Molecular characterisation and structural assessment of an RXLR effector from Phytophthora palmivora, the coconut bud rot pathogen

Phytophthora species are phytopathogenic oomycetes that damage a wide variety of crops. Phytophthora delivers effectors, which are secretory proteins, into the host cells. Effectors promote infection by reprogramming the host cellular machinery and are key determinants of oomycete virulence. The major class of Phytophthora effector proteins contains the RXLR motif. In this study, we have carried out the molecular and structural characterisation of an RXLR effector (RXLR6744) from a virulent P. palmivora isolated from bud rot disease-affected coconut palm. The open reading frame (ORF) of the RXLR6744, amplified using RT-PCR, had a length of 411 bp. The gene was found to encode a predicted protein of 136 amino acids and had a molecular weight of 15.52 kDa. Phylogenetic analysis of the amino acid sequence revealed that it was closely related to RXLR proteins from P. palmivora (causing black pod disease in cocoa) and related species P. megakarya. Topology analysis revealed that the protein was composed of six α-helices. The structural prediction was undertaken by computer-aided homology modelling. From the Ramachandran plot analysis, it could be observed that the majority (96.3%) of amino acids were present in the preferred region, 3.7 per cent of amino acid residues were present in the allowed region, and no residues were observed in the disallowed region. The structure showed an average quality of 94.4 per cent, indicating it to be a high-quality structure. This study provides the detailed characterisation of an RXLR effector from P. palmivora. It will aid the elucidation of its role in pathogenesis and facilitate further refined investigations of the structure/function relationships of oomycete effectors

Characterization of gibberellin 2-oxidase isoforms in coconut (Cocos nucifera L.)

Gibberellins (GAs) are plant hormones that are essential for many developmental processes in plants, including seed germination, stem elongation, leaf expansion, trichome development, pollen maturation and the induction of flowering. Gibberellin 2-oxidase (GA2-ox) regulates plant growth by inactivating endogenous bioactive GAs through 2β-hydroxylation. There is no information about GA2-ox encoding genes or their functions in coconut. In this study, we have identified 10 transcripts encoding different isoforms of GA2-ox from coconut leaf transcriptome data. Sequence comparison and phylogenetic analysis revealed that these 10 transcripts represented different types of GA2-ox. The secondary structure, three dimensional structure and active sites of these 10 isoforms were predicted. Docking studies of different active GAs with these isoforms was also carried out

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Not AvailableGibberellins (GAs) are plant hormones that are essential for many developmental processes in plants, including seed germination, stem elongation, leaf expansion, trichome development, pollen maturation and the induction of flowering. Gibberellin 2-oxidase(GA2-ox) regulates plant growth by inactivating endogenous bioactive GAs through 2β-hydroxylation. There is no information about GA2-ox encoding genes or their functions in coconut. In this study, we have identified 10 transcripts encoding different isoforms of GA2-ox from coconut leaf transcriptome data. Sequence comparison and phylogenetic analysis revealed that these 10 transcripts represented different types of GA2-ox. The secondary structure, three dimensional structure and active sites of these 10 isoforms were predicted. Docking studies of different active GAs with these isoforms was also carried out.Not Availabl

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Not AvailableFruit and crown rot, diseases caused by Phytophthora meadii in arecanut (Areca catechu L.), leads to huge yield losses to the growers and remains one of the greatest challenges to arecanut productivity. Developing an arecanut variety with Phytophthora resistance, therefore, is of prime significance. Areca triandra and A. concinna, which are close relatives of arecanut, are possible candidates to develop disease resistant inter-specific hybrids. It is thus imperative to have an effective screening assay for disease resistance. A detached leaf assay is often used to characterize disease susceptibility and screening for resistance to a particular pathogen. Influence of external biotic and abiotic factors in these assays pose potential challenge apart from the biosecurity risks which it carries. In the present study, two approaches were followed to screen for phytophthoral resistance in A. triandra and A. concinna. In the first approach, challenge inoculation was carried out in embryo cultured plantlets, whereas in latter, it was done on leaf segments of adult field grown plants cultured in Eeuwens Y3 media. Results indicate that in vitro assay, involving challenge inoculation on either entire plantlet or leaf segment, is an easy and rapid for disease screening.Not Availabl

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Insights into the population dynamics of phytophthora species associated with arecanut fruit rot disease

Phytophthora, a ubiquitous filamentous oomycete, causes huge yield losses and is fatal to arecanut palms in case of severe infection. From 2014 onwards, a severe fruit rot epidemic has been recorded during the South-West monsoon season in major arecanut-growing states of South India. We have assessed the diversity and delineated the population structure of Phytophthora isolates infecting arecanut in India using a combination of morphological traits, multi-gene profiling, haplotype analysis, and pathogenicity. Ninety-eight Phytophthora isolates were obtained from infected samples collected from disease-endemic regions of Karnataka, Kerala, and Goa states in southern India from 2014 to 2019. Morphological traits coupled with phylogenetic analysis using the loci ITS, β-tub, TEF-1α, and Cox-II identified the A2 mating type of P. meadii as the predominant species together with two isolates of homothallic P. heveae. Linkage disequilibrium analysis revealed significant diversity in the form of single nucleotide polymorphisms (SNPs) from the concatenated sequence dataset. Population structure analysis using 590 SNPs demonstrated the existence of four population groups (sub-populations at K = 4 and K = 2) and significant diversity in the geo-distant Phytophthora populations. In summary, 22 haplotypes were identified from the representative 40 isolates with higher haplotype diversity of 1.23 and relatively varying haplotype frequency. Pathogenicity assays confirmed that both the species of Phytophthora were pathogenic to arecanut, while P. meadii coastal region isolates exhibited more virulence compared to others