Molecular Plant Pathology Laboratory

Lucknow, India

Molecular Plant Pathology Laboratory

Lucknow, India
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Prabhakar C.S.,CSK Himachal Pradesh Krishi Vishvavidyalaya | Prabhakar C.S.,Bhabha Atomic Research Center | Sood P.,CSK Himachal Pradesh Krishi Vishvavidyalaya | Sood P.,Crop Research Sub Station | And 2 more authors.
Biochemical Systematics and Ecology | Year: 2013

The population genetic structure of the pumpkin fruit fly, Bactrocera tau, a fruit fly pest that causes significant losses to cucurbit cultivations, has been studied in Himachal Pradesh (India) using mitochondrial cytochrome oxidase I (mtCOI) gene sequences. Levels of differentiation (genetic distances and FST values) among samples from different locations are minimal, suggesting the local occurrence of a large and geographically undifferentiated population, with the possible exception of population Solan. Nevertheless, overall genetic variability is substantial, with 10 different haplotypes detected in 16 individuals and only one of these - likely the original one as it occupies a central position in the network and is found at a relatively high frequency -shared between multiple populations. The phylogenetic analysis of local B. tau samples in the context of the different sibling species that constitute the B. tau complex in its South-East Asia region of origin revealed that local B. tau is closely related to B. tau species A from Thailand. This should be taken into account in any intervention aimed at the control of this pest, e.g. area wide integrated pest management (AW IPM). The marked local genetic uniformity and predominance of one single species of the species complex further suggest that the sterile insect technique (SIT) may be a viable option. © 2013 Elsevier Ltd.

Prabhakar C.S.,CSK Himachal Pradesh Krishi Vishvavidyalaya | Prabhakar C.S.,Indian International Crops Research Institute for the Semi Arid Tropics | Sood P.,CSK Himachal Pradesh Krishi Vishvavidyalaya | Sood P.,Crop Research Sub Station | And 4 more authors.
Phytoparasitica | Year: 2013

Gut bacteria of fruit fly, Bactrocera tau (Walker) (Diptera: Tephritidae), were isolated and the isolates attractive to B. tau adults were characterized using morphological, biochemical and 16S rRNA analyses to determine their taxonomic position. Based upon morphological, biochemical and 16S rRNA sequences (on the basis of closest match), five gut bacterial species of B. tau were characterized as Delftia acidovorans, Pseudomonas putida, Flavobacterium sp., Defluvibacter sp. and Ochrobactrum sp., of which four bacterial isolates, viz., Delftia acidovorans, Flavobacterium sp., Defluvibacter sp. and Ochrobactrum sp. are new records from guts of the fruit fly species. © 2012 Springer Science+Business Media Dordrecht.

Prabhakar C.S.,CSK Himachal Pradesh Krishi Vishvavidyalaya | Prabhakar C.S.,Research Complex for Eastern Region Research Center | Mehta P.K.,CSK Himachal Pradesh Krishi Vishvavidyalaya | Sood P.,CSK Himachal Pradesh Krishi Vishvavidyalaya | And 5 more authors.
Genetica | Year: 2012

Population genetic structure of melon fly analysed with mitochondrial cytochrome oxidase I gene suggested that melon fly populations across the globe is homogeneous with non-significant variation of 0. 000-0. 003 base substitutions per site. Test isolates representing various geographic situations across the world were placed in 26 mitochondrial haplotypes based on variations associated with a maximum of three mutational steps and the predominant haplotype i. e. H1 was present in all melon fly populations except Hawaiian population. Evolution of mtCOI gene suggested that the fly could have originated some 0. 4 million years ago. The present study also indicated that the B. cucurbitae population expansion is an event of post Pleistocene warm climatic conditions with small number of founder population. The invasion of B. cucurbitae in Hawaii was associated with the large population size and the global presence of the fly is associated with human mediated dispersal. The very low genetic variation suggested that the fly management might be possible by large scale sterile insect techniques programme. © 2012 Springer Science+Business Media B.V.

Lima L.S.,Molecular Plant Pathology Laboratory | Gramacho K.P.,Molecular Plant Pathology Laboratory | Pires J.L.,Molecular Plant Pathology Laboratory | Clement D.,Molecular Plant Pathology Laboratory | And 12 more authors.
Tree Genetics and Genomes | Year: 2010

In this study, we report results of the detection and analysis of SSR markers derived of cacao-Moniliophthora perniciosa expressed sequence tags (ESTs) in relation to cacao resistance to witches' broom disease (WBD), and we compare the polymorphism of those ESTs (EST-simple sequence repeat (SSR)) with classical neutral SSR markers. A total of 3,487 ESTs was used in this investigation. SSRs were identified in 430 sequences: 277 from the resistant genotype TSH 1188 and 153 from the susceptible one Catongo, totalizing 505 EST-SSRs with three types of motifs: dinucleotides (72.1%), trinucleotides (27.3%), and tetranucleotides (0.6%). EST-SSRs were classified into 16 main categories; most of the EST-SSRs belonged to "Unknown function" and "No homology" categories (45.82%). A high frequency of SSRs was found in the 5'UTR and in the ORF (about 27%) and a low frequency was observed in the 3'UTR (about 8%). Forty-nine EST-SSR primers were designed and evaluated in 21 cacao accessions, 12 revealed polymorphism, having 47 alleles in total, with an average of 3.92 alleles per locus. On the other hand, the 11 genomic SSR markers revealed a total of 47 alleles, with an average of 5.22 alleles per locus. The association of EST-SSR with the genomic SSR enhanced the analysis of genetic distance among the genotypes. Among the 12 polymorphic EST-SSR markers, two were mapped on the F2 Sca 6×ICS 1 population reference for WBD resistance. © 2010 Springer-Verlag.

Lee I.-M.,Molecular Plant Pathology Laboratory | Polashock J.,U.S. Department of Agriculture | Bottner-Parker K.D.,Molecular Plant Pathology Laboratory | Bagadia P.G.,University of Maryland University College | And 3 more authors.
European Journal of Plant Pathology | Year: 2014

The identity of the presumed phytoplasmal pathogen associated with cranberry false-blossom disease has never been fully clarified. In the present study a molecular-based procedure was employed to determine the identity of the phytoplasma. Tissues of cranberry plants exhibiting cranberry false-blossom symptoms were collected from multiple bogs on each of three randomly selected commercial cranberry farms in New Jersey. Leafhoppers, including the known vector Limotettix vaccinii (Van Duzee) (=Scleroracus vaccinii, Euscellis striatulus) and the sharp-nosed leafhopper Scaphytopius magdalensis (Provancher), a known vector of blueberry stunt disease, were collected from two different farms in New Jersey. Nested PCR assays and RFLP analysis of 16S rRNA gene sequences were employed for the detection and identification of the associated phytoplasmas. All of 20 cranberry plants sampled and five out of 14 batches of leafhoppers tested positive for phytoplasma. Virtual RFLP and sequence analyses revealed that all the associated phytoplasmas were members or variants of a new subgroup, 16SrIII-Y. Phylogenetic analysis of 16S rRNA sequences indicated that cranberry false-blossom phytoplasma strains represented a lineage distinct from other 16SrIII subgroups. This is the first report confirming that a new phytoplasma (designated as a new subgroup 16SrIII-Y) is associated with cranberry false-blossom disease and associated with both leafhopper species in New Jersey. © 2014 US Government.

Fu S.,Southwest University | Fu S.,Molecular Plant Pathology Laboratory | Fu S.,Lingnan Normal University | Shao J.,Molecular Plant Pathology Laboratory | And 2 more authors.
BMC Genomics | Year: 2016

Background: Huanglongbing (HLB) and tristeza, are diseases of citrus caused by a member of the α-proteobacteria, 'Candidatus Liberibacter asiaticus' (CaLas), and Citrus tristeza virus (CTV) respectively. HLB is a devastating disease, but CTV strains vary from very severe to very mild. Both CaLas and CTV are phloem-restricted. The CaLas-B232 strain and CTV-B6 cause a wide range of severe and similar symptoms. The mild strain CTV-B2 doesn't induce significant symptoms or damage to plants. Results: Transcriptome profiles obtained through RNA-seq revealed 611, 404 and 285 differentially expressed transcripts (DETs) after infection with CaLas-B232, CTV-B6 and CTV-B2. These DETs were components of a wide range of pathways involved in circadian rhythm, cell wall modification and cell organization, as well as transcription factors, transport, hormone response and secondary metabolism, signaling and stress response. The number of transcripts that responded to both CTV-B6 and CaLas-B232 was much larger than the number of transcripts that responded to both strains of CTV or to both CTV-B2 and CaLas-B232. A total of 38 genes were assayed by RT-qPCR and the correlation coefficients between Gfold and RT-qPCR were 0.82, 0.69, 0.81 for sweet orange plants infected with CTV-B2, CTV-B6 and CaLas-B232, respectively. Conclusions: The number and composition of DETs reflected the complexity of symptoms caused by the pathogens in established infections, although the leaf tissues sampled were asymptomatic. There were greater similarities between the sweet orange in response to CTV-B6 and CaLas-B232 than between the two CTV strains, reflecting the similar physiological changes caused by both CTV-B6 and CaLas-B232. The circadian rhythm system of plants was perturbed by all three pathogens, especially by CTV-B6, and the ion balance was also disrupted by all three pathogens, especially by CaLas-B232. Defense responses related to cell wall modification, transcriptional regulation, hormones, secondary metabolites, kinases and stress were activated by all three pathogens but with different patterns. The transcriptome profiles of Citrus sinensis identified host genes whose expression is affected by the presence of a pathogen in the phloem without producing symptoms (CTV-B2), and host genes whose expression leads to induction of symptoms in the plant (CTV-B6, CaLas-B232). © 2016 Fu et al.

Sharma P.,Indian Agricultural Research Institute | Sharma P.N.,Molecular Plant Pathology Laboratory | Kapil R.,Molecular Plant Pathology Laboratory | Sharma S.K.,Molecular Plant Pathology Laboratory | Sharma O.P.,Molecular Plant Pathology Laboratory
Indian Journal of Virology | Year: 2011

Four strains (NL-1, NL-1n, NL-7 and NL-7n) of Bean common mosaic virus (BCMV) prevalent on common bean (Phaseolus vulgaris) in Himachal Pradesh, a north-western Himalayan state of India were compared at the 3′ terminal region of the viral genome to elucidate variation and relationship among these strains. 3′ terminal region comprising of partial nuclear inclusion b (NIb) (171-233 bp), complete coat protein (CP) (860 bp) and 30 untranslated region (UTR) (208-244 bp) shared 96-98% nucleotide and 96-99% amino acids identity among various strains of BCMV. Multiple alignment as well as cluster dendrograms of the 3′ terminal region placed the test isolates in BCMV species of genus Potyvirus. Phylogenetic analysis of complete CP as well as 3′ UTR also showed Indian strains to be distinct strains. Sequence homology, multiple alignment and evolutionary divergence of 3′ terminal region could not differentiate the pathogenic strain groups, thereby establishing least role of this region in strain characterization of the virus. Comparisons of CP and 3′ UTR region of BCMV strains and other members of genus Potyvirus clearly indicated the little utility of 3′ terminal region in distinction of virus strains. Implications of coat protein region in viral strain distinction are also discussed. © Indian Virological Society 2011.

Gupta V.K.,University of Rajasthan | Pandey A.,Molecular Plant Pathology Laboratory | Kumar P.,Molecular Plant Pathology Laboratory | Pandey B.K.,Molecular Plant Pathology Laboratory | And 4 more authors.
African Journal of Biotechnology | Year: 2010

Twenty-five isolates of Colletotrichum gloeosporioides causing mango anthracnose were collected from different agroclimatic zones of India. The isolates were evaluated for their pathogenic variability on mango seedlings and genetic characterization using random amplified polymorphic DNA (RAPD molecular techniques). The random primers OPA-1, 3, 5, 9, 11, 15, 16 and 18 were used and the twentyfive isolates were grouped into two. The amplified DNA fragments (amplicons) obtained was compared by agarose gel electrophoresis. Isolate specific RAPD fingerprints were obtained. Out of eight primers in RAPD, OPA-1, 3 and 18 were able to produce reproducible banding pattern. Each of these primers generated a short spectrum of amplicons, located between 661 and 2291-bp markers, indicative of genetic polymorphism. Dendogram revealed more than 75% level of similarity. 4.36% polymorphism was also found in individual isolates that was not statistically significant (P > 0.05) among the sample, it also indicates that all the isolates tested had approximately same genetic identity. The data suggest that RAPD may be of value by virtue of its rapidity, efficiency and reproducibility in generating genetic fingerprints of C. gloeosporioides isolates. © 2010 Academic Journals.

Gupta V.K.,University of Rajasthan | Gupta V.K.,National University of Ireland | Misra A.K.,Molecular Plant Pathology Laboratory
Archives of Phytopathology and Plant Protection | Year: 2011

Wilt of Psidium guajava L., incited by Fusarium oxysporum f. sp. psidii and Fusarium solani is a serious soil-borne disease of guava in India. Forty-two isolates each of F. oxysporum f. sp. psidii (Fop) and F. solani (Fs) collected from different agro climatic zones of India showing pathogenicity were subjected to estimate the genetic and molecular characterisation in terms of analysis of microsatellite marker studies. Out of eight microsatellite markers, only four microsatellite markers, viz. MB 13, MB 17, RE 102 and AY212027 were amplified with single band pattern showing the character of identical marker for molecular characterisation and genetic identification. Microsatellite marker MB 13 was amplified in F. oxysporum f. sp. psidii and F. solani isolates. Product size of 296 bps and 1018 bps were exactly amplified with a single banding pattern in all the isolates of F. oxysporum f. sp. psidii and F. solani, respectively. Microsatellite markers, viz. MB 17, RE 102 and AY212027 were also exactly amplified with a single banding pattern. MB 17 was amplified in F. oxysporum f. sp. psidii isolates with a product size of 300 bp. RE 102 and AY212027 were amplified in F. solani isolates with the product size of 153 bp and 300 bp, respectively. Therefore, amplified microsatellite marker may be used as identifying DNA marker. © 2011 Copyright Taylor and Francis Group, LLC.

Gupta V.,University of Rajasthan | Misra A.,Molecular Plant Pathology Laboratory | Gupta A.,Mahatma Jyotiba Phule Rohilkhand University | Pandey B.,Molecular Plant Pathology Laboratory | Gaur R.,University of Rajasthan
Journal of Plant Protection Research | Year: 2010

The paper concerns randomly amplified polymorphic DNA RAPD-PCR analysis of seven Trichoderma species isolates and their in vitro antagonism against wilt pathogens of Psidium guajava L. viz. Fusarium oxysporum f. sp. psidii (F. o. f. sp. psidii) and Fusarium solani. Out of 10 RAPD oligonucleotides (OPA 1-OPA 10) tested, seven markers OPA 1, 3, 5, 7, 8, 9 and 10 efficiently differentiated the isolates of Trichoderma and showed reproducible banding patterns. A total of 248 bands were obtained from these markers along with a 61.84% per cent similarity among the seven isolates of Trichoderma. From the seven isolates of Trichoderma, isolates T. harzianum, T. virens and T. viride were evaluated for in vitro efficacy against F. o. f. sp. psidii and F. solani. Per cent inhibition was maximum by direct use of Trichoderma spp. in dual cultures against both pathogens, F. o. f. sp. psidii and F. solani. Two species, T. virens and T. viride were superior in inhibiting the growth of both Fusarium spp. Fusarium isolates showed intra species variability.

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