Infection by Pseudocercospora musae leads to an early reprogramming of the Musa paradisiaca defense transcriptome.

Deep sequencing technologies such as RNA sequencing can help unravel mechanisms governing defense or resistance responses in plant-pathogen interactions. Several studies have been carried out to investigate the transcriptomic changes in Musa germplasm against Yellow Sigatoka disease, but the defense response of Musa paradisiaca has not been investigated so far. We carried out transcriptome sequencing of M. paradisiaca var. Kachkal infected with the pathogen Pseudocercospora musae and found that a vast set of genes were upregulated while many genes were downregulated in the resistant cultivar as a result of infection. After transcriptome assembly and differential gene expression analysis, 429 upregulated and 156 downregulated genes were filtered out (considering fold change ± 2, p < 0.01). Functional annotation of the differentially expressed genes (DEGs) enriched the upregulated genes into 49 gene ontology (GO) classes of biological processes (BP), 20 classes of molecular function (MF) and 9 classes of cellular component (CC). Similarly, the downregulated genes were classified into 35 GO classes of BP, 28 classes of MF and 6 classes of CC. The KEGG enrichment analysis revealed that the upregulated genes were most highly represented in 'metabolic' and 'biosynthesis of secondary metabolites' pathways. Additionally, 'plant hormone signal transduction', 'plant-pathogen interaction' and 'phenylpropanoid biosynthesis' pathways were also significantly enriched indicating their involvement in resistance responses against the pathogen. The RNA-seq analysis also depicts that a range of important defense-related genes are modulated as a result of infection, all of which are responsible for either mediating or activating resistance responses in the host. We studied and validated the expression profiles of ten important defense-related genes potentially involved in conferring resistance to the pathogen through qRT-PCR. Almost all the selected defense-related genes were found to be highly and significantly upregulated within 24 h post inoculation (hpi) and for some genes, the expression remained consistently high till the later time point of 72 hpi. These results, thus, indicate that the infection by P. musae leads to a rapid reprogramming of the defense transcriptome of the resistant banana cultivar. The defense-related genes identified to be modulated in response to infection are important not only for pathogen recognition and perception but also for activation and persistence of defense in the host. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03245-9.

Purification and characterization of phenoloxidase from the hemolymph of healthy and diseased Antheraea assamensis Helfer (Lepidoptera: Saturniidae): Effects of certain biological components and chemical agents on enzyme activity.

In the current study, a dimeric phenoloxidase (PO) from the hemolymph of healthy and diseased (pebrine infected) larvae of Antheraea assamensis Helfer was extracted and purified. The protein was subjected to purification using Sephacryl S-100 and CM Sepharose chromatography. The enzyme comprised of two subunits of ~76.8 and 76 kDa that showed PO activity in 6 mM l-3,4-dihydroxyphenylalanine (L-DOPA) and 8 mM catechol but not in hydroquinone. Optimum temperature for PO activity was 30°C in l-DOPA and 37°C in catechol. Optimum pH ranged from 6.8 to 7.0 in L-DOPA and 7.0-7.2 in catechol. Specific activity of the purified PO from healthy larvae was 53.9 µM/min per mg of protein per ml in L-DOPA and 50.77 µM/min per mg of protein per ml in catechol. Specific activity of PO from diseased larvae was 30.0 µM/min per mg of protein per ml in L-DOPA and 28.55 µM/min per mg of protein per ml in catechol. Purification fold was 3.27-4.21 for healthy and 2.38-2.56 for diseased fractions. The enzyme showed the Michaelis constant (Km ) of 2.46-2.85 mM for healthy and diseased fractions in L-DOPA. In catechol Km of 9.23-17.71 mM was observed. Peptidoglycan was the best activator of purified PO from both healthy and diseased fractions. Interactions between controls and activators appeared statistically significant (F = 767.5; df = 3; P < 0.0001). Na+ , K+ , and Cu2+ increased, whereas Ca2+ , Zn2+ , Mg2+ , and Co2+ decreased PO activity. The overall interactions appeared highly significant (F = 217.0; df = 27; P < 0.0001). Kojic acid, dithiothreitol, thiourea, phenylthiourea, carbendazim, N-bromosuccinimide, N,N,N',N'-tetraacetic acid, and diethyldithiocarbamate inhibited PO activity.

Growth inhibitory effect of phenolic extracts of Ziziphus jujuba Mill. in dengue vector Aedes aegypti (L) in parent and F1 generation.

OBJECTIVE: To evaluate the lethal and emergence inhibitory effect of alkaloid, phenolic and terpenoid extracts of Ziziphus jujuba (Rhamnaceae) against Aedes aegypti (Diptera: Culicidae), and to explore the effect of the most effective fraction on developmental and biochemical parameters of the dengue vector. METHODS: The fourth instar larvae of Ae. aegypti were exposed to alkaloid, phenolic and terpenoid extracts from Z. jujuba leaves to test their toxicity and emergence inhibitory effects. Phenolic extract, being the most effective was further tested against the mosquitoes for their growth inhibitory effect supported by biochemical changes in the parent and F1 generation. RESULTS: While the different secondary metabolite fractions i.e., alkaloid, phenolics and terpenoid caused mortality at larval and pupal stages, the LC50 value was the lowest for phenolic fraction. Further study carried out with the phenolic fraction revealed that it affected growth by decreasing adult life span, fertility and fecundity of the mosquitoes. The reduction in growth was also accompanied by decrease in carbohydrate and lipid levels. CONCLUSIONS: It is concluded that the phenolic extract of the leaves of Z. jujuba is a potential candidate for control of Aedes mosquitoes.