Chemical characterization and the intrusion through elicitation and Agrobacterium rhizogenes mediated hairy root transformation in Saussurea costus C.B. Clarke.

Saussurea costus (Asteraceae) commonly known as kuth, is an important medicinal plant with a rich repository of medicinally valuable compounds. During the present study, pharmacologically important sesquiterpene lactones namely costunolide, dehydrocostus lactone, betulinic acid and syringin were isolated from different plant extracts. Furthermore, the elicitation effect of jasmonic acid (JA) and different light regiments on the accumulation of secondary metabolites (costunolide and dehydrocostus lactone) was evaluated using HPLC. There was an increase in amount of costunolide and dehydrocostus lactone compared to control after 96 h of treatment with JA and continuous light. The amount of costunolide after 96 h was maximum 6.47 mg/g DW in response to JA as compared to control which was found to be 1.7 mg/g DW. Similarly, the concentration of dehydrocostus lactone after 96 h showed maximum accumulation of compound 4.7 mg/g DW in response to continuous light. The in vitro response in MS medium augmented with BAP (4 mg/l) produces friable and creamish coloured callus, however, number of days increased from 10 to 22 days with 70% culture response. Also, Agrobacterium rhizogenes strain LBA9402 was found to be most effective strain for the establishment of hairy root cultures among all the strains used. The genomic DNA was used as template in PCR to amplify rolB gene which confirmed the efficient transformation of the roots. Additionally, total metabolite content of in vitro raised hairy roots of S. costus was significantly higher than the field grown plants. The production of secondary metabolites through elicitation and hairy roots can serve as a potential tool for the conservation action programme in S. costus. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01270-9.

Hepatoprotective effect of seabuckthorn leaf-extract in lead acetate-intoxicated Wistar rats.

The present study investigated the hepatoprotective effects of seabuckthorn (Hippophae rhamnoides) leaf-extract (SLE) supplementation in lead acetate-intoxicated Wistar rats. Adult male Wistar rats (n = 35) were divided into five equal groups as per completely randomized design. Group I was kept as a control, group II received 250 ppm lead acetate in drinking water, and group III received SLE at 100 mg/kg body weight per os. Animals in group IV and group V received lead acetate at 250 ppm in drinking water for the first 45 days. Additionally, group IV received SLE at 100 mg/kg body weight per os throughout the experiment, whereas group V received SLE at 100 mg/kg body weight per os during the last 15 days of the trial. Blood samples were collected on day zero and at the 45th and 60th day to study the liver function enzymes. Lead exposure caused increase in aspartate aminotransferase (AST), alanine aminotransferase (ALT), acid phosphatase (ACP), and alkaline phosphatase (ALP) activity, and liver weight, and hepatic oxidative stress in lead acetate-intoxicated groups (groups II, IV, and V) as compared to group I. SLE on simultaneous supplementation with lead acetate (group IV) exerted a protective effect against lead toxicity. SLE supplemented after the establishment of lead acetate-toxicity (group V) also reduced the AST, ALT, and ALP activity and hepatic oxidative stress, indicating its ameliorative effect. SLE supplementation at 100 mg/kg body weight per os protects against hepatic damage caused by 250 ppm lead acetate in the drinking water of Wistar rats.

Ecological Characterisation of Native Isolates of Heterorhabditis indica from Viti Levu, Fiji Islands.

Entomopathogenic nematodes (EPNs) in the families Steinernematidae and Heterorhabditidae are obligate parasites of soil inhibiting insects. EPNs are being widely researched as promising biocontrol agents for a wide range of agricultural pests. It is known that strains of EPNs isolated from different geographical regions differ in their attributes, such as host-finding ability, host range, infectivity, reproduction, and environmental stress tolerance. A precise knowledge of these factors is therefore an essential pre-requisite for devising successful strategies to use these nematodes in biological control programmes. Thus, ecological characterisation of the EPN Heterorhabditis indica (Rhabditida: Heterorhabditidae) newly isolated and representing the only species of EPN reported from the island of Viti Levu, Fiji was carried out using Galleria mellonella larvae (L) (Pyralidae: Galleriinae) as hosts to allow comparisons between bioassays conducted in different laboratories around the world. Temperature data showed that native isolates of H. indica are warm-adapted nematodes with thermal range for infectivity between 15˚C and 35˚C and can reproduce between 20˚C and 30˚C. They are highly virulent with LC50 values against G. mellonella ranging from 2.8 IJ to 3.8 IJ/larva. However, they showed poor desiccation tolerance and fail to infect hosts in soil with moisture levels below 8%. They showed a moderate level of hypoxic tolerance and can be stored at 15˚C for 4 months. Results also showed great variability within the selected native isolates of H. indica. Beneficial traits for selected isolates were added up to identify a superior candidate. The current study also suggested that the thermal niche breadth for infection can differ among conspecific strains of an EPN species. The results of this experimental study on ecological aspects of these native isolates of H. indica should form a basis for their potential use in biological control of insect pests in Fiji.

Modeling the potential global distribution of suitable habitat for the biological control agent Heterorhabditis indica.

Entomopathogenic nematode (EPN) Heterorhabditis indica is a promising biocontrol candidate. Despite the acknowledged importance of EPN in pest control, no extensive data sets or maps have been developed on their distribution at global level. This study is the first attempt to generate Ecological Niche Models (ENM) for H. indica and its global Habitat Suitability Map (HSM) for H. indica to generate biogeographical information and predicts its global geographical range and help identify of prospective areas for its exploration and to help identify the suitable release areas for biocontrol purpose. The aim of the modeling exercise was to access the influence of temperature and soil moisture on the biogeographical patterns of H. indica at the global level. Global Heterorhabditis indica ecosystems. CLIMEX software was used to model the distribution of H. indica and assess the influence of environmental variable on its global distribution. In total, 162 records of H. indica occurrence from 27 countries over 25 years were combined to generate the known distribution data. The model was further fine-tuned using the direct experimental observations of the H. indica's growth response to temperature and soil moisture. Model predicts that much of the tropics and subtropics have suitable climatic conditions for H. indica. It further predicts that H. indica distribution can extend into warmer temperate climates. Examination of the model output, predictions maps at a global level indicate that H. indica distribution may be limited by cold stress, heat stress, and dry stresses in different areas. However, cold stress appears to be the major limiting factor. This study highlighted an efficient way to construct HSM for EPN potentially useful in the search/release of target species in new locations. The study showed that H. indica which is known as warm adapted EPN generally found in tropics and subtropics can potentially establish itself in warmer temperate climates as well. The model can also be used to decide the release timing of EPN by adjusting with season for maximum growth. The model developed in this study clearly identified the value and potential of Habitat Suitability Map (HSM) in planning of future surveys and application of H. indica.

Protective and ameliorative effect of sea buckthorn leaf extract supplementation on lead induced hemato-biochemical alterations in Wistar rats.

AIM: To evaluate the protective and ameliorative effect of aqueous sea buckthorn leaf extract (SLE) on hemato-biochemical profile in lead intoxicated Wistar rats. MATERIALS AND METHODS: An experiment was conducted for 60 days. 36 adult male Wistar rats with a mean body weight of 177.8±12.6 g were divided into five groups and were subjected to various daily oral treatment regimens. Group I served as a negative control receiving only feed and water, Group II (positive control for lead) received lead acetate at 250 ppm in drinking water, and Group III (positive control for SLE) received SLE at 100 mg/kg b.wt. Animals in Group IV received a combination of lead acetate at 250 ppm in drinking water for the first 45 days and SLE at 100 mg/kg b.wt. throughout the experimental period of 60-day, and in Group V for the last 15 days of the trial after the administration of lead acetate until the first 45 days of the trial to study the protective and ameliorating effects of SLE, respectively. Blood samples were collected from retro-orbital fossa of each rat on 0th, 45th, and 60th day of the experiment for hemato-biochemical analysis including hemoglobin (Hb), packed cell volume (PCV), serum total protein, albumin, globulin, albumin:globulin ratio, cholesterol, urea, and creatinine. RESULTS: Significantly (p<0.01) lower levels of serum total proteins and albumin, and a significantly (p<0.01) higher serum cholesterol, urea and creatinine levels were observed in Group II (lead intoxicated group) in comparison to Group I (negative control). Administration of SLE at 100 mg/kg body wt. to lead intoxicated Wistar rats resulted in normalization of almost all the biochemical parameters studied in both the treatment Groups, i.e., IV and V (protective and ameliorative). However, the effects were more pronounced in the protective group. No effects of SLE supplementation were observed on Hb levels. PCV levels improved in protective groups, but no effect was observed in ameliorative group in comparison to lead intoxicated groups. CONCLUSION: SLE administration at 100 mg/kg b.wt. to lead intoxicated Wistar rats may be used to protect/ameliorate lead induced biochemical alterations in Wistar rats.