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Transcript dynamics at early stages of molecular interactions of MYMIV with resistant and susceptible genotypes of the leguminous host, Vigna mungo.

Kundu A, Patel A, Paul S, Pal A - PLoS ONE (2015)

Bottom Line: A significant fraction of modulated transcripts are of unknown function indicating participation of novel candidate genes in restricting this viral pathogen.T9 is perhaps due to the poor execution of these transcript modulation exhibiting remarkable repression of photosynthesis related genes resulting in chlorosis of leaves followed by penalty in crop yield.In addition to inflate the existing knowledge base, the genomic resources identified in this orphan crop would be useful for integrating MYMIV-tolerance trait in susceptible cultivars of V. mungo.

View Article: PubMed Central - PubMed

Affiliation: Division of Plant Biology, Bose Institute, Kolkata 700054, West Bengal, India.

ABSTRACT
Initial phases of the MYMIV-Vigna mungo interaction is crucial in determining the infection phenotype upon challenging with the virus. During incompatible interaction, the plant deploys multiple stratagems that include extensive transcriptional alterations defying the virulence factors of the pathogen. Such molecular events are not frequently addressed by genomic tools. In order to obtain a critical insight to unravel how V. mungo respond to Mungbean yellow mosaic India virus (MYMIV), we have employed the PCR based suppression subtractive hybridization technique to identify genes that exhibit altered expressions. Dynamics of 345 candidate genes are illustrated that differentially expressed either in compatible or incompatible reactions and their possible biological and cellular functions are predicted. The MYMIV-induced physiological aspects of the resistant host include reactive oxygen species generation, induction of Ca2+ mediated signaling, enhanced expression of transcripts involved in phenylpropanoid and ubiquitin-proteasomal pathways; all these together confer resistance against the invader. Elicitation of genes implicated in salicylic acid (SA) pathway suggests that immune response is under the regulation of SA signaling. A significant fraction of modulated transcripts are of unknown function indicating participation of novel candidate genes in restricting this viral pathogen. Susceptibility on the other hand, as exhibited by V. mungo Cv. T9 is perhaps due to the poor execution of these transcript modulation exhibiting remarkable repression of photosynthesis related genes resulting in chlorosis of leaves followed by penalty in crop yield. Thus, the present findings revealed an insight on the molecular warfare during host-virus interaction suggesting plausible signaling mechanisms and key biochemical pathways overriding MYMIV invasion in resistant genotype of V. mungo. In addition to inflate the existing knowledge base, the genomic resources identified in this orphan crop would be useful for integrating MYMIV-tolerance trait in susceptible cultivars of V. mungo.

No MeSH data available.


Related in: MedlinePlus

qPCR expression profiles of 16 transcripts that were differentially expressed on challenging compatible (T9: Light bars) and incompatible (VMR84: Dark bars)) plants with MYMIV.Relative expression values were estimated using 2-Δ(ΔCt) method after normalization with actin. Expression analyses were done in five sampling points (6, 12, 24, 36 and 48 hpi) for HSP90 (heat shock protein 90); SGT1; CaM (calmodulin); WRKY transcription factor; GST (glutathione S transferase); APOX (ascorbate peroxidase); Csp (cysteine protease); RuAc (rubisco activase); PAL (phenylalanine ammonia lyase); TRX (thioredoxin); SOD (superoxide dismutase); MET (metallotheionein); PR1 (pathogenesis related protein 1); MAPK (MAP kinase); TS (tryptophan synthase); UBL (ubiquitin ligase) MADS (MADS box protein); ARF (auxin response factor) and OEC (oxygen evolving complex) identified from SSH enriched libraries of either susceptible or resistant blackgram plants. Bars represent mean ± standard deviation; bars followed by different letters indicate significant differences at p ≤ 0.05 according to DMRT.
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pone.0124687.g004: qPCR expression profiles of 16 transcripts that were differentially expressed on challenging compatible (T9: Light bars) and incompatible (VMR84: Dark bars)) plants with MYMIV.Relative expression values were estimated using 2-Δ(ΔCt) method after normalization with actin. Expression analyses were done in five sampling points (6, 12, 24, 36 and 48 hpi) for HSP90 (heat shock protein 90); SGT1; CaM (calmodulin); WRKY transcription factor; GST (glutathione S transferase); APOX (ascorbate peroxidase); Csp (cysteine protease); RuAc (rubisco activase); PAL (phenylalanine ammonia lyase); TRX (thioredoxin); SOD (superoxide dismutase); MET (metallotheionein); PR1 (pathogenesis related protein 1); MAPK (MAP kinase); TS (tryptophan synthase); UBL (ubiquitin ligase) MADS (MADS box protein); ARF (auxin response factor) and OEC (oxygen evolving complex) identified from SSH enriched libraries of either susceptible or resistant blackgram plants. Bars represent mean ± standard deviation; bars followed by different letters indicate significant differences at p ≤ 0.05 according to DMRT.

Mentions: It is of special interest to explore the expression kinetics of transcripts that clearly discriminate susceptible from resistant reactions. Since SSH does not provide any quantitative estimation of gene expression, therefore qPCR analyses of selected ESTs were undertaken to elucidate the dynamic alterations of gene expression over time. qPCR analyses was carried out at 6, 12, 24, 36 and 48 hpi of different biological replicates (Fig 4). Levels of the accumulated transcripts were normalized against the expression of the VmACT gene (JZ078743) that was shown earlier by us as the choice of internal standard for V. mungo under MYMIV-stress condition [21]. Amplification efficiencies for the primer combinations were observed to be around 2, with a 0.97 correlation in the dilution curve analyses (data not shown). The obtained qPCR expression data of the selected genes are consistent with the results of SSH analyses. Transcripts coding for SGT1 and HSP90, the two functional components of R-protein complex were quantified, maximum expression level noted was 7 and 10 folds at 48 hpi, respectively in VMR84 (Fig 4). SAR indicator gene PR1 was increased ten times at 48 hpi, triggering the orthodox defence responses against MYMIV.


Transcript dynamics at early stages of molecular interactions of MYMIV with resistant and susceptible genotypes of the leguminous host, Vigna mungo.

Kundu A, Patel A, Paul S, Pal A - PLoS ONE (2015)

qPCR expression profiles of 16 transcripts that were differentially expressed on challenging compatible (T9: Light bars) and incompatible (VMR84: Dark bars)) plants with MYMIV.Relative expression values were estimated using 2-Δ(ΔCt) method after normalization with actin. Expression analyses were done in five sampling points (6, 12, 24, 36 and 48 hpi) for HSP90 (heat shock protein 90); SGT1; CaM (calmodulin); WRKY transcription factor; GST (glutathione S transferase); APOX (ascorbate peroxidase); Csp (cysteine protease); RuAc (rubisco activase); PAL (phenylalanine ammonia lyase); TRX (thioredoxin); SOD (superoxide dismutase); MET (metallotheionein); PR1 (pathogenesis related protein 1); MAPK (MAP kinase); TS (tryptophan synthase); UBL (ubiquitin ligase) MADS (MADS box protein); ARF (auxin response factor) and OEC (oxygen evolving complex) identified from SSH enriched libraries of either susceptible or resistant blackgram plants. Bars represent mean ± standard deviation; bars followed by different letters indicate significant differences at p ≤ 0.05 according to DMRT.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401676&req=5

pone.0124687.g004: qPCR expression profiles of 16 transcripts that were differentially expressed on challenging compatible (T9: Light bars) and incompatible (VMR84: Dark bars)) plants with MYMIV.Relative expression values were estimated using 2-Δ(ΔCt) method after normalization with actin. Expression analyses were done in five sampling points (6, 12, 24, 36 and 48 hpi) for HSP90 (heat shock protein 90); SGT1; CaM (calmodulin); WRKY transcription factor; GST (glutathione S transferase); APOX (ascorbate peroxidase); Csp (cysteine protease); RuAc (rubisco activase); PAL (phenylalanine ammonia lyase); TRX (thioredoxin); SOD (superoxide dismutase); MET (metallotheionein); PR1 (pathogenesis related protein 1); MAPK (MAP kinase); TS (tryptophan synthase); UBL (ubiquitin ligase) MADS (MADS box protein); ARF (auxin response factor) and OEC (oxygen evolving complex) identified from SSH enriched libraries of either susceptible or resistant blackgram plants. Bars represent mean ± standard deviation; bars followed by different letters indicate significant differences at p ≤ 0.05 according to DMRT.
Mentions: It is of special interest to explore the expression kinetics of transcripts that clearly discriminate susceptible from resistant reactions. Since SSH does not provide any quantitative estimation of gene expression, therefore qPCR analyses of selected ESTs were undertaken to elucidate the dynamic alterations of gene expression over time. qPCR analyses was carried out at 6, 12, 24, 36 and 48 hpi of different biological replicates (Fig 4). Levels of the accumulated transcripts were normalized against the expression of the VmACT gene (JZ078743) that was shown earlier by us as the choice of internal standard for V. mungo under MYMIV-stress condition [21]. Amplification efficiencies for the primer combinations were observed to be around 2, with a 0.97 correlation in the dilution curve analyses (data not shown). The obtained qPCR expression data of the selected genes are consistent with the results of SSH analyses. Transcripts coding for SGT1 and HSP90, the two functional components of R-protein complex were quantified, maximum expression level noted was 7 and 10 folds at 48 hpi, respectively in VMR84 (Fig 4). SAR indicator gene PR1 was increased ten times at 48 hpi, triggering the orthodox defence responses against MYMIV.

Bottom Line: A significant fraction of modulated transcripts are of unknown function indicating participation of novel candidate genes in restricting this viral pathogen.T9 is perhaps due to the poor execution of these transcript modulation exhibiting remarkable repression of photosynthesis related genes resulting in chlorosis of leaves followed by penalty in crop yield.In addition to inflate the existing knowledge base, the genomic resources identified in this orphan crop would be useful for integrating MYMIV-tolerance trait in susceptible cultivars of V. mungo.

View Article: PubMed Central - PubMed

Affiliation: Division of Plant Biology, Bose Institute, Kolkata 700054, West Bengal, India.

ABSTRACT
Initial phases of the MYMIV-Vigna mungo interaction is crucial in determining the infection phenotype upon challenging with the virus. During incompatible interaction, the plant deploys multiple stratagems that include extensive transcriptional alterations defying the virulence factors of the pathogen. Such molecular events are not frequently addressed by genomic tools. In order to obtain a critical insight to unravel how V. mungo respond to Mungbean yellow mosaic India virus (MYMIV), we have employed the PCR based suppression subtractive hybridization technique to identify genes that exhibit altered expressions. Dynamics of 345 candidate genes are illustrated that differentially expressed either in compatible or incompatible reactions and their possible biological and cellular functions are predicted. The MYMIV-induced physiological aspects of the resistant host include reactive oxygen species generation, induction of Ca2+ mediated signaling, enhanced expression of transcripts involved in phenylpropanoid and ubiquitin-proteasomal pathways; all these together confer resistance against the invader. Elicitation of genes implicated in salicylic acid (SA) pathway suggests that immune response is under the regulation of SA signaling. A significant fraction of modulated transcripts are of unknown function indicating participation of novel candidate genes in restricting this viral pathogen. Susceptibility on the other hand, as exhibited by V. mungo Cv. T9 is perhaps due to the poor execution of these transcript modulation exhibiting remarkable repression of photosynthesis related genes resulting in chlorosis of leaves followed by penalty in crop yield. Thus, the present findings revealed an insight on the molecular warfare during host-virus interaction suggesting plausible signaling mechanisms and key biochemical pathways overriding MYMIV invasion in resistant genotype of V. mungo. In addition to inflate the existing knowledge base, the genomic resources identified in this orphan crop would be useful for integrating MYMIV-tolerance trait in susceptible cultivars of V. mungo.

No MeSH data available.


Related in: MedlinePlus