Browsing by Author "Chanda Kushwaha"

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Association of melanin content with conidiogenesis in Bipolaris Sorokiniana of barley (Hordeum vulgare L.)
(2010) B.M. Bashyal; Ramesh Chand; Chanda Kushwaha; Devyani Sen; L.C. Prasad; A.K. Joshi
Sixty-seven isolates of Bipolaris sorokiniana of barley, belonging to three groups (black, white and mixed) were studied to find an association of melanin with the spore production of the fungus. Conidiogenesis in black, white and mixed subpopulation of B. sorokiniana was positively correlated with melanin content/g of mycelium. Primary hyphae of black and mixed subpopulation differentiated into secondary hyphal structures which subsequently produced conidiophores and conidia. Primary hyphae could not differentiate into secondary hyphae and subsequently conidiophores and conidia in white subpopulation. A melanin containing mutant developed from white subpopulation regained its ability to differentiate into secondary hyphae, conidiophores and conidia. Results showed that melanization of mycelia B. sorokiniana mycelia is an important factor for conidia production. © 2009 Springer Science+Business Media B.V.
Identification and characterisation of mlo genes in pea (Pisum sativum L.) vis-à-vis validation of Mlo gene-specific markers
(Scientific and Technical Research Council of Turkey, 2016) Chinmayee Mohapatra; Ramesh Chand; Vinay Kumar Singh; Anil Kumar Singh; Chanda Kushwaha
Pea is infected by a number of pathogens, and among them powdery mildew (PM) caused by Erysiphe pisi is an important fungal disease. This work was undertaken in silico to characterise the identified PM resistant genes of pea and design specific primers to be used in marker-assisted selection (MAS) of resistant and susceptible pea genotypes. Mlo gene sequences of A. thaliana and O. sativa were retrieved from different databases and used for a homology search of P. sativum EST databases. Each identified gene was used for a similarity search and phylogenetic classification with different crops. Four gene-specific primers were designed from the identified Mlo nucleotides and were amplified in different pea genotypes. Pea Mlo1 analogue of the ‘er’ gene was related to AtMlO2, AtMlO6, AtMlO12, and AtMlO3. Pea Mlo2 was related to AtMlo14 and Pea Mlo4. Pea Mlo3 gene was related to AtMlO15, AtMlO13, AtMlO4, and AtMlO11. The Pea Mlo4 gene was closely related with AtMlO8 and AtMlO7. In phylogenetic classification, two main clusters were formed; the first cluster possessed the 4 identified pea Mlo genes and the core conserved motifs. Validation of pea Mlo primers confirmed the effectiveness of PsMlo2 primer for use in MAS. © TÜBİTAK.
Identification and evaluation of a critical time for assessment of slow rusting in pea against Uromyces fabae
(2007) Chanda Kushwaha; C.P. Srivastava; Ramesh Chand; B.D. Singh
Field and polyhouse studies were conducted to determine the appropriate time for the assessment of slow rusting in pea to Uromyces fabae (Pers de Bary). Early and late assessments of rust disease led to differential estimation of resistance under field conditions. The critical time occurred when disease severity on the susceptible (check) genotype HUVP 1 had crossed 90% but was <20% on the resistant (check) genotype FC 1. The rust response of germplasm lines at the critical time were in accordance with their known performances both in field and polyhouse. The disease assessment at critical time revealed precise differentiation between resistance and susceptible reactions in the F2 generation of the cross HUVP 1/FC 1. Moreover, AUDPC estimates based on only two disease assessments, including one assessment at the critical time provided comparable information to those based on six assessments. Reduction in 100-seed weight of inoculated F3 progeny rows showed high correlation with rust severity at the critical time and AUDPC based on two assessments in the field. Significant reduction in 100-seed weight was observed only for susceptible lines whereas, reductions in moderately resistant and resistant lines were not significant. © 2007 Elsevier B.V. All rights reserved.
Identification of RAPD markers linked to the rust (Uromyces fabae) resistance gene in pea (Pisum sativum)
(2005) S. Vijayalakshmi; Kusum Yadav; Chanda Kushwaha; S.B. Sarode; C.P. Srivastava; Ramesh Chand; B.D. Singh
Two RAPD markers linked to gene for resistance (assayed as pustule number cm-2 leaf area) to rust [Uromyces fabae (Pers.) de Bary] in pea (Pisum sativum L.) were identified using a mapping population of 31 BC 1F1 [HUVP 1 (HUVP 1 × FC 1] plants, FC 1 being the resistant parent. The analysis of genetics of rust resistance was based on the parents, F1, F2, BC1F1 and BC 1F2 generations. Rust resistance in pea is of non-hypersensitive type; it appeared to be governed by a single partially dominant gene for which symbol Ruf is proposed. Further, this trait seems to be affected by some polygenes in addition to the proposed oligogene Ruf. A total of 614 decamer primers were used to survey the parental polymorphism with regard to DNA amplification by polymerase chain reaction. The primers that amplified polymorphic bands present in the resistant parent (FC 1) were used for bulked segregant analysis. Those markers that amplified consistently and differentially in the resistant and susceptible bulks were separately tested with the 31 BC1F1 individuals. Two RAPD makers, viz., SC10-82 360 (primer, GCCGTGAAGT), and SCRI-71 1000 (primer, GTGGCGTAGT), flanking the rust resistance gene (Ruf) with a distance of 10.8 cM (0.097 rF and LOD of 5.05) and 24.5 cM (0.194 rF and a LOD of 2.72), respectively, were identified. These RAPD markers were not close enough to Ruf to allow a dependable maker-assisted selection for rust resistance. However, if the two makers flanking Ruf were used together, the effectiveness of MAS would be improved considerably. © Springer 2005.
Importance of aecial cups/pustule for selection for slow rusting in pea (Pisum sativum) against Uromyces fabae
(2010) Chanda Kushwaha; Ramesh Chand; C.P. Srivastava; A.K. Singh; R.A.I. Rashmi; B.D. Singh
Field and polyhouse studies indicated wide variation in number of aecial cups/pustule in pea (Pisum sativum L.) against Uromycesfabae Pers de Bary. Size of pustules varied from 0.6 mm to 2.9 mm. Correlation coefficient (r) between pustule size and AUDPC was -0.19. The coefficient of determination indicated that 3.5% of variation in AUDPC was due to pustule size. Correlations between pustule size and number of pustules/leaf were negative in resistant categories and positive in the susceptible categories, indicating compensatory effects between these traits. Size of aecial cups did not vary significantly, but the number of aecial cups/pustule varied across the genotypes. A single gene seems to govern number of aecial cups/pustule and high number of aecial cups/pustule seems to be dominant over low number. Number of aecial cups/pustule may be useful as a criterion for selection slow rusting in pea.
Improved in vitro technique for screening of barley (hordeum vulgare) genotypes against toxin produced by spot blotch pathogen bipolaris sorokiniana
(2009) B.M. Bashyal; Ramesh Chand; Chanda Kushwaha; L.C. Prasad; A.K. Joshi
An improved technique was developed for screening of barley genotypes in response to fungal toxin of B. sorokiniana. Sixteen genotypes of barley were screened against the toxin in vivo and in vitro conditions. Chlorosis symptom appeared at particular time interval in in vivo condition was correlated with the symptom appeared in in vitro with similar ranking order. Similar results for polyhouse and laboratory conditions were observed at different temperature treatments. Results of in vitro test were found consistant and with higher infiltration efficiency. Similar host response to the toxin in laboratory condition and standing plants open up the possibility of screening large number of genotypes against the earlier methods.
Lignification and early abortive fungal colonies as indicators of partial resistance to rust in pea
(Springer International Publishing, 2016) Chanda Kushwaha; Ramesh Chand; Anil K. Singh; Rashmi Rai; Chandra P. Srivastava; Brahma D. Singh; Chinmayee Mohapatra
Ten recombinant inbred lines (RILs) of pea were selected, on the basis of rust (Uromyces fabae) reaction under screenhouse conditions and molecular makers associated with rust resistance, to study the association of lignification and early abortive fungal colonies in response to attack of U. fabae. The present investigation indicated that partial resistance to rust in pea, as measured by the area under disease progress curve (AUDPC), is negatively influenced by lignification (r = −0.48). Histological studies indicated a higher number of early abortive fungal colonies and delayed development of colonies in resistant RILs than susceptible ones. Furthermore, investigations into the association of phenylalanine ammonia lyase activity and total phenolics during the partial resistance reaction to U. fabae in pea revealed a very low correlation with AUDPC in the RILs. This study infers that lignification plays a major role and is the best indicator of partial resistance towards U. fabae in pea, by influencing colony size and the number of early abortive colonies. © 2016, Sociedade Brasileira de Fitopatologia.
Role of aeciospores in outbreaks of pea (Pisum sativum) rust (Uromyces fabae)
(2006) Chanda Kushwaha; Ramesh Chand; Chandra Prakash Srivastava
Aeciospores in Uromyces fabae were found to be repeating spores and play an important role in pea rust outbreaks in the North Eastern Plain Zone (NEPZ) of India. Experiments conducted on pea rust from 2001 to 2004 revealed the dominance of aeciospores at all growth stages of pea in this region. Urediospore production was erratic and was only observed in a few samples of stems and tendrils (5-10%). Inoculation of pea plants either by aeciospores or urediospores resulted in the production of aeciospores. Production of aeciospores was observed at a temperature range of 10-25°C, with a maximum at 25 ± 2°C. Among the different growth stages of pea, the pod formation stage was highly susceptible and produced the maximum number (744) of aecidia/leaf at 20-25°C. Significant effects of growth stages and temperature were also noticed for pustule number. Urediospore production mainly coincided with the senescence of the pea plants. Maximum germination (2%) of aeciospores was observed at 25°C, whereas maximum urediospore germination (3.5%) was at 15°C. Temperatures > 15°C decreased urediospore germination. A relative humidity (RH) of 100% was favourable for aeciospore germination while 98% RH favoured urediospore germination. Typical histo-pathological behaviour of the aeciospores was observed. © Springer 2006.
Role of melanin in release of extracellular enzymes and selection of aggressive isolates of Bipolaris sorokiniana in barley
(Springer New York LLC, 2014) Ramesh Chand; Manoj Kumar; Chanda Kushwaha; Kavita Shah; Arun K. Joshi
Eighteen barley isolates of Bipolaris sorokiniana belonging to wild and clonal type of black, mixed and white subpopulations were quantitatively assayed for their melanin content and aggressiveness with respect to production of some of the extracellular enzymes such as cellulase, pectinase, amylase and protease. Cellulase and pectinase constituted major portion of the enzymes recovered from the black, mixed and white isolates. Enzyme production and aggressiveness were relatively higher in melanin devoid or low melanin isolates. The melanin deficient isolates were also differentiated from black and mixed isolates on the basis of variation in internal transcribed spacer region of the ribosomal DNA. Higher enzyme productions positively correlated with area under disease progress curve (AUDPC) and lesion development. Melanin content was negatively correlated with extracellular enzymes and aggressiveness of the isolates. Based on melanin content, lesion size, AUDPC and extracellular enzymes, the isolates were grouped in two major clusters (I and II) with further division of cluster II into two sub-clusters (II-A and II-B). The results appears to indicate a possible role of melanin in release of extracellular enzymes and hence in evolution and selection of aggressive isolates of B. sorokiniana in barley. © 2014 Springer Science+Business Media.
Rust (Uromyces viciae-fabae Pers. de-Bary) of Pea (Pisum sativum L.): Present Status and Future Resistance Breeding Opportunities
(MDPI, 2023) Anil Kumar Singh; Chanda Kushwaha; Kumari Shikha; Ramesh Chand; Gyan P. Mishra; Harsh Kumar Dikshit; Jyoti Devi; Muraleedhar S. Aski; Shiv Kumar; Sanjeev Gupta; Ramakrishnan M. Nair
Uromyces viciae-fabae Pers. de-Bary is an important fungal pathogen causing rust in peas (Pisum sativum L.). It is reported in mild to severe forms from different parts of the world where the pea is grown. Host specificity has been indicated in this pathogen in the field but has not yet been established under controlled conditions. The uredinial states of U. viciae-fabae are infective under temperate and tropical conditions. Aeciospores are infective in the Indian subcontinent. The genetics of rust resistance was reported qualitatively. However, non-hypersensitive resistance responses and more recent studies emphasized the quantitative nature of pea rust resistance. Partial resistance/slow rusting had been described as a durable resistance in peas. Such resistance is of the pre-haustorial type and expressed as longer incubation and latent period, poor infection efficiency, a smaller number of aecial cups/pustules, and lower units of AUDPC (Area Under Disease Progress Curve). Screening techniques dealing with slow rusting should consider growth stages and environment, as both have a significant influence on the disease scores. Our knowledge about the genetics of rust resistance is increasing, and now molecular markers linked with gene/QTLs (Quantitative Trait Loci) of rust resistance have been identified in peas. The mapping efforts conducted in peas came out with some potent markers associated with rust resistance, but they must be validated under multi-location trails before use in the marker-assisted selection of rust resistance in pea breeding programs. © 2023 by the authors.
Screening technique for pea (Pisum sativum) genotypes against rust disease (Uromyces fabae)
(2004) R. Chand; C.P. Srivastava; Chanda Kushwaha
Twenty-three diverse genotypes of pea (Pisum sativum L.) were screened against rust disease (Uromyces fabae Pers. de Bary) under field and poly-house conditions during 1999-2000 and 2000-2001. Single pustule isolates were multiplied on a susceptible cultivar 'HFP 4'; the aeciospores produced were used for inoculation of the test genotypes in the field and poly-house conditions. Results under poly-house were consistent for both the years but those of field varied. Based on area under disease progress curve, the test genotypes were categorized as slow and fast rusting. None of the test genotypes were immune to rust disease.
Studies on endophytic colonization ability of two upland rice endophytes, rhizobium sp. and burkholderia sp., using green fluorescent protein reporter
(2009) Manoj Kumar Singh; Chanda Kushwaha; Ramesh Kumar Singh
Colonization ability of the two endophytic bacteria, isolated from surface sterilized roots of upland cultivated rice viz., Rhizobium sp. and Burkholderia sp., was compared after genetically tagging them with a constitutively expressing green fluorescent protein gene (gfp/gusA). Confocal laser scanning microscopy (CLSM) of gnotobiotically grown seedlings of Narendradhan 97, inoculated with gfp/gusA-tagged endophytes, revealed that both Rhizobium sp. and Burkholderia sp. colonized the intercellular spaces in the root cortex when inoculated separately. Colonization by gfp/gusA-tagged Rhizobium sp. was severely inhibited when co-inoculated with an equal number (106 cfu ml-1) of wild type Burkholderia sp. Burkholderia sp. was a more aggressive endophytic colonizer of rice than Rhizobium sp. The potential of using gfp/gusA reporter and CLSM as tools in evaluating competitive ability of colonization among endophytes is demonstrated in this study. © 2009 Springer Science+Business Media, LLC.