Objective:To evaluate the growth inhibition activity of the crude extract of Cyperus aromaticus (C. aromaticus) cultured cells against the 3rd instar larvae of Aedes aegypti (Linn.) and Aedes albopictus Skuse (Ae. albopictus) under laboratory conditions, and determine the sublethal effects (EI50) of the crude extract of C. aromaticus cultured cells on some biological and morphological parameters of both Aedes mosquito species during two generations as well.
Methods:The cell suspension cultures of C. aromaticus were activated from five callus lines (P4, Pa, Z1, Z6 and Ml) derived from the root explants of in vitro plantlets. The cultured cells were extracted in chloroform and used as plant material for the present study. For detection of juvenile hormone III, the crude extracts were analyzed by HPLC. Then the crude extracts of the three C. aromaticus cultured cell lines which contained varied amounts of juvenile hormone III [high level (P4 cell line), medium level (Z1 cell line) and low level (Ml cell line)] were tested against Aedes mosquito species. Laboratory evaluation was performed against late third instar larvae of the Vector Control Research Unit strains of Ae. aegypti and Ae. albopictus using the standard WHO method. The effects of EI50 of the C. aromaticus cultured P4 cells on fecundity, fertility, growth period, sex ratio, adult size and longevity of Aedes mosquitoes were assessed.
Results:Bioassay tests presented the remarkable growth inhibition activity of the crude extracts of C. aromaticus cultured cells against the two Aedes mosquitoes. Between the two mosquito species, Ae. albopictus was more susceptible to the crude extracts with lower EI50 values. EI50 of the crude extract of C. aromaticus cultured cells (P4) increased the sterility indices in the parental generation females in both Aedes mosquito species. A significant delay in the pupal formation and adult emergence were observed in the parental generation of the both mosquito species. The sex ratio of the adult population either parental or F1 generation of the Aedes mosquito species was not significantly affected by the EI50 dosage of the crude extract of C. aromaticus cultured P4 cells. A significant decrease in the wing length of the treated adult (female and male) of Aedes aegypti as well as the treated female of Ae. albopictus were observed. Longevity of the adult female of the parental generation of both Aedes mosquitoes as well as females of F1 generation of Ae. albopictus were significantly decreased.
Conclusions: The present study revealed the potential of the crude extract of C. aromaticus cultured cells in controlling vector mosquito populations in the effort to reduce the transmission of vector borne diseases.

Aims: The purpose of this experiment was to determine the artificial symbiosis interaction of Herbaspirillum seropedicae (Z78) on oil palm embryogenic calli. Methodology and results: For this purpose, symbiotic associations were established between Z78 and embryogenic calli of oil palm tissue cultured. A total of five treatments involved, in particular: i) + 3.0 mg/L 2,4-D + 100% N MS medium (control), ii) + Z78 pellet cells (1 mL) + 25% N MS medium, iii) + Z78 supernatant (1 mL) + 25% N MS medium, iv) + Z78 broth culture (1 mL) + 25% N MS medium, and v) + Z78 sonicated cells (1 mL) + 25% N MS medium. All treatments were supplied with minimal N sources (25% N), ammonium nitrate and potassium nitrate, while the control was treated with 100% N sources. Treated samples were harvested on D80 and observed for biomass and diameter increment (%), formation of embryoids, and Z78 colonization. The results showed embryogenic calli in the inoculated treatments that contained depleted N produced similar result to the control treatment which contained 100% N nutrients. Positive interactions occurred between the diazotroph and host plant tissues as viewed under FESEM and EFTEM. Among the treatments, Z78 sonicated cell showed better growth of embryogenic calli compared to others. Conclusion, significance and impact study: The in vitro nitrogen-depleted artificial symbiosis environment allowed the diazotroph (Z78) to be expressed and provide the nitrogen sources and indole-3-acetic acid for cell growth. This study represents beneficial co-culture interaction effects of different inocula of diazotrophic bacterial cells with in vitro embryogenic calli of oil palm.