Dietary Syzygium cumini leaf extract influences growth performance, immunological responses and gene expression in pathogen-challenged Cyprinus carpio.

This study evaluated the effects of jamun leaf extract (JLE) as a feed supplement on growth performance, haemato-immunological, oxidative stress-related parameters, and cytokine gene expression in Cyprinus carpio challenged with Aeromonas hydrophila.. Diets containing four different JLE concentrations, that is, 0 (basal diet), 5 (JLE5), 10 (JLE10), and 15 g kg-1 (JLE15), were fed to carp (6.17 ± 0.43 g) for eight weeks. Growth performance was significantly higher in JLE10. Haemato-immunological and antioxidant parameters were determined in fish at 48 h post-challenge with A. hydrohila. The cumulative survival was highest in JLE10 (69.69%) 14 days post-challenge. Serum protein (2.18 ± 0.06 g dL-1), lysozyme (32.38 ± 1.2 U mL-1), alternative complement pathway (70.43 ± 1.61 U mL-1), phagocytic activity (21.18 ± 0.48%), respiratory burst activity (0.289 ± 0.09 OD630nm), and immunoglobulin levels (6.67 ± 0.36 U mg mL-1) were significantly higher in JLE10 than in the control. Serum alanine aminotransferase (44.06 ± 1.62 Unit mL-1), aspartate aminotransferase (31.58 ± 1.82 Unit mL-1), and malondialdehyde (2.57 ± 0.26 nmol mL-1) levels were lower in JLE10 than in the control (p < 0.05), whereas myeloperoxidase activity was significantly higher in JLE5 and JLE10 than in the control. Superoxide dismutase levels in the serum were higher (p < 0.05) in JLE5 and JLE10 than in the other groups. Gene expression analysis revealed that the mRNA expression of pro-inflammatory cytokines TNF-α and IL-1ß was upregulated (p < 0.05) in the liver, head-kidney, and intestine of challenged carp in JLE10. The signalling molecule NF-κB p65 was upregulated in lymphoid organs in JLE10 but not in the liver. The anti-inflammatory cytokine IL-10 was significantly downregulated in challenged carp in JLE10 compared with that in the control. Quadratic regression analysis showed that optimal dietary JLE was estimated to be 9.03-10.15 g kg-1 to maximize the growth performance. Results of the present study revealed that dietary JLE at 10 g kg-1 can significantly improve the immunity and disease resistance of C. carpio. Thus, JLE is a promising food additive for carp aquaculture.

Genome-wide characterization of DcHsp90 gene family in carnation (Dianthus caryophyllus L.) and functional analysis of DcHsp90-6 in heat tolerance.

Plant heat shock protein 90 (Hsp90) participates in various physiological processes including protein folding, degradation, and signal transduction. However, the DcHsp90 gene family in carnation (Dianthus caryophyllus L.) has not been systematically analyzed. We thoroughly examined and comprehensively analyzed the carnation DcHsp90 gene family in this study and discovered 9 DcHsp90 genes. Based on the phylogenetic examination, DcHsp90 proteins may be divided into two groups. DcHsp90 structural features were similar but varied between groups. Promoter analysis revealed the presence of many cis-acting elements, most of which were connected to growth and development, hormones, and stress. DcHsp90 genes may play distinct functions in heat stress response, according to gene expression analyses. The DcHsp90-6 was isolated, and its role in the reaction to heat stress was studied. Thermotolerance and superoxide dismutase activity in transgenic seedlings were enhanced by Arabidopsis overexpression of DcHsp90-6. After heat stress, transgenic plants' electrolyte leakage and malondialdehyde levels were much lower than wild-type plants. Furthermore, overexpression of DcHsp90-6 altered the expressions of stress-responsive genes such as AtHsp101, AtHsp90, AtGolS1, AtRS4/5, and AtHsfB1. This study provides comprehensive information on the DcHsp90 gene family and suggests that overexpressed DcHsp90-6 positively regulates thermotolerance highlighting the adaptation mechanism of carnation under heat stress.

Genomic insights into rapid speciation within the world's largest tree genus Syzygium.

Species radiations, despite immense phenotypic variation, can be difficult to resolve phylogenetically when genetic change poorly matches the rapidity of diversification. Genomic potential furnished by palaeopolyploidy, and relative roles for adaptation, random drift and hybridisation in the apportionment of genetic variation, remain poorly understood factors. Here, we study these aspects in a model radiation, Syzygium, the most species-rich tree genus worldwide. Genomes of 182 distinct species and 58 unidentified taxa are compared against a chromosome-level reference genome of the sea apple, Syzygium grande. We show that while Syzygium shares an ancient genome doubling event with other Myrtales, little evidence exists for recent polyploidy events. Phylogenomics confirms that Syzygium originated in Australia-New Guinea and diversified in multiple migrations, eastward to the Pacific and westward to India and Africa, in bursts of speciation visible as poorly resolved branches on phylogenies. Furthermore, some sublineages demonstrate genomic clines that recapitulate cladogenetic events, suggesting that stepwise geographic speciation, a neutral process, has been important in Syzygium diversification.

The clove (Syzygium aromaticum) genome provides insights into the eugenol biosynthesis pathway.

The clove (Syzygium aromaticum) is an important tropical spice crop in global trade. Evolving environmental pressures necessitate modern characterization and selection techniques that are currently inaccessible to clove growers owing to the scarcity of genomic and genetic information. Here, we present a 370-Mb high-quality chromosome-scale genome assembly for clove. Comparative genomic analysis between S. aromaticum and Eucalyptus grandis-both species of the Myrtaceae family-reveals good genome structure conservation and intrachromosomal rearrangements on seven of the eleven chromosomes. We report genes that belong to families involved in the biosynthesis of eugenol, the major bioactive component of clove products. On the basis of our transcriptomic and metabolomic findings, we propose a hypothetical scenario in which eugenol acetate plays a key role in high eugenol accumulation in clove leaves and buds. The clove genome is a new contribution to omics resources for the Myrtaceae family and an important tool for clove research.

Effect of clove on improving running ability in aging mice.

In this study, a D-galactose-induced aging mouse model was established, and Syringa oblata Lindl. extract (SOLE) was administered orally to observe the effect and mechanism of SOLE on the running ability of aging mice. The role of SOLE was evaluated by H&E histopathological observation, detection of serum biochemical indices, and detection of mRNA expression levels by qPCR experiments. The experimental results showed that SOLE could increase the exhaustive running time of aging mice and reduce the oxidative aging of the liver and kidney. At the same time, the levels of BUN, lactic acid, GOT, GPT, MDA, iNOS, and TNF-α in the serum of the mice were decreased, and the relative mRNA expression levels of nNOS, iNOS, TNF-α and syncytin-1 in the liver tissue and skeletal muscle tissue of the mice were downregulated. In addition, it increased the levels of CAT, GSH-Px, and T-SOD in mouse serum and upregulated the relative mRNA expression of Cu/Zn-SOD, Mn-SOD, and CAT in mouse liver tissue and muscle tissue. The analysis results showed that SOLE mainly contained rutin, isoquercitrin, ferulic acid, dihydroquercetin, and quercitrin. In summary, SOLE can enhance the running ability and exercise ability of aging mice and slow down aging. PRACTICAL APPLICATIONS: Clove is used as health tea or traditional Chinese medicine in China, but there is no relevant research and application on the improvement of human exercise ability. This study found this new function of clove, which accumulated a theoretical basis for further popularizing its application.

De Novo Transcriptome Study Identifies Candidate Genes Involved in Resistance to Austropuccinia psidii (Myrtle Rust) in Syzygium luehmannii (Riberry).

Austropuccinia psidii, causal agent of myrtle rust, was discovered in Australia in 2010 and has since become established on a wide range of species within the family Myrtaceae. Syzygium luehmannii, endemic to Australia, is an increasingly valuable berry crop. Plants were screened for responses to A. psidii inoculation, and specific resistance, in the form of localized necrosis, was determined in 29% of individuals. To understand the molecular basis underlying this response, mRNA was sequenced from leaf samples taken preinoculation, and at 24 and 48 h postinoculation, from four resistant and four susceptible plants. Analyses, based on de novo transcriptome assemblies for all plants, identified significant expression changes in resistant plants (438 transcripts) 48 h after pathogen exposure compared with susceptible plants (three transcripts). Most significantly up-regulated in resistant plants were gene homologs for transcription factors, receptor-like kinases, and enzymes involved in secondary metabolite pathways. A putative G-type lectin receptor-like kinase was exclusively expressed in resistant individuals and two transcripts incorporating toll/interleukin-1, nucleotide binding site, and leucine-rich repeat domains were up-regulated in resistant plants. The results of this study provide the first early gene expression profiles for a plant of the family Myrtaceae in response to the myrtle rust pathogen.

Binding affinity and in vitro cytotoxicity of harmaline targeting different motifs of nucleic acids: An ultimate drug designing approach.

The work focuses towards interaction of harmaline, with nucleic acids of different motifs by multispectroscopic and calorimetric techniques. Findings of this study suggest that binding constant varied in the order single-stranded (ss) poly(A) > double-stranded calf thymus (CT) DNA > double-stranded poly(G)·poly(C) > clover leaf tRNAPhe . Prominent structural changes of ss poly(A), CT DNA, and poly(G)· poly(C) with concomitant induction of optical activity in the bound achiral alkaloid molecule was observed, while with tRNAPhe , very weak induced circular dichroism perturbation was seen. The interaction was predominantly exothermic, enthalpy driven, and entropy favored with CT DNA and poly(G)·poly(C), while it was entropy driven with poly(A) and tRNAPhe . Intercalated state of harmaline inside poly(A), CT DNA, and poly(G)·poly(C) was shown by viscometry, ferrocyanide quenching, and molecular docking. All these findings unequivocally pointed out preference of harmaline towards ss poly(A) inducing self-structure formation. Furthermore, harmaline administration caused a significant decrease in proliferation of HeLa and HepG2 cells with GI50 of 28µM and 11.2µM, respectively. Nucleic acid fragmentation, cellular ultramorphological changes, decreased mitochondrial membrane potential, upregulation of p53 and caspase 3, generation of reactive oxygen species, and a significant increase in the G2 /M population made HepG2 more prone to apoptosis than are HeLa cells.

Impact of Postharvest Nitric Oxide Treatment on Lignin Biosynthesis-Related Genes in Wax Apple (Syzygium samarangense) Fruit.

The role of nitric oxide (NO) during storage in wax apple through NO (10 µL/L) fumigate fruit was investigated. Wax apple fruit treated with NO had a significantly lower rate of weight loss, a softening index, and loss of firmness during storage. The transcriptional profile of 10 genes involved in lignin biosynthesis has been analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). The qRT-PCR analysis showed nine genes regulated in the wax apple (p < 0.05) upon NO fumigation, which coincided with the enzyme activity results (NO group lower than control group in peroxidase, phenylalanine ammonia-lyase, and 4-coumarate-CoA ligase), whose total lignin content decreased upon treatment with NO. These results indicate that NO treatment can effectively delay the softening and senescence of wax apple fruit and play an important regulatory role in lignin biosynthesis.

De novo assembly of Eugenia uniflora L. transcriptome and identification of genes from the terpenoid biosynthesis pathway.

Pitanga (Eugenia uniflora L.) is a member of the Myrtaceae family and is of particular interest due to its medicinal properties that are attributed to specialized metabolites with known biological activities. Among these molecules, terpenoids are the most abundant in essential oils that are found in the leaves and represent compounds with potential pharmacological benefits. The terpene diversity observed in Myrtaceae is determined by the activity of different members of the terpene synthase and oxidosqualene cyclase families. Therefore, the aim of this study was to perform a de novo assembly of transcripts from E. uniflora leaves and to annotation to identify the genes potentially involved in the terpenoid biosynthesis pathway and terpene diversity. In total, 72,742 unigenes with a mean length of 1048bp were identified. Of these, 43,631 and 36,289 were annotated with the NCBI non-redundant protein and Swiss-Prot databases, respectively. The gene ontology categorized the sequences into 53 functional groups. A metabolic pathway analysis with KEGG revealed 8,625 unigenes assigned to 141 metabolic pathways and 40 unigenes predicted to be associated with the biosynthesis of terpenoids. Furthermore, we identified four putative full-length terpene synthase genes involved in sesquiterpenes and monoterpenes biosynthesis, and three putative full-length oxidosqualene cyclase genes involved in the triterpenes biosynthesis. The expression of these genes was validated in different E. uniflora tissues.

Development and characterization of new microsatellites for Eugenia dysenterica DC (Myrtaceae).

Microsatellite markers were developed for population genetic analyses of the Neotropical tree Eugenia dysenterica DC (Myrtaceae), after construction of a shotgun genomic library for microsatellite discovery. Nine primers were designed, of which 5 yielded amplified product. These primers were polymorphic for 97 individuals collected in 3 distinct localities. The number of alleles per locus (primer) ranged from 3 to 11 and expected heterozygosities varied from 0.309 to 0.884. The probability of locus identity was ~1.88 x 10(-4) and the probability of paternity exclusion was ~0.9367. The 5 microsatellite primer pairs may be suitable for population genetic studies such as parentage and fine-scale genetic analyses of this species.

Identification of microRNAs from Eugenia uniflora by high-throughput sequencing and bioinformatics analysis.

BACKGROUND: microRNAs or miRNAs are small non-coding regulatory RNAs that play important functions in the regulation of gene expression at the post-transcriptional level by targeting mRNAs for degradation or inhibiting protein translation. Eugenia uniflora is a plant native to tropical America with pharmacological and ecological importance, and there have been no previous studies concerning its gene expression and regulation. To date, no miRNAs have been reported in Myrtaceae species. RESULTS: Small RNA and RNA-seq libraries were constructed to identify miRNAs and pre-miRNAs in Eugenia uniflora. Solexa technology was used to perform high throughput sequencing of the library, and the data obtained were analyzed using bioinformatics tools. From 14,489,131 small RNA clean reads, we obtained 1,852,722 mature miRNA sequences representing 45 conserved families that have been identified in other plant species. Further analysis using contigs assembled from RNA-seq allowed the prediction of secondary structures of 25 known and 17 novel pre-miRNAs. The expression of twenty-seven identified miRNAs was also validated using RT-PCR assays. Potential targets were predicted for the most abundant mature miRNAs in the identified pre-miRNAs based on sequence homology. CONCLUSIONS: This study is the first large scale identification of miRNAs and their potential targets from a species of the Myrtaceae family without genomic sequence resources. Our study provides more information about the evolutionary conservation of the regulatory network of miRNAs in plants and highlights species-specific miRNAs.

Effect of fluoride pollution on genetic diversity of a medicinal tree, Syzygium cumini.

Syzygium cumini Linn. (Myrtaceae) is a medicinal tree (Jamun) used worldwide in treatment of diabetes. However, no molecular data is available on genetic polymorphism and its relationship, if any with fluoride pollution. In the present study, the genetic variability of two populations of S. cumini growing in fluoride rich soils and normal soils located in Rajasthan and Haryana regions of India, respectively was determined using random amplified polymorphic DNA (RAPD) markers. Different measures of diversity in Rajasthan populations: Shannon's index of phenotypic diversity (I) = 0.440; Nei's genetic diversity (h) = 0.292; effective number of alleles per locus (Ne) = 1.497; total species diversity (Hsp) = 0.307 and within population diversity (Hpop) = 0.158 showed high diversity in comparison to Haryana populations. Thus, it seems that Rajasthan population responds with increased genetic variation resulting possibly from new mutation that affect allele frequencies as a consequence of adaptation to contaminated environment. This may imply that the increased diversity levels may act as a buffer to combat fluoride stress. Cluster analysis and principal component analysis (PCA) results showed mixing between the populations.

Structural partitioning, paired-sites models and evolution of the ITS transcript in Syzygium and Myrtaceae.

The internal transcribed spacers (ITS) of nuclear ribosomal DNA are widely used for phylogenetic inference. Several characteristics, including the influence of RNA secondary structure on the mutational dynamics of ITS, may impact on the accuracy of phylogenies estimated from these regions. Here, we develop RNA secondary structure predictions for representatives of the angiosperm family Myrtaceae. On this basis, we assess the utility of structural (stem vs. loop) partitioning, and RNA-specific (paired-sites) models for a 76 taxon Syzygium alignment, and for a broader, family-wide Myrtaceae ITS data set. We use a permutation approach to demonstrate that structural partitioning significantly improves the likelihood of the data. Similarly, models that account for the non-independence of stem-pairs in RNA structure have a higher likelihood than those that do not. The best-fit RNA models for ITS are those that exclude simultaneous double substitutions in stem-pairs, which suggests an absence of strong selection against non-canonical (G.U/U.G) base-pairs at a high proportion of stem-paired sites. We apply the RNA-specific models to the phylogeny of Syzygium and Myrtaceae and contrast these with hypotheses derived using standard 4-state models. There is little practical difference amongst relationships inferred for Syzygium although for Myrtaceae, there are several differences. The RNA-specific approach finds topologies that are less resolved but are more consistent with conventional views of myrtaceous relationships, compared with the 4-state models.