studied using 40 random deca primers. The results revealed high genetics variability ranging from 10 to 48%. The data generated by RAPD polymorphism was partly corresponded to the geographical origin of the isolates.
Based on the host pathogen reaction on a set of 19 Canadian differentials twenty two isolates of ... more Based on the host pathogen reaction on a set of 19 Canadian differentials twenty two isolates of Ustilago segetum f.sp. tritici collected from north India were grouped into four distinct races. Race T1 was detected in Himachal Pradesh; two races T7 and T11 were observed in Haryana isolates, races T7 and T4 were found in Rajasthan and T1 and T11 were observed in Punjab.
Bipolaris sorokiniana is a devastating fungal pathogen causing spot blotch of wheat. We report he... more Bipolaris sorokiniana is a devastating fungal pathogen causing spot blotch of wheat. We report here the first draft genome of Bipolaris sorokiniana strain BS_112 from India using sequence reads from the Ion Torrent, Illumina HiSeq, and Nanopore platforms. The genome size was estimated at 35.64 Mb with an average G+C content of 50.20%.
Tilletia indica is an important quarantined fungal pathogen inciting Karnal bunt disease of wheat... more Tilletia indica is an important quarantined fungal pathogen inciting Karnal bunt disease of wheat. In fungal pathogens, the Hog1-type MAPK is involved in response to various stresses and virulence. In the present study,we identified, characterized and analyzed functional expression of the TiHog1-type gene. As HOG1 homologous genes are salt-inducible, the TiHOG1 gene (1104 bp) was amplified in 1 M NaCl stressed condition in T. indica; cloned in pJET2.0 cloning vector. The TiHOG1 gene was subcloned in expression vector (pRS426Gpd plasmid) and sequences were analyzed. The TiHOG1 in pRS426GPD (pRS426GPD-TiHOG1) was expressed functionally in heterologous system (Saccaromyces cerevisiae). TiHOG1 protein exhibits high amino acid similarity with the homologous protein sequences in Tilletia caries, Ustilago maydis and Puccinia graminis f. sp. tritici. Functional expression of TiHOG1 in S. cerevisiae ∆hog1 mutant showed that TiHOG1 was restored salinity stress tolerance up to 1 M NaCl, 1 M KCl, 0.25 CsCl2, 0.5 M MgCl2 and 0.5 M CaCl2 and osmostress tolerance up to 1 M sorbitol and 1 M sucrose. The expression of TiHOG1 was elicited up to 19.81-fold in response to a susceptible host, whereas resistant host factors showed 9.65-fold transcripts. This first study revealed involvement of a TiHOG1 gene in stress tolerance and may have a role in pathogenesis of T. indica (Karnal bunt of wheat).
Four Indian bread wheat cultivars viz., DL788-2, GW322, Raj3765 and UP2425 were genetically analy... more Four Indian bread wheat cultivars viz., DL788-2, GW322, Raj3765 and UP2425 were genetically analyzed against three selected pathotypes 21 (9G5), 40A (62G29) and 117A (36G2) of Puccinia graminis tritici at Indian Agricultural Research Institute, New Delhi. The segregation pattern in seedlings of F2 derived from crosses with Agra Local as susceptible parent showed the presence of three dominant independent genes for stem rust resistance in DL788-2 and Raj 3765, and two dominant independent genes in each GW322 and UP2425 when analyzed with pathotype 21 (9G5). These findings were supported by segregation of F2 seedlings of reciprocal crosses (AL x parent), in same pattern and reconfirmed by analyses of BC1 and BC2 progenies to test pathotypes. The F2 seedlings of all the diallele crosses did not segregate for susceptibility to pathotype 21 (9G5). It indicated that resistance gene(s) may be common in test cultivars, however, some of the diallele crosses showed segregation to pathotypes 40A (62G29) and 117A (36G2) showing dissimilarity of resistance. Genes Sr24in DL788-2, Sr1 1 in GW 322 and Sr31in UP2425 were validated based on pedigree and infection types of cultivars, and confirmed by test of allelism. An adult plant resistance gene Sr2was also identified in all these test cultivars based on mottling effect in the seedlings.
Host-pathogen responses on 10 differential hosts distinguished populations of six isolates into t... more Host-pathogen responses on 10 differential hosts distinguished populations of six isolates into three pathotypes, pathotype I(KB-1,-2 and-4), pathotype II(KB-3 and-5) and pathotype III(KB-6). Isolate KB-2, which was the most virulent, showed the highest content of lipid( ...
Burkholderia multivorans V2 (BMV2) isolated from soil was found to produce an extracellular solve... more Burkholderia multivorans V2 (BMV2) isolated from soil was found to produce an extracellular solvent tolerant lipase (6.477 U/mL). This lipase exhibited maximum stability in n-hexane retaining about 97.8% activity for 24h. After performing statistical optimization of medium components for lipase production, a 2.2-fold (14 U/mL) enhancement in the lipase production was observed. The crude lipase from BMV2 was partially purified by ultrafiltration and gel permeation chromatography with 24.64-fold purification. The K(m) and V(max) values for partially purified BMV2 lipase were found to be 1.56 mM and 5.62 micromoles/mg min. The metal ions Ca(2+), Mg(2+) and Mn(2+) had stimulatory effect on lipase activity, whereas Cu(2+), Fe(2+) and Zn(2+) strongly inhibited the lipase activity. EDTA and PMSF at 10mM concentration strongly inhibited the lipase activity. Non-ionic and anionic surfactants stimulated the lipase activity. BMV2 lipase was proved to be efficient in synthesis of ethyl butyrate ester under non-aqueous environment.
Archives Of Phytopathology And Plant Protection, 2011
Genetic variability in 23 monosporidial lines developed from five isolates of Tilletia indica cau... more Genetic variability in 23 monosporidial lines developed from five isolates of Tilletia indica causing Karnal bunt of wheat isolated from four wheat growing states of India was determined by using 19 rapid amplified polymorphic DNA (RAPD) markers. Amplification ...
Tilletia indica, commonly called Karnal bunt, is an internationally quarantined wheat fungal path... more Tilletia indica, commonly called Karnal bunt, is an internationally quarantined wheat fungal pathogen which affects commercial seed trading as well as the quality of wheat grain for consumption. The teliospores of Tilletia indica surviving in soil serve as the primary source of inoculum and play a major role in disease development. Proper identification and detection of T. indica teliospores based on morphological features and germination of teliospores is time consuming and tedious. In this study, we validated PCR based species-specific primer which amplified 570 bp fragments using ITSKB primers. Further, the real time PCR assay was developed and standardized for detection and quantification of teliospores in soil. The (R) correlation coefficient (0.994) between CT values and DNA concentrations showed the accuracy of qPCR based quantification. The sensitivity of qPCR marker was 100 fg. Thirteen field soil samples were assessed by qPCR for quantification of teliospore DNA. Low funga...
Molecular characterization of 18 Chaetomium isolates collected from India based on the internal t... more Molecular characterization of 18 Chaetomium isolates collected from India based on the internal transcribed spacer (ITS) region of the rRNA gene sequences was done. Phylogenetic analysis of full length ITS region showed that Chaetomium globosum isolates, Cg1, Cg2, Cg6, Cg11 and Cg15, Chaetomium spp. isolates, C16, C17 and a Chaetomium perlucidum isolate formed a group with American isolates of Chaetomium spp., SW287, SW271 and CL024, thereby supporting the close relationships among these isolates. Other C. globosum isolates, Cg3, Cg4, Cg5, Cg10, Cg12, Cg13 and Cg14 clustered with European isolates, UOA/HCPF 9215 and UOA/HCPF 9860 and an Australian isolate NC1. Isolates Cg7, Cg8, and Cg9 were closely related to the Australian isolates but distantly related to the isolates from New Zealand. However, all these isolates clustered in the same Australia group as evident in the evolutionary history analysis using parsimony method. European isolate MU-2009 and Australian isolates NA26 were ...
Genetic variability among the isolates of Ustilago segetum f.sp. tritici collected from north wes... more Genetic variability among the isolates of Ustilago segetum f.sp. tritici collected from north west plain zone was studied using 40 random deca primers. The results revealed high genetics variability ranging from 10 to 48%. The data generated by RAPD polymorphism was partly corresponded to the geographical origin of the isolates.
Based on the host pathogen reaction on a set of 19 Canadian differentials twenty two isolates of ... more Based on the host pathogen reaction on a set of 19 Canadian differentials twenty two isolates of Ustilago segetum f.sp. tritici collected from north India were grouped into four distinct races. Race T1 was detected in Himachal Pradesh; two races T7 and T11 were observed in Haryana isolates, races T7 and T4 were found in Rajasthan and T1 and T11 were observed in Punjab.
Bipolaris sorokiniana is a devastating fungal pathogen causing spot blotch of wheat. We report he... more Bipolaris sorokiniana is a devastating fungal pathogen causing spot blotch of wheat. We report here the first draft genome of Bipolaris sorokiniana strain BS_112 from India using sequence reads from the Ion Torrent, Illumina HiSeq, and Nanopore platforms. The genome size was estimated at 35.64 Mb with an average G+C content of 50.20%.
Tilletia indica is an important quarantined fungal pathogen inciting Karnal bunt disease of wheat... more Tilletia indica is an important quarantined fungal pathogen inciting Karnal bunt disease of wheat. In fungal pathogens, the Hog1-type MAPK is involved in response to various stresses and virulence. In the present study,we identified, characterized and analyzed functional expression of the TiHog1-type gene. As HOG1 homologous genes are salt-inducible, the TiHOG1 gene (1104 bp) was amplified in 1 M NaCl stressed condition in T. indica; cloned in pJET2.0 cloning vector. The TiHOG1 gene was subcloned in expression vector (pRS426Gpd plasmid) and sequences were analyzed. The TiHOG1 in pRS426GPD (pRS426GPD-TiHOG1) was expressed functionally in heterologous system (Saccaromyces cerevisiae). TiHOG1 protein exhibits high amino acid similarity with the homologous protein sequences in Tilletia caries, Ustilago maydis and Puccinia graminis f. sp. tritici. Functional expression of TiHOG1 in S. cerevisiae ∆hog1 mutant showed that TiHOG1 was restored salinity stress tolerance up to 1 M NaCl, 1 M KCl, 0.25 CsCl2, 0.5 M MgCl2 and 0.5 M CaCl2 and osmostress tolerance up to 1 M sorbitol and 1 M sucrose. The expression of TiHOG1 was elicited up to 19.81-fold in response to a susceptible host, whereas resistant host factors showed 9.65-fold transcripts. This first study revealed involvement of a TiHOG1 gene in stress tolerance and may have a role in pathogenesis of T. indica (Karnal bunt of wheat).
Four Indian bread wheat cultivars viz., DL788-2, GW322, Raj3765 and UP2425 were genetically analy... more Four Indian bread wheat cultivars viz., DL788-2, GW322, Raj3765 and UP2425 were genetically analyzed against three selected pathotypes 21 (9G5), 40A (62G29) and 117A (36G2) of Puccinia graminis tritici at Indian Agricultural Research Institute, New Delhi. The segregation pattern in seedlings of F2 derived from crosses with Agra Local as susceptible parent showed the presence of three dominant independent genes for stem rust resistance in DL788-2 and Raj 3765, and two dominant independent genes in each GW322 and UP2425 when analyzed with pathotype 21 (9G5). These findings were supported by segregation of F2 seedlings of reciprocal crosses (AL x parent), in same pattern and reconfirmed by analyses of BC1 and BC2 progenies to test pathotypes. The F2 seedlings of all the diallele crosses did not segregate for susceptibility to pathotype 21 (9G5). It indicated that resistance gene(s) may be common in test cultivars, however, some of the diallele crosses showed segregation to pathotypes 40A (62G29) and 117A (36G2) showing dissimilarity of resistance. Genes Sr24in DL788-2, Sr1 1 in GW 322 and Sr31in UP2425 were validated based on pedigree and infection types of cultivars, and confirmed by test of allelism. An adult plant resistance gene Sr2was also identified in all these test cultivars based on mottling effect in the seedlings.
Host-pathogen responses on 10 differential hosts distinguished populations of six isolates into t... more Host-pathogen responses on 10 differential hosts distinguished populations of six isolates into three pathotypes, pathotype I(KB-1,-2 and-4), pathotype II(KB-3 and-5) and pathotype III(KB-6). Isolate KB-2, which was the most virulent, showed the highest content of lipid( ...
Burkholderia multivorans V2 (BMV2) isolated from soil was found to produce an extracellular solve... more Burkholderia multivorans V2 (BMV2) isolated from soil was found to produce an extracellular solvent tolerant lipase (6.477 U/mL). This lipase exhibited maximum stability in n-hexane retaining about 97.8% activity for 24h. After performing statistical optimization of medium components for lipase production, a 2.2-fold (14 U/mL) enhancement in the lipase production was observed. The crude lipase from BMV2 was partially purified by ultrafiltration and gel permeation chromatography with 24.64-fold purification. The K(m) and V(max) values for partially purified BMV2 lipase were found to be 1.56 mM and 5.62 micromoles/mg min. The metal ions Ca(2+), Mg(2+) and Mn(2+) had stimulatory effect on lipase activity, whereas Cu(2+), Fe(2+) and Zn(2+) strongly inhibited the lipase activity. EDTA and PMSF at 10mM concentration strongly inhibited the lipase activity. Non-ionic and anionic surfactants stimulated the lipase activity. BMV2 lipase was proved to be efficient in synthesis of ethyl butyrate ester under non-aqueous environment.
Archives Of Phytopathology And Plant Protection, 2011
Genetic variability in 23 monosporidial lines developed from five isolates of Tilletia indica cau... more Genetic variability in 23 monosporidial lines developed from five isolates of Tilletia indica causing Karnal bunt of wheat isolated from four wheat growing states of India was determined by using 19 rapid amplified polymorphic DNA (RAPD) markers. Amplification ...
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