In vitro proliferated sentang shoots were cultured onto half-strength Mnrashige and Skoog (MS) medium containing combinations of 1-naphthyleneacetic acid (NAA) and indole-3-butyric acid (IBA). Sentang shoots were unable to root in the absence of both auxins. A combination of 0.5 mg NAA per and 1 mg IBA per I induced the most shoots to form roots. With the addition of 2.5 g activated charcoal per 1 into half-strength MS medium containing 0.5 mg NAA per l and 1 mg IBA per 1, roots were more numerous and longer. Substances like gelrite and phloroglucinol and sugar content which would commonly influence in vitro rooting were inhibitory to adventitious root formation of sentang. Maximal rooting of 100% was achieved in "Culture Pack," made of fluorocarbon polymer film containing charcoal-free medium with 0.5 mg NAA per 1 and 1 mg IBA per 1. Rooted shoots were acclimatized for 4 wk. Overall survival was 80%. These findings suggest the use of Culture Pack as the culture vessels, with 0.5 mg NAA per 1 and 1 mg IBA per 1 in half-strength MS media to effectively induce roots in sentang shoots.

Analizamos el efecto de varios “reveladores” en la biomasa celular y la producción de alcaloides en células del árbol asiático Eurycoma longifolia. Las suspensiones celulares fueron preparadas por inoculación 0,5g FW (peso fresco) de células extraídas de suspensiones celulares de primer procesamiento preparadas de callos celulares en medio líquido MSBs. Agregamos quitosan, NaH2PO4, Na2CO3 y polivinilpirrolidona (PVP) como reveladores para aumentar la biomasa celular y la producción de alcaloides. El medio MSBs con 100mg/L chitosan induce un incremento significativo en la biomasa celular mientras que un aumento de quitosan (150mg/L) induce una alta producción de 9-hidroxicanthina-6-uno (0,44%), pero no de 9-metoxicanthina-6-uno. La adición de 2mg/L y 20mg/L NaH2PO4 aumenta la biomasa celular y la producción de alcaloides respectivamente. Sin embargo, la adición de Na2CO3 y polivinilpirrolidona muestran un efecto inhibitorio en el crecimiento celular y en la producción de alcaloides no hay un aumento significativo.

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 (EI 50 ) 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 EI 50 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 EI 50 values. EI 50 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 EI 50 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.

An efficient micropropagation protocol was established for Cryptocoryne wendtii and Cryptocoryne becketti using shoot tips explants. Multiple shoots were induced from shoot tip explants of both species cultured on agar-gelled as well as liquid MS medium supplemented with 0.5 mg/L BA and 0.2 mg/L IBA (proliferation medium). The multiple shoots of both the species formed on agar-gelled as well as liquid medium were vigorously growing with well-developed roots and leaves after 4 weeks of culture. Highest number of multiple shoots was obtained from shoot tip explants of both the species cultured in liquid proliferation medium after 4 weeks of culture. The shoot tip explants of C. wendtii and C. becketti, that were cultured in liquid proliferation medium (2 weeks) followed by culturing on agar-gelled proliferation medium (2 weeks) also produced the multiple shoots. Shoot tips cultured on agar-gelled medium produced the least number of multiple shoots after 4 weeks of culture. Histological study did not show any abnormalities in the leaves of in vitro plantlets of both the species, cultured in agar-gelled and liquid proliferation medium. The leaves of the in vitro plantlets formed normal mesophyll cells and vascular bundles. More than 95% of the acclimatized plantlets grew vigorously without any morphological abnormalities.

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