ABSTRACT
Aim: The existence of Orchid Mycorrhizal Fungi (OMF) has a role to stimulate growth and support the supply of orchid nutrition as a biofertilizer agent. This study aimed to determine the association of mycorrhizal with Phalaenopsis amabilis (L.) Blume which was carried out through the effectiveness test of two Indonesian orchid mycorrhizal isolates i.e. Ceratorhiza and Trichoderma. Study Design: This study consisted of 4 treatments. Each treatment was repeated 3 times, each repetition of 5 plantlets, so that the total plantlet used was 60. Place and Duration of Study: Laboratory of Plant Biotechnology, Department of Biology, Universitas Gadjah Mada, Indonesia, between June 2017 and April 2018. Methodology: The method of inoculating orchid mycorrhizal by placing a plantlet in a petri dish containing orchid mycorrhizal for 1, 2, 3, and 4 days. Then plantlets are grown on sterile moss growing media and acclimatized in a greenhouse. Observation of each treatment is carried out every day for the next month. Observation variables include the number of initial and final roots, the number of live and dead roots, and the number of living and dead plants. Results: The results of the orchid mycorrhizal induction test showed that the Ceratorhiza inoculation treatment showed a fluctuation in the mean increase in the number of final roots, live roots, dead roots, and dead plantlets that were higher than the Trichoderma inoculation treatment. The results also showed that the best inoculation time on Ceratorhiza and Trichoderma was day 3 and 4. The adaptation process had the effect of increasing the number of dead roots in weeks 1 and 2. The adaptation process stopped at the beginning of week 4 with the number of new roots appearing a lot. Conclusion: Orchid mycorrhizal Ceratorhiza shows the value of effectiveness test compared with Trichoderma. The results of this study are expected to be basic information in efforts to cultivate natural orchids in Indonesia.
ABSTRACT
The aim of this work was to study the standardization of conditions to obtain and regenerate Epulorhiza repens and Ceratorhiza sp. protoplasts. For E. repens, the largest number of protoplasts (8.0 × 10(6) protoplasts/mL) was obtained in 0.6 M KCl, using 15 mg/mL of Lysing Enzymes, and 2-day-old fungal mycelium. When 0.5 M sucrose was used as osmotic stabilizer, the highest frequency of regeneration was achieved (8.5 percent); 80.0 percent of protoplasts were nucleated, and 20.0 percent anucleated. For Ceratorhiza sp., the largest number of protoplasts (4.0 × 10(7) protoplasts/mL) was achieved in 0.6 M NaCl, when 15 mg/mL of Lysing Enzymes and 15mg/mL of Glucanex, with 2-day-old fungal mycelium were used. The highest frequency of regeneration was 6.7 percent using 0.5 M sucrose as osmotic stabilizer; 88.8 percent of protoplasts were nucleated, and 11.2 percent anucleated.
O objetivo deste trabalho foi padronizar as condições de obtenção e regeneração de protoplastos de Epulorhiza repens e Ceratorhiza sp. Para o fungo E. repens, a maior produção de protoplastos, 8.0 x 10(6) protoplastos/mL, foi obtida em KCl 0.6 M, na presença de 15 mg/mL de "Lysing Enzymes" e micélio fúngico com 2 dias de idade. A maior freqüência de regeneração obtida foi de 8,5 por cento quando sacarose 0.5 M foi utilizada como estabilizador osmótico. Do total de protoplastos obtidos, 80 por cento eram nucleados e 20 por cento anucleados. Para Ceratorhiza sp., a maior produção de protoplastos, 4,0 x 10(7) protoplastos/mL, foi obtida em NaCl 0.6 M, na presença de 15 mg/mL de "Lysing Enzymes" e 15mg/mL de Glucanex, e micélio fúngico com 2 dias de idade. A maior freqüência de regeneração obtida foi de 6.7 por cento utilizando sacarose 0.5 M como estabilizador osmótico. Do total de protoplastos obtidos, 88.8 por cento eram nucleados e 1.2 por cento anucleados. O estabelecimento de protocolo otimizado para obtenção e regeneração de protoplastos dos fungos E. repens e Ceratorhiza sp. é importante, permitindo o estabelecimento de técnicas de transformação genética, o isolamento de mutantes, a determinação de cariótipo eletroforético e o cruzamento de linhagens.