Mycelium Growth and Yielding of Black Poplar Mushroom-Agrocybe aegerita （Brig.） Sing. on Different Substrates

Mycelial growth and yield of two strains ofA. aegerita on different substrates was investigated. Mycelial growth on agar media （PDA, standard, wheat, MEA, CYM, PCA, sawdust extracts： alder and mixture of beech and birch （1：1）） and sawdust substrates （birch, beech, oak, maple, alder） was investigated. Petri dishes （Ф 9 cm） for agar media and biological tubes （18 cm long and Ф 2.5 cm） for sawdust substrates were used. Yield and morphological features were studied on birch, beech and mixture of beech and alder （1：1） sawdust substrates. The temperature of inoculation for agar media and sawdust substrates was 25 ℃. For yielding, when mycelium has completely overgrown the substrate the temperature was decreased to 15-17 ℃ to initiate primordia formation. The cultivation was enlightened 10 h/d with daylight lamps （500 Ix）. One crop was harvested after five weeks. The carpophores of black poplar mushrooms were picked up in clusters. There was no statistically important difference between the mycelial growths of the investigated strains. The best growing agar media were PDA （7.3 cm）, MEA （7.2 cm） and wheat （7.1 cm）, both strains showed the slowest mycelium growth on CYM （5.2 cm）. The mycelial growth on sawdust was the best on the beech （7.1 cm） and birch （6.8 cm） sawdust. The best substrate for cultivation ofA. aegerita was mixture of beech and alder sawdust （39.5 g/100 substrate DM）. The dry yield of carpophores were the highest on beech and alder substrate （3.2 g/100 substrate DM）, but dry matter content was the highest on beech sawdust （15.7%）. The heaviest carpophores were harvested from birch sawdust （3.7 g）; the biggest caps from beech and alder sawdust （3.3 cm）. There were no statistically important differences between the mycelium growth and yielding between both investigated strains.

Adjuvant effects mediated by the carbohydrate recognition domain ofAgrocybe aegerita lectin interacting with avian influenza H9N2 viral surface glycosylated proteins

Objective: To evaluate the potential adjuvant effect of Agrocybe aegerita lectin(AAL), which was isolated from mushroom, against a virulent H_9N_2 strain in vivo and in vitro. Methods: In trial 1, 50 BALB/c male mice(8 weeks old) were divided into five groups(n=10 each group) which received a subcutaneous injection of inactivated H_9N_2(control), inactivated H_9N_2+0.2%(w/w) alum, inactivated H_9N_2+0.5 mg recombinant AAL/kg body weight(BW), inactivated H_9N_2+1.0 mg AAL/kg BW, and inactivated H_9N_2+2.5 mg AAL/kg BW, respectively, four times at 7-d intervals. In trial 2, 30 BALB/c male mice(8 weeks old) were divided into three groups(n=10 each group) which received a subcutaneous injection of inactivated H_9N_2(control), inactivated H_9N_2+2.5 mg recombinant wild-type AAL(AAL-wt)/kg BW, and inactivated H_9N_2+2.5 mg carbohydrate recognition domain(CRD) mutant AAL(AAL-mut R63H)/kg BW, respectively, four times at 7-d intervals. Seven days after the final immunization, serum samples were collected from each group for analysis. Hemagglutination assay, immunogold electron microscope, lectin blotting, and coimmunoprecipitation were used to study the interaction between AAL and H_9N_2 in vitro. Results: Ig G, Ig G1, and Ig G2 a antibody levels were significantly increased in the sera of mice co-immunized with inactivated H_9N_2 and AAL when compared to mice immunized with inactivated H_9N_2 alone. No significant increase of the Ig G antibody level was detected in the sera of the mice co-immunized with inactivated H_9N_2 and AAL-mut R63 H. Moreover, AAL-wt, but not mutant AAL-mut R63 H, adhered to the surface of H_9N_2 virus. The interaction between AAL and the H_9N_2 virus was further demonstrated to be associated with the CRD of AAL binding to the surface glycosylated proteins, hemagglutinin and neuraminidase. Conclusions: Our findings indicated that AAL could be a safe and effective adjuvant capable of boosting humoral immunity against H_9N_2 viruses in mice through its interaction with the viral surface glycosylated proteins, hemagglutinin and neuraminidase.