First report of Athelia rolfsii (=Sclerotium rolfsii) associated with foot rot disease of Chrysanthemum morifolium in India.

Chrysanthemum morifolium L. is an important flower crop grown in different parts of Karnataka for its striking cut flowers and international market value. During a field survey (Mysore district, Karnataka, February, 2022), chrysanthemum fields were found infected with foot rot disease. The presence of white mycelial structures with sclerotia were recorded near the stem-soil interface. The disease incidence ranged 10-12% measured in an area of approximately 10 hectares. The infected plants showed quick wilt, yellowing and toppling of the entire plant. Infected plants from Doddamaragowdanahally and Rayanahally (n=15) were collected and associated fungal pathogen isolated after surface sterilization with NaOCl (1%) on potato dextrose agar (PDA) amended with chloramphenicol (50 mg/L). Fungal mycelia developed from the infected tissues were inoculated on to fresh PDA plates to obtained pure cultures for further identification. Fungal colonies with dense, aerial whitish-cottony mycelia with uniformly globoid sclerotia (0.284.2 mm) were observed after 15 days of incubation (28 ± 2°C). Sclerotia were white in the beginning and turned brown at maturity. The average number of sclerotia produced per plate ranged from 240 to >480 (n = 10). To further to confirm the identity of the isolates, two representative isolates (CmSr1 and CmSr2) was subjected to molecular identification based on ITS-rDNA sequences. Briefly, genomic DNA was isolated from 12 day old cultures using the CTAB method and ITS-rDNA was amplified using ITS1-ITS4 primers (White et al., 1990). An expected amplicon of >650 bp (ITS) was obtained and later sequenced from both the directions. The consensus sequences were analysed through nBLAST search which revealed that 100% sequence similarity with reference sequences of Athelia rolfsii (S. rolfsii) from GenBank database (MT127465, MN974137, KC292637; identity 656/656; 0 gaps). A phylogenetic tree obtained by the neighbor-joining method using MEGAX shared a common clade with the reference sequences retrieved and computed, thus confirming the identification based on sequence analysis and molecular phylogeny. The representative sequence of A. rolfsii isolates CmSr1 and CmSr2 isolates deposited in GenBank with Accession nos. ON456153 and ON456154, respectively. Based on etiology, morphological, cultural and molecular data the pathogen was identified as Athelia rolfsii (Curzi) Tu & Kimbrough (Syn: Sclerotium rolfsii Sacc.) (Mordue, 1974; Mahadevakumar et al., 2016, 2018). Plants (n=60) were inoculated with sclerotial bodies (2 sclerotia/plant) near stem soil interface under green house and covered with polythene bags (at 27 ± 2°C and 80% RH). Non-inoculated plants (n=20) served as controls. The development of foot rot disease was observed eight days after inoculation. A total of 48 plants showed the foot rot symptoms and 12 inoculated plants and control plants remained healthy. The identity of the fungus was confirmed by morphological and cultural characters after re-isolation. C. morifolium is an important flower crop in Karnataka. S. rolfsii is known to be associated with blight and collar rot of Chrysanthemum spp. from Kerala (Beena et al., 2002) but no species (host) identity provided. Therefore, to the best of our knowledge, this is the first report of foot rot disease caused by Athelia rolfsii on C. morifolium in India. Early diagnosis of this disease will help the farmers to adopt suitable management practices to avoid loss.

Effect of Telmisartan on Arsenic-Induced (Sub-chronic) Perturbations in Redox Homeostasis, Pro-inflammatory Cascade and Aortic Dysfunction in Wistar Rats.

An experimental study was conducted in male Wistar rats to explore the antioxidant potential of telmisartan (an AT1 receptor blocker) to overcome arsenic ('As')-induced perturbations in redox homeostasis pro-inflammatory cytokines, prostaglandin-E2 levels and aortic dysfunction in Wistar rats. Wistar rats were randomly divided into four groups of six each. Group-I served as untreated control, while group-II received sodium (meta) arsenite (NaAsO2) (10 mg/kg b.wt. p.o) for a period of 60 days. Experimental rats in group-III received treatment similar to group-II, but in addition received telmisartan (with 1% aqueous solution of Tween 80) @ 10 mg/kg b.wt. (p.o) for a similar duration, while rats in group-IV received telmisartan alone. Arsenic exposure resulted in significant (p < 0.05) elevation in the levels of superoxide anion ([Formula: see text]) radicals (control: 768.20 ± 126.77 vs group-II: 1232.75 ± 97.85 pmol of NBT reduced/min/mg protein). Telmisartan administration showed significant (p < 0.05) reduction in [Formula: see text] generation (815.34 ± 43.41 pmol of NBT reduced/min/mg protein). Sub-chronic exposure to 'As' significantly (p < 0.05) decreased the activities of SOD, CAT, GPx and GR activity and GSH levels in the aorta, thus induced lipid peroxidation (LPO) measured as measured in terms of thiobarbituric acid reactive substances (TBARS) called as malondialdehyde (MDA). However, the administration of telmisartan effectively countered the LPO (24.03 ± 1.18 nmol of MDA/g) on account of restoring the levels of aforesaid antioxidant defense system. Telmisartan administration effectively attenuated the 'As'-induced surge in pro-inflammatory cytokines (viz., IL-1ß, IL-6 and TNF-α) levels, as well as countered the activity of cyclooxygenase (COX2) as indicated by a significant (p < 0.05) decrease in PGE2 level in the aorta. In addition to it, there was a significant (p < 0.05) decrease in plasma angiotensin II (Ang-II) levels in experimental rats receiving telmisartan. Quantitative RT-PCR studies revealed that sub-chronic exposure to 'As' upregulated the Nox2 mRNA expression, but there was a 1.2-fold reduction in expression level upon co-administration of telmisartan. Histopathological examination revealed marked recovery from 'As'-induced disruption of tunica adventitia and loss of connective tissue in experimental rats receiving telmisartan. The study concludes that telmisartan can overcome aortic dysfunction induced by sub-chronic exposure to arsenic through drinking water in experimental rats through restoration of redox balance, attenuation of pro-inflammatory cytokines and mediators and downregulation of Nox2 mRNA expression.

Anticancer efficacy of 6-thioguanine loaded chitosan nanoparticles with or without curcumin.

6-Thioguanine encapsulated chitosan nanoparticles (6-TG-CNPs) has formulated by the ionic-gelation method. Morphologically, the 6-TG-CNPs were spherical and showed mean size, PDI, zeta potential, and entrapment efficiency of 261.63 ± 6.01 nm, 0.34 ± 0.10, +15.97 ± 0.46 mV and 44.27%, respectively. The IR spectra confirmed the 6-TG complex with chitosan. The in vitro drug release profile of 6-TG-CNPs revealed an increase in sustained-release (91.40 ± 1.08% at 48 h) at pH 4.8 compared to less sustained-release (73.96 ± 1.12% at 48 h) at pH 7.4. The MTT assay was conducted on MCF-7 and PA-1 cell lines at 48 h incubation to determine % cell viability. The IC50 values of 6-TG, 6-TG-CNPs, and curcumin for MCF-7 were 23.09, 17.82, and 15.73 µM, respectively. Likewise, IC50 values of 6-TG, 6-TG-CNPs, and curcumin for PA-1 were 5.81, 3.92, and 12.89 µM, respectively. A combination of 6-TG-CNPs (IC25) with curcumin (IC25) on PA-1 and MCF-7 showed % cell viability of 43.67 ± 0.02 and 49.77 ± 0.05, respectively. The in vitro cytotoxicity potential in terms of % cell viability, early apoptosis, G2/M phase arrest, and DNA demethylating activity of 6-TG-CNPs alone and combination with curcumin proved to be more effective than that of 6-TG on PA-1 cells.

Studies on the luteolytic, oestrogenic and follicle-stimulating hormone like activity of plant Rhaphidophora pertusa (Roxb.).

The phytochemical, pharmacological and toxicological feature of plant Rhaphidophora pertusa (Roxb.) was done. Phytosteroids, flavonoids, tannins and glucosides were detected in the plant extracts. In cross-bred (Zebu X Holstein-Friesian or Jersey) dairy cows, subsequent to prostaglandin (PG) induced oestrus, to each group (n=4), cloprostenol (PG control) 100 microg i.m. on day 10, the rice gruel (vehicle) was fed on day 10 or the fresh stem (1 kg/animal/day) in rice gruel on day 9, or days 9 and 10, or days 9-11 of the oestrous cycle. Each group received subcutaneously either 5% gum acacia suspension or the plant ethyl acetate or methanol extract (1g in gum acacia) on days 8 (to bannur ewes) or 10 (to dairy cows) of the oestrous cycle. In PG control cows or ewes, there was induction of oestrus in 48 h and a fall in serum progesterone concentration. The feeding of fresh stem in the rice gruel or the s.c. administration of the plant extract did not induce oestrus or significantly (P>0.05) alter the serum progesterone, bilirubin, calcium, creatinine, phosphorus, magnesium and glucose concentrations or the total erythrocyte and leucocyte count, differential leucocyte count and haemoglobin concentration. The plant did not cause any toxicity in the cow or ewe. In immature rats, the aqueous or methanol (hot or cold) extract did not cause any follicle-stimulating hormone (FSH)-like activity. The methanol extract increased the uterine weight in ovariectomised rats. This suggested the presence of oestrogenic activity in the plant. In conclusion, the present study revealed the presence of oestrogenic activity in the plant and the absence of luteolytic or FSH-like or toxic activity.