Optimization of the sterilization method for leaf explant Robusta BP 308 coffee in vitro

Asmono, SL and Wardana, R and Rahmawati, Rahmawati (2022) Optimization of the sterilization method for leaf explant Robusta BP 308 coffee in vitro. In: IOP Conference Series: Earth and Environmental Science.

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IOP Conference Series: Earth and Environmental Science PAPER • THE FOLLOWING ARTICLE ISOPEN ACCESS Optimization of the sterilization method for leaf explant Robusta BP 308 coffee in vitro S L Asmono1, R Wardana1 and Rahmawati1 Published under licence by IOP Publishing Ltd IOP Conference Series: Earth and Environmental Science, Volume 980, The 4th International Conference on Food and Agriculture (ICOFA), 6-7 November 2021, Jember, Indonesia Citation S L Asmono et al 2022 IOP Conf. Ser.: Earth Environ. Sci. 980 012001 DOI 10.1088/1755-1315/980/1/012001 DownloadArticle PDF References Download PDF 236 Total downloads Turn on MathJax Share this article Share this content via email Share on Facebook (opens new window) Share on Twitter (opens new window) Share on Mendeley (opens new window) Article and author information Abstract The research aimed to know the optimal sterilizing agent for the success of the coffee callus inoculation from young leaf explant. This research was conducted at the Plant Tissue Culture Laboratory for 2 months. The explants are used from young leaves of Robusta clones BP 308. This experiment used a nonfactorial completely randomized design with three sterilization methods and was repeated 10 times. The treatments tested were as follows Method 1: Fungicide 2 gr/L, Bactericide 2 gr/L, Erythromycin 4 gr/L (20 min); 70% alcohol (3 sec); NaOCl 0.525% (10 min); NaOCl 1.05% (10 min). Method 2: 70% alcohol (3 sec), 0.1% HgCL2 (5 min). Method 3: Fungicide 2 gr/L, Bactericide 2 gr/L, Erythromycin 4 gr/L (20 min); 70% alcohol (3 sec); NaOCl 0.525% (10 min); NaOCl 1.05% (10 min); 0.1% HgCL2 (5 min). The results showed that the sterilization method used suppressed the rate of bacterial and fungal contamination, but there was no significant difference between the methods. A very significant effect was seen in the percentage of browning explants. In sterilizing agent treatment 1, there was no browning explant, while the other methods caused browning to reach 62.5%. Browning also causes differences in callus regeneration ability. The explants which were sterilized in method 1 showed a high percentage of callus (85%) and were also appear to callus faster at 11 days after inoculation.

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