Sanehdeep Kaur

Enhanced Resistance to Spodoptera litura in Endophyte Infected Cauliflower Plants

ABSTRACT Endophytic fungi, which live within host plant tissues without causing any visible symptom of disease, are important mediators of plant-herbivore interactions. These endophytes enhance resistance of host plant against insect herbivores mainly by productions of various alkaloid based defensive compounds in the plant tissue or through alterations of plant nutritional quality. Two endophytic fungi, i.e., Nigrospora sp. and Cladosporium sp., were isolated from Tinospora cordifolia (Thunb.) Miers, a traditional indian medicinal plant. Cauliflower (Brassica oleracea L.) plants were inoculated with these two endophytic fungi. The effect of endophyte infected and uninfected cauliflower plants were measured on the survival and development of Spodoptera litura (Fab.), a polyphagous pest. Endophyte infected cauliflower plants showed resistance to S. litura in the form of significant increase in larval and pupal mortality in both the fungi. Inhibitory effects of endophytic fungi also were observed on adult emergence, longevity, reproductive potential, as well as hatchability of eggs. Thus, it is concluded that antibiosis to S. litura could be imparted by artificial inoculation of endophytes and this could be used to develop alternative ecologically safe control strategies.

An alpha-glucosidase inhibitor from an endophytic Cladosporium sp. with potential as a biocontrol agent.

This study highlights the importance of alpha-glucosidase inhibitors as mechanisms for endophyte-mediated resistance to insect pests. One of the major benefits which endophytes confer on plants is providing resistance against insect pests. This built-in defense mechanism of the plant can be used for exploring ecofriendly strategies for pest control. In the present study, 34 endophytic fungi were isolated from Tinospora cordifolia and screened for their ability to produce alpha-glucosidase inhibitors. Maximum inhibitory activity was observed in an isolate from T. cordifolia (TN-9S), identified to be Cladosporium sp. The inhibitor was purified using chromatographic techniques. The insecticidal activity of the purified inhibitor was evaluated against Spodoptera litura. The inhibitor induced a significant mortality in the larvae of S. litura and adversely affected its survival and development. It also inhibited the activity of α-glycosidases in vivo in the gut of the larvae. The purified inhibitor was determined to be a phenolic compound with amine groups, demonstrating a noncompetitive type of inhibition in vitro. The production of the inhibitor was optimized. Response surface methodology (RSM) analysis revealed a significant interaction between dextrose and malt extract, with first-order effect of pH.

Antifeedent and toxic activity of endophytic Alternaria alternata against tobacco caterpillar Spodoptera litura

Fungal endophytes play a significant role in plant resistance to insects by altering the nutritional quality of plants or by production of various alkaloid-based defensive compounds. Ethyl acetate extract of endophytic Alternaria alternata isolated from Azadirachta indica A. Juss was tested for its bioactivity against Spodoptera litura (Fab.). Effect of different concentrations of ethyl acetate extract of A. alternata was assessed on growth and development of S. litura larvae. The results indicate higher larval mortality on diet supplemented with the fungal extract than control. Adverse effects of A. alternata were also observed on development of the insect. Ethyl acetate extract of the fungus also significantly reduced the adult emergence, longevity and reproductive potential of S. litura. Larvae feeding on diet supplemented with fungal extract showed significant reduction in relative growth and consumption rate, as well as efficiency of ingested and digested food. The antifeedant and toxic effects of A. alternata may be due to production of bioactive molecules by this fungus.

Detrimental effects of endophytic fungus Nigrospora sp. on survival and development of Spodoptera litura

Abstract Endophytes have been known to confer resistance to host plants against insect herbivores mediated by fungal alkaloids. In this study we have isolated an endophytic fungus Nigrospora sp. (Ascomycota: Sordariomycetes) from a native plant Tinospora cordifolia. To assess anti-insect potential, we tested to what extent the survival and development of Spodoptera litura (Lepidoptera: Noctuidae), a polyphagous pest, was affected when fed on artificial diet amended with different concentrations of ethyl acetate extract of Nigrospora sp. In feeding assay, the insect suffered significantly higher mortality and showed prolonged development period on amended diet as compared to unamended diet. A significant reduction in relative growth and consumption rate, efficiency of conversion of ingested and digested food and approximate digestibility indicated deterrent as well as toxic effects of the fungal extract. Significant adverse effects were also observed on adult emergence, longevity and reproductive potential of S. litura. This preliminary information on insecticidal properties of Nigrospora sp. may further be used for imparting resistance in plants against insects.

Pathogenicity of bacteria isolated from gut of Spodoptera litura (Lepidoptera: Noctuidae) and fitness costs of insect associated with consumption of bacteria.

Gut microbes contribute to the health of insects and perturbations in the composition or location of gut microbiota can lead to pathological states and host mortality. We explored the culturable bacterial community in the gut of Spodoptera litura (Fab.) larvae, which is a polyphagous pest. Bacterial isolates were identified as Microbacterium arborescens (SL6), Enterococcus casseliflavus (SL10) and Enterobacter cloacae (SL11) by using culture dependent technique based on 16S rRNA gene sequencing. Screening of these three isolates for insecticidal potential against the same host i.e. S. litura indicated the highest larval mortality in E. cloacae (73.33%). Further, we assessed the effect of E. cloacae (SL11) infection on growth and development of S. litura. A significant effect of E. cloacae was observed on various biological parameters viz. larval and pupal period, total development period and reproductive potential of S. litura. E. cloacae significantly influenced the immune response of S. litura. A marked decrease in total hemocyte count was observed in larvae infected with E. cloacae whereas lysozyme and phenoloxidase activity increased initially followed by a decline. The gut microbial diversity in larvae infected with E. cloacae differed from control larvae. The population of E. cloacae in the gut of infected larvae exceeded over the other two microbes and resulted in pathogenicity and death of S. litura larvae. This indicates that E. cloacae can have the potential to be used as a promising biological control agent.

Insecticidal potential of an endophytic Cladosporium velox against Spodoptera litura mediated through inhibition of alpha glycosidases

Alpha glucosidase inhibitory activity was exhibited by partially purified fractions obtained from an endophytic Cladosporium velox, isolated from Tinospora cordifolia. Taking into account the increasing importance of digestive enzyme inhibitors as insecticidal agents, the entomopathogenic potential of the fractions obtained was evaluated against Spodoptera litura (Fab.), a polyphagous pest. Considerable mortality was obtained when the larvae were fed on diet supplemented with the partially purified extract. All the concentrations of the extract significantly prolonged the overall developmental period of S. litura. At higher concentrations, the extract influenced the longevity of females as well as their reproductive potential. Phytochemical analysis revealed the presence of phenolic compounds in the active fraction. The phenolic compound responsible for the bioactivities was purified and identified to be chlorogenic acid using HPLC and MS analysis. The content of chlorogenic acid in the extract was quantified to be 250μg/ml. The purified compound also demonstrated inhibition of alpha glycosidases in vivo. The present study indicates that the endophyte imparted resistance to the insects in the plants could be mediated through chlorogenic acid targeting the alpha glycosidases present in the gut of the insect. The isolate obtained can be exploited for the production of chlorogenic acid, which has the potential to be exploited as a biocontrol agent against S. litura.

Effect of Alternaria alternata on the coccinellid pest Henosepilachna vigintioctopunctata and its implications for biological pest management

Twenty-eight spotted potato ladybird beetle (Henosepilachna vigintioctopunctata) is a polyphagous pest that infests various crop plants of the family Solanceae. It extensively infests Withania somnifera, a high-value medicinal plant of this family. Both the beetle and W. somnifera leaf spot pathogen (Alternaria alternata) are individually found to be closely associated with the foliar damage and may rarely infest the same leaf simultaneously. Beetles fed on diseased leaves showed high pupal and larval mortality. Furthermore, the development of the larvae into pupa and from pupa to adult was also delayed, which in turn delayed the overall development of the insect. Adult emergence from pupae is also reduced in the insects fed on diseased leaves. Since the fungal pathogen is host specific to W. somnifera and does not infect the other solanaceous crops, therefore this fungal stain can be used in designing bio-control strategies for the other solanaceous plants frequently infested by the beetle.

Impact of sesquiterpenes from Inula racemosa (Asteraceae) on growth, development and nutrition of Spodoptera litura (Lepidoptera: Noctuidae)

BACKGROUND The use of botanical pesticides for protecting crops from insect pests has assumed greater importance all over the world owing to growing awareness of harmful effects of indiscriminate use of synthetic pesticides. Inula racemosa Hook. f. (Asteraceae), a medicinally important perennial herb, is rich in sesquiterpenes with many biological activities. The present studies were conducted with the objective to evaluate the sesquiterpenes isolated from I. racemosa for insecticidal activity against Spodoptera litura (F.). RESULTS Alantolactone and isoalantolactone isolated from I. racemosa exerted growth inhibitory effects on S. litura. Addition of both the sesquiterpenes to larval diet extended the development period and reduced pupation as well as adult emergence. The dietary utilisation experiments on third-instar larvae of S. litura revealed reduction in consumption and growth rates of larvae as well as efficiency of conversion of ingested and digested food owing to alantolactone and isoalantolactone. CONCLUSION The root extract of I. racemosa, which is rich in two sesquiterpenes, i.e. alantolactone and isoalantolactone, has the potential for management of S. litura. However, there is a need to understand the specific mechanism of action of these compounds.

Endophyte-mediated interactions between cauliflower, the herbivore Spodoptera litura, and the ectoparasitoid Bracon hebetor

AbstractFungal endosymbionts in plants may influence interactions among plants, herbivores and their parasitoids through the production of secondary metabolites. We used a lepidopteran pest and its generalist parasitoid to test the effect of endophyte-infected plants on a third trophic level. Endophytic fungi, Aspergillus flavus and Aspergillus niger, isolated from Acacia arabica, were used to infect cauliflower plants. We found that the presence of the endophyte in the plants significantly extended the development period of Spodoptera litura (Fab.) larvae. Feeding of the host on endophyte-infected plants further adversely affected the development and performance of its parasitoid, Bracon hebetor (Say). A negative impact was also recorded for longevity and fecundity of endophyte-naive parasitoid females due to the parasitization of host larvae fed on endophyte-infected plants. The presence of endophytes in the diet of the host larvae significantly prolonged the development of the parasitoid. A strong detrimental effect was also recorded for larval survival and emergence of parasitoid adults. The longevity and parasitism rate of female wasps were reduced significantly due to the ingestion of endophyte-infected cauliflower plants by S. litura larvae. Overall, we found that both endophytic fungi had a negative impact on the parasitoid.

Suppression of Cellular Immune Response in Spodoptera litura (Lepidoptera: Noctuidae) Larvae by Endophytic Fungi Nigrospora oryzae and Cladosporium uredinicola

ABSTRACT Endophytic fungi are a group of microbial plant symbionts that occur in living tissues of plants without causing visible disease symptoms. The mechanism underlying the anti-insect properties of endophytes is mainly attributed to the production of various alkaloid-based defensive compounds. The current study reveals the cytotoxic effects of endophytic Nigrospora oryzae- and Cladosporium uredinicola-infected cauliflower plants on larval hemocytes of Spodoptera litura (F.). Larvae fed on leaves of endophyte-infected plants exhibited abnormalities such as change in shape, extensive vacuolization, and necrosis in significantly higher percentage of hemocytes. The percentage of abnormal hemocytes increased with increase in feeding duration. The cytotoxic effect of endophytic fungi on the hemocytes ultimately leads to adverse effects on survival and fitness of the insect.