R. Aswati Nair

Bio: R. Aswati Nair is an academic researcher from Central University of Kerala. The author has contributed to research in topics: Zingiber zerumbet & Pythium myriotylum. The author has an hindex of 8, co-authored 22 publications receiving 172 citations. Previous affiliations of R. Aswati Nair include Rajiv Gandhi Centre for Biotechnology & National Institute of Technology Calicut.

Isolation, characterization and expression studies of resistance gene candidates (RGCs) from Zingiber spp.

Abstract: Ginger (Zingiber officinale Rosc.) production is seriously affected by many fungal and bacterial diseases to which no resistant source is available in the cultivated germplasm. Degenerate primers based on conserved motifs of plant resistance (R) genes were used to isolate analogous sequences called resistance gene candidates (RGCs) from cultivated and wild Zingiber species. Cloning and sequence characterization identified 42 Zingiber RGCs, which could be classified into five classes following phenetic analysis. Deduced amino acid sequences of Zingiber RGCs showed strong identity, ranging from 16 to 43%, to non-toll interleukin receptor (non-TIR) R-gene subfamily. Non-synonymous to synonymous nucleotide substitution (dN/dS) ratio for the NBS domains of Zingiber RGC classes showed evidence of purifying selection. RT-PCR analysis with 15 Zingiber RGC-specific primers demonstrated 8 of the 15 Zingiber RGCs to be expressed. The present study reports for the first time the isolation and characterization of RGCs from ginger and its wild relatives, which will serve as a potential resource for future improvement of this important vegetatively propagated spice crop.

Correlation of Phenylalanine ammonia lyase (PAL) and Tyrosine ammonia lyase (TAL) activities to phenolics and curcuminoid content in ginger and its wild congener, Zingiber zerumbet following Pythium myriotylum infection

Abstract: The biochemical basis of resistance exhibited by a wild Zingiber species, Zingiber zerumbet (L.) Smith, towards the economically devastating soft-rot disease caused by necrotrophic Pythium myriotylum was investigated. Quantification of phenolic compounds revealed higher total phenolic (TP), total flavonoid (TF) and total tannin (TT) content in the uninfected susceptible ginger (Z. officinale) cultivar compared to the resistant taxon. However systemic induction in activities of rate-limiting enzymes of phenolic biosynthetic pathway, phenylalanine ammonia lyase (PAL) and tyrosine ammonia lyase (TAL), were observed in the resistant wild taxon. In the ginger cultivar, even though the inherent PAL specific activity was observed to be higher (24.2 ± 1.9 U mg−1) compared to the wild taxon (4.2 ± 0.8 U mg−1), a subsequent gradual decrease in both PAL and TAL activities were observed following infection of rhizomes with P. myriotylum. This was in contrast to the gradual increase in PAL (13.1 ± 0.8 U mg−1) and TAL (442.5 ± 35.1 U mg−1) specific activity after 5 days post infection (dpi) in the wild taxon. Subsequent HPLC analysis of rhizomes showed an increase in total curcuminoid content in the wild taxon compared to the ginger cultivar. Results are indicative of phenylpropanoid pathway regulation in a manner such that the induced defense metabolites contribute to restrict pathogen invasion in the resistant wild taxon.

Molecular Characterization of an Oomycete-Responsive PR-5 Protein Gene from Zingiber zerumbet

Abstract: The tropical spice crop ginger (Zingiber officinale Roscoe) is highly susceptible to soft rot disease caused by the necrotrophic oomycete Pythium aphanidermatum (Edson) Fitzp. However, Zingiber zerumbet (L.) Smith, a wild relative of ginger, is resistant to P. aphanidermatum and has been proposed as a potential donor for soft rot resistance to Z. officinale. We identified a member of the pathogenesis-related protein group 5 (PR-5) gene family in Z. zerumbet that is expressed constitutively but upregulated in response to infection by P. aphanidermatum. Expression of this gene was upregulated as early as 1.5 h post inoculation (hpi) with the pathogen, peaked at 6 hpi, declined by 9 hpi, and again peaked at 15 hpi before declining at 48 hpi. A cDNA of this PR-5 gene, designated as ZzPR5, encodes a 226-amino-acid predicted protein with a calculated pI of 5.05. The N terminus of this protein contains a 22-amino-acid signal peptide, suggesting that the protein may show apoplastic accumulation like other acidic PR-5 proteins. Phylogenetic analysis revealed high similarity between ZzPR5 and PR-5 proteins reported from other plant species, especially from other Zingiberales. Molecular modeling of ZzPR5 protein revealed an acidic surface cleft, a feature characteristic of glycoside hydrolases and antifungal PR-5 proteins. In molecular docking studies, a linear polymeric molecule of (1,3)-β-d-glucan, a major constituent of the oomycete cell wall, fitted favorably into the surface cleft of ZzPR5 and interacted with acidic amino acids known to be involved in glucan hydrolysis, suggesting a potential antioomycete activity for ZzPR5 protein. Elucidation of the molecular mechanism of ZzPR5 may provide important insight toward engineering soft rot resistance into the obligatory asexual ginger.

Metabolic profiling of Zingiber zerumbet following Pythium myriotylum infection: investigations on the defensive role of the principal secondary metabolite, zerumbone.

Abstract: Induced biosynthesis of bioactive secondary metabolites constitutes one of the mechanisms of plant basal innate immunity to fungal infection. Metabolic changes were studied in rhizomes of Zingiber zerumbet, a wild congener of ginger, after infection with soft rot-causative necrotrophic phytopathogen, Pythium myriotylum, by gas chromatography-mass spectrometry (GC-MS) analysis. Infection triggered a considerable alteration in the relative content of zerumbone and α-caryophyllene (humulene) with enhancement in zerumbone content (81.59%) and that of α-caryophyllene (11.91%) compared to 9.97 and 1.11%, respectively, in uninfected rhizomes. While zerumbone is the principal secondary metabolite in Z. zerumbet, α-caryophyllene is its immediate precursor. Principal component analysis (PCA) identified the correlations between metabolite changes in Z. zerumbet rhizomes and P. myriotylum infection. Radial diffusion assay with zerumbone indicated a concentration-dependent P. myriotylum growth inhibition with 93.75% inhibition observed at 700 μg and 50% maximal effective concentration (EC50) value of 206 μg. Scanning electron microscopy (SEM) analysis revealed that the mechanistic basis of zerumbone's antagonistic action on P. myriotylum growth involved the induction of aberrant morphology including severe hyphal deformities and membrane disruption. Results are discussed highlighting the critical role played by sesquiterpenoid zerumbone in affording resistance in Z. zerumbet and could expedite the development of appropriate strategies for biocontrol of Pythium spp., thus reducing the usage of broad-spectrum fungicides.

TIR-NBS-LRR genes are rare in monocots: evidence from diverse monocot orders

Abstract: Plant resistance (R) gene products recognize pathogen effector molecules. Many R genes code for proteins containing nucleotide binding site (NBS) and C-terminal leucine-rich repeat (LRR) domains. NBS-LRR proteins can be divided into two groups, TIR-NBS-LRR and non-TIR-NBS-LRR, based on the structure of the N-terminal domain. Although both classes are clearly present in gymnosperms and eudicots, only non-TIR sequences have been found consistently in monocots. Since most studies in monocots have been limited to agriculturally important grasses, it is difficult to draw conclusions. The purpose of our study was to look for evidence of these sequences in additional monocot orders. Using degenerate PCR, we amplified NBS sequences from four monocot species (C. blanda, D. marginata, S. trifasciata, and Spathiphyllum sp.), a gymnosperm (C. revoluta) and a eudicot (C. canephora). We successfully amplified TIR-NBS-LRR sequences from dicot and gymnosperm DNA, but not from monocot DNA. Using databases, we obtained NBS sequences from additional monocots, magnoliids and basal angiosperms. TIR-type sequences were not present in monocot or magnoliid sequences, but were present in the basal angiosperms. Phylogenetic analysis supported a single TIR clade and multiple non-TIR clades. We were unable to find monocot TIR-NBS-LRR sequences by PCR amplification or database searches. In contrast to previous studies, our results represent five monocot orders (Poales, Zingiberales, Arecales, Asparagales, and Alismatales). Our results establish the presence of TIR-NBS-LRR sequences in basal angiosperms and suggest that although these sequences were present in early land plants, they have been reduced significantly in monocots and magnoliids.

Genome-wide analysis of NBS-encoding disease resistance genes in Cucumis sativus and phylogenetic study of NBS-encoding genes in Cucurbitaceae crops

Abstract: Plant nucleotide-binding site (NBS)-leucine-rich repeat (LRR) proteins encoded by resistance genes play an important role in the responses of plants to various pathogens, including viruses, bacteria, fungi, and nematodes. In this study, a comprehensive analysis of NBS-encoding genes within the whole cucumber genome was performed, and the phylogenetic relationships of NBS-encoding resistance gene homologues (RGHs) belonging to six species in five genera of Cucurbitaceae crops were compared. Cucumber has relatively few NBS-encoding genes. Nevertheless, cucumber maintains genes belonging to both Toll/interleukine-1 receptor (TIR) and CC (coiled-coil) families. Eight commonly conserved motifs have been established in these two families which support the grouping into TIR and CC families. Moreover, three additional conserved motifs, namely, CNBS-1, CNBS-2 and TNBS-1, have been identified in sequences from CC and TIR families. Analyses of exon/intron configurations revealed that some intron loss or gain events occurred during the structural evolution between the two families. Phylogenetic analyses revealed that gene duplication, sequence divergence, and gene loss were proposed as the major modes of evolution of NBS-encoding genes in Cucurbitaceae species. Compared with NBS-encoding sequences from the Arabidopsis thaliana genome, the remaining seven TIR familes of NBS proteins and RGHs from Cucurbitaceae species have been shown to be phylogenetically distinct from the TIR family of NBS-encoding genes in Arabidopsis, except for two subfamilies (TIR4 and TIR9). On the other hand, in the CC-NBS family, they grouped closely with the CC family of NBS-encoding genes in Arabidopsis. Thus, the NBS-encoding genes in Cucurbitaceae crops are shown to be ancient, and NBS-encoding gene expansions (especially the TIR family) may have occurred before the divergence of Cucurbitaceae and Arabidopsis. The results of this paper will provide a genomic framework for the further isolation of candidate disease resistance NBS-encoding genes in cucumber, and contribute to the understanding of the evolutionary mode of NBS-encoding genes in Cucurbitaceae crops.

Plant-Based Phytochemicals as Possible Alternative to Antibiotics in Combating Bacterial Drug Resistance

Abstract: The unprecedented use of antibiotics that led to development of resistance affect human health worldwide. Prescription of antibiotics imprudently and irrationally in different diseases progressed with the acquisition and as such development of antibiotic resistant microbes that led to the resurgence of pathogenic strains harboring enhanced armors against existing therapeutics. Compromised the treatment regime of a broad range of antibiotics, rise in resistance has threatened human health and increased the treatment cost of diseases. Diverse on metabolic, genetic and physiological fronts, rapid progression of resistant microbes and the lack of a strategic management plan have led researchers to consider plant-derived substances (PDS) as alternative or in complementing antibiotics against the diseases. Considering the quantitative characteristics of plant constituents that attribute health beneficial effects, analytical procedures for their isolation, characterization and phytochemical testing for elucidating ethnopharmacological effects has being worked out for employment in the treatment of different diseases. With an immense potential to combat bacterial infections, PDSs such as polyphenols, alkaloids and tannins, present a great potential for use, either as antimicrobials or as antibiotic resistance modifiers. The present study focuses on the mechanisms by which PDSs help overcome the surge in resistance, approaches for screening different phytochemicals, methods employed in the identification of bioactive components and their testing and strategies that could be adopted for counteracting the lethal consequences of multidrug resistance.