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        Purification and characterization of prophenoloxidase from cotton bollworm, Helicoverpa armigera

        Hanumanth G. Goudru,Sathish KUMAR,Senigala K. JAYALAKSHMI,Chandish R. BALLAL,Hari C. SHARMA,Kuruba SREERAMULU 한국곤충학회 2013 Entomological Research Vol.43 No.1

        Phenoloxidases are oxidative enzymes, which play an important role in both cell mediated and humoral immunity. Purification and biochemical characterization of prophenoloxidase from cotton bollworm, Helicoverpa armigera (Hübner) were carried out to study its biochemical properties. Prophenoloxidase consists of a single polypeptide chain with a relative molecular weight of 85 kDa as determined by SDS–PAGE, MALDI–TOF MS and LC–ESI MS. After the final step, the enzyme showed 71.7 fold of purification with a recovery of 49.2%. Purified prophenoloxidase showed high specific activity and homology with phenoloxidase subunit‐1 of Bombyx mori and the conserved regions of copper binding (B) site of phenoloxidase. Purified prophenoloxidase has pH optima of 6.8 and has high catalytic efficiency towards the dopamine as a substrate in comparison to catechol and L‐Dopa. The PO activity was strongly inhibited by phenylthiourea, thiourea, dithiothreitol and kojic acid.

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        Treatment of millet crop plant (Sorghum bicolor) with the entomopathogenic fungus (Beauveria bassiana) to combat infestation by the stem borer, Chilo partellus Swinhoe (Lepidoptera: Pyralidae)

        Narasimha P. Reddy,Akbar P. Ali Khan,Uma K. Devi,Hari C. Sharma,Annette Reineke 한국응용곤충학회 2009 Journal of Asia-Pacific Entomology Vol.12 No.4

        Experiments were done to test if Beauveria bassiana can become an endophyte in sorghum and confer protection from stem borer. Four-week-old sorghum seedlings were treated with B. bassiana. The plants were examined for endophytic presence of B. bassiana, 30 and 60 days after treatment. Stem cultures from treated plants showed growth of B. bassiana. PCR amplification using fungal specific primers for a conserved region of β tubulin gene yielded identical 360 bp products from both B. bassiana and treated sorghum plants. In a subsequent experiment, B. bassiana treated and untreated (control) sorghum plants were artificially infested with stem borer (Chilo partellus) larvae 15 days post treatment and the extent of damage was compared. About 40% of the control plants developed dead heart while no plant in the B. bassiana treated plot did. In the surviving control plants, stem tunneling by shoot borer was significantly higher compared to B. bassiana treated sorghum plants. Experiments were done to test if Beauveria bassiana can become an endophyte in sorghum and confer protection from stem borer. Four-week-old sorghum seedlings were treated with B. bassiana. The plants were examined for endophytic presence of B. bassiana, 30 and 60 days after treatment. Stem cultures from treated plants showed growth of B. bassiana. PCR amplification using fungal specific primers for a conserved region of β tubulin gene yielded identical 360 bp products from both B. bassiana and treated sorghum plants. In a subsequent experiment, B. bassiana treated and untreated (control) sorghum plants were artificially infested with stem borer (Chilo partellus) larvae 15 days post treatment and the extent of damage was compared. About 40% of the control plants developed dead heart while no plant in the B. bassiana treated plot did. In the surviving control plants, stem tunneling by shoot borer was significantly higher compared to B. bassiana treated sorghum plants.

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