Novel report of Acinetobacter johnsonii as an indole-producing seed endophyte in Tamarindus indica L.

Plant-microbe associations have been regarded as an exciting topic of research due to their potential as environment friendly alternatives for stimulating crop growth and development. Seeds of Tamarindus indica L. have been chosen for the present study as seed endophytes prefer larger or nutritive cotyledon and hard seed coats for their colonization. The main objectives of our study were to isolate and identify the seed endophytes, their bioefficacy, and responsible chemical compounds. In a dose-dependent experiment, tamarind seed exudates (TSE) showed plant growth-promoting properties on Oryza sativa (53-81%), Daucus carota (10-31%), and Raphanus sativa (21-42%). Identification of the bacterial load in TSE through 16S rRNA sequencing revealed the existence of two bacterial species, Acinetobacter johnsonii and Niallia nealsonii. This is the first report of these two bacteria as seed endophytes of Tamarindus indica L. HRLC-MS analysis of TSE confirmed the presence of indole derivatives, primarily indole-3-lactic acid (ILA). The quantitative phytochemical estimation of bacterial culture filtrates revealed that indole-like substances were present in the extracts only in A. johnsonii at a concentration of 0.005 mg/ml of indole acetic acid equivalent. Experimental results suggested that the stimulatory activity of TSE was caused by the presence of A. johnsonii, a potential plant growth-promoting bacteria that produced indole-like compounds. This study suggests tamarind seed exudates with its endophytic microbiota as a potent plant growth-promoting agent that may find use as a cheap and sustainable source of metabolites useful in the agro-industries.

Synthesis, antimicrobial, anticancer evaluation and molecular docking with Bax and MDM2 of dibromosterculic acid.

Dibromosterculic acid [8-(1,2-dibromo-2-octylcyclopropyl)-octanoic acid], a new synthetic derivative was prepared by bromination of sterculic acid. This synthetic derivative showed strong fungicidal activity against two pathogenic fungal species namely Penicillium chrysogenum and Aspergillus niger with minimum inhibitory concentration (MIC) value of 0.007 mg/ml and good bactericidal activity against Bacillus subtilis and Xanthomonas sp. with MIC value of 0.015 mg/ml. Cytotoxic activity on both normal (MCF-10A) and cancerous (MDA-MB-468) cell lines revealed that the survivability percentage of normal cells was unaffected, whereas cancerous cells were decreased greatly by dibromosterculic acid with 50% survivability at 9 µg/ml concentration. Molecular-docking using AutoDock 4.2 with Bax exhibited strong pi-sigma interaction with PHE-93, pi-alkyl and alkyl interaction with TRP-139, ARG-89 and PHE-92 whereas MDM2 revealed strong hydrogen bond interaction with GLN-59 and pi-alkyl interaction with PHE-55. All experimental parameters suggested that this synthetic derivative would be valuable for target-specific drug development with nominal side effects.

Global transcriptome analysis reveals fungal disease responsive core gene regulatory landscape in tea.

Fungal infections are the inevitable limiting factor for productivity of tea. Transcriptome reprogramming recruits multiple regulatory pathways during pathogen infection. A comprehensive meta-analysis was performed utilizing previously reported, well-replicated transcriptomic datasets from seven fungal diseases of tea. The study identified a cumulative set of 18,517 differentially expressed genes (DEGs) in tea, implicated in several functional clusters, including the MAPK signaling pathway, transcriptional regulation, and the biosynthesis of phenylpropanoids. Gene set enrichment analyses under each pathogen stress elucidated that DEGs were involved in ethylene metabolism, secondary metabolism, receptor kinase activity, and various reactive oxygen species detoxification enzyme activities. Expressional fold change of combined datasets highlighting 2258 meta-DEGs shared a common transcriptomic response upon fungal stress in tea. Pervasive duplication events caused biotic stress-responsive core DEGs to appear in multiple copies throughout the tea genome. The co-expression network of meta-DEGs in multiple modules demonstrated the coordination of appropriate pathways, most of which involved cell wall organization. The functional coordination was controlled by a number of hub genes and miRNAs, leading to pathogenic resistance or susceptibility. This first-of-its-kind meta-analysis of host-pathogen interaction generated consensus candidate loci as molecular signatures, which can be associated with future resistance breeding programs in tea.

Alternative splicing shapes the transcriptome complexity in blackgram [Vigna mungo (L.) Hepper].

Vigna mungo, a highly consumed crop in the pan-Asian countries, is vulnerable to several biotic and abiotic stresses. Understanding the post-transcriptional gene regulatory cascades, especially alternative splicing (AS), may underpin large-scale genetic improvements to develop stress-resilient varieties. Herein, a transcriptome based approach was undertaken to decipher the genome-wide AS landscape and splicing dynamics in order to establish the intricacies of their functional interactions in various tissues and stresses. RNA sequencing followed by high-throughput computational analyses identified 54,526 AS events involving 15,506 AS genes that generated 57,405 transcripts isoforms. Enrichment analysis revealed their involvement in diverse regulatory functions and demonstrated that transcription factors are splicing-intensive, splice variants of which are expressed differentially across tissues and environmental cues. Increased expression of a splicing regulator NHP2L1/SNU13 was found to co-occur with lower intron retention events. The host transcriptome is significantly impacted by differential isoform expression of 1172 and 765 AS genes that resulted in 1227 (46.8% up and 53.2% downregulated) and 831 (47.5% up and 52.5% downregulated) transcript isoforms under viral pathogenesis and Fe2+ stressed condition, respectively. However, genes experiencing AS operate differently from the differentially expressed genes, suggesting AS is a unique and independent mode of regulatory mechanism. Therefore, it can be inferred that AS mediates a crucial regulatory role across tissues and stressful situations and the results would provide an invaluable resource for future endeavours in V. mungo genomics.

Deciphering the role of alternative splicing as modulators of defense response in the MYMIV-Vigna mungo pathosystem.

Alternative splicing (AS) is a crucial regulatory mechanism that impacts transcriptome and proteome complexity under stressful situations. Although its role in abiotic stresses is somewhat understood, our understanding of the mechanistic regulation of pre-messenger RNA splicing in plant-pathogen interaction is meager. To comprehend this unexplored immune reprogramming mechanism, transcriptome profiles of Mungbean Yellow Mosaic India Virus (MYMIV)-resistant and susceptible Vigna mungo genotypes were analyzed for AS genes that may underlie the resistance mechanism. Results revealed a repertoire of AS-isoforms accumulated during pathogenic infestation, with intron retention being the most common AS mechanism. Identification of 688 differential alternatively spliced (DAS) genes in the resistant host elucidates its robust antiviral response, whereas 322 DAS genes were identified in the susceptible host. Enrichment analyses confirmed DAS transcripts pertaining to stress, signaling, and immune system pathways have undergone maximal perturbations. Additionally, a strong regulation of the splicing factors has been observed both at the transcriptional and post-transcriptional levels. qPCR validation of candidate DAS transcripts with induced expression upon MYMIV infection demonstrated a competent immune response in the resistant background. The AS-impacted genes resulted either in partial/complete loss of functional domains or altered sensitivity to micro-RNA-mediated gene silencing. A complex regulatory module, miR7517-ATAF2, has been identified in an aberrantly spliced ATAF2 isoform that exposes an intronic miR7517 binding site, thereby suppressing the negative regulator to enhance the defense reaction. The present study establishes AS as a noncanonical immune reprogramming mechanism that operates in parallel, thereby offering an alternative strategy for developing yellow mosaic-resistant V. mungo cultivars.

Characterization and metabolomic profiling of two pigment producing fungi from infected fruits of Indian Gooseberry.

Two pigment producing fungi, Talaromyces atroroseus and Penicillium choerospondiatis, were isolated and identified from infected fruits of Phyllanthus emblica L. based on amplification and sequencing of internal transcribed spacer region and beta-tubulin gene. This is the first occurrence report of these two fungi from fruits of P. emblica. Culture extract containing metabolites of T. atroroseus and P. choerospondiatis contained phenolics of 26.35 mg and 30.89 mg GAE/g dry extract respectively; whereas no significant amount of flavonoids and tannins were detected. P. choerospondiatis metabolites extract showed higher DPPH and ABTS activity with IC50 values of 21.94 mg/ml and 27.03 mg/ml respectively than T. atroroseus. LC-HRMS analysis of metabolites extract of T. atroroseus revealed presence of trimethyl-isopropyl-butanamide, perlolyrine, N-hexadecanoylpyrrolidine etc. whereas P. choerospondiatis displayed presence of tangeraxanthin, ugaxanthone, daphniphylline, etc. Therefore, fungal metabolites are rich natural sources of diversified compounds that can be utilized in dyeing industries, cosmetics and novel drug development.

Ecophysiological traits differentially modulate secondary metabolite accumulation and antioxidant properties of tea plant [Camellia sinensis (L.) O. Kuntze].

Owing to the diverse growing habitats, ecophysiology might have a regulatory impact on characteristic chemical components of tea plant. This study aimed to explore natural variations in the ecophysiological traits within seasons and the corresponding multifaceted biochemical responses given by the gene pool of 22 tea cultivars. Leaf temperature and intercellular carbon concentration (Ci), which varies as a function of transpiration and net photosynthesis respectively, have significant impact on the biochemical traits of the leaf. Occurrence of H2O2, in leaves, was associated to Ci that in turn influenced the lipid peroxidation. With the increment of Ci, total phenolics, epicatechin gallate (ECG), reducing power, and radical scavenging activity is lowered but total catechin and non-gallylated catechin derivatives (e.g. epicatechin or EC, epigallocatechin or EGC) are elevated. Leaf temperature is concomitantly associated (p ≤ 0.01) with phenolics, flavonoids, proanthocyanidin, tannin content, reducing power, iron chelation and free radical scavenging activities. Increased phenolic concentration in leaf cells, conceivably inhibit photosynthesis and moreover, gallic acid, thereafter conjugated to catechin derivatives. This study shed light on the fundamental information regarding ecophysiological impact on the quality determining biochemical characteristics of tea, which on further validation, might ascertain the genotype selection paradigm toward climate smart cultivation.

Phylogenetic inference of Ericales based on plastid genomes and implication of cp-SSRs.

Ericales is an ancient eudicot order encompassing numerous species of economic and ornamental values. Despite several phylogenomic studies, the evolutionary relationship among certain families of this group remains uncertain. The present study assessed a multilocus species tree of Ericales based on 107 chloroplast genomes. The plastome derived microsatellite motifs were also simultaneously explored to check their dynamicity in corroboration of species phylogeny and systematics. In addition to resolving the usual hierarchy, the present phylogenetic analysis enabled to resolve the persisting lineage disparity with valid statistical support. Accordingly, divergence incongruences of Primulaceae, Ebenaceae, and Sapotaceae from earlier reports were reinstated in presently inferred phylogeny, which further supported the latest transcriptome-based relationship of the corresponding group. Various SSR motif characteristics emerged following the recognition of the evolutionary pathway. Numerical variation in tetranucleotide repeats showed even intraspecific or varietal differences in Camellia sinensis. Validation of plastome microsatellite-based polymorphism among the related taxa might pave the way for future phylogenetic and population studies of this economically important group.

Genome-wide SNP discovery from Darjeeling tea cultivars - their functional impacts and application toward population structure and trait associations.

Genotyping by sequencing and identification of functionally relevant nucleotide variations in crop accessions are the key steps to unravel genetic control of desirable traits. Elite cultivars of Darjeeling tea were undergone SNP genotyping by double-digest restriction-site associated DNA sequencing method. This study reports a set of 54,206 high-quality SNP markers discovered from ~10.4 GB sequence data, encompassing 15 chromosomes of the reference tea genome. Genetic relatedness among the accessions conforms to the analyses of Bayesian clustering, UPGMA, and PCoA methods. Genomic positions of the discovered SNPs and their putative effect on annotated genes designated a thoughtful understanding of their functional aspects in tea system biology. A group of 95 genes was identified to be affected by high impact variants. Genome-wide association analyses of 21 agronomic and biochemical phenotypes resulted in trait-linked polymorphic loci with strong confidence (p < 0.05 and 0.001).

Temporal depletion of packaged tea antioxidant quality under commercial storage condition.

Shelf life studies play a significant role in determination of time duration for the retention of product quality after packaging. Assessment of tea shelf life in terms of antioxidant quality, a prime health benefit trait of tea would substantiate its marketing and consumption preference to the trade and end users. In shelf life analysis of tea with respect to its antioxidant potentialities, both antioxidant activity and incidences of secondary metabolites are responsible. A temporal analysis with regular intervals since 1 year of said characteristics has been carried out in four types of processed teas. To be precise, the overall initial antioxidant concentrations and activities were almost maintained up to 90-120 days and thereafter declination appeared. Beyond 180 days, rapid declination occurs and beyond 330 days, depletion recorded up to 60-75% of the initial activity. Black tea showed maximum ferrous ion chelating activity initially and white tea commenced with slight lower value but it maintained a similar trend up to 150 days while a rapid declination occurred in such activity of black and green tea after 30 days only. It is observed that total tannins or proanthocyanidins amount highest in white tea among all other three types. The preservation of metal chelating activity of white tea was observed as comparable to its stability in tannin composition (r2 = 0.869, P ≤ 0.01) during the storage period.

Groundwater inhabited Bacillus and Paenibacillus strains alleviate arsenic-induced phytotoxicity of rice plant.

Arsenic contamination in agricultural soil now imposing a major threat to crop productivity and causing several hazardous health effects through percolation in food chain. Bioremediation, an efficient way of soil health restoration toward sustainability offered by some soil-borne microorganisms, has been reported. The present work deals with application of two potent arsenic-tolerant bacterial strains (Bacillus thuringiensis A01 and Paenibacillus glucanolyticus B05), obtained from natural sources in modulating overall growth and antioxidant defense against arsenic-treated rice plants. Between the two, former could reduce arsenic uptake up to 56% (roots) and 85% (shoots), and the preceding one up to 31% (roots) and 65% (shoots) in a hydroponic environment. Germination percentage was noted to be enhanced significantly (p ≤ 0.05). Expression of oxidative stress defensive enzymes such as superoxide dismutase, peroxidase and catalase have been augmented at seedling stages (21 days) toward detoxification of arsenic imposed excess ROS generation. Increment of leaf Thiobarbituric acid reactive substances due to arsenic exposure have been ameliorated by both the bacterial application. Phenolic and flavonoid mediated free radical scavenging ability of the test plants elevated significantly (p ≤ 0.05). The present work revealed that, selected bacterial strains can perform efficient bioremediation against arsenic pollutant rice cultivation.

MIPS: Functional dynamics in evolutionary pathways of plant kingdom.

The myo-inositol biosynthesis pathway triggering protein MIPS is best known for its necessity, ubiquitous nature and occurrence throughout all living kingdom. However, the functional disparity of MIPS genes in green plant is still viable. The present work considered a comprehensive genome-wide analysis from sequenced plants to identify MIPS homologs in respective organisms and their genomic architecture. Variation of MIPS gene expression in twelve different species in diverse conditions has also been analysed. All MIPS genes share a conserved sequence property in most of its coding region, but its regulatory elements, gene structure and expression network vary significantly. Phylogenetic inference confirms the evolution of MIPS from a single common algal ancestor to seed plants and acquiring functional variation through genomic control. This paper represents MIPS as a model for studying gene duplication, functional divergence and diversification events in plant lineages.

Extrapolative microRNA precursor based SSR mining from tea EST database in respect to agronomic traits.

Tea (Camellia sinensis, (L.) Kuntze) is considered as most popular drink across the world and it is widely consumed beverage for its several health-benefit characteristics. These positive traits primarily rely on its regulatory networks of different metabolic pathways. Development of microsatellite markers from the conserved genomic regions are being worthwhile for reviewing the genetic diversity of closely related species or self-pollinated species. Although several SSR markers have been reported, in tea, the trait-specific Simple Sequence Repeat (SSR) markers, leading to be useful in marker assisted breeding technique, are yet to be identified. Micro RNAs are short, non-coding RNA molecules, involved in post transcriptional mode of gene regulation and thus effects on related phenotype. Present study deals with identification of the microsatellite motifs within the reported and predicted miRNA precursors that are effectively followed by designing of primers from SSR flanking regions in order to PCR validation. In addition to the earlier reports, two new miRNAs are predicting here from tea expressed tag sequence database. Furthermore, 18 SSR motifs are found to be in 13 of all 33 predicted miRNAs. Trinucleotide motifs are most abundant among all followed by dinucleotides. Since, miRNA based SSR markers are evidenced to have significant role on genetic fingerprinting study, these outcomes would pave the way in developing novel markers for tagging tea specific agronomic traits as well as substantiating non-conventional breeding program.