Journal Article•
Molecular cloning, characterization and transcriptional variability study of resistance gene candidates from wild Curcuma spp. for resistance against Pythium aphanidermatum.
TL;DR: A PCR mediated approach has been made using degenerate primers designed on conserved region (P-loop and GLPL) of the NBS domain from R-genes that provides the source for cloning analogous sequences called resistance gene candidates (RGCs) from three wild turmeric plants resistant or susceptible to Pythium aphanidermatum.
Abstract: The genetic base of preferred turmeric (Curcuma longa L.) genotypes has eroded due to their continuous domestication through exclusive vegetative routes thus making them highly susceptible to various pests and pathogens. Molecular cloning of resistance related sequences from wild genotypes can result in efficient turmeric improvement by evolving more effective resistance specificities compared to cultigens. In this study, a PCR mediated approach has been made using degenerate primers designed on conserved region (P-loop and GLPL) of the NBS domain from R-genes that provides the source for cloning analogous sequences called resistance gene candidates (RGCs) from three wild turmeric- Curcuma aromatica, Curcuma angustifolia and Curcuma zedoaria. Twenty-one wild turmeric RGCs were isolated and grouped into four phenetic classes. A strong amino acid identity ranging from 41 to 53% together with presence of internal conserved motifs provided evidence that the isolated RGCs belong to the non-toll interleukin receptor (non-TIR) NBS-LRR R-gene sub-family. Southern hybridization showed a high copy representation of turmeric RGCs. Expression variability of wild turmeric RGCs was analyzed through reverse transcription PCR in root tissues of the three wild turmeric plants resistant or susceptible to Pythium aphanidermatum. Cap12 and Can12 showed a constitutive expression in both resistant and susceptible plants of Curcuma aromatica and Curcuma angustifolia respectively while Czp11 expression was realized only in Pythium aphanidermatum resistant lines of Curcuma zedoaria as well as Curcuma longa L. This result can pave way towards the identification and characterization of a potential Pythium aphanidermatum resistance gene in turmeric.
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TL;DR: Structural and phylogenetic analyses grouped CzR1 within the non-TIR (homology to Toll/interleukin-1 receptors) subclass of NBS-LRR R-genes and semi quantitative RT-PCR analysis showed constitutive expression of CZR1 which gets significantly upregulated in response to infection by different strains of P. aphanidermatum.
18 citations
TL;DR: In this article , degenerate primers were used for cloning and characterization of NBS-LRR type resistant gene from betelvine against anthracnose disease, and a strong amino acid identity ranging from 90.38 to 97.75% and the presence of internal conserved motifs, provided evidence that the identified resistance gene analogs (RGAs) belong to the non-TIR NBS LRR class of R genes.
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TL;DR: A cluster analysis based on the neighbor-joining method was carried out using Curcuma NBS analogs together with several resistance gene analogs and known R-genes, which classified them into two distinct subclasses, corresponding to clades N3 and N4 of non-TIR-NBS sequences described in plants.
Abstract: Turmeric (Curcuma longa), an important asexually reproducing spice crop of the family Zingiberaceae is highly susceptible to bacterial and fungal pathogens. The identification of resistance gene analogs holds great promise for development of resistant turmeric cultivars. Degenerate primers designed based on known resistance genes (R-genes) were used in combinations to elucidate resistance gene analogs from Curcuma longa cultivar surama. The three primers resulted in amplicons with expected sizes of 450-600 bp. The nucleotide sequence of these amplicons was obtained through sequencing; their predicted amino acid sequences compared to each other and to the amino acid sequences of known R-genes revealed significant sequence similarity. The finding of conserved domains, viz., kinase-1a, kinase-2 and hydrophobic motif, provided evidence that the sequences belong to the NBS-LRR class gene family. The presence of tryptophan as the last residue of kinase-2 motif further qualified them to be in the non-TIR- NBS-LRR subfamily of resistance genes. A cluster analysis based on the neighbor-joining method was carried out using Curcuma NBS analogs together with several resistance gene analogs and known R-genes,
19 citations
"Molecular cloning, characterization..." refers result in this paper
...In our earlier work, we had isolated five non-TIR NBS-LRR RGCs from Curcuma longa cultivar surama (Joshi et al., 2010)....
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...No TIR-RGCs were obtained in wild Curcuma genotypes in the present study, which is in accordance with the earlier reports on the absence of such groups among the Rgenes/RGCs of Zingiberaceae (Nair and Thomas, 2007; Joshi et al., 2010; Joshi et al., 2012)....
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Journal Article•
TL;DR: The structure and development of male and female gametophytes of ginger were studied and stigmatic and stylar incompatibility exists in ginger which may contribute to the failure of sexual reproduction.
Abstract: The structure and development of male and female gametophytes of ginger were studied. Pollen-pistil interactions and structural incompatibility were sorted out. Stigmatic and stylar incompatibility was found predominant in ginger cultivars. Assisted pollination methods were attempted. Varying degrees of pollen tube growth initiation were found in ginger cultivars. Stigmatic and stylar incompatibility exists in ginger which may contribute to the failure of sexual reproduction.
12 citations
TL;DR: This will provide a starting point towards characterization of candidate resistance genes in Kaempferia and can act as a source pool for disease resistance development in other asexually reproducing plants.
Abstract: Majority of the plant disease resistance genes expresses cytoplasmic receptor-like proteins characterized by an N-terminal nucleotide-binding site (NBS) and a leucine-rich repeat (LRR) domain. Degenerative primers based on these conserved motifs were used to isolate NBS type sequences in Kaempferia galanga. Cloning and sequencing identified 12 Kaempferia NBS-type sequences called resistance gene candidates (RGCs) classified into four classes. The amino acid sequences of the RGCs detected the presence of conserved domains, viz., kinase-1a, kinase-2, and hydrophobic GLPL, categorizing them with the NBS–LRR class gene family. Structural and phylogenetic characterization grouped the RGCs with the non-toll interleukin receptor (non-TIR) subclasses of the NBS sequences. Reverse transcription PCR with 10 Kaempferia RGC specific primers revealed 7 out of 10 Kaempferia RGCs to be expressive. The isolation and characterization of Kaempferia RGCs has been reported for the first time in this study. This will provide a starting point towards characterization of candidate resistance genes in Kaempferia and can act as a source pool for disease resistance development in other asexually reproducing plants.
11 citations
"Molecular cloning, characterization..." refers background or result in this paper
...No TIR-RGCs were obtained in wild Curcuma genotypes in the present study, which is in accordance with the earlier reports on the absence of such groups among the Rgenes/RGCs of Zingiberaceae (Nair and Thomas, 2007; Joshi et al., 2010; Joshi et al., 2012)....
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...…identified in other plant species also showed a comparable range of identities of Rgenes/RGCs such as 30-45% in Phaseolus vulgaris (Rivkin et al., 1999), 31-51% in grape (Di Gaspero and Cipriani, 2002), 27-50% ginger (Nair and Thomas, 2007) and 38 to 82% in Kaempferia galanga (Joshi et al., 2012)....
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...Joshi et al (2012) also found significant sequence identity of 11 Kaempferia RGCs with dicot sequences....
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...Many numbers of RGC classes have also been identified in other plant species such as six classes in apricot (Soriano et al., 2005), five classes in ginger (Nair and Thomas, 2007) and four classes in Kaempferia galanga (Joshi et al., 2012)....
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